Starch branching enzymes (SBE) and granule-bound starch synthase (GBSS) are two important enzymes for starch biosynthesis. SBE mainly contributes to the formation of side branches, and GBSS mainly contributes for the synthesis of amylose molecules. However, there are still gaps in the understanding of possible interactions between SBE and GBSS.Nineteen natural rice varieties with amylose contents up to 28% were used. The molecular structure, in the form of the chain-length distribution  was measured after enzymatic debranching, using fluorophore-assisted carbohydrate electrophoresis for amylopectin and size- exclusion chromatography for amylose. The resulting distributions were fitted to two mathematical models based on the underlying biosynthetic processes, which express the CLDs in terms of parameters reflecting relevant enzyme activities.Finding statistically valid correlations between the values of these parameters showed that GBSSI and SBEI compete for substrates during rice starch biosynthesis, and synthesis of amylose short chains involves several enzymes including GBSSI, SBE and SSS (soluble starch synthase). Since the amylose CLD is important for a number of functional properties such as digestion rate, this knowledge is potentially useful for developing varieties with improved functional properties. Amylose and amylopectin are the two main components of starch, which is a complex branched glucose polymer. Amylose has moderate molecular weight with a small number of long-chain branches, and amylopectin has large molecular weight with a vast number of short-chain branches , starch branching enzymes (SBE), starch debranching enzymes (DBE) and ADP-glucose pyrophosphorylase polypeptide (AGPase), are involved in starch biosynthesis in cereal endosperms  and β(ii) are calculated based on CLDs of region1, β(iii) and β(iv) are calculated based on those of region 2, and β(v) and β(vi) on those of region 3. For the biosynthesis parameters representing the relative rates of chain elongation, h(iii/i) is the ratio of region 2 height to region 1 height, while h(v/i) is the ration of region 3 height to region 1 height.The amylopectin CLDs are shown in Fig. X) close to 100. Those for ≲ 100 are amylopectin chains and those with X ≳100 are amylose chains. While SEC is needed for the amylose CLDs, the SEC amylopectin CLDs are not used here because FACE can give a much better resolution for DP ≲ 180. Amylose CLDs were fitted with the biosynthesis-based models described previously  which is the relative activity of the dominant SS in that region  significantly negatively correlates with β(iii), and strongly negatively correlates with β(v). βAm,(iii) represents long amylose chains, and β(iii) and β(v) represent intermediate and long amylopectin chains. This suggests that the synthesis of long amylose chains competes with the syntheses of intermediate and long amylopectin chains, especially the long amylopectin chains, and perhaps these two syntheses could happen at the same site in starch granules. It has been shown that GBSSI is the dominant enzyme controlling long amylose chain synthesis  and h(v/i) have no significant correlation with hAm,(iii), which indicates that the interaction between SSIII and GBSSI is not significant. SBEI can transfer long glucosyl chains (DP > 20) to form long amylopectin branches, while GBSSI can elongate amylose chains with long glucosyl chains (DP > 20)  is not significantly correlates with β(i) and β(ii), which shows that GBSSI has little interaction with SBEIIb in rice. Future experiments are needed to analyze the activity of different enzyme isoforms  and the interactions between different isoforms.It is noted that the complete chain-length distribution can be used to calculate the degree of branching through the formula  and βAm,(ii) and amylopectin structural parameters. Earlier studies have shown that, besides GBSSI, some branching enzymes and soluble starch synthases are also involved in short amylose chain biosynthesis  and β(v) show negative correlation with hAm,hAm,(ii) and hAm,(iii). The amylose h parameters are of course strongly positively correlated with amylose content (Table (iii) and β(v) values. These results might indicate that some elongation of amylose chains happens inside of starch granules, where the synthesis of amylose and amylopectin can compete for substrates and enzymes.It is seen that βnt Table . GBSSI i(iv) is positively correlated with βAm,(iii), hAm, (i), hAm,(ii) and hAm,(iii), the opposite of what is seen with β(iii) and β(v). β(iv) is negatively correlated with β(iii), which is an expected result because the amylopectin model assumes  has a weak negative correlation with hAm, (i) and hAm,(ii). If this weak correlation is indeed the case, this might be because same SBE isoforms participate in the synthesis of both intermediate amylopectin and short amylose chains. It also could be a statistical coincidence, because it is only a weak correlation and h(iii/i) does not have a significant correlation with hAm,(iii), while the latter is strongly correlated with hAm, (i) and hAm,(ii). The internal correlations between amylopectin structural parameters, and internal correlations between amylose structural parameters, are not discussed here as they are not germane to the aim of the present study, which is to look at which enzymes are involved in the synthesis of both amylopectin and amylose.It is seen that Quantitative analysis of the structural parameters for amylopectin and amylose chain length distributions of nineteen natural rice samples indicate that GBSSI competes with SBEI for substrates during starch synthesis in rice. This might be of use for developing rice varieties with desirable amylose contents and CLDs. The results also confirm that GBSSI, SBE and SS are all involved in biosynthesis of short amylose chains.Carbohydrate Labelling and Analysis Kit, was purchased from BeckmanCoulter . Other chemical reagents were analytical grade and used as received.Nineteen natural rice varieties were harvested from the same large field in Sanya, Hainan Province, China; details are listed in Table − 1 following the method of  were precooled in liquid nitrogen for 5 min before being ground to rice flour using a cryo-grinder . Grinding at a low temperature reduces the damage of the molecular and granular structure of starch. All samples were ground for 2 min at 30 s− 1), acetate buffer solution  and isoamylase , and incubated at 37 °C for 3 h. The debranching process was stopped by adding NaOH solution (0.1 M) to adjust the pH to 7, and the solution was incubated at 80 °C for 1 h. All the samples were freeze-dried overnight and stored in a desiccator for SEC and FACE analysis.Rice starch was extracted following method of  X using the Mark–Houwink equation: NA is Avogadro’s constant, K and α for linear starch chains in the eluent of DMSO/LiBr at 80 °C are 1.5 × 10− 4 dL g–1 and 0.743, respectively (Vilaplana and Gilbert w (logX). Although of course SEC signals vary with elution times in different runs, such variation is explicitly taken into account by the SEC calibration used here; that is why one must never present SEC data as a function of elution volumes  was dissolved in 1 mL DMSO/LiBr  at 80 °C overnight, and the solution was transferred into SEC vials for SEC analysis. The SEC separation was carried out with a Shimazu LC20 system  with a combination of Gram pre-column, Gram 1000 column and Gram 100 column  in sequence. The eluent used in this study was DMSO-LiBr solution , and the flow rate was set at 0.6 mL/min. The column temperature was set at 80 °C and the differential refractive index (DRI) detector temperature was set at 45 °C. The DRI signal was calibrated with a series of pullulan standards ranging from 342 to 2.35 × 10X where there is a clear division between short amylopectin chains and long amylose chains, at X ~ 100) to the AUC of the entire starch composition  of amylose composition , the number distribution of chains containing X monomer units after debranching. The relation between the number and weight distributions is w (logX) = X2Nde(X)  was labeled with 1.5 μL APTS solution  and 1.5 μL sodium cynoborohydride, and incubated in a thermomixer at 60 °C for 4 h in the dark. Then 80 μL deionized water was added to the sample, and the solution was vortexed at a low speed to until dissolution was complete. 50 μL of the solution was transferred to FACE micro-vials for analysis, using a P/ACE MDQ plus system . The analysis was conducted following the method of  were fitted with the Wu-Gilbert model , β(ii), β(iii), β(iv), β(v), β(vi), h(iii/i) and h(v/i). The fit uses publicly available code , βAm,(ii), βAm,(iii), hAm, (i), hAm,(ii) and hAm,(iii). Again the fitting was performed with publicly available code  and model-fitted  number chain-length distributions (Nde(X), arbitrary normalization) of debranched amylopectin (DP < 100) from sample 1 to 9. Figure S1-b. Experiment  and model-fitted  number chain-length distributions (Nde(X), arbitrary normalization) of debranched amylopectin (DP < 100) from samples 10 to 19. Figure S2-a. Experiment  and model-fitted (“fit”) weight chain-length distributions (w (logX), arbitrary normalization) of debranched amylose (DP > 100) from samples 2 to 10. Figure S2-b. Experiment  and model-fitted (“fit”) weight chain-length distributions (w (logX), arbitrary normalization) of debranched amylose (DP > 100) from samples 11 to 19.


In this study, GLXN and GTR were cultivated in the dark only in deionized H2O, and their shoot and root growth, starch in situ degradation, and starch property changes were investigated during seedling growth. Compared with GLXN, GTR showed a significantly slow seedling growth, which was not due to the embryo size and vitality. The slow degradation of starch in the seed restrained the seedling growth. GLXN starch was completely degraded gradually from the proximal to distal region of the embryo and from the outer to inner region in the endosperm, but GTR starch in the peripheral region of the endosperm was not completely degraded, and the starch residual was located in the outside of the compound starch though its degradation pattern was similar to GLXN. During seedling growth, GLXN starch had the same A-type crystallinity and a similar ordered structure, but the crystallinity changed from the CA-type to B-type and the ordered structure gradually increased in the GTR starch. The above results indicated that GTR had a heterogeneous starch distributed regionally in the endosperm. The starch in the peripheral region of the endosperm had a B-type crystallinity, which was located in the outside of the compound starch and significantly increased the resistance to in situ degradation, leading to the seedling slow growth.High-resistant starch cereal crops with the inhibition of the starch branching enzyme (SBE) have been widely studied. However, the effects of the inhibition of SBE on waxy cereal crops are unclear. A transgenic rice line (GTR) derived from a  Starch is the major storage carbohydrate in cereal endosperm and provides energy and nourishment for humans. In normal cereal crops, starch mainly consists of amylose and amylopectin. The content and structure of amylose and amylopectin determine the physicochemical properties of starch and influence the applications of cereal seeds ,2. Amylosbe I mutant with changed amylopectin structure and amylose content, seed starch is resistant to amylase hydrolysis during grain germination and seedling growth, leading to the inhibition of seedling growth [2O, the slow degradation of seed starch, which is due to the high resistance of amylopectin long branch-chains and B-type crystallinity to in situ degradation, restrains the growth of the shoot and root [For cereal crops, endosperm starch provides energy and nutrition for grain germination and seedling growth. For normal cereal crops, seed starch can be homogeneously degraded gradually from the embryo and aleurone layer during seedling growth ,13,14. Hg growth ,15. Its and root  thought So far, the high-RS cereal crops with the inhibition of SBE expression or mutation of SBE genes all have high amylose content . The higjaponica waxy rice cultivar Guang-ling-xiang-nuo (GLXN) has been developed through antisense RNA inhibition of both SBEI and SBEIIb [SBEIIb to also play a role in SBEIIa expression due to the sequence similarity [2O. Our objective was to reveal the in situ degradation and characterization of endosperm starch in waxy rice with inhibition of SBE during seedling growth.A transgenic rice line (GTR) derived from a d SBEIIb . The defd SBEIIb . In GTR,d SBEIIb . The redmilarity ,19. In t2O according to the method by Shaik et al. [2O. The grain germination began from 2 days after imbibition (DAI) and was slightly slower in GTR than in GLXN. The seedling growth, including shoot height and root length, was significantly slower in GTR than in GLXN from 4 DAI. The shoot height, shoot weight, and root weight were further quantitatively measured on the 30 seed basis and on the same weight basis of pre-germinated dry seeds  showed that the decreased seed weight and seedling growth had a highly positive relationship, and the degraded seed materials were used for seedling growth. Compared with GLXN, the slow growth of the GTR seedling in the dark only in deionized H2O resulted from the slow degradation of seed materials. Similar phenomena are also observed in high-amylose rice and barley with an inhibition of SBE expression [The seed dry weight was measured during seedling growth on a 30 seed basis and on the same weight basis of pre-germinated dry seed A,D. The pression ,14.R ≥ 0.991) also indicated that the degraded starch was used for seedling growth . Duringndosperm . In the Residual starch in seed was isolated during seedling growth. The iodine absorption spectrum of the starch is shown in A-type starch. With the seedling growth, the shoulder peak at 18° 2θ gradually becomes weak and vanishes, the peak at 23° 2θ gradually widens and becomes two peaks at 22° and 24° 2θ, indicating that the starch changed from the CA-type to the B-type with the growth of the seedling. A similar phenomenon has been reported in high-amylose rice with the inhibition of the SBE expression [The X-ray diffraction (XRD) patterns of starches are shown in pression . The shopression ,14.−1 are associated with the ordered and amorphous regions in starch, respectively [−1 gradually decreased  spectra of starches are shown in ectively . For GLXecreased B. The prjaponica waxy rice cultivar GLXN and its derived transgenic rice line GTR were used in this study. The GTR was generated by the antisense RNA inhibition of both SBEI and SBEIIb [The d SBEIIb . They we2O at 28 °C in the dark for 2 days with changes of water three times a day. The germinated grains were transferred into a 96-well plate with the embryo up and it continued to grow at 28 °C in the dark with the lower two-thirds of the grain immersed in deionized H2O. During seedling growth, the water was changed every day. The seedlings were taken out at 1, 4, 8, 12, and 16 DAI, and the grains at 1 DAI were used as the control.Rice grains were imbibed in deionized HThe length from the culm base to the tip of the longest leaf of the seedling was measured as the shoot height. The shoot and root were separated from the grain and dried at 110 °C for 3 h and 80 °C for 2 days in an oven. The germinated grains with the shoot and root removed were freeze-dried, and then they were dehulled carefully to obtain the seeds. The dry weights of the shoot, root, and seed were weighed. The seeds were ground extensively into flour and filtered using a 100-mesh sieve. The starch content in the flour was measured using a Total Starch Assay Kit , and then converted to the weight of the endosperm starch.The mature embryo was in vitro cultured exactly following the method by Pan et al. . BrieflyThe section of the whole seed was prepared following the method by Zhao et al.  with somg for 10 min. The starch precipitate was washed 3 times with deionized H2O and 2 times with absolute ethanol, freeze-dried, and filtered through a 100-mesh sieve.The starch was isolated from the seeds following the method by Pan et al. . BrieflyThe starch-iodine absorption spectrum was measured following the method by Lin et al. . BrieflyThe starch was analyzed using an X-ray powder diffractometer  as previously described . The samThe short-range ordered structure of the starch was analyzed using a Varian 7000 Fourier transform infrared (FTIR) spectrometer exactly following the method by Pan et al. .The data reported in all Figures and Tables were mean values and standard deviations. Analysis of variance (ANOVA) by Tukey’s test was evaluated using the SPSS 16.0 Statistical Software Program .A-type to B-type and the ordered structure gradually increased in GTR starch. Therefore, the B-type crystallinity located in the outer part of the compound starch had a high resistance to in situ degradation in GTR endosperm and inhibited seedling growth.Compared with GLXN, GTR showed a significantly slow seedling growth. The slow degradation of starch in GTR seed restrained the seedling growth. GLXN starch was homogeneous in endosperm and completely degraded gradually from the proximal to the distal region of the embryo and from the outer to the inner part in the endosperm. However, GTR starch was heterogeneous in the endosperm, and the starch located in the outer layer of compound starch in the peripheral region of the endosperm was not completely degraded. During seedling growth, GLXN starch had the same A-type crystallinity and similar ordered structure, but the crystallinity changed from the C


International Journal of Telerehabilitation (IJT) positioned among journals in the telemedicine and telehealth space? IJT is an online, open  journal that focuses on telerehabilitation -- a segment of telehealth and telemedicine that encompasses many rehabilitation associated disciplines. Because successful telerehabilitation often depends upon interprofessional collaborations, much of the work published within IJT is by at least two authors from different disciplines. Authors in the current issue include persons with collective expertise in the disciplines of architecture, occupational therapy, medicine, physical therapy, and/or software engineering.Where is the An examination of the subject matter published in IJT since its inception in 2008 provides insight into the evolution of telerehabilitation. Readers will note that several current issue papers describe telerehabilitation services delivered in the home environment, with one investigating in-home modifications. Two articles studied the provision of telehealth to persons post-stroke. The therapeutic use of virtual environments is a relatively newly prominent theme.We posit that multiple factors influence emerging research themes: reimbursement for services rendered, regulations, and most importantly in the case of research articles, trends in research funding. International submissions to the IJT have also shifted, sometimes between continents. The international geographic trends are interesting to follow.In the current issue, readers will note cautions related to safety and patient selection. Moreover, since some telerehabilitation protocols are best hybrid , IJT welcomes articles that include research conducted in authentic, hybrid conditions. Appropriate patient selection for telerehabilitation services is broached in this issue, and remains a challenge that requires future research.Sincerely,Ellen R. Cohn, PhD, CCC-SLP, ASHA-F, IJT EditorJana Cason, DHSc, OTR/L, FAOTA, Senior Associate Editor


This study was conducted to test the hypothesis that heat-bonding with a liner positively affects the bond strength and fracture resistance of an implant-supported glass–ceramic crown bonded to a zirconia abutment produced by a computer-aided design/computer-aided milling (CAD/CAM) procedure. Lithium disilicate-reinforced Amber Mill-Q glass ceramic blocks were bonded to 3 mol% yttria stabilized tetragonal zirconia polycrystal (3Y-TZP) blocks by heat-bonding with a liner or cementation with a dual-cure self-adhesive resin cement for a microtensile bond strength test. CAD/CAM implant-supported glass ceramic crowns were produced using Amber Mill-Q blocks and bonded to a milled 3Y-TZP zirconia abutments by heat-bonding or cementation for a fracture test. A statistical analysis was conducted to investigate the significant differences between the experimental results. The mode of failure was analyzed using high-resolution field emission scanning electron microscopy. Chemical bonding was identified at the interface between the zirconia ceramic and liner. The mean tensile bond strength of the liner-bonded group was significantly higher than that of the cement-bonded group. The initial chipping strength of the liner-bonded group was significantly higher than that of the cement-bonded group, although no statistically significant difference was found for the fracture strength. The mode of failure was mixed with cohesive fracture through the liner, whereas the cement-bonded group demonstrated adhesive failure at the interface of bonding. Implant-supported crowns restore oral functions and aesthetics without affecting the integrity of adjacent teeth. These crowns are usually connected to the implants by means of titanium abutments. However, the use of a metallic abutment may compromise the gingival aesthetics of implant crowns, particularly in patients presenting with a thin biotype gingival architecture. The grayish metallic color can be pronounced with a light reflected from the metallic surface of the abutment ,2. In adCeramic abutments are biocompatible and aesthetic by mimicking the color of natural teeth ,5. In adZirconia abutments are commonly designed as one-piece system to support implant crowns and establish the implant–abutment interface with a zirconia ceramic. This system eliminates the negative effects of titanium abutments related to aesthetics. However, the implant–abutment interface may experience an excessive wear under occlusal loading due to dissimilarities between the zirconia ceramic and the titanium implant ,14. CompThe two-piece system of zirconia abutments was designed to avoid the possible consequences related to the use of the one-piece system. This system consists of a ceramic core and a titanium link to establish the implant–abutment interface with titanium-to-titanium without compromising the aesthetics of the ceramic abutment. The ceramic core is customized using CAD/CAM technology and is bonded to the titanium base with adhesive resin cement . This twHowever, problems may occur when bonding a final prosthesis to a zirconia ceramic abutment ,21,22. TA lithium disilicate-reinforced liner has recently been developed to overcome the weak linkage of the final prosthesis of a glass–ceramic to a zirconia abutment designed to replace single anterior and posterior teeth. However, no scientific research has been conducted to elucidate the bond characteristics of the liner. The objective of this in vitro study was to investigate the effect of heat-bonding with the liner on the bond strength and fracture resistance of an implant-supported glass–ceramic crown bonded to a two-piece system zirconia abutment produced by a CAD/CAM procedure. The null hypothesis was set to test no significant difference between the liner-bonding and the cement-bonding of the implant-supported ceramic crown to the zirconia abutment on the initial chipping and fracture strengths of crowns, as well as the microtensile bond strength.Three CAD/CAM 3Y-TZP zirconia ceramic blocks  were machined to fabricate 6 zirconia ceramic specimens with dimensions of 10 × 10 × 5 mm, and they were sintered in an electric furnace  at 1450 °C. In addition, lithium disilicate-reinforced Amber Mill-Q glass ceramic blocks  were machined to fabricate 6 glass ceramic specimens matching the zirconia ceramic specimens (10 mm × 10 mm × 5 mm).2O, 55–65 wt.% SiO2, and 5–25 wt.% other trace elements of oxides and colorants was applied on the surfaces of 3 zirconia ceramic specimens. Then, heat-bonding with 3 lithium disilicate-reinforced glass ceramic specimens was conducted at 800 °C. For bonding with the liner, the liner , consisting of 5–15 wt.% LiTM Sil, 3M/ESPE, Seefeld, Germany) coated. The surfaces of 3 zirconia ceramic specimens were roughened using an air-borne particle abrasion technology with 50 µm alumina particles  under the pressure of 3 atm at a distance of 10 mm [TM plus, BISCO, Schaumburg, IL, USA) was coated on the surface of zirconia ceramic specimens [TM U200, 3M/ESPE, Neuss, Germany) with an equal amount of base and catalyst pastes mixed for 20 s. The cement was applied to the prepared ceramic surface and a pressure of 49 N was applied to the ceramic specimens under a constant-load device. After the removal of excess cement, the resin cement was photopolymerized using an light emitting diode (LED) curing unit  for 20 s from each of the four directions for a total of 80 sec under a light intensity of 550 mW/cm2 at a distance of 2 mm. The specimens were kept under pressure for additional 10 min, relieved from the constant-load device and immersed in distilled water for 24 h.For bonding with a resin cement, 3 lithium disilicate-reinforced glass ceramic specimens were acid-etched with 9.5% hydrofluoric acid gel  for 30 s, washed with distilled water, dried, and silane primer  to produce a 1 mm thick plate, and the sliced plate was cut using a low-speed diamond cutting machine  after rotating 90° for second set of cuts. Twelve specimens (1.0 mm × 1.0 mm × 10 mm) were prepared with 3 ceramic blocks per each group by selecting 4 specimens from the middle of a bonded ceramic block (10 mm × 10 mm × 10 mm). Thus, a total of 24 specimens (12 × 2 groups) were used for the bond strength test. 2) of each test specimen.The prepared ceramic specimens were attached to the grip of metal holders, mounted, and subjected to tensile force in a universal testing machine  at a crosshead speed of 0.5 mm/min ,26. The The mandibular right first molar was scanned using a D900L scanner  to create an implant-supported crown. Twenty ceramic crowns were produced by milling a lithium disilicate-reinforced glass ceramic block using CAD/CAM milling machine  to make 10 samples per group for the liner-bonded and cement-bonded groups. The internal aspect of each crown was acid-etched for bonding to a zirconia abutment presenting with a diameter of 4 mm and length of 8 mm. The bonding was performed using the same method as described in the section of the bond strength test for each liner-bonded and cement-bonded group. The zirconia abutments were connected to the titanium link abutments  with a dual-cure resin cement after applying an MDP containing primer. The prepared crowns were coated with a thin layer of e.max Ceram glaze paste  and glazed in an electric furnace (Programat EP3000/G2). The temperature was raised to 730 °C at a rate of 35 °C/min and held for 1 min. For the liner-bonded group , the ceramic crowns were glazed following the bonding with the zirconia abutments. However, for the cement-bonded group , the ceramic crowns were glazed prior to bonding to the abutments.TM Z350 XT, 3M ESPE, MN, USA), and the prepared specimens were immersed in distilled water at 37 °C for 24 h. The prepared ceramic crown/abutment specimens were connected to titanium implant analogs  with titanium abutment screws and tightened to 30 N·cm twice at 30 seconds interval using a torque gauge  . The scrThe crowns were supported and secured to a base retainer and loaded axially to fracture in a universal testing machine . The compression load was transferred through a 5 mm diameter steel ball positioned in the central fossa of the crown. The load was applied at a crosshead speed of 0.5 mm/min and continued to the point of fracture, and the fracture load of each ceramic crown was recorded ,29. The High-resolution field emission scanning electron microscopy  was used to investigate the topography of the liner-bonded surface, cross-sectional bonded interfaces, and fractured surfaces after etching with a 9.5% hydrofluoric acid gel for 30 s. The liner layer was removed with a 5% hydrofluoric acid solution for 30 min to identify the effects of liner treatment on zirconia ceramic. The distribution of the chemical elements at the bonded interface was analyzed using an energy dispersive X-ray spectrometer , and the crystal structure on the liner-bonded surface was investigated by X-ray diffraction .p < 0.05). A Weibull analysis was performed with the experimentally measured load-at-failure values for each group of bonded ceramic specimens.The statistical analysis was conducted by SPSS software . A Student’s t-test was conducted to investigate significant differences between the experimental results for the 2 test groups (p < 0.05). In addition, the Weibull distribution showed a matched tendency with a single mode (r2 > 0.958) (The null hypothesis on the microtensile bond strength was rejected. > 0.958) . The heat-bonding with the liner was found to induce a chemical interaction with the milled zirconia ceramic abutment. A large number of pores was found in the reaction layer of liner bonding, altering the surface topography of the zirconia ceramic a,b. The 2SiO3) and zirconium silicate (ZrSiO4), as well as zirconia (ZrO2) and silica (SiO2)  .The interfacial layer created by the liner-bonding was found to be more consistent than that created by the cement-bonding. The liner-bonded interfacial layer was thicker with a higher mean value and standard deviation (33.6 ± 5.2 µm) than the cement-bonded interfacial layer (13.3 ± 1.6 µm) . NeitherWhen the debonded surfaces of the liner-bonded and liner-bonded groups were visually evaluated with high magnification images of HR FE-SEM, the mode of failure of the liner-bonded group was mixed with cohesive fracture propagated through the liner . In contp < 0.05), although no significant difference was found in the fracture strength.The null hypothesis was rejected on the initial chipping and fracture strengths but accepted on the fracture strength. The mean values and standard deviations of the initial chipping strength and fracture strength of implant-supported glass ceramic crowns bonded to zirconia ceramic abutments were 843.8 ± 317.5 N and 1929.6 ± 191.1 N for the liner-bonded (MLG) groups and 341.0 ± 90.2 N and 1711.1 ± 275.4 N for the resin cement-bonded (MGC) group . The iniWhen the fractured surfaces of the lithium disilicate-reinforced glass ceramic crown and the zirconia ceramic abutment were visually evaluated with a high magnification images of HR FE-SEM, the mode of failure of the MLG group was mixed with adhesive and cohesive fractures propagated through the liner b–d. HoweThe liner-bonded ceramic crowns were superior to the cement-bonded ceramic crowns in developing a higher resistance to fracture. The bond strength of the lithium disilicate-reinforced glass ceramic to the zirconia ceramic was higher when the bonding was established with the lithium-disilicate liner than with an adhesive cement. The mean values of microtensile bond strength for liner bond and cement bond were 19.6 and 47.7 MPa, respectively, in this study. As an ideal biomaterial would have bonding forces included in the interval of 5–50 MPa, even if these values are mostly theoretical, both the liner and cement bond tested in this study exhibited bond strength within these limits . The linThe bonding of ceramic restorations commonly involves acid etching and/or sandblasting procedures ,33. In tThe one-piece system of zirconia abutments was found to present with some degree of misfit at the implant–abutment interface because of the challenge of the manufacturing process of the ceramic abutment ,42. UndeThe two-piece system of zirconia abutments involves using a titanium base to establish an interfacial connection with similar materials to the implant to avoid possible complications resulting from the use of dissimilar materials ,22. ThesThe fracture resistance was higher when the zirconia abutment was designed to receive the support of the titanium base . Gehrke Sandblasting is a common method of surface treatment used to bond zirconia ceramic restorations. The effect of this mechanical method, however, is limited to creating desired undercut features and increasing bond strength because of the dense polycrystalline structure of the zirconia ceramic ,50. HoweThe mode of failure was mixed with cohesive fracture propagated through the liner in the MLG group, whereas the failure was adhesive at the bonded interface of the ceramic and resin cement in the MGC group. The cement layer was thin at the interface and contained numerous pores and gaps. Meanwhile, no gap or pore was noted at the interface of the heat-bonding with the liner. These characteristics of bonding and mode of failure might have played a critical role in developing a higher fracture resistance of the implant-supported glass ceramic crown combined with a two-piece system of a zirconia abutment produced by a CAD/CAM procedure. However, a clinical study is yet to be conducted to determine the clinical success of the glass ceramic crown bonded to the zirconia abutment by means of heat-bonding with the liner.The bond strength and fracture resistance of a milled lithium disilicate-reinforced glass ceramic bonded to a milled zirconia ceramic are affected by the modification of bonding procedures. The bond strength and fracture resistance were significantly higher when the bonding was established by means of heat-bonding with the lithium-disilicate liner than by a resin cement. The pattern of interfacial failure for the liner-bonded group was mixed with the fracture propagated through the liner. However, after visual inspection, the cement-bonded group demonstrated an adhesive failure at the interface of bonding. The results of this study suggest that heat-bonding with the liner can be an alternative to bond CAD/CAM-produced glass ceramic crowns to zirconia ceramic abutments in order to reduce the risk of crown dislodgement and fracture.


Although starch based materials have an array of fascinating industrial applications, the native starches do not show good mechanical strength, thermal stability, and rheological properties for their use in the mainstream processing industry. For example, the use of starches for producing controlled release fertilizers is a new research endeavor with detailed knowledge still to come. The thermal processing of native starches with water as a plasticizer results in poor physical and pasting properties of the final product. Therefore in this study, corn starch was thermally processed with urea and borate in a water medium. The pure starch (PS), starch-urea (SU), starch-borate (SB), and starch-urea-borate (SUB) samples were prepared and characterized for their rheological traits. The PS sample exhibited a peak viscosity of 299 cP after 17 min of thermal processing. Further heating of the suspension caused a decrease in viscosity of 38 points due to thermal cracking of the starch granules. A similar trend was depicted in the viscosity measurements of SU, SB, and SUB adhesives. However, the viscosity of these samples remained slightly higher than that for PS. Also, the reduction in viscosity after the peak value was not as notable as for PS. The modified starch behaved like a gel and its storage modulus was significantly higher than the loss modulus. The lower magnitudes of storage and loss moduli revealed that the modified starch was in the form of a weak gel and not a solid. The PS is more fluid in nature with dominating loss modulus at lower angular frequencies. Starch is a tasteless white-colored carbohydrate powder, which occurs in the form of amylose and amylopectin in tubers, seeds, and other parts of plants. Amylose, a linear polymer, is the simplest form of starch while amylopectin is its branched form. Applications of pure and modified starches have been reported by many researchers during the past few decades ,2. The sStarch derivatives are usually produced by reacting some other components with the hydroxyl groups in the native starch molecules. The modified starch shows notably different physico–chemical properties from the parent starch without compromising the biodegradability ,11. SubsOwing to their unique thermal processing properties, starch-based polymers are much more complex than their conventional counterparts . MultiplSince the decomposition temperature of starches is reported to be lower than their melting temperature , the staIn this study, corn starch was thermally processed with urea and borate. The obtained adhesives were highly viscous and non-Newtonian in character, and therefore tested for their rheological traits. The urea was believed to work as a plasticizer other than the water and to facilitate the movement of polymer chains. The borate ions, produced during the dissociation of di-sodium tetraborate, were assumed to be the reactive species for cross-linking . The cro2B4O7·10H2O) of Analytical Reagent (AR) grade . The starch with degree of substitution of 0.81 was reacted with urea and borate in water medium. The degree of substitution of a polymer is defined as the average number of substituent groups attached with a base unit or a monomeric unit. Six samples of the modified starch were prepared and characterized for their physical and rheological traits. Details of the formulation of theses starch based adhesives are given in The food grade corn starch was purchased from the local market  and dried under sunlight to removal the moisture content. The plasticizer was 98% de-ionized water whereas the other constituents were urea and borate  sample was left to cool overnight at room temperature. For the starch-urea (SU) sample, first 5 g of starch was added to the hot water. The starch dispersion was heated for 10 min for better gelatinization. Thereafter, the solution was supplemented with 1 g of urea and heated further for 20 min to complete the reaction. A similar procedure was repeated to prepare the starch-borate (SB) and starch-urea-borate (SUB) samples by adding 1 g of each urea and borate in the starch dispersion at a processing temperature of 85 °C. The whole procedure was repeated again to modify the starch with 2 g of urea and borate and to study the effect of the modifier weight on the rheological traits.The effect of the modifiers on the properties of the modified starch samples was evaluated using different characterization techniques. The gelatinization time, viscosity, as well as storage modulus and loss modulus of the prepared samples were evaluated as a function of temperature and heating time. The controlled experiments were performed by dividing the samples into two groups: Pure starch control (PS) and experimental group of six modified starch compositions. The reliability of the results of viscosity, surface tension, density and other fluid flow parameters was checked by comparison between the controlled and uncontrolled measurements.TM operated viscometer  was used for viscosity measurements. It is a portable viscometer for advanced research for the most required fluid parameters. This fully automated viscometer was capable of measuring the viscosity of adhesives, polymers, stimulation fluids, drilling fluids, completion fluids, and cements at varying temperature, shear rate, shear stress and time. In the present case, the sample temperature and shear rate were varied from 20 to 100 °C and from 0 to 1000 s−1, respectively. For high accuracy in the measurements, the test temperature was achieved and maintained for 2 min before each run.An ORCADAThe surface tension measurements were carried out using a spinning drop tensiometer. This tensiometer was suitable for very low to high surface tension measurements. The equipment was fitted with a controlled temperature measuring cell. This removable measuring cell included a water coolable high dynamic measuring drive. Inside the measuring cell, helium gas was used as heat carrier to raise the temperature of the measuring capillary to the required level before recording the surface tension of the sample.Fourier transform infrared spectroscopy (FTIR) was used to produce infrared spectra and to understand the gelatinization mechanism of the pure and modified starches. FTIR is used to identify the bonds in a sample by producing an infrared absorption spectrum. It is also an effective analytical instrument for detecting functional groups and characterizing bond information. An Anton Paar density meter  was used to measure the density of the prepared samples under similar conditions set for both viscosity and surface tension measurements. The density measurements were based on the oscillating U-tube principle, which ensured high accuracy of the recorded data.The temperature at which the starch particles undergo a change in state by forming a gel is called the gelatinization temperature. This temperature provides useful information about the heterogeneity of the starch granules. The past reports reveal that most of the starches gelatinize at temperatures ranging from 80 to 90 °C ,18,19. IThe industrial applications of the starch based materials involve the suspension of the starches in water at temperatures slightly above their gelatinization temperature . SometimA further comparison of the PS and those modified with 2 g of the urea and borate is provided in The SEM micrographs of pre- and post-cracking of the corn starch are shown in The typical viscosity profile revealed the possibility of change in shape of the particles during thermal processing of the corn starch. The inset provides a photographic view of the thermally processed starch for 30 min. The viscosity breakdown in the time based curve of the pure starch, as suggested in A linear increase in viscosity during the early stage of heating is attributed to the swelling and amylose leaching from the starch particles. The viscosity profile attains the peak value after the majority of the particles have swelled out. A further heating of the suspension results in a decrease in viscosity due to break-down of the starch particles under the shear field of the instrument. The mechanical action of the instrument ruptures the swelled particles, in particular the native starch particles. After break-down the amylopectin molecules start to shear and solubilize by reducing the molecular weight of the amylopectin, subsequently, the adhesive viscosity ,28. In l−1. A new peak appeared in the FTIR spectrum of the SUB sample at a wavenumber of 1555 cm−1. This peak shows the presence of secondary amide in the SUB sample. In a secondary amide, the amido-group (nitrogen) is directly bonded to two carbon atoms. This secondary amide was produced during a polymerization reaction between urea and starch in the presence of borate. In response, molecular weight, density, viscosity and surface tension of the adhesive were improved in the presence of urea and borate in the starch suspension [Since modified starch samples revealed an almost similar trend in viscosity profiles, only PS and SUB samples were compared and discussed further. In SUB complex, it is believed that the di-sodium tetraborate molecules dissociate into sodium and borate ions . FTIR tespension .The viscosity of the concentrated dispersions, when subjected to the stress or shear rate, does not reach a steady state for some time due to the instability of the internal network structures. The structures, fragmented by the shearing, take time to rebuild. A steady state would appear in the respective viscosity plot soon after establishment of an equilibrium between the breakdown and rebuilding of the structures. The frequency sweep response of PS and SUB adhesives at 1% strain is compared in The low magnitudes, as depicted from the frequency sweep curves, indicate that the modified starch was in the form of a weak gel but not a solid. The PS was more fluid in nature with the viscous component being more dominant at the lower angular frequencies. One can say that in spray coating applications, these modified starch samples would produce larger droplets with steady jet breakup. Finally, the elastic and viscous components of the PS showed an almost similar increasing trend over changing angular frequency. Both parameters were in good agreement at higher angular frequencies. The reported literature reveals that thermally processed starches show shear-thickening response when the quantity of the modifier in the dispersion is high enough to cause particle crowding ,30.−1. This peak showed the presence of secondary amide, which was produced during the polymerization reaction between urea and starch in the presence of borate. In response, molecular weight, density, viscosity, and surface tension of the SUB adhesive were improved in the presence of urea and borate in the starch suspension.The presence of small amounts of urea and borate in a starch dispersion can significantly alter the physical properties of the final product. Urea works as a plasticizer other than water and facilitates the movement of the polymer chains. The unmodified starch attained peak viscosity after 17 min of heating at a fixed temperature of 85 °C. With further heating, the viscosity exhibited a slight decrease of 38 points over time. Although a similar trend was depicted in time based viscosity plots of modified starches, the peak viscosities of these samples were reported to be slightly higher than the pure starch. Also, the reduction in viscosity after the peak value was not as notable as for pure starch. A small difference between the peak point and end point viscosities of the modified starch samples suggested that the urea and borate modifiers significantly reduced the cracking of the starch particles. Therefore, the granules in modified starch suspensions retained their tightness and the breakdown was not as bad as in the case of PS. In the FTIR spectrum of SUB, a new peak appeared at a wavenumber of 1555 cmThe modified starches showed good stability whereas pure starch exhibited poor stability over time. A sharp decrease in storage modulus of pure starch after 300 s suggested an unstable gel structure, which breaks down after a certain interval of time. The modified starches behaved like weak gels and their storage modulus was significantly higher than the loss modulus. Contrarily, the pure starch showed dense fluid like character. The lower magnitudes, as depicted from the frequency sweep curves, revealed that the modified starch was in the form of a weak gel and not a solid. The pure starch was more fluid in nature with the viscous component being more dominant at lower angular frequencies.


Ara h 2, a peanut 2S albumin, is associated with severe allergic reactions, but a homologous protein, soybean 2S albumin, is not recognized as an important allergen. Structural difference between these proteins might explain this clinical discrepancy. Therefore, we mapped sequential epitopes and compared the structure of Ara h 2, Soy Al 1, and Soy Al 3 (Gly m 8) to confirm whether structural differences account for the discrepancy in clinical responses to these two proteins. Commercially synthesized peptides covering the full length of Ara h 2 and two soybean 2S albumins were analyzed by peptide microarray. Sera from 10 patients with peanut and soybean allergies and seven non-atopic controls were examined. The majority of epitopes in Ara h 2 identified by microarray are consistent with those identified previously. Several regions in the 2S albumins are weakly recognized by individual sera from different patients. A comparison of allergenic epitopes on peanut and soybean proteins suggests that loop-helix type secondary structures and some amino acids with a large side chain including lone electron pair, such as arginine, glutamine, and tyrosine, makes the peptides highly recognizable by the immune system. By utilizing the peptide microarray assay, we mapped IgE epitopes of Ara h 2 and two soybean 2S albumins. The use of peptide microarray mapping and analysis of the epitope characteristics may provide critical information to access the allergenicity of food proteins. The ability of some proteins to induce an allergic reaction in susceptible individuals is well established. However, it is not clear what characteristics confer various proteins the ability to induce allergic responses or why some proteins are more allergenic than others. Many factors have been suggested to contribute to the overall allergenicity of any given protein, such as the number of IgE-binding epitopes, resistance to heat and proteolysis, the glycosylation status and enzymatic activity ,2. The p2S albumins are known to be intrinsically more allergenic than many other proteins ,7,8. ThePeanut allergen Ara h 2, soybean 2S albumin 1 , and soybean 2S albumin 3 , which belong to the 2S albumin family, are evolutionarily close to each other. Sequence homology between peanut and soybean 2S albumins is about 40% and all of them are stabile during proteolytic digestion; however, their allergenic behaviors are very different ,19. Ara In the present study, we mapped sequential epitopes using peptide microarrays and compared the structure of Ara h 2, Soy Al 1, and Soy Al 3 at different levels such as primary, secondary and tertiary structure. Bioinformatics tools were used to investigate associations with the frequent discrepancies in clinical responses to these two proteins.In order to compare the sequential epitopes identified on peanut Ara h 2 with soybean 2S albumins, sera from peanut and soybean allergic subjects were screened against three sets of 15 overlapping peptides; each set encompasses the entire Ara h 2, Soy Al 1, and Soy Al 3 protein sequences. The microarray used in this study is a modified method documented previously as a reliable tool to identify sequential IgE epitopes on peanut allergens ,21. In oIn soybean 2S albumins, eight regions (a–h) of Soy Al 1 and Soy Al 3 are recognized by patients’ IgE . EpitopeAmino acid sequence-based structure analysis shows that the degree of similarity (identity) between Ara h 2 and Soy Al 1 and Soy Al 3 is 44.2% (35.6%) and 41.5% (34.8%), respectively. The composition of the helix-type secondary structure of Ara h 2, Soy Al 1, and Soy Al 3 are 51.28%, 54.19%, and 56.33%. The compositions of the loop type of Ara h 2, Soy Al 1, and Soy Al 3 are 48.72%, 45.81%, and 43.67%, respectively. Overall, the three proteins have similar primary and secondary values, which suggests that their structures are similar. Three-dimensional images of the three proteins are shown in Based on the comparison of structure, the secondary and tertiary structures are not good enough to explain the difference in reactivity to IgE between Ara h 2 and Soybean albumins. The number of amino acids comprising Ara h 2 epitopes is 105. Amino acids in Ara h 2 epitopes and equivalent peptides of soybean 2S albumins were analyzed for chemical characteristics. The relative contribution of each of the 20 amino acid types to the Ara h 2 epitopes and corresponding peptide in soybean 2S albumins are presented in The characterization of IgE epitopes of food allergens is fundamental to the understanding of mechanisms responsible for food allergies. In this study, we were interested in determining which structural characteristics of peptides are important for determining IgE binding epitopes. We chose Ara h 2 and soybean 2S albumins due to their apparent difference in clinical allergenicity and similarity in protein stability and sequence. First, we tried to identify the allergenic epitopes of Ara h 2 and soybean 2S albumins utilizing a microarray technology, which employed a sensitive Dendrimer detection system. Then, we analyzed the structural features of the epitopes on the two proteins to determine which structural characteristics might endow the protein with its allergenic properties. Patients clinically reactive to both soybean and peanut concomitantly were used so that IgE binding was comparable. We found eight areas on soybean 2S albumins that are recognized by IgE and some structural features of epitopes on Ara h 2 that differ from Soy Al 1 and Soy Al 3. This analysis suggests that loop-helix type secondary structures and some amino acid such as Arginine, glutamine, proline, and tyrosine on the food protein surface may be important factors in determining the IgE binding sites.In Ara h 2, twelve sequential IgE binding epitopes, three of which are immunodominant epitopes, are identified. These results coincide with allergenic epitopes reported in previous studies ,23,24. Ietc. [The characteristics that actually endow an antigenic or allergenic property are the center of much debate ,25,26,27etc. .et al. show that 10 sequential B-cell epitopes on Ara h 2 are primarily displayed on the molecular surface, especially the three immunodominant epitopes, which are located in a protruding extra loop [et al. mapped the epitopes of Brazil nut allergen, Ber e 1, onto a helix-loop-helix region [In our studies, most epitopes include the loop-helix structure, which is consistent with previous studies ,30,31,32tra loop . Alcocerx region . These rThe epitopes of peanut are enriched with polar amino acids. These results are similar to previous reports, which show that polar residues are commonly found on surfaces and within epitopes ,35. ArgiA peptide microarray assay allows characterization of large numbers of individual patient samples and examination of thousands of peptides on a single slide ,37. SimuThis study has some limitations. First, the main focus of the study was on identifying some characteristics of food allergy area; therefore, the study result might not be enough to be generalized to other area. Second, the study lacks in identification of epitopes that are associated clinical reactions because of absence of sensitized patients who tolerate peanuts . For theA/L were considered predictive of reactivity [Ten patients, who were allergic to soybean and peanut concomitantly, were included in the study . Diagnosactivity . Sera fr® technique by JPT Peptide Technologies . Peptides were printed in two sets of duplicates on SuperEpoxy glass slides  using the NanoPrint™ Microarrayer 60 . Additional array elements included protein printing buffer  as negative control spots for background normalization.A library of overlapping peptides, consisting of 15 amino acids with an offset of 3, corresponding to the primary sequence of Ara h 2, Soy Al 1 and Soy Al 3, were commercially synthesized using the PepStarImmunolabeling was performed, as previously described, but with some modifications ,29,44. I®Gx . Images were saved as a TIF format and the fluorescence signal was digitized with ScanArray Express Microarray Analysis System (PerkinElmer), exported as comma-delimited (CSV) files and transformed to z score as previously described [Immunolabeled slides were scanned using a ScanArrayescribed ,43,45. Bescribed , an epitA comparison of amino acid similarity was performed using Vector NTI Advance 10 software program (Invitrogen). PredictProtein Server was used to predict the consensus secondary structure and the relative solvent accessibility . SWISS-MIn this study, we have mapped epitopes of Ara h 2, Soy Al 1, and Soy Al 3 using a peptide microarray system, which made it possible to compare the characteristics of various epitopes. Loop-helix type structures and some amino acids with a large side chain including electron lone pair, such as arginine, glutamine and tyrosine, on the surface of the tertiary structure appear to comprise peptides with a higher potential for immune recognition; this seems to play an important role in antigen–antibody interactions. The overlapping peptide microarray system will greatly facilitate screening large numbers of individuals for epitope recognition. In turn, analysis of the characteristics of epitopes will give critical information to assess the allergenicity of various proteins. Our finding in this study might be limited in the area of food allergy and suggested one factor to determine antigen–antibody interaction. Therefore, further studies to identify the characteristics of epitopes and the factors to determine of antigen–antibody interaction are needed.


The basic elements can be adapted to consider shorteror longer time intervals, according to the area of knowledge.Bibliometric indicators are qualitative methods that allow the evaluation of scientificproduction through statistical techniques, and one of the most important indicators isthe impact factor. The basis for the impact factor is the Journal Citation Report (JCR).Available from Clarivate Analytics, it was developed by Eugene Garfield in 1958, whoseidea was to allow a practical analysis of the propagation of scientific information intheir respective areas of knowledge. To do this, Garfield determined a numerator: thenumber of citations in the current year for any items published in the journal in theprevious two years; and the denominator: the number of articles published in those twoyears,4. However, the impact factor is also being used toevaluate research performance and is applied at all organizational levels: authorship,institution, country/region, research field or journal, which distorts its application.To compensate for this, in 2018 the JCR reviewed journal profiles with a richer datacontext. One example is the bar chart that provides the value of the impact factor inthe Percentile Rank in Category item, allowing the quartile to be viewed quickly.The metric was developed for responsible use in journal management, allowing librariansto evaluate and select which journals to sign and/or discard, and editors to track andcheck the evolution of their journalsWhen analyzing the impact factor data from Brazilian Journal of Cardiovascular Surgery(BJCVS), they were promising: the impact factor was 0.805  and the BJCVS believes it is on the right track by prioritizing the quality of its publicationsand a structure that protects the authenticity of published work, what is reflected inthe journal indicators.This issue of BJCVS presents a blind peer-reviewed selection of 20 papers that wereselected by order of acceptance: 11 original papers, 1 review article, 4 selectedcase reports and 1 letter to the editor.Paulo Roberto B. Evora https://orcid.org/0000-0001-9631-946X1Editor-in-Chief Interim - BJCVS Faculdade de Medicina deRibeirão Preto da Universidade de São Paulo (FMRP-USP), RibeirãoPreto, SP, Brazil.Andreia C. Feitosa do Carmo https://orcid.org/0000-0002-0387-79462Hospital São Paulo, Escola Paulista de Medicina da UniversidadeFederal de São Paulo, (EPM - UNIFESP), São Paulo, SP, Brazil.Camila Sáfadi Alves Gonçalves https://orcid.org/0000-0003-0611-72293Sociedade Brasileira de Cirurgia Cardiovascular, São Paulo, SP, Brazil.Domingo M. Braile https://orcid.org/0000-0001-7704-22584Editor-in-Chief - BJCVS Faculdade de Medicina de SãoJosé do Rio Preto (FAMERP), São José do Rio Preto, SP, Brazil andUniversidade de Campinas (UNICAMP), Campinas, SP, Brazil.


Betatropis formosana, two new species , Plectoderini sp. and Paracatonidia sp., were sequenced for the first time through next-generation sequencing. The five mitogenomes ranged from 15,214 to 16,216 bp in length, with the typical gene content and arrangement usually observed in Hexapods. The motif “ATGATAA” between atp8 and atp6 was found in all the analyzed species. An overlap “AAGCTTA” between trnW and trnC was observed in the mitogenomes of most Fulgoroidea. The structural and compositional analyses of 26 Fulgoroidea mitogenomes, including the gene rearrangement of five tRNAs , the A + T content and AT-skew of the whole mitogenomes, and the nuclear acid and amino acid compositions of the protein-coding genes (PCGs), revealed family-level differences between Delphacidae and other families . Phylogenetic analyses of 13 protein-coding genes from 26 Fulgoroidea species by maximum likelihood and Bayesian Inference were consistent and well supported the basal position of Delphacidae, a close affinity among the families Flatidae, Issidae and Ricaniidae, and a close relationship between Achilidae and Fulgoridae.In the present study, the complete mitochondrial genomes (mitogenomes) of five Achilidae (Hemiptera: Fulgoroidea),  Family Achilidae is a medium-sized group of the superfamily Fulgoroidea (order Hemiptera), called planthoppers. They are worldwide distributed, mainly tropical and sub-tropical regions, and have higher regional endemism than most species in Ricaniidae and Cixiidae of Fulgoroidea, Aphidoidea and Coccoidea of Sternorrhyncha, and Heteroptera . Like otnad1-6 and nad4l for NADH dehydrogenase subunits one to six, and 4l; atp6 and atp8 for ATP synthase subunits six and eight; cox1-3 for cytochrome c oxidase subunits one to three; cob for cytochrome b), 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs: rrnS and rrnL). In addition, it also includes at least one sequence known as the A + T-rich region of variable length, which plays an important role in the process of transcription initiation and replication regulation  is generally a highly conserved, double-stranded circular molecule, ranging in length from approximately 14 to 36 kb . It contgulation . Becausegulation .In the last 40 years, first-generation sequencing technology, which is also called Sanger sequencing, has been a widely used technology. However, poor quality in the first 15–40 bases of the sequence due to primer binding, deteriorating quality of sequencing traces after 700–900 bases, and the base composition bias (high AT or GC content) and particular structure (poly and stem loop structures) of template DNA fragments have become common challenges of DNA sequencing and main reasons for lengthening research time. In recent years, with the rapid development of sequencing technologies, next-generation sequencing (NGS) technology has become a fast, low-cost and high-throughput method to generate mitogenomic data sets for phylogenetics and other biological questions .cox1, cob and rrnL) in Achilidae are available, hindering molecular phylogenetic analyses among Achilidae genera and species by using molecular data.Hemiptera has as many as 92,000 described species and exhibits the highest diversity among hemimetabolous insects . Up to dBetatropis formosana Matsumura, 1914 (B. formosana), Magadhaideus luodiana sp. nov (M. luodiana), Peltatavertexalis horizontalis sp. nov , Plectoderini sp. (Pl. sp.) and Paracatonidia sp. (Pa. sp.), from southwestern China using the NGS technology. In order to reveal the mtDNA molecular features, we also compared Achilidae mitogenomes obtained in the present study with other Fulgoroidea insects. Gene rearrangement, A + T content and AT-skew of 26 whole mitogenomes, and nucleotide and amino acid compositions of 13 PCGs revealed family-level differences between Delphacidae and the other five families . The nucleotide sequences of the 13-PCGs of these five species and 21 other Fulgoroidea insects were used for the maximum likelihood (ML) and Bayesian inference (BI) to investigate the phylogenetic relationships among Achilidae and the other Fulgoroidea subfamilies. Both ML and BI trees well supported the basal position of Delphacidae, a close relationship of Achilidae and Fulgoridae, and an affinity of Ricaniidae, Flatidae and Issidae.In this study, we sequenced the complete mitogenomes of five Achilidae species, Adults of these five Achilidae insects were originally collected from Guizhou and Yunnan province in 2017. All samples were preserved in absolute ethanol immediately and stored at −80 °C in the Institute of Entomology, Guizhou University, Guiyang, China before morphological identification and DNA extraction.The morphological identification of these five insects were performed by Shiyan Xu. Light microscope images were obtained by a Leica M125 stereomicroscope equipped with a Nikon D7000 digital camera, and were digitally processed with Helicon Focus and Adobe Photoshop CS6 software . Sex of http://zoobank.org/. The LSID for this publication is: urn:lsid:zoobank.org:pub:D1E6581B-15A0-4277-B00E-0564ABD04DE1. The online version of this work is archived and available from the following digital repositories: PeerJ, PubMed Central and CLOCKSS.The electronic version of this article in portable document format will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new names contained in the electronic version are effectively published under that Code from the electronic edition alone. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank Life Science Identifiers (LSIDs) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix http://hannonlab.cshl.edu/fastx_toolkit/index.html), respectively. The clean data was then used for mitochondrial genome reconstruction by SOAPdenovo  according to the manufacturer’s instructions. Quality of the extracted DNA was checked on 1% agarose gel, sheared to 250–300 bp segments, A- and B- tailed and ligated to Illumina paired-end (PE) adapters. About 350 bp ligated fragments were quality selected on agarose gel and amplified to yield the corresponding short-insert libraries. Subsequently, HiSeq X 10 was used to sequence PE reads and the length of each read was 150 bp. Raw data was adapter clipped and qualified using tools fastx clipper and fastq_quality_filter in FASTAX-Toolkit (APdenovo  and GapFAPdenovo  with defhttp://mitos.bioinf.uni-leipzig.de/index.py)  . PCGs wendex.py) .http://lowelab.ucsc.edu/tRNAscan-SE) with invertebrate codon predictors (The identification and structure prediction of tRNAs were performed using MITOS and tRNAScan-SE Search Server available online (edictors . While tedictors . The preedictors .MH324927–MH324931 for Betatropis formosana (B. formosana), Magadhaideus luodiana sp. nov (M. luodiana), Peltatavertexalis horizontalis sp. nov , Plectoderini sp. (Pl. sp.) and Paracatonidia sp. (Pa. sp.), respectively.The base composition of nucleotide sequences was described by skewness and was measured according to the following formulas : AT-skewPomponia linearis and Meimuna opalifera in Cicadoidea, and Cosmoscarta sp. and Aeneolamia contigua in Cercopoidea as outgroups (https://www.phylo.org/)  . Four sitrnW, trnC and trnY; trnT and trnP) order of Delphacidae mitogenomes differed from the putative ancestral gene order observed in Achilidae, Flatidae, Fulgoridae, Issidae and Ricaniidae mitogenomes  with the remaining 14 genes on the light-strand (L-strand).Each of all five newly sequenced mitogenomes was a circular double-stranded DNA molecule , containogenomes . In contPa. sp.) to 16,216 bp (Pl. sp.)  , of whicM. luodiana to 77.7% in B. formosana. The high AT-skew values among analyzed mitogenomes (0.091–0.284) indicated the occurrence of more As than Ts, which was also observed in other examined Fulgoroidea mitogenomes  have been frequently reported in the mitogenomes of other insects. In the five newly sequenced mitogenomes, except for nylation .The average nucleotide composition analysis of the PCGs of Fulgoroidea mitogenomes showed that the PCGs in the H-strand had much higher AT-skew than those in the L-strand . MoreoveThe amino acid composition analysis of the proteins  coded byTheoretically, 20 amino acids in the mitochondria corresponding to 22 tRNAs and 62 codons, indicated that each tRNA must recognize at least two types of codon. However, RSCU indicated strong biases of codon usage and amino acid composition . The codtrnS1, which lacked dihydrouridine (DHU) arm and formed a simple loop, all the tRNAs could be folded into canonical clover-leaf secondary structure with an aminoacyl arm of seven base pairs and anticodon arm of five base pairs  by the L-strand. The length of tRNAs ranged from 55 to 72 bp. Except for the se pairs . Moreoveipterans . The WattrnK had the highest %INUC (81.7%), and four tRNAs  on the H-strand and three tRNAs  on the L-strand showed a higher %INUC (>70%) in five mitogenomes, while the %INUC of trnS1 located on the H-strand was only 25%. Moreover, in these conserved regions, six and five G–U wobble base pairs were observed in acceptor and DHU arm, respectively; while three  mismatched base pairs were found. The above results indicated a high-level conservation of tRNAs encoded on the H-strand.The percentage of identical nucleotides (%INUC) in the five mtDNAs was calculated . The trnrrnL) and small ribosomal RNA (rrnS) genes were oriented on the L-strand of the five mitogenomes, with a high A + T content (>70%), a negative AT-skew and a positive GC-skew  occurred between trnF and nad5 in Pa. sp. mitogenome. A seven-bp overlap (ATGATAA) between atp8 and atp6 found in other insect mitogenomes was also observed in the mitogenomes analyzed in this study  between trnW and trnC was detected in five newly sequenced mitogenomes, which was absent in species of Delphacidae  was much longer than those in the other four Achilidae species  were found in the A + T-rich region of l region . HoweverBecause of limited mitogenome sequences of Fulgoroidea, we included in the phylogenetic analyses only 26 species to understand the evolutionary relationships of Achilidae with the other Fulgoroidea families. Phylogenetic trees of ML and BI analyses were constructed based on 13 PCGs nucleotides sequences from 30 species ; Fig. 10In both ML and BI topologies, the basal position of Delphacidae was well supported with high node values (BPP/BS = 1/100), which was also supported by many previous studies based on morphological characters and molecular data . In bothPa. sp. to 1,850 bp in Pl. sp., than other regions. Furthermore, as the length of control region decreases among taxa, so it does the number of repeat units and their copy numbers. This feature indicated that characteristics of these regions in Achilidae were taxon-specific, and the different sizes or copy numbers of repeat units had some influences on the size of the control regions and also the mitogenomes. Except for nad4 and nad5, the length differences of each PCGs among five newly sequenced mitogenomes were no more than 20 bp, indicating relatively conserved features of protein-coding genes. The limited length variations in all tRNAs and rRNAs were also observed among different species, which was mainly due to their stable secondary structures.This study presents the description of five complete mitochondrial genomes of Achilidae species. Among these five mitogenomes, the A + T-rich region exhibited more variation in length, varying from 908 bp in The calculation of A + T content, AT-skew and GC-skew of 26 Fulgoroidea mitogenomes indicated a significantly biased toward A + T nucleotide, which was considered to be due to the abundance of adenines caused by the high ATP concentration , the higIn general, T was the most common nucleotide at second codon position (avg. 43%). The frequently usage of the fourfold degenerated codon NNA and the rarity of NNG in Fulgoroidea mitogenomes were also observed in other hemipterans , and mayThe length of the tRNAs is affected by the size of the TψC and the DHU arms which may have aberrant loop and very short stems varying in length within different tRNAs . The antIn both the ML and BI trees, the basal position of Delphacidae were supported with high probabilities. Placement of this family is consistent with previous hypotheses , and is The monophyly of a number of Fulgoroidea families recovered as monophyletic were supported by both morphological and molecular datas, but relatively limited taxonomic or biogeographic sampling could have contribution to the higher BS values, particularly for the families Achilidae or Delphacidae, which have greater taxonomic diversity in comparison to other planthopper families and occur in several biogeographic regions. The representative mitogenomes from Fulgoroidea are still limited, while phylogenetic analyses will be more reliable and convincing as more mitogenomes and genomes of the fulgoroidea species are available in databases.The phylogenetic structure of 26 Fulgoroidea species supported in most previous studies and this study  might beAlthough the first five mitochondrial sequences of Achilidae were sequenced, it was still insufficient in comparison to a total of 161 genera and nearly 520 species in this family. Furthermore, only 26 mitogenomes downloaded from Genbank were included in the phylogenetic analyses. Considering the limited representatives of Fulgoroidea mitogenomes, a denser taxon sampling is still needed for further structural and compositional analyses and also for molecular classification of Fulgoroidea.atp8 and atp6 was found in 26 Fulgoroidea species. Additionally, an 8-bp overlap “AAGCCTTA” between trnW and trnC was found for the first time in the mitogenomes of several Fulgoroidea taxa. In the 26 analyzed Fulgoroidea mitogenomes: the gene rearrangement of five tRNAs , the A + T content and AT-skew of the whole mitogenomes, and the nucleotide and amino acid compositions of the PCGs, revealed some family-level differences between family Delphacidae and the other five families . The phylogenetic relationships constructed by both ML and BI methods were consistent and supported the monophyly of Fulgoroidea, a close affinity among the families Flatidae, Issidae and Ricaniidae, and a close relationship between Achilidae and Fulgoridae. We believe that the mitogenomes of the five Achilidae insects will be useful for further understanding of the evolutionary phylogenetic relationship within Fulgoroidea or even Hemiptera.In the present study, we firstly sequenced and analyzed the complete mitochondrial genomes of five Achilidae species (Hemiptera: Fulgoroidea). The five mitogenomes ranged from 15,214 to 16,216 bp in length, with the typical gene content and similar arrangement of genes usually observed in Hexapods. The 7-bp overlap “ATGATAA” between 10.7717/peerj.6659/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.6659/supp-2Supplemental Information 2Click here for additional data file.


This paper presents a comparison on the effects of blending chitin and/or starch with poly(lactic acid) (PLA). Three sets of composites  with 92%, 94%, 96% and 98% PLA by weight were prepared. The percentage weight (wt.%) amount of the chitin and starch incorporated ranges from 2% to 8%. The mechanical, dynamic mechanical, thermal and microstructural properties were analyzed. The results from the tensile strength, yield strength, Young’s modulus, and impact showed that the PLA–chitin–starch blend has the best mechanical properties compared to PLA–chitin and PLA–starch blends. The dynamic mechanical analysis result shows a better damping property for PLA–chitin than PLA–chitin–starch and PLA–starch. On the other hand, the thermal property analysis from thermogravimetry analysis (TGA) shows no significant improvement in a specific order, but the glass transition temperature of the composite increased compared to that of neat PLA. However, the degradation process was found to start with PLA–chitin for all composites, which suggests an improvement in PLA degradation. Significantly, the morphological analysis revealed a uniform mix with an obvious blend network in the three composites. Interestingly, the network was more significant in the PLA–chitin–starch blend, which may be responsible for its significantly enhanced mechanical properties compared with PLA–chitin and PLA–starch samples. Polymers can be categorized into synthetic and non-synthetic, or biodegradable and non-biodegradable . The serStarch is a natural biopolymer which is abundantly available at low cost ,10. A blChitin is abundantly available in shrimps, crabs ,14, and Literature established that for the blending of PLA, the percentage of the polymer blend should be a ratio 1:9 by weight of the blend to ensure good miscibility . SeveralObviously, starch and chitin have been differently used by several researchers to enhance the mechanical and biodegradability of PLA ,22,23,242, respectively. The average molecular weight of PLA (4032D) is 100,000 g/mole. It has a specific gravity of 1.24, melt flow rate (MFR), g/10 min  of 7 and a melt density of 1.08 g/cc at 230 °C. Commercially available chitin was used  with a 90% degree of deacetylation. On the other hand, commercial high amylose starch from corn was supplied by Sigma Aldrich . The particle size distribution is presented in Commercial grade poly(lactic acid) (4032D) was supplied from Nature Works  in pellet form. PLA (4032D) has excellent grease and oil resistance as well as optical, machinability, twist and dead fold. PLA has a melting point between 150–170 °C and a glass transition temperature of 60–65 °C. The tensile strength, yield strength, elastic modulus and impact strength of the PLA used are 53.5 MPa, 60 Mpa, 3500 Mpa and 2.99 kJ/m3/h. The PLA powder was mixed with chitin and starch in the proportions shown in Poly(lactic acid) from Nature Works was ground into a powder using an AA 150 Power granulator . The ground PLA was then dried for 4 h at 60 °C in a Thermo Electron Luxor 50 Motan Desiccant Dryer  with dry air at 50 mThe composition variation for the PLA–starch composite was done based on previous studies ,22. It wThe composition variation for the percentage proportion by weight of PLA–chitin–starch in this study is shown in Tensile samples were prepared according to the American Society for Testing and Materials ASTM D3039 method for plastic using the Carver Press machine. The tensile test method was used with standard dimensions and loaded using an Instron Universal Testing Machine (model 5966)  with a force capacity of 10 kN for five replicates per sample. 2). The dimensions of an ASTM D256 standard specimen was used. The impact test was done using Ceast Resil Impactor 7181 , Akron, OH, USA), which uses an instrumented hammer. The test is to determine the energy absorbed in breaking. The energy required to break the specimen is obtained from the loss of energy in the pendulum. This energy is simply the difference in potential energy of the hammer before and after the impact. The values are reported in terms of absorbed energy per unit cross-sectional area at the notch  . Sample shape and dimension were prepared according to the ASTM D4065 standard for polymer composites at a frequency of 10 Hz and a temperature range of −50 °C to 150 °C.The thermal properties of the samples were measured using a PerkinElmer TG-IR-GCMS Interface Q500, TA Instruments . The result was plotted and analyzed with TA universal analysis software . Sample quantities between 20 mg and 21 mg were heated at a rate of 10 °C/min from room temperature to 600 °C under air.The microstructure of the fractured surface (after the tensile test) of samples were thoroughly investigated using SEM. The samples were carbon-coated, and microstructural examination was performed on carbon-coated samples using a JEOL Field Emission Scanning Electron Microscope .The yield strength of the composite samples of the blend of the three polymers (PLA–chitin–starch) has the highest value compared with PLA–starch and PLA–chitin. PLA–starch and PLA–chitin show the same trend as that of the tensile result. This result of yield strength for PLA–chitin is in accordance with . HoweverThe value of the elongation shows that the value for PLA–chitin–starch falls between that of PLA–chitin and PLA–starch as shown in The Young’s modulus value is a critical factor for high strength applications. The value of the Young’s modulus follows a different trend from the tensile strength and elongation. There is no significant trend with variation in composition for PLA–starch–chitin and PLA–starch, but samples have the highest values at 96% PLA–2% chitin–2% starch and 96% PLA–4% starch. However, it is worthy of note that the value of Young’s modulus for the PLA–chitin composite increases with decreasing chitin content. The results shown in  and 25]25] have The impact strength is a measure of the resistance of a material to the sudden force. The result of the impact strength for each of the samples is shown in The result of the dynamic mechanical analysis for the twelve samples is shown in The loss modulus indicates heat dissipated per unit deformation, and its peak is referred to as the glass temperature, Tg. The loss modulus and loss factor curve b,c show The Thermogravimetry Analysis (TGA) and Derivative Thermogravimetry Analysis (DTA) results for the twelve samples are shown in The change in composition can be seen to affect the melting peaks in the DTA curves and it is more obvious in the 94% PLA and 98% PLA set of samples, but lower for the 96% samples. As evident in The miscibility can be explained using the nature of the polymer blends. PLA is hydrophobic, whereas starch is hydrophilic, which makes them thermodynamically immiscible . HoweverThe fractured surface images of the samples are presented in The SEM images shows an uneven distribution in the 92% PLA–4% chitin–4% starch sample a, which The comparative analysis of the effect of PLA blending with chitin and starch shows that PLA–chitin–starch composites are miscible with each other without segregation of the particles. The composite, molded using two-step mixing and a Carver Press, results in a more uniform composite. The mechanical properties of the polymer composites increased significantly compared to pure PLA for all samples. The mechanical strength result of PLA–chitin–starch revealed a significant improvement compared with binary blends such as PLA–chitin and PLA–starch. However, the dynamic mechanical properties of the composites showed notable improvement with an increase in chitin content and decrease in starch content. Also, the DMA result shows the high amorphous flow of the polymer blend with excellent mechanical strength over a range of temperatures. The TGA/DTA result shows one single degradation curve with a single glass temperature, which conforms with the DMA result and indicates miscibility and compatibility of the three polymers. The SEM images show excellent blending properties with a dispersed network of starch and chitin. Overall, the combined effect of PLA–chitin–starch gave a better result than PLA–chitin or PLA–starch.


Patients with chronic obstructive pulmonary disease (COPD) have elevated cardiovascular risk, and cardiovascular disease is a major cause of death in COPD. The current literature indicates that changes in cardiovascular risk during pulmonary rehabilitation (assessed using aortic stiffness) are heterogeneous suggesting that there may be sub-groups of patients who do and do not benefit.To investigate the characteristics of COPD patients who do and do not experience aortic stiffness reduction during pulmonary rehabilitation, examine how changes relate to physical activity and exercise capacity, and assess whether changes in aortic stiffness are maintained at 6 weeks following rehabilitation.We prospectively measured arterial stiffness (aortic pulse-wave velocity), exercise capacity  and physical activity (daily step count) in 92 COPD patients who started a six week pulmonary rehabilitation programme, 54 of whom completed rehabilitation, and 29 of whom were re-assessed six weeks later.p = 0.34), 56% patients responded with a significant reduction in aortic stiffness. Change in aortic stiffness (absolute and/or percentage) during rehabilitation was associated with both increased physical activity  and change in exercise capacity , but in multivariable analysis most closely with physical activity. 92% of the responders who attended maintained this response six weeks later.Whilst on average there was no influence of pulmonary rehabilitation on aortic stiffness (pre- vs. post pulse-wave velocity 11.3 vs. 11.1 m/s Elevated aortic stiffness in COPD is potentially modifiable in a subgroup of patients during pulmonary rehabilitation and is associated with increased physical activity.ClinicalTrials.gov Identifier: NCT03003208. Registered 26/12/ 2016. Chronic obstructive pulmonary disease (COPD) is a leading cause of global morbidity and mortality . PatientIt is known that physical activity (PA) reduces CV risk. In coronary artery disease, exercise programmes reduce aPWV . ReducedWe approached 106 consecutive patients enrolling on the Central and North West London NHS Foundation Trust PR classes held at the Peckwater Center, and St. Pancras Hospital in London, UK. A total of 102  with a confirmed diagnosis of COPD (post-bronchodilator FEV1/ FVC < 0.70 and appropriate exposure history) were recruited  PR guidelines . The firComprehensive data were gathered from each participant including demographic and clinical information. COPD exacerbation frequency was defined as the number of events treated with oral antibiotics and or corticosteroids in the previous year. Breathlessness, quality of life, and anxiety & depression were assessed using CAT COPD , MRC dysThe following measurements were made at the start and end of PR, and the duration between these times and number of classes attended were recorded.Arterial stiffness was determined by measuring aPWV between the carotid and femoral arteries using Vicorder  equipment. Based on the manufacturer’s instructions, the participant was asked to lie at 45°. Then, both of the carotid and femoral cuffs were attached. The carotid cuff was positioned over the carotid palpation area and the femoral cuff was positioned around the upper right thigh. Next, the participant lay supine to measure the distance between the suprasternal notch and each of the femoral and carotid arteries. After the distance was recorded, the bed was raised back to 45° to start the aPWV measurement, expressed in meters per second. After 10 min rest, one aPWV set was measured for each participant (three readings a set). The mean of those measures was used. We have previously reported that aPWV is stable over time in patients with COPD  and thusParticipants were referred to PR with spirometry results confirming COPD. However, to ensure contemporaneous lung function results, we performed post-bronchodilator hand-held spirometry using a Micro 1 Handheld Spirometer  which conform to the requirements of the ATS/ERS standards  and thesThe ISWT was conducted in accordance to the ERS/ATS guidelines . ParticiWe asked participants to wear a step counter pedometer on their waist all the time whether inside or outside the PR class (except when sleeping and showering) and to record the total daily physical activity on a diary card. This was for the six-week duration of the class and for the subsequent six-weeks after PR completion. We used a Yamax SW-200 electronic pedometer, which has previously been shown to be a reliable and valid device –33. One Participants who completed the PR programme were asked to attend again six weeks after the end of PR for re-evaluation. Exercise was not supervised during this period; however, physical activity was monitored by providing a step counter pedometer. At the end of the six weeks, patients were re-assessed, completing the same evaluations described above.Data were assessed using histograms and tested for normality using the Kolmogorov–Smirnov test. Data are expressed as mean (SD) for normally distributed data or median (IQR) for non-normally distributed data as appropriate. We examined the change in aPWV pre- and post-exercise and classified our participants as responders (reduction of ≥0.5 m/s) or non-responders (less benefit than this).To complete a power calculation we contacted the authors of the largest previous study , who rep1 was 1.23 L (50% predicted). Table We approached 106 consecutive patients enrolling on two PR programmes in Camden, London, UK and 104 agreed to take part. Spirometry did not confirm COPD in two of these. The CONSORT diagram is illustrated as Fig. p < 0.001), CAT questionnaire  and mMRC dyspnoea score (3(2–4) vs. 3(2–3), p < 0.001).First, we wanted to confirm that the PR programme met standard goals. In the 54 completers there was a significant improvement from baseline to completion in mean (SD) ISWT  and physical activity (assessed using average step count).p = 0.020, Fig. p = 0.042, Fig. Change in aPWV related to both change in exercise capacity and to physical activity. In simple correlation analysis, there was a significant association between larger reduction in aPWV and greater change in ISWT  and that a greater proportion of the non-responders were already prescribed anti-hypertensive medication . The blood pressure in the responders reduced by a clinically and statistically significant degree: the mean fall in systolic BP was 6.30 mmHg (p = 0.043), diastolic BP was 4.94 mmHg (p = 0.041) and mean arterial pressure was 5.39 mmHg (p = 0.020). There were no alterations in any of the participant’s medications during the rehabilitation programme.We noted that the starting blood pressure was higher in the responders than the non-responders . There was not an independent effect of change in exercise capacity on pulse wave reduction (p = 0.19).We performed multivariable regression analysis to better understand the relationship between physical activity, exercise capacity and change in aortic stiffness during PR. For every 1000 additional steps walked during PR, adjusted for baseline aPWV, aPWV decreased by 0.2 m/s , and of the 13 people in this group only one patient experienced a subsequent rise in PWV of ≥0.5 m/s. The beneficial aortic stiffness reduction was therefore maintained in 92% of the responders who attended. Non-responders included those who had a change in aPWV within 0.5 m/s of baseline (the “no change” group) and those who had a > 0.5 m/s increase (the “increased” group) during PR. We subdivided our non-responders and showed that the “no change” group maintained similar aortic stiffness  and exercise capacity  at six weeks following PR completion. In contrast, the six patients who appeared to have an increase in aPWV during the PR programme showed a significant reduction in aPWV at six weeks , associated with a significant increase in exercise capacity during this period , perhaps suggesting a delayed response to PR in this group.We specifically wanted to examine whether those patients who experienced aortic stiffness reduction with PR maintained this response at six weeks. The mean aPWV was not different in the responders from the end of PR to the six week visit  and change in exercise capacity  suggesting that patients who maintain activity and gains in exercise capacity are those who maintain the aortic stiffness response achieved during PR.Finally, we wanted to assess over this second six-week period whether PA and change in exercise capacity still related to change in aPWV. In simple correlation analysis aPWV related to both physical activity . The responders had a clinically meaningful and statistically significant reduction in blood pressure. Change in aPWV was related to both change in exercise capacity during PR, and to physical activity outside of the class. Multivariable analysis confirmed that physical activity was the more important of these two factors. Six weeks later, 92% of the responders who attended maintained this benefit, with both average physical activity and change in exercise capacity in the six weeks following PR related to change in aortic stiffness over this period. Change in aortic stiffness was not restricted to those with or without a previous history of cardiovascular diseases or recognised cardiac risk factors. There was no alteration in potential confounders such as anti-hypertensive medication during PR. Our results emphasise the importance of physical activity on reduction of aortic stiffness in COPD.We have recently published a systematic review summarising studies examining the effect of PR on CV risk reduction measured by aPWV in COPD . This id2 max during a cardiopulmonary exercise testing test was independently associated with arterial stiffness in COPD subjects without CVD [People with COPD are known to be less active than healthy controls . Physicahout CVD .In our study, 56% of patients had a beneficial aortic stiffness response during PR, defined as a reduction of 0.5 m/s or greater in aPWV. Meta-analysis of > 12 000 subjects has demonstrated that a difference of ≥0.5 m/ s in aPWV corresponds to a 7.5% reduction in CV risk, such that a 0.5 m/s reduction in aPWV can be considered clinically meaningful . We therThe question of whether elevated aortic stiffness is modifiable in COPD is clinically important. Our study builds on studies suggesting that CV risk in COPD may be modifiable with respiratory interventions such as long-acting bronchodilators and inhaled corticosteroids , and notThe strengths of this study include the fact that patients with and without CV history were approached sequentially, and therefore that our subjects are representative of those attending PR classes in the UK. The completers were similar to the non-completers, except that they had slightly elevated aortic stiffness and COPD severity. All testing was performed to established guidelines and quality assurance, and arterial stiffness was determined by aPWV which is a gold standard method for measuring arterial stiffness. Our study was adequately powered, with an a priori power calculation and statistical plan. One of the limitations of our study is the relatively high dropout rate (36%). However, this is a recognised problem in pulmonary rehabilitation programmes and our rate is lower than the UK national COPD Audit data reporting a 38% dropout rate . The rean = 90, age 72.07 ± 9.92 years and FEV1 1.28 ± 0.54 L) monitored aortic stiffness in COPD patients over time in the absence of PR. In this study the proportion of participants who had a decrease in aortic stiffness of > 0.5 m/s was 30% compared to 56% in our study with the PR intervention. In our current study, non-responders were defined as those who did not achieve > 0.5 m/s reduction in aortic stiffness. This included those with no change  and those with greater than a 0.5 m/s increase in aortic stiffness. Among the 90 patients in the previous study with no PR intervention, the proportion of those with an increase of more than 0.5 m/s was 23/90 (26%) compared to 10/54 (18%) of our participants who attended PR. This indicates a downward shift in the whole population in aPWV in patients undergoing PR compared to those assessed over time with no intervention.A previous study in our department  recruitiOur results have important implications for clinical practice. First, they emphasise the value of encouraging people with COPD to maintain physical activity, and that encouraging this during PR may be associated with optimal, holistic benefits from PR in reducing aortic stiffness. Second, our study increases the evidence base for PR in that we have shown that PR can reduce aortic stiffness in a proportion of patients with COPD, likely through an effect on reducing blood pressure. Importantly this benefit was not restricted to those with (or without) established CV disease, and we provide the first evidence that a non-pharmacological COPD-targeted intervention may be able to modify aortic stiffness in COPD for at least six weeks.We report that elevated aortic stiffness in COPD is potentially modifiable in a subgroup of patients through pulmonary rehabilitation and that this reduction is related to physical activity. This improvement was maintained for at least six weeks following PR. Our results widen the accepted benefits of PR to include reduction of aortic stiffness, suggest that PR programmes should focus on maximising physical activity, and demonstrate that elevated aortic stiffness in COPD is modifiable with PR.


Twenty years have passed since the first number of The Journal of Headache and Pain Fig. . I can pWith a continuous daily teamworking we have waited until indexing services noticed us, we dared to choose the open access option when no one would have put a penny on it, we continued our run by continuously introducing innovative initiatives and procedures. In one word, we unceasingly tested our limits.Apart from the progressive, constant and therefore solid growth of the Impact Factor, I particularly care about how our readers have rewarded the Journal with about 1 million papers downloaded in 2019, and the increasingly permeating social activity.ça va sans dire together with SpringerNature, that has made available, always and in preview, its immense technological know-how and, certainly not least, together with the affiliated Scientific Societies, the European Headache Federation and Lifting The Burden, who have given us the privilege of hosting their best publications. A special thank to the reviewers, the real engine of the Journal, who have been able to add speed to the quality of the reports.We savor a moment of joy for this birthday together with the many, many scientists who over time have opted to publish their scientific works here, have shared the activities of the Boards of the Journal and Lastly, we will keep thinking The Journal of Headache and Pain as a privileged cradle for new ideas, other than the Thematic Series, Greppi Awards, EHF-School of Advanced Studies reviews, European Migraine Registry reports.Thanks to all of you we have built a new stylistic feature within our scientific community.