Korean Journal of Women Health Nursing (KJWHN), that the PubMed Central (PMC) Full-Participation Agreement and PMC banner for the journal were dispatched to PMC, and PMC received them on July 30, 2022. I was thrilled to hear the news because this is the first case of PMC accepting a non-English life science journal published in Korea. Seeing a Korean-language journal be accepted by PMC has always been my goal since August 2006, when I succeeded in producing PMC XML files [Child Health Nursing Research, which only publishes in English [I heard from Dr. Sue Kim, the editor-in-chief of the ML files . It is aIn retrospect, I was invited as a speaker at the 15th Summer Seminar of the Korean Society of Women Health Nursing on May 27, 2010. I suggested changing the journal’s language to English-only to add the journal to PMC. I made this suggestion because biomedical journals published in Korea began to be listed in PMC in 2008, and at the time, PMC only accepted English-language journals . FurtherHowever, PMC changed its language policy in 2019, and it now accepts non-MEDLINE, non-English journals. PMC began to review these journals if their primary content was largely in English. Although the exact amount required for “largely in English” is not fixed, more than half of the journal content would be a reasonable target.KJWHN stated in June 2021 that it would continue to value Korean-language submissions to serve the Korean scholarly community while simultaneously offering high-quality English-language articles from researchers in Korea and abroad . This stTaehan Yongsang Uihakhoe Chi . Therefore, these journals can serve as excellent examples for other non-MEDLINE, non-English journals in Korea. More than 20 Korean-language nursing journals are published in Korea, only two of which are published in English: Asian Nursing Research and Child Health Nursing Research. Although both journals are open-access, Child Health Nursing Research was only included in PMC in January 2022 [KJWHN is the first case of a non-MEDLINE, non-English journal in Korea to be indexed in PMC. Its inclusion makes it possible to disseminate valuable nursing information on women’s health from Korea to the world. Another non-MEDLINE, non-English journal is at the forthcoming stage: Journal of the Korean Medical Association (https://jkma.org/), published in Korean by the Korean Medical Association (KMA), may be able to survive for more than 100 years. The reason for this is that it is the official journal of the KMA, the official organization of all physicians in Korea, and the editorial board invites most articles. This journal’s scope mainly relates to education and training for self-employed physicians, general practitioners, medical residents, and the general public. However, the reality of many Korean-language journals is that the number of submissions has decreased, even for training purposes. In other words, researchers want to be recognized for a higher-valued achievement if they spend time writing papers. Therefore, it is challenging to recruit authors unless the journal is listed in international indexing databases. Korean-language life science journals have gradually converted to English-only since 2008 and are usually added to PMC. For the promotion of the journal to a top-tier international level, many editors have changed the journal’s language to English. However, KJWHN’s success in PMC inclusion has now shown that it is possible for Korean-language journals to be added to PMC if the amount of English articles is more than 50% per issue. The survival of Korean-language journals is essential to maintain Korean as a scientific language, although English has already been positioned as the main scientific language. Even while keeping the original language, expanding the number of English articles, although difficult, will be essential preparation for the promotion of academic society journals.The https://www.scimagojr.com) was 0.731; while that of nine PubMed journals was 1.703  . Out of as 1.703 , excludihttps://www.equator-network.org/), for example, CONSORT for randomized controlled trials, STROBE for observational studies, PRISMA for systematic reviews and meta-analyses, and TREND for quasi-experimental studies, and COREQ for qualitative studies; statistical power should be verified through study size estimation [Now that KJWHN will be searchable in PubMed and PMC, article quality offers its best chance to compete with other international journals. Therefore, the editor's job is to recruit high-quality manuscripts from researchers both in Korea and worldwide. What are high-quality manuscripts? A manuscript should be scientifically sound by adopting the following style and format: the study design should be specified; the hypothesis could be defined and answered; manuscripts should be described according to the appropriate reporting guidelines , like many other scientific journals in Korea [https://dataverse.harvard.edu/) [KJWHN has adopted most publishing policies adequately by adhering to the in Korea . The jourd.edu/) ,8. It reThe Journal Impact Factor, manually calculated from Web of Science and cites per document (2 years) from Scopus, is presented in KJWHN aims to be “a core resource for cutting-edge advancements and clinical applications of new nursing practice, therapeutic protocols for managing health problems in women, and innovative research on gender-based issues that impact treatment and nursing care.” By being indexed in PMC, the journal will be able to meet worldwide readers and nurses to provide them with invaluable nursing experiences. This is another way to complete the aims and mission of the journal. The past and present editors have done their best to refine manuscripts and publishing policies to reach the present accomplishments for the journal. I applaud them for their enormous and endless devotion and sacrifice for this academic society journal that is small, yet like a gem in its contribution to women’s health nursing in Asia.


There was no infection, graft failure, or flexion restriction. Arthroscopic ACLR with an adjustable-loop suspensory device is a successful fixation method for femoral-side graft fixation and offers a similar functional outcome as with fixed-loop devices.Arthroscopic anatomic anterior cruciate ligament reconstruction (ACLR) is the gold standard treatment for an ACL tear and requires the use of fixed or adjustable-loop devices to fix a femoral-side graft. Although the adjustable mechanism is designed to provide one-way tensioning, there is a concern that the adjustable loop will loosen and lengthen during cyclic loads, creating graft laxity. The present paper is a retrospective study of patients who underwent ACLR with the fixation of a hamstring graft with an adjustable loop on the femoral side from November 2016 to October 2018. The knee’s functional outcome was evaluated using an International Knee Documentation Committee (IKDC) score, Lysholm score, Lachman test, and pivot shift test. The patients were assessed preoperatively and finally postoperatively after two years of surgery. Thirty-two patients were analyzed. Significant improvement was obtained in the final clinical outcome of the patients. Twenty-seven patients (84.4%) were Lachman negative, and twenty-eight patients (87.5%) were pivot shift test negative, the mean Lysholm score was 96.91, and the IKDC score was 91.47 ( An anterior cruciate ligament (ACL) injury is the most common major sports trauma to the knee, and the arthroscopic anatomical reconstruction of it is considered the gold standard treatment. Anterior cruciate ligament reconstruction (ACLR) is performed on individuals with functionally unstable knees to restore normal knee kinematics. The primary objective is to obtain a secure initial graft fixation for tendon–bone healing and graft integration into the tibial and femoral tunnels. In addition, the fixation should be able to withstand slip during cyclic-loading circumstances to prevent loosening in the early post-operative phase. Popular femoral graft-fixing methods are the use of compression (interference screws), transfixation pins, or cortical-suspension-loop device.Although the best femoral fixation system is controversial, conventional fixed-loop cortical-suspension devices provide excellent and consistent results and are widely used around the world . Fixed-lAlthough the adjustable mechanism is designed to provide one-way tensioning, there is a concern that the loop will loosen and lengthen during cyclic loads, creating graft laxity. Several recent biomechanical studies have shown considerable loosening of the adjustable-loop system under conditions of cyclic-loading stress compared to a fixed-loop device, which could affect clinical and radiological outcomes following ACLR ,2,5,6,7.This was a retrospective study on subjects with an ACL injury who visited our hospital from November 2016 to October 2018. Subjects in the age group of 18 to 50 years with an ACL tear or ACL tear with meniscal injury complaining of instability confirmed clinically and on MRI were included in the study. The exclusion criteria included patients operated with a fixed-loop device and patients with inadequate follow-up .Thirty-two patients with an ACL injury were included. For femoral fixation, we employed an adjustable-length loop device TR , and for tibial side-graft fixation, we used a bioabsorbable screw. The study was conducted in accordance with the Declaration of Helsinki, and approved by the Institutional Ethics Committee. All patients provided informed consent. A senior surgeon performed all the operations. Tourniquet was used in all the cases. The ipsilateral semitendinosus and gracilis tendons were harvested after confirming the diagnosis of ACL tear arthroscopically. In patients with a meniscal tear, partial meniscectomy was performed, and we did not perform the meniscal repair in any of the patients. After hyper-flexing the knee, a femoral tunnel was created through the anteromedial portal. A tibial tunnel was made with a tibial jig at 55°. The femoral tunnel was drilled according to the size of the quadrupled hamstring graft. TR was used for the fixation of the femoral side-graft and the bio-absorbable screw for the fixation of the graft to the tibia. Cycling was performed 20 times to remove any graft creep. A reassessment of the laxity of the graft was performed under arthroscopic visualization. Re-tensioning was performed until the ACL graft was completely tense and firmly placed in the femoral socket by alternate dragging of the white strands. Notch impingement and graft tightness were assessed by flexing and extending the knee. Three doses of antibiotic prophylaxis were administered during the perioperative period as per our institutional policy .On the second postoperative day, full weight bearing with a crutch/walker was allowed. Patients were discharged on postoperative day three. Knee flexion was increased by 30° each week. A knee brace was advised for four weeks after surgery and then discontinued according to the patient’s comfort. After six to eight months, sports activities were allowed .The functional outcome of the patients was evaluated using the International Knee Documentation Committee (IKDC) scoring method, Lysholm scoring method, Lachman test, and the pivot shift test ,12,13. WWe evaluated the demographic information as well as the preoperative and postoperative functional scores .Lachman and pivot shift tests were performed on each patient preoperatively . In the After two years of surgery, we performed a final evaluation of the patients. There has been a significant improvement in the scoring and laxity assessments. There were five patients (15.6%) with positive Lachman grade 1 and four patients (12.5%) with positive pivot shift grade 1. The postoperative Lysholm score was 96.9 ± 1.69 (95–100) (<0.001) and the IKDC score was 91.4 ± 3.6 (87.4–96.6) (<0.001). The average wasting of quadriceps was reduced to 1.0 ± 0.5 cm (0.0–2.5) (<0.001).There were no infections, graft failure, or flexion restrictions in any of the patients.Adjustable-loop devices are the newer version of femoral cortical suspension devices, offering multiple advantages. There is no need to pre-calculate the loop length, and over-drilling of the femoral tunnel is not required. Even in a short femoral tunnel, the maximum amount of graft can be pulled into the tunnel, avoiding multiple fixed-loop sizes in the inventory .In our study, most of the patients required reconstruction as they were in grades 2 and 3 instability by the pivot shift and Lachman tests preoperatively. In most patients, we restored knee stability as 84.4 % tested negative for the Lachman test, and 87.5 percent of patients had a negative pivot shift test. Plaweski et al.  observedIn our study, a significant improvement in the functional results was observed in terms of the Lysholm (96.91) and the IKDC (91.47) scores. In the study conducted by Khan et al. , the meaSome in vitro experiments have suggested that an adjustable-loop device is inferior to a fixed-loop device, and raises concerns about the propensity of the former to elongate during cyclical loads, thus compromising functional graft length. This is crucial during the initial 8–12 weeks after surgery during the early recovery phase when the graft is healing. Lengthening in this phase results in functional knee instability due to impaired tendon–bone healing. Graft failure criteria are widely accepted as >3 mm elongation or side-to-side difference . In one Despite all these facts, other researchers claim that there is no significant variation between the two devices ,25. In oThe results of most biomechanical studies do not reflect clinical studies. This may be because laboratory tests do not provide a good enough simulation of the patient’s biomechanical and physiological fixation environment. Altered angles of ACL loading in vivo ,5, the dThe present study had several limitations. To begin with, rather than using an instrument-assisted knee laxity testing  method, subjective evaluations were employed to determine knee stability. We used the International Knee Documentation Committee (IKDC) scoring method, Lysholm scoring method, Lachman test, and the pivot shift test for the outcome measures. In our study, all the measurements were performed by a senior professor in the department, having clinical experience of more than 20 years, who was blinded to the type of surgery. Therefore, the subjective variations were minimized. The validity and reliability of these tests have been confirmed in previous studies . FurtherIn patients with an ACL injury, arthroscopic anatomic reconstruction of the ACL with an adjustable-loop suspensory device is an efficient fixation technique for the attachment of the graft to the femur. It produces comparable outcomes in terms of knee stability, IKDC score, and Lysholm score. Despite biomechanical studies showing graft slippage with these devices, our study revealed that using an adjustable-loop device to attach the graft to the femur during ACLR is an efficient method. To confirm our results, more studies are needed using randomization between fixed and adjustable-loop devices, large sample sizes, and longer follow-ups.


The purposes of this study were to (1) detect the intra- and inter-reliabilities of the lumbar erector spinae stiffness by MyotonPRO among participants with chronic lower back pain (CLBP); (2) compare the muscle stiffness between pain and non-pain sides during different positions; (3) explore the correlation between intensity of pain and muscle stiffness.Twenty participants with CLBP were recruited and the stiffness measurements were carried out by two experienced physiotherapists (operators Ⅰ and Ⅱ). Each participant was evaluated by the operatorⅠ in different postures (static prone and sitting). After a 5-day interval, the same participant was reassessed by the operatorⅠ in the static prone posture. For the inter-rater reliability test, each participant was quantified by both operators once, with 30 minutes between the measurements on the same day. The intensity of pain was evaluated using a 0–10 visual analog scale (VAS).The intra- and inter-rater reliabilities were excellent (ICC = 0.88–0.99). The MDC values ranged from 25.03 to 86.26 N/m. Examples of Bland-Altman plots showed good agreement. The erector spinae stiffness on the painful sides was higher with a marked increase in the sitting position (P < .05) when compared with the non-painful side. However, there was no significant difference in the prone position (P > .05). The intensity of pain among adults with CLBP was not associated with muscle stiffness of the lumbar erector spinae muscle.Our findings indicated that the MyotonPRO is a feasible device in quantifying the stiffness of the lumbar erector spinae muscle in patients with CLBP. Meanwhile, the erector spinae stiffness on the painful sides was higher in the sitting when compared with the non-painful side. Chronic low back pain (CLBP) is prevalent musculoskeletal discomfort and the principal cause of a decreased quality of life and disability worldwide . From 19Biomechanical characteristics of muscles such as stiffness are objective parameters that manifest tissue condition  and are Many new technologies such as magnetic resonance elastography (MRE)  and sheaFurthermore, poor posture might be an occupational risk factor for CLBP. Alessa and Ning  reportedThe objectives of this study were to (1) detect the intra- and inter-reliabilities of the lumbar erector spinae stiffness by MyotonPRO among adults with CLBP; (2) compare the stiffness of lumbar erector spinae between pain and non-pain sides and the changes of stiffness from prone to sitting; (3) explore the correlation between intensity of pain and lumbar erector spinae stiffness.The study was conducted after receiving approval from the ethics committee of Henan Provincial Luoyang Orthopedic Hospital and in accordance with the Helsinki Declaration (KY2019-001-01). This study was registered in International Clinical Trials Registry Platform (ChiCTR1900025471). All subjects provided written informed consent.0) in this study was 0.5, and the expected ICC value (ρ1) was 0.8. Based on α = 0.05, β = 0.20, an 80% power, and 3 replicate measurements, the minimum required sample size is 15. Twenty volunteers with CLBP between the ages of 18 and 77 years were recruited to participate in this study. All the candidates were hospitalized patients with diagnosis-confirmed by an experienced orthopaedic surgeon. The participants were recruited from Henan Provincial Luoyang Orthopedic Hospital between May and June 2019. In the experiments, every participant had a unique code that could identify individual participants during or after data collection. The inclusion criteria were participants over 18 years of age with unilateral CLBP  lasting 3 months or more [2 or greater [According to the minimum sample size calculation method established in the study of Walter et al. , the acc or more . Unilate or more . The exc greater .amax ∙ mprobe/Δl, where amax is the acceleration of the damped oscillation, mprobe is the mass of the measurement mechanism, and Δl is the maximal displacement of the tissue.The stiffness of the lumbar erector spinae muscles was assessed using a small, noninvasive hand-held MyotonPRO device . The method of measurement is based on recording damped natural oscillation of soft tissue in the form of an acceleration signal. First, the probe of the MyotonPRO is positioned perpendicular and stable on the skin surface of the belly of the lumbar erector spinae muscle. Second, the preloaded pressure (0.18 N) was applied to slightly compress subcutaneous superficial tissue, and a mechanical impulse was released quickly (0.4N), causing muscle tissue damped oscillation. The signal of the oscillation pattern recorded by the accelerometer was used to calculate the muscle stiffness (N/m). The calculation formula is: S = 4 . Both operators had used MyotonPRO as part of daily scientific research for 2–3 years and practiced using the device on the lumbar erector spinae muscle prior to the start of the investigation. To assess the intra-rater reliability of the erector spinae stiffness-related measurements using the MyotonPRO, repeated measurements were obtained by operator YPL according to the above protocol. Two measurements were taken from the same participants each day in 5-day intervals, and the average of these two measures was used for intra-rater reliability calculations. For the inter-rater reliability test, each participant was quantified by both operators (YPL and ZJZ) once, with 30 minutes between the measurements on the same day. The mean of three measurements was used for statistical analysis.The intensity of pain was evaluated using a 0–10 visual analog scale (VAS), with 0 meaning no pain and 10 meaning worst pain .t test was conducted to compare the erector spinae stiffness between the painful side and non-painful side in different postures and the changes of stiffness from prone to sitting in the painful and non-painful sides. Spearman rank correlation tests were used to examine the VAS and the erector spinae stiffness on the painful side.Statistical analyses were performed using SPSS 16.0 software . Descriptive statistics were used to summarize all demographic data as means and standard deviations (SDs). Intraclass correlation coefficients (ICCs) with 95% confidence interval (CI) were used to assess intra-rater within-session reliability (one-way random model) and between-rater reliability (two-way random model). For intra-rater reliability, the association within session was analyzed using a one-way random model consistency. Inter-rater reliability was analyzed using a two-way random model consistency. The following classification was used to interpret reliability: excellent = 0.90–1.00; good = 0.70–0.89; moderate = 0.50–0.69; poor < 0.49 . The staTwo experienced physiotherapists were recruited for the study. All participants  completed the experimental protocol with no reports of medical problems, discomfort, or adverse reactions related to the study.A summary of the results for intra- and inter-rater reliability of the erector spinae stiffness using the MyotonPRO is shown in The Bland-Altman plot for the erector spinae stiffness is shown in P < .01) compared with prone (P > .05) positions . On the non-painful sides, muscle stiffness increased slowly   .r = -0.078, P > .05 and r = -0.107, P > .654, respectively) (ctively) .The primary purpose of the study was to detect the intra- and inter-reliabilities of lumbar erector spinae stiffness by MyotonPRO among adults with CLBP in the prone position. Furthermore, we were able to compare erector spinae stiffness between painful and non-painful sides in different postures and the changes of stiffness from prone to sitting in painful and non-painful sides. This study also explored the correlation between intensity of pain and lumbar erector spinae stiffness among adults with CLBP. Overall reliability estimates were excellent with ICCs ranging from 0.88 to 0.99. The erector spinae stiffness on the painful sides was higher, with a marked increase in the sitting position when compared with the non-painful side. However, the intensity of pain among adults with CLBP was not associated with muscle stiffness of the lumbar erector spinae muscles.1 to L5 levels in their study. Our study added to the comparisons of erector spinae stiffness between the painful side and the non-painful side in different postures, which may be one of the important causes of CLBP occurrence. In a previous study, Lohr et al. [3–L4 in healthy adults. Kelly et al. [In the present study, the ICCs (ICC = 0.99) were excellent for inter-rater reliability. In reference to intra-rater reliability, the ICCs (from 0.88 to 0.91) were good to excellent. The ICC values indicate the degree of consistency between the testing sessions. Within the highly reliable values, between-day reliability was lower than within-day reliability. A possible reason for the results may be attributed to the changes of muscle stiffness between days due to physical activity. Our reliability results are consistent with a previous study using MyotonPRO in young adults with CLBP, where ICC scores ranged between 0.86 and 0.96 . In addir et al.  demonstry et al.  reportedThe SEM was used to estimate how repeated measures tend to be distributed around the “true” score, and the MDC reflects the smallest amount of true change rather than measurement error inherent in the score . For intBland-Altman analyses were conducted to identify systematic bias and compare the 95% limits of agreement. A high agreement, providing a range of error that may relate to clinical acceptability, was shown with less systematic bias . In othe4 levels). However, this study did not distinguish between the painful side and the non-painful side. Compared with the prone posture, pain was primarily featured by the presence of a palpable taut band within the erector spinae muscle in a sitting posture. It may be that pain transmitted through the nerves negatively affects local soft tissues. In our previous study, Zhang et al. [The results of this study suggest that participants with unilateral CLBP had more lumbar erector spinae stiffness on the painful side than on the non-painful side in sitting. The painful side is 13.31% larger than the non-painful side . However, there was no significant difference in the lumbar erector spinae muscle stiffness in the prone position. Our results were in accordance with previous studies using SWE in patients with CLBP, where CLBP is associated with muscle stiffness of the lumbar multifidus muscle in young and middle-aged medical workers . In a reg et al.  demonstrg et al.  indicateInterestingly, we demonstrated that the erector spinae stiffness was significantly increased from prone to sitting, especially on the painful side. Becker et al.  reported4 rather than the painful region. For example, Zhang et al. [In our study, there was no significant correlation between the lumbar erector spinae muscle stiffness and pain intensity in different postures on the painful side . In contg et al.  found thThe current research has a certain positive effect on guiding clinical practice. In a healthy state, the human musculoskeletal system is in a balanced state of left-right symmetry. When some bad postures appear, this balance is gradually broken and a morbid state appears. In our previous study of healthy people, there was no difference in the erector spinae stiffness on both sides in prone, sitting, and standing positions . However4 rather than the pain location. In addition, self-reported VAS score has certain subjectivity. We recognized that relationships between VAS and the erector spinae stiffness on painful side may be affected. Third, the muscle stiffness of the elders might be different with the youngers according to previous studies [We realize that there are some limitations to this study. First, the measurement sites were defined at particular area of the muscle that doesn’t represent the condition of the whole muscle. In this study, we presumed that painful and non-painful sides within the same participants was considered as separate individuals. However, there could be pathological cross-effects from painful side. Second, the lumbar erector spinae muscles consist of multiple small muscles interspersed between verifying fascial planes. In our study, the measurement of muscle stiffness was acquired on the palpable muscle belly one-finger breadth from the spinous process at level L studies , 40. TheIn the current study, we indicated that the MyotonPRO device has excellent intra- and inter-rater reliabilities in quantifying the stiffness of the lumbar erector spinae muscles in patients with CLBP. Meanwhile, the erector spinae stiffness on the painful sides was higher with a marked increase in sitting when compared with the non-painful side. Hence, the MyotonPRO can be used in clinical assessment and research in patients with CLBP, although there was no significant correlation between the lumbar erector spinae muscle stiffness and pain intensity.S1 Data(DOCX)Click here for additional data file.S1 Checklist(DOCX)Click here for additional data file.


Primula, an important ornamental taxon, is more limited than the information on their nuclear and plastid counterparts, which has hindered the comprehensive understanding of Primula mitogenomic diversity and evolution. The present study reported and compared three Primula mitogenomes and discussed the size expansion of mitogenomes in Ericales.Although knowledge of the sizes, contents, and forms of plant mitochondrial genomes (mitogenomes) is increasing, little is known about the mechanisms underlying their structural diversity. Evolutionary information on the mitogenomes of Primula taxa and were compared with publicly available Ericales mitogenomes. The three mitogenomes contained similar gene contents and varied primarily in their structures. The Primula mitogenomes possessed relatively high nucleotide diversity among all examined plant lineages. In addition, high nucleotide diversity was found among Primula species between the Mediterranean and Himalaya-Hengduan Mountains. Most predicted RNA editing sites appeared in the second amino acid codon, increasing the hydrophobic character of the protein. An early stop in atp6 caused by RNA editing was conserved across all examined Ericales species. The interfamilial relationships within Ericales and interspecific relationships within Primula could be well resolved based on mitochondrial data. Transfer of the two longest mitochondrial plastid sequences (MTPTs) occurred before the divergence of Primula and its close relatives, and multiple independent transfers could also occur in a single MTPT sequence. Foreign sequence [MTPTs and mitochondrial nuclear DNA sequences (NUMTs)] uptake and repeats were to some extent associated with changes in Ericales mitogenome size, although none of these relationships were significant overall.Mitogenome master circles were sequenced and successfully assembled for three Primula mitogenomes. Relatively high nucleotide diversity was found in the Primula mitogenomes. In addition, mitogenomic genes, collinear gene clusters, and locally collinear blocks (LCBs) all showed phylogenetic signals. The evolutionary history of MTPTs in Primula was complicated, even in a single MTPT sequence. Various reasons for the size variation observed in Ericales mitogenomes were found.The present study revealed relatively conserved gene contents, gene clusters, RNA editing, and MTPTs but considerable structural variation in The online version contains supplementary material available at 10.1186/s12864-022-08983-x. The mitochondrion is one of three genetic compartments in plant cells that play a fundamental role in encoding several essential mitochondrial electron transfer chains , 2. OnlyViscum scurruloideum Barlow  contained two (trnW-CGA and trnA-UGC) and one (trnM-CAU) plastid-derived gene, respectively.Among the 29 conserved gene clusters identified in angiosperms , the thr<) Fig. S. Eight ames Fig. ; Fig. S7. First, Primula mitogenomes exhibited relatively high nucleotide diversity (π: 0.009) among all examined plant lineages, which was 10-fold higher than the lowest diversity, found in Zea (π: 0.0009)  and the two alpine Primula taxa . The π value between the two alpine Primula taxa was 0.004, while the π value between the taxa from the two regions  was 0.013 (Table SThe nucleotide diversity (π) of LCBs was calculated for different plant lineages Table S. First, ) Table S. The π va Table S. Second,y π: 0.00 among alMonotropa hypopitys Crantz, Rhododendron simsii Planch., Actinidia argute Miq, Actinidia eriantha Benth., and Camellia sinensis (L.) Kuntze mitogenome sizes were 810 kb, 802 kb, 792 kb, 772 kb, and 707 kb, respectively, which were much larger than those of Vaccinium macrocarpon Aiton (459 kb), V. microcarpum Miyabe & T. Miyake (468 kb), Ae. corniculatum (425 kb), and the three Primula taxa (349–426 kb) , P. smithiana (181 kb), M. hypopitys (178), and C. sinensis (121 kb) were significantly larger than those of the remaining mitogenomes (each < 20 kb)  from NCBI, with the aim of revealing the potential causes of the observed mitogenome size variation at the order level Table S. The Mon Miyake 48 kb, Ae. Miyake 48 kb, Ae. Miyake 48 kb, Ae.Ac. arguta (53 kb)  Fig. b. Additi05 Fig. S. The rels Table S, which i arguta 5 kb  and P. valentiniana (Sect. Amethyatina) exhibited the closest relationship  and Bayesian inference trees were reconstructed based on the 13 shared protein-coding genes . Both trees suggested that hip Fig. . The intPrimula mitochondrial contigs into a master circle. These master circles indicated highly homologous recombination (HR) frequencies, as no long collinear sequences were shared within Primula . y Table S, which wPrimula and among plant lineages. First, Primula probably presents an elevated mutation rate in the mitogenome, as shown in several plant lineages  as references. tRNAs were identified by using tRNAscan-SE v2.0.7 [https://www.ncbi.nlm.nih.gov/orffinder/). The GC count per 500 bp was calculated with an in-house Python3 script. The depth of sequencing coverage per locus was calculated using the genomecov command in bbMap (https://www.sourceforge.net/projects/bbmap/) with the parameter –d.Protein and rRNA genes were annotated using the GeSeq online server  and locaE v2.0.7 . The staPrimula mitogenomes was visualized using Circos v0.69 [Repeat regions were identified by alignment against each assembly using BLASTN  with a mos v0.69 .ccmB and nad4L in P. smithiana were calculated using ProtScale [P. smithiana protein-coding genes was calculated based on the method described by Kyte and Doolittle [RNA editing sites in the three mitogenomes were predicted using the PRET-Mt server  with a crotScale . In contoolittle .atp6 and ccmFc gene sequences were extracted from the eight publicly available Ericales and Primula mitogenomes and were aligned using MAFFT v.7.4 [atp6 and ccmFc were inferred using PSIPRED [Ac. arguta (SRR3823655), Ae. Corniculatum (SRR1688722), C. sinensis (SRR20083852), M. hypopitys (SRR10159707), R. simsii (SRR10415549), and V. macrocarpon (SRR18449568) were downloaded from the NCBI SRA database and mapped to the atp6 and ccmFc gene sequences using BWA v0.7 [The FT v.7.4 . The sec PSIPRED . To valiBWA v0.7 . EditingPrimula taxa through visual checking, as described by Kan et al. [The 29 gene clusters in the plant mitogenomes described by Richardson et al.  were iden et al. .Primula mitogenomes was aligned against the others to identify LCBs using the nucmer command in MUMMER4 [Each pair of  MUMMER4  under th MUMMER4  and then MUMMER4 . Nucleot MUMMER4 .Primula plastid genomes were assembled using GetOrganelle [Primula mitogenome against the P. palinuri plastid genome (after removing one inverted repeat) using local BLASTN with a minimum length of 100 bp and an e-value of 1e-20. Putative MTPTs, including atp1/atpA, rrn26/rrn23, and rrn18/rrn16, were excluded because they simultaneously occurred in the mitogenome and plastid genome [Three rganelle  and annorganelle . Transfed genome , 66.Two MTPTs longer than 500 bp  were extracted and validated using read mapping and PCR amplification. Each clean read was mapped against the 100 bp up- and downstream regions of MTPT in the mitogenome and its plastid genome counterpart for read mapping. MTPT7 and MTPT10 were aligned with the other 30 publicly available plastid genomes Table S after vaPrimula mitogenome against the P. veris nuclear genome (GCA_000788445.1) using BLASTN with a minimum length of 50 bp and an e-value of 1e-20. The nuclear-shared regions in other Ericales mitogenomes were also identified against their corresponding nuclear genomes. This reference nuclear genome information can be found in Table STo account for gene transfer events between the mitogenome and nuclear genome, the nuclear-shared sequences were identified by aligning each Repeat regions and MTPTs were also identified among Ericales mitogenomes. Pearson’s correlation was calculated between repeats, NUMTs, MTPTs, and mitogenome size in Ericales to reveal the underlying causes of mitogenome size variation .Primula mitogenomes  were included in the phylogenetic analysis, with Platycodon grandiflorus A. DC.  and Solanum lycopersicum L.  as outgroups. A total of 13 coding sequences (CDSs) among the mitogenomes were aligned with MAFFT v7.4 [7 generations with four incrementally heated chains, starting from random trees and sampling one out of every 1000 generations. The stability of the Markov chain was ascertained by plotting likelihood values against the number of generations (effective sample size > 200) using Tracer v1.7 [The above eight publicly available Ericales mitogenomes and three FFT v7.4  and concFFT v7.4 . ML phylFFT v7.4  with 500FFT v7.4  under thcer v1.7  and by sAdditional file 1: Figure S1. Electrophoretic gel visualization of the amplified fragments of the three draft mitogenome assemblies and MTPTs. M1 is the DL2000 DNA marker, whereas M2 is the DL1000 DNA marker. The first two wells in each gel represent the corresponding primer pairs for MTPT7 and MTPT10 in each Primula mitogenome, whereas the other primer pairs were used for assembly validation in each draft mitogenome  NUMT length and Ericales mitogenome size after removing the Aegiceras corniculatum mitogenome; (b) repeat length and Ericales mitogenome size after removing the kiwifruit mitogenomes; (c) MTPT length and Ericales mitogenome size after removing the Rhododendron simsii and Monotropa hypopitys mitogenomes.Additional file 10: Table S1. Assembly information of three Primula taxa. Table S2. The number of protein-coding genes in the three Primula mitogenomes. Table S3. Tandem repeats in the Primula smithiana mitogenome. Table S4. Tandem repeats in the Primula palinuri mitogenome. Table S5. Tandem repeats in the Primula valentiniana mitogenome.Table S6. Information on MTPTs within the three Primula mitogenomes. Table S7. The shared nuclear sequences within the three Primula mitogenomes. Table S8. The nucleotide diversity of mitogenomes within different plant lineages. Table S9. The nucleotide diversity of mitogenomes among Primula taxa. Table S10. The general features of the 11 mitogenomes used for this study. Table S11. The primers used for Primula palinuri, Primula smithiana, and Primula valentiniana. Table S12. The 30 publicly available plastid genomes used for MTPT analysis. Table S13. The publicly available nuclear genomes used for NUMT analysis.


Ascophyllum nodosum seaweed flour (AF) and corn starch (CS) on the interactions between polyphenols and starch was studied in this paper. These methods comprised the blending of AF with native starch (NT) with previously gelled starch gel (GL) and promoting the gelling of corn starch in the presence of AF (CGL). Different AF–CS (g/g) ratios (from 1:0.5 to 1:25) were studied. The liquid phase was chemically characterized by polyphenols (TPC) and carbohydrates content. The antioxidant activity of the liquid phase after achieving the solid–liquid equilibrium was determined by DPPH, ABTS, and FRAP methods. The solid phase was characterized by FT-IR and SEM techniques. The Halsey model successfully fitted the equilibrium TPC in liquid and polyphenols adsorbed/retained by the solid phase of tested systems. NT samples showed lower polyphenols sorption than gelled samples. The differences found between samples obtained with GL and CGL methods suggested different interactions between polyphenols and starch. Specifically, physisorption is predominant in the case of the GL method, and molecular trapping of polyphenols in the starch gel structure is relevant for the CGL method. Results allowed us to determine the enhancement of the retention of polyphenols to achieve starchy foods with high bioactivity.The effect of several blending procedures between  The demand for gluten-free products is increasing since celiac disease affects around 1% of the world population. However, the food industry seems to be more focused on technological than nutritional quality . A gluteAscophyllum nodosum (A. nodosum) brown seaweed are potential food additives that may act as competitive inhibitors of digestive α–amylase and α–glucosidase enzymes, inhibiting the hydrolysis polysaccharides and thus reducing the glucose absorption levels [The main polyphenols from seaweeds, named phlorotannins, are notorious antioxidant, anti-diabetic, and anti-hypertensive natural compounds . They arn levels . Ingesten levels .In vitro studies are easy, rapid, widespread, and reproducible methods to evaluate inhibitory phlorotannins capacities ,9. The iPhlorotannins are easily affected by oxygen, pH, ions, light, and temperature . As repoThese interactions are affected not only by the specific polyphenol structure but also by the different experimental conditions and/or the nature of the polymer . Starch A. nodosum seaweeds powder (AF) followed by the sorption/complexation process between polyphenols and starch.There is extensive literature reporting mechanisms of sorption, complexation, interaction, or encapsulation of polyphenols on several starchy materials , using a wide variety of bioactive sources  and using different immobilization methodologies ,10. HoweThe use of whole raw material as a bioactive compounds source involves a reduction in time, costs, and energy consumption of processing since extraction and purification processes are omitted . ConversAscophyllum nodosum seaweed flour (AF) bioactive molecules and corn starch (CS) interactions were studied in this paper, employing three different methods . The NT method consisted of blending AF and native CS. The GL method consisted of blending AF and pregelatinized CS. Finally, in the CGL procedure, CS was gelatinized in the presence of AF. Different proportions of AF and CS were proposed , and their aqueous phases were chemically characterized by bioactive compound content (polyphenols and carbohydrates) and by antioxidant activities . Solid was characterized by FT-IR and SEM techniques.tris (2-pyridyl)-S-triazine (TPTZ), potassium hydroxide, potassium chloride, iron (III) chloride, sodium chloride, potassium dihydrogen phosphate, sodium phosphate, sodium azide, phloroglucinol, and sodium acetate were from Millipore Sigma . 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), sodium carbonate, Folin–Ciocalteau reagent, chlorohydric acid, glucose, Trolox, sodium hydroxide, and methanol were from Panreac .All reagents used for characterization were analytical grade. Sulfuric acid, phenol, and iron sulfate were supplied by Merck . 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,4,6-Ascophyllum nodosum seaweed (A. nodosum) from Galicia’s coasts (NW of Spain) was harvested in November 2019, supplied by Mar de Ardora S.L. company , dried in a hot air convective dryer  at 50 °C, with a constant relative humidity of 30% and air velocity at 2 m/s. Dried A. nodosum was ground in an ultra-centrifugal mill . A. nodosum seaweed powder (AF) was stored at 4 °C with a final moisture content of 10.0% ± 0.1% (d.b) until its use.Food-grade native corn starch  of 95% purity (20.3% amylose content), 8.1% ± 0.2% (d.b) moisture content was used. Fresh Three methods , Starch gelatinization was performed according to the method reported by Wang et al. . AqueousW/gAF (g of water/g of seaweed). AF and CS content varied to obtain different A. nodosum flour-to-cornstarch blending ratios  corresponding to 4, 50, 100, and 200 gW/gCS, (g of water/g of starch), respectively. A ratio of 1:25 was used as the average proportion studied by bakery products [The liquid-solid ratio was set at 100 gproducts . The phyPE/L). TPC values provided by AF in the AF–CS blends were calculated using Equation (1):AF–CS is the polyphenols content measured in the liquid phase of AF–CS blends and TPCCS is the corresponding polyphenols content of CS control samples.Chemical characterizations were carried out with a spectrophotometer  at least in triplicate. Total polyphenol content (TPC) was determined using phloroglucinol as standard following the method proposed by Singleton and Rossi  based onGE/L). The antioxidant activity of the aqueous phases was determined using DPPH, FRAP, and ABTS methods, expressing the results as the equivalent activity of the Trolox standard in micromole units (µMTE). DPPH scavenging activity determinations were performed following the methodology proposed by Brand–Williams et al. [The total content of carbohydrates (CHOs) was spectrophotometrically determined at 485 nm, applying the method reported by Dubois et al. , using gs et al. . The scas et al.  method, s et al.  procedurP (%), of polyphenols was evaluated by Equation (2), and the polyphenols adsorbed by CS, q (mgPE/gCS), were determined by Equation (3):AF is the polyphenol content (gPE/L) of the aqueous phase corresponding to AF control samples, V is the liquid volume (L), and mCS is the final CS mass (g), evaluated by means of Equation (4):CSi is the initial CS mass (g) and CHOs the carbohydrate content (gGE/L) of the liquid phase released from starch (adsorbent).Equilibrium sorption yield, Y–1. Samples were blended with KBr and compressed into disks. FT-IR spectra treatment was carried out with Omnic 7.1 software .AF–CS samples at the intermediate ratio (1:1) were analyzed by FT-IR to characterize the AF–CS interactions. FT-IR spectra were recorded with a Bruker FT–MIR model Vertex 70 V spectrometer. The wave number range was set in the range of 4000 to 50 cmAF–CS (1:1 ratio) samples were freeze-dried for 36 h (−55 °C and 50 Pa), and their microstructure was analyzed by scanning electron microscopy (SEM). Samples were sputtered with iridium using a vacuum metallizer/shader model Q150T S  with a thickness layer of 5–10 nm was deposited. Samples micrographs were obtained using a scanning electron microscope  at 3 Kv using a SE/InLens secondary electron detector.p < 0.05) using a Duncan test. The experimental results were treated and plotted on Microsoft Excel . All experimental results were expressed as mean ± standard deviation of triplicate experiments (n = 3).Statistical analysis was carried out by IBM SPSS statistics 27  software. A one-way analysis of variance (ANOVA) was assessed based on a confidence interval of 95% (Ascophyllum nodosum seaweed powder (AF), corn starch (CS), and AF–CS blends were chemically characterized, determining polyphenols content (TPC), carbohydrates content (CHOs), and their antioxidant activities by DPPH, ABTS, and FRAP methods. TPC values (A), sorption yields, YP (B), and CHOs values (C) of AF , CS  control samples and AF–CS blending methods of NT , GL , and CGL  samples are summarized in p < 0.05) differences among samples. For the CS control, it was necessary to evaluate according to CS structures, consequently, native (NT samples) and gelled CS (GL and CGL samples) controls were employed in each case.The aqueous phase from A. nodosum control sample in solution (TPCAF) was 396.9 ± 28.6 mgPE/L. The TPC evaluation of corn starch control samples was performed to correct the values of total polyphenols for some reducing power that starch-derived carbohydrates might have on the Folin–Ciocalteau reagent. Starch concentration almost did not affect the TPC values obtained (p < 0.05) higher. Slight differences were found between TPC from native CS (from 1.9 ± 0.1 to 94.9 ± 6.8 (mgPE/L) and gelled CS (from 2.9 ± 0.2 to 143.4 ± 10.3). The control samples evaluation showed that starch, either gelled or not, did not significantly contribute to final TPC values, but it was not negligible for 0:25 samples. The obtained TPC values from both control samples separately (AF and CS) rendered higher values than those measured with the corresponding blends, PE/L, GL samples showed lower content, from 0.02 ± 0.01 to 192.9 ± 13.9 mgPE/L, and CGL showed the lowest values (at constant AF–CS ratio) with a maximum value of 82.8 ± 6.0 for 0:0.5 ratio (p < 0.05) polyphenols content in relation to NT. The different available sorption surface areas between native starch (granular structure) and gel starch could explain these differences [The average total polyphenols content of the .5 ratio A. These A. nodosum particles and extracted polyphenols can be trapped within the gel structure during gels formation and subsequent retrogradation [The blending procedure that involves the gelling of starch in the presence of seaweeds polyphenols (CGL) caused a dramatic reduction in TPC values. The NT and GL methods involved the direct contact between AF and CS (native or gelled) at rt for 15 min; meanwhile, CGL put them in contact during the gelatinization process carried out at 100 °C (20 min), a cooling step (20 min), and the final dispersion and homogenization in water. During CS gelling with the presence of AF, both the radation . Anotherradation ,27 or thradation .PE/L) was lower than those employed by Wang et al. [Wang et al.  studied g et al.   of the samples (P varied between 29.3 ± 1.8 (NT 1:0.5) and 99.8 ± 5.9 (both GL and CGL 1:25). YP values showed higher retention capacities of the gelled corn starch (GL and CGL), with similar trends for the three methods tested, where the higher the CS content, the higher the polyphenols retention yield. NT samples showed lower sorption yield values to the lower active surface area of native starch to interact with polyphenols [P  were calculated, and the corresponding values are presented in TPC from the aqueous phase of AF–CS blends and AF and CS control samples were used to determine the sorption yield  was used. These values indicated that the use of starch gel increased almost twice the polyphenols retained in comparison to native starch. These values could be related to the higher surface area developed in the gel structure in comparison to the granular structure. When the formation of starch gel took place in the presence of seaweeds, flour increased by almost 3 and 1.54 times regarding the NT method and GL methods, respectively. This notorious increase could be explained by additional retention of polyphenols beyond adsorption, such as entrapment or encapsulation of bioactive molecules in the gel structure. As the amount of starch increased (1:1 and 1:2 ratios), the differences decreased due to the respective increase in sorption yields.gPE/gCS), determined by means of Equation (3) against TPC values of the liquid phase, are plotted in PE/gCS, equilibrium is achieved at TPC of 240, 115, and 35 mgPE/L of TPC for NT, GL, and CGL samples. These results confirmed that low sorption surface area was available when native corn starch granules were employed in comparison to the area developed by starch gel structures in GL and CGL samples. In all cases, the shapes, convex to the abscissa axis over the tested range, of equilibrium adsorption/retention isotherms of polyphenols on starch allowed their classification as III according to the BET classification [Polyphenols retained by CS, q (mfication . An empification .(5)q =−2R > 0.98) and root mean square error (ERMS < 3.6), the goodness of fitting can be considered acceptable; −1 varied from almost linear (1.1) for NT, quadratic (1.9) for GL, and cubic (3.0) for CGL.GE/L) values  ranged from 213.2 ± 15.4 to 1223 ± 88 for 1:0.5 to 1:25 AF–CS ratios, respectively. CHOs significant (p < 0.05) differences stressed the notorious effect that gelatinization had on CHOs signals.CHOs (mg) values C were nop < 0.05) higher in GL (495.1 ± 35.6 to 955.1 ± 68.8) in comparison to CGL (252.0 ± 18.2 and 804.6 ± 57.9) and samples (332.7 ± 24.0 to 431.9 ± 31.1). Higher CHOs values of GL samples were associated with the leaching of some polysaccharides (mainly amylose) during starch gelatinization and, in addition, with the extracted carbohydrates from AF, increased the total carbohydrates content; this effect was clearly evidenced in assays performed at 1:25 ratio. Low CHOs values determined in CGL samples can be again explained by the gel trapping effect. The formation of CS gel in the presence of AF limited the release of carbohydrates from algae to the liquid phase, which notoriously (p < 0.05) reduced CHOs in AF–CS samples. Moreover, thermal deterioration during gelatinization could also partially alter sugars [A. nodosum carbohydrates have demonstrated relevant phytochemical features.At constant AF–CS ratios, CHOs values were significantly  between DPPH and TPC values, without distinctions among employed blending procedure, was found, indicating that scavenging activities of polyphenols measured by this method depend only on TPC, and the possible deterioration of bioactive compounds is not detected. Nevertheless, to achieve suitable linear correlations (R2 > 0.91) for ABTS-TPC results, experimental data had to be separated into two sets according to AF thermal treatment, subjected (CGL) and non-subjected (NT and GL) to the heating process, and consequently employing this more sensitive method in different antioxidant activities trends with TPC could be successfully established. Finally, a linear correlation for FRAP-TPC data was only found for NT samples (R2 = 0.93) because FRAP values corresponding to GL and, particularly, CGL methods were very low. This last result demonstrated that the most bioactive polyphenols became notoriously trapped by CS gel, remaining with low bioactivity in the liquid phase. Other researchers also reported linear correlations in polyphenol–starch blends [An acceptable linear correlation  from NT, GL, and CGL methods. FT-IR can provide relevant structural information and was previously employed in polyphenols–starch systems ,33.–1 that corresponded with the vibrational stretching of inter- and intra-molecular hydroxyl groups (–OH), –1, AF and CGL showed two peaks not observed in other samples corresponding to hydrogen bridge bonds. Indeed, intensity values of this region are often used as a criterion to measure the amount (concentration) of hydrogen bridge bonds [All samples showed a notorious peak at 3400 cm–1 corresponds to the characteristic peak of aromatic rings (stretching vibrations of C=C bonds). In this region, significantly (p < 0.05) different quantitative signals among samples with and without AF were found and were straightly related to polyphenols (phlorotannins) content (AF > CGL > GL > NT > CS samples). Particularly, CGL’s high peak intensity indicated the increasing retention by entrapment of polyphenols by CS, even after gelatinization treatment [The region around 1600 cmreatment . NT and reatment ,29,33, ireatment ,36. No nSEM micrographs of CS and AF–CS blends (1:1 ratio) were carried out to observe the proposed physical bonding  and trapping (CGL) interactions between AF and CS . CS contA. nodosum particles. NT  with starch could be necessary to understand better their antioxidant, anti-diabetic, and bioactive behavior during digestion.


Due to the high clinical heterogeneity of neuronal intranuclear inclusion disease (NIID), it is easy to misdiagnose this condition and is considered to be a rare progressive neurodegenerative disease. More evidence demonstrates that NIID involves not only the central nervous system but also multiple systems of the body and shows a variety of symptoms, which makes a clinical diagnosis of NIID more difficult. This review summarizes the clinical symptoms in different systems and demonstrates that NIID is a multiple-system intranuclear inclusion disease. In addition, the core triad symptoms in the central nervous system, such as dementia, parkinsonism, and psychiatric symptoms, are proposed as an important clue for the clinical diagnosis of NIID. Recent studies have demonstrated that expanded GGC repeats in the 5′-untranslated region of the NOTCH2NLC gene are the cause of NIID. The genetic advances and possible underlying mechanisms of NIID  are briefly summarized in this review. Interestingly, inflammatory cell infiltration and inflammation were observed in the affected tissues of patients with NIID. As a downstream pathological process of NIID, inflammation could be a therapeutic target for NIID. Neuronal intranuclear inclusion disease (NIID) was first described in 1968 and is considered to be a rare neurodegenerative disease . With thRepeated GGC expansion in the 5′UTR of the NOTCH2NLC gene was identified as the causative mutation for NIID in Japan and China in 2019 . These fHundreds of NIID cases have been reported so far, and the most common feature of NIID is symptom heterogeneity; different families and individuals have different symptoms of NIID. In early clinical studies, the major clinical manifestations of previously described cases were cerebral cortical dysfunction and extrapyramidal symptoms, including cognitive dysfunction and dementia , PD-likeDementia was one of the main symptoms described in a patient with NIID in 1978 . Since tParkinson’s disease-like symptoms are regarded as another common and typical representative symptom in patients with NIID . PD-likeIt is worth noting that a significant proportion of patients with NIID have adverse changes in terms of mood and personality , but dueLimb weakness was found in many patients with NIID and is one of the key phenotypes of NIID . A previAlthough progressive symptoms, such as cognitive impairment and parkinsonism, are key features of NIID as a neurodegenerative disease, episodic attacks are also found in some patients with NIID, such as encephalitis-like, vestibular migraine-like attacks, or MELAS-like or epileptic episodes .Several ophthalmological manifestations have been reported in patients with NIID, such as abnormal pupillary functions, miosis, oculogyric crisis , reducedExcept for the nervous system, many studies have indicated that other systems are involved in the pathology of NIID . The nonA recent study showed that ubiquitin and p62-positive cells are found in the lung tissues of patients with NIID, indicating the presence of intranuclear inclusion bodies in the lung . By inteCardiomyopathy and coronary atherosclerosis are reported to be associated with circulatory symptoms in patients with NIID. In previous studies, intranuclear inclusions were found in the kidneys of patients with NIID , but uriDigestive system symptoms in patients with NIID commonly manifest as severe nausea and vomiting, which can occur alone, but in most cases, nausea and vomiting occur together. A case report showed that a patient with NIID repeatedly vomited for 7 years before the apparent abnormality in DWI was found, suggesting that periodic vomiting could be the only symptom of NIID in the early stages of NIID . GastroiThe patients with familial NIID were reported to have erectile dysfunction beginning in the first or second decade of life . Chen etAlthough different systems and organs are involved in NIID, the sequences of involvement in different systems may vary in individual patients according to previous case reports; for example, the existence of intranuclear inclusions and symptoms in systemic organs, such as the stomach and kidneys, precedes the onset of nervous system symptoms in NIID case reports . TherefoImaging examination is of great value in the clinical diagnosis of NIID. The earlier reported MRI findings were atrophy of the cerebellar hemispheres and no specific imaging for NIID . With thNeuronal nuclear inclusions (NIIs) in the skin or biopsy samples of other tissues are the characteristic histopathologic findings of NIID. NIIs are not limited to NIID, but are also present in a variety of multiple neurodegenerative diseases, such as fragile X-associated tremor/ataxia syndrome (FXTAS), distal ophthalmopharyngeal myopathy, and oropharyngeal myopathy, which have overlapping clinical symptoms and similar pathological outcomes, and even have some commonalities in genetic diagnosis . The exiThe NIID was defined as an intranuclear inclusion disease when it was first described in These inclusion bodies are circular, 1.5–10 μm in diameter, and are located near nucleoli, and ubiquitin/p62 staining, but not tau epitope, can be seen around NIIs . ElectroExcept for the NIIs in the different tissues, in the pathological examination of NIID tissues, diffuse inflammatory cell infiltration near the intranuclear inclusions, such as neutrophil monocytes and macrophages, was found in the affected tissues, including the brain, lung, bladder, and prostate gland , indicatIn 2019, studies reported GGC repeat expansion at the 5′ region of NOTCH2NLC as the genetic cause of NIID . NOTCH2NBased on a multiethnic cohort of gene-confirmed patients with NIID from Southeast Asia, it was suggested that the presence of GGA interruptions in the repeated expansion of GGC may play a role in modifying the disease in terms of age at symptom onset . Based oA previous study demonstrated that the NIIs in sporadic and familial NIID contained Sumo-1 and SUMOylation substrate PML and HDAC4 . Based oAlthough GGC repeat expansion in the NOTCH2NLC gene has been proven to be the cause of NIID, the molecular mechanisms underlying NIID remain unclear. Similar to FXTAS (GGC repeats in the FMR1 gene), Huntington’s disease (CAG repeats in the HTT gene), amyotrophic lateral sclerosis/FTD (GGGGCC repeats in the C9ORF72 gene), and myotonic dystrophy , NIID is one kind of nucleotide repeat expansion disorder or microsatellite repeat expansion disorder with expanded GGC repeats in the NOTCH2NLC gene. Therefore, NIID may share some similar mechanisms to these microsatellite repeat expansion disorders. Here, we briefly propose some possible mechanistic models of NIID at the DNA, RNA, and protein levels . For morUnconventional translation, named repeat-associated non-AUG translation (RAN), of expanded repeats in toxic proteins has been identified in a variety of microsatellite disorders . For exaPrevious studies have demonstrated that repeat expansion-containing RNAs from the non-coding regions of genes linked to diverse human diseases, such as myotonic dystrophy type 1 and FXTAS, can form intramolecular hairpin secondary structures, bind to many RNA-binding proteins, and form RNA-protein aggregates in the pathology of these neurodegenerative diseases, resulting in toxic gain-of-function of these mRNAs . NOTCH2NExtended GGC repeats may cause the pathology of neurodegenerative diseases not only at the protein level and the RNA level but also at the DNA level. A previous study indicated that GC enrichment in the extended GGC repeats of the FMR1 gene increases the propensity of the formation of RNA-DNA hybrid R-loops . These cIn early case reports, autopsy, rectal biopsy, and nerve biopsy were mostly used to diagnose patients with NIID . Sone etDue to the high heterogeneity and multisystem symptoms of this disease, it is difficult to reach the diagnosis of NIID in clinical practice. The diagnosis of NIID can be considered from typical MRI signals, clinical manifestations, pathology of NIIs, and genetic tests. DWI high-intensity signals in head MRI are the strong indicators for the diagnosis of NIID. If patients had no typical MRI changes, the triad of clinical manifestations such as dementia, PD-like behaviors, and personality changes is a good cue for the diagnosis of NIID. For patients without the triad of NIID and DWI high-intensity signals, NIIs in the pathological findings from skin or other tissues are also a cue to suspect the diagnosis of NIID. Once the diagnosis of NIID is suspected, a genetic test for GGC repeats in the NOTCH2NLC gene should be performed to confirm or rule out the diagnosis of NIID, as shown in Because NIID is a symptom-heterogeneous disease and shares similar imaging features with some diseases, NIID has many differential diagnoses from other diseases.Because FXTAS and NIID are both autosomal-dominant GGC trinucleotide repeat expansion diseases in different genes  , FXTAS iNeuronal intranuclear inclusion disease should be distinguished from Creutzfeldt–Jakob disease and some diseases with leukoencephalopathy or dementia. Creutzfeldt–Jakob disease has progressive cognitive impairment and myoclonus in the extremities, as well as characteristic lacy high-intensity signals in the cortex, caudate nucleus, or putamen on brain DWI. A recent study proposed that brain DWI is the key to differentiate from Creutzfeldt–Jakob disease and different leukoencephalopathy grades .Middle cerebellar peduncle lesions are considered to be a characteristic finding of NIID , but theCurrently, there is no treatment to cure or slow down the process of NIID, but medications that control symptoms, such as muscle weakness, impaired consciousness, abnormal behavior, and sensory impairment, can improve a patient’s quality of life . ClinicaRecently, different therapeutic strategies have been proposed for NIID, for example, antisense oligonucleotide therapy, RNA interference, small molecule RNA drugs, and CRISPR-based therapy . AlthougWith an increasing number of reported cases of NIID, the clinical manifestations of NIID are well characterized. Early studies showed that NIID mainly has neurological symptoms; the latest studies demonstrated that there are a variety of symptoms and signs in various systems, indicating the high heterogeneity of clinical symptoms and signs of NIID. This increases the difficulty in the clinical diagnosis of NIID. Although patients with NIID present heterogeneous symptoms, they also show some core symptoms, such as triad symptoms in the central nervous system . Before the NOTCH2NLC gene was linked to the etiology of NIID, tissues/skin biopsy and DWI high-intensity signals along the corticomedullary junction were strong clues for the diagnosis of NIID; now, genetic tests for expanded GGC repeats in the NOTCH2NLC gene have become the gold standard for NIID.Expanded GGC repeats in the NOTCH2NLC gene have been confirmed as the cause of NIID, but how expanded GGC repeats in the NOTCH2NLC gene cause pathological changes and the accumulation of NII in the nucleus remains unclear. As a nucleotide repeat expansion disorder, NIID may share similar mechanisms to other nucleotide repeat expansion disorders, such as expanded nucleotide repeat-induced DNA damage, RNA toxicity, and abnormal encoded protein toxicity. Although recent studies have demonstrated the presence of polyG-NOTCH2NLC protein in the animal models and tissues of patients with NIID, the mechanism by which the polyG-NOTCH2NLC protein causes toxicity still needs more investigation. RNA toxicity and DNA instability theory need to be identified in the mechanisms of NIID. How do expanded GGC repeats in the NOTCH2NLC gene induce the formation of NIIs? What are the components in expanded GGC repeat-induced RNA foci? Which RNA-binding proteins are involved in the pathogenesis of NIID? These questions await more investigation, and the answers to these questions will help to guide the treatment of NIID. To date, no effective treatment is available. Even though different gene therapies, such as antisense oligonucleotide therapy and RNA interference, have been proposed, many great challenges must be overcome, such as the specificity and out-of-target nature of RNA drugs. Recent findings have shown that inflammation is involved in the pathological changes of NIID and may be downstream of expanded GGC repeat-induced RNA/protein toxicity. Therefore, the role of inflammation should be further identified in the pathology of NIID, and anti-inflammation could be a promising therapeutic strategy for NIID.XX, MS, and BoW designed and revised the manuscript. YL and HL collected the literature and made the first draft. XL, BiW, HY, BoW, and MS discussed and interpreted the analysis. All authors contributed to the article and approved the submitted version.


Objectives There is a lack of information regarding the longevity of prefabricated crowns on primary teeth. Therefore, the objective of the present study was to evaluate the clinical success rate of NuSmile pediatric Zirconia crowns in both primary anterior and posterior teeth up to 2 years in terms of gingival health, levels of plaque, restoration failure, and marginal integrity.Materials and Methods This interventional study evaluated the clinical performance of NuSmile primary Zirconia crowns on 232 (172 primary incisors and 60 primary molars) pulp-treated primary teeth. Statistical analysis included independentt-test and Kaplan–Meier survival method for which the level of significance was set up atp < 0.05.Results Gingival and plaque index of the teeth restored with NuSmile Zirconia crowns (test tooth) compared with adjacent and antagonist teeth (control tooth) showed no statistically significant difference at all time intervals (p > 0.05). The Kaplan–Meier survival graph indicated that only 34% of NuSmile Zirconia incisor crowns and 86% of NuSmile Zirconia molar crowns have survived at the end of 24 months. Of the 172 NuSmile Zirconia incisor crowns placed, only 82 teeth survived at the end of 2 years and the majority of the crowns completely lost the Zirconia crown. The Kaplan–Meier graph also indicated that more than two-thirds of crowns, i.e., 90% incisor crowns and 76% of molar crowns, had good marginal integrity at the end of 24 months.Conclusions Zirconia pediatric crowns preserve and maintain gingival health and have long-term survival rates with good retention and marginal integrity, indirectly preventing secondary caries. Hence, Zirconia pediatric crowns are a well-suited restoration of choice for primary teeth rehabilitation. In today's modern society, both parents and children are cosmetically conscious and increasingly concerned about the esthetics of their teeth. The child's parents often influence the dental professionals treating the children in selecting dental restoration, and parents are getting more involved in clinical decision-making than ever before.Management of extensive carious lesions and traumatized primary teeth has gradually shifted from extraction to full-coverage restorations. Extra-coronal full-coverage crowns are indicated in primary teeth with developmental defects, multi-surface caries, patients with high caries risk, fractured teeth, teeth where direct restoration tends to fail, after pulpal therapy, as abutments for space maintainers, and teeth with excessive wear.Traditionally, composite strip crowns and stainless steel crowns (SSC) have been used as full-coverage restorations for grossly decayed and pulp-treated primary anterior teeth and molars. Strip crowns are esthetically better but are technique sensitive. Moisture and hemorrhage control is essential with strip crowns as it could lead to resin placement failure. SSC crowns have been successful for many years in terms of durability, retention, and function. The most significant problem was their poor esthetics limiting their use to the posterior segment only.Pre-veneered stainless steel crowns (PSSC) combine conventional SSC's durability with the esthetic appeal of a composite resin veneer. However, the significant concerns with PSSC were removing additional coronal tooth structure, inability to crimp the margins of the crowns before cementation, and loss of the esthetic acrylic facing.Prefabricated pediatric Zirconia crowns offer an excellent alternative to full-coverage crowns when restoring deciduous teeth with a sizeable carious defect. These crowns have high flexural strength, allowing them to resist crack propagation. The additional benefits of these crowns are the ability to replace metals due to extremely high strength and toughness, higher resistance to chemicals, and superior erosion resistance.Since the introduction of primary Zirconia crowns, several studies have reported their clinical success. Severely mutilated primary anterior teeth showed that NuSmile Zirconia crowns  offered superior esthetics and durability with remarkable gingival responses up to 24 months.This interventional study was done between 2014 and 2017 in accordance with the Declaration of Helsinki (as revised in Edinburgh 2000). It evaluated the clinical performance of NuSmile primary Zirconia crowns on 75 children  aged 2 to 8 years. The study included healthy and cooperative children managed by non-pharmacological and pharmacological behavioral management techniques. Children with no history of systemic illness and parents who signed the consent on behalf of their children and were willing to follow-up during the entire course of the study formed the study cohort. Inclusion criteria for the study were primary teeth with sufficient tooth structure and expected to survive for at least 2 years. Grossly carious non-restorable clinical crown, primary tooth with root resorption, tooth exfoliated within 1 year of crown placement, special need children, and parents not willing to come for follow-up were excluded from the study.Pulpal status was assessed clinically and confirmed by pre-operative periapical radiographs. Around 232 (172 primary incisors and 60 primary molars) pulp-treated primary teeth received a prefabricated Zirconia crown . The stThe same operator who placed the crown recorded clinical parameters such as gingival index, plaque index, restoration failure, and marginal integrity at 6, 12, 18, and 24 months. Both operators were standardized to evaluate index scores. The gingival index was recorded using a blunt periodontal probe, and a plaque disclosing swab was used to assess the plaque index. The gingival health and plaque scores were evaluated and compared with adjacent and opposing teeth. The evaluation of restoration failure and marginal integrity was done clinically with visual assessment of restoration, according to the United States Public Health Service criteria .Both operators prepared pulp-treated teeth for NuSmile Zirconia crowns as per the manufacturer's guidelines on crown placement protocol. There was no formal training or standardization between the two operators for crown placements except following technical guide instructions mentioned and recommended for use and general information on NuSmile Zirconia crowns. Before the tooth preparation, the operator chose a well-fitting prefabricated crown based on the pulp-treated tooth's original size. Local anesthesia was given using lidocaine 2% with epinephrine , especially when placing crowns without general anesthesia. Tooth structure of 1–1.5 mm was removed from the natural occlusal contours followed by 2 mm sub-gingival reduction with a special crown cutting kit provided by NuSmile. This opened up the interproximal contact areas and reduced the entire clinical crown by 20% (or 0.5–1.25 mm). All line angles of the prepared tooth were rounded. The crown's fit was checked, and crowns were cemented with Ketac Cem luting cement. Excess cement was removed, and crowns were held in place until the cement had been set.t-test. The Kaplan–Meier survival method was used to analyze restoration failure and marginal integrity of Zirconia crown for 2 years. The 95% confidence interval (CI) was included for stating the relevance of the finding, and the level of significance was set up atp < 0.05. The exfoliated teeth restored with NuSmile Zirconia crowns were evaluated for restoration failure and marginal integrity.Data were analyzed for descriptive data by SPSS version 20.0. Inferential statistics to compare gingival health scores/plaque level scores over the period between test tooth (NuSmile Zirconia crowns) and control tooth (adjacent/ antagonist tooth) was done using independent samplesp > 0.05).The prefabricated Zirconia crowns were assessed for restoration failure every 6 months for a 2-year follow-up period . The inOut of 172 anterior Zirconia crowns that were followed up at 6 months, 19 crowns were lost completely . At 24 The 2-year marginal integrity calculated with the Kaplan–Meier method revealed the NuSmile Zirconia crowns incisor group to be 94.5% (95% CI: 92.5–96.5). Kaplan–Meier 2-year marginal integrity estimate of the NuSmile Zirconia crowns incisor group was 92% (95% CI: 96.5–98) and the molar crowns group was 84.5 % (95% CI: 86.5–96) survival probability. The data show that incisor and molar significantly influence the time taken to lose marginal integrity, individually . PrefabPediatric dentistry has undergone a paradigm shift from clinical decision-making to a patient-sensitive treatment plan.The preformed crown is a promising method of providing substantial shelter for endodontic-treated primary teeth.Any metric to be justifiably validated in dentistry needs to be comprehensively drafted to evaluate its survivability, marginal integrity, and effect on surrounding oral structures. Hence, the purpose of this work was to efficiently investigate the longitudinal detailing of NuSmile Zirconia crowns' clinical performance for 2 years. Zirconia pediatric crown requires extensive subgingival preparation; hence, assessing its biocompatibility with gingiva is of paramount importance. In this work, better gingival scores consistently over 2 years were evidenced. This finding was similar to other studies reported earlier.The biological outcome measured in terms of mean gingival health scores corresponded to the results of plaque level scores for 2 years. This was in line with previous findings that polished and smooth surfaces result in less plaque accumulation and minor gingival irritation.The Zirconia crowns were cemented with conventional glass ionomer cement due to the limited control of hemostasis as the preparation for Zirconia crowns is 1–2 mm subgingival. However, as per manufacturer recommendations, the luting cement of choice for prefabricated zirconium crowns is bioactive resin-based glass ionomer cement which requires good gingival bleeding control prior to cementation. Numerous published studies have highlighted the success of resin-reinforced glass ionomers,The survival rate measured in terms of restoration failure for NuSmile prefabricated Zirconia restoration in this work reported no chipping or fracturing. Only loss of crowns was evaluated at each follow-up visit. The endodontic failures were not evaluated and, hence, not included. The absence of porcelain-veneered restoration and monolithic Zirconia preparation, which was said to exhibit higher resistance to fracture loads than layered Zirconia crowns and metal-ceramic crowns.Marginal integrity is an integral component of clinical performance. It has many subsets of factors—the first being marginal fit. The excellent finish line of the study group could have contributed to a high marginal fit in this study. The second factor is the vertical gap between the restoration and the prepared tooth abutment. Creating a favorable area for bacterial accumulate in the marginal gap jeopardizes the longevity of the restoration, leading to the development of secondary caries. No evidence of secondary caries in this study substantiates good marginal fit and a minimal vertical gap in this pediatric crown.In the present study, traumatized teeth that were infra-occluded and restored with the Zirconia crowns re-erupted along with the natural tooth, spontaneously suggesting a favorable prognosis.The strength of this research work is a sufficient sample size and comprehensive clinical performance evaluation longitudinally. Limitations of the present study were that both patients under general and local anesthesia formed the study cohort that could have resulted in mixed clinical outcomes challenging the external validity. Since it is a retrospective study, potential confounding factors such as patient selection, operator bias, and lack of standardization between operators can have an effect on the results. Moreover, further long-term studies among different commercially available Zirconia crowns can help us understand the differences in survival and biological outcomes in each group and foster amendments as needed. To summarize, pediatric Zirconia crowns have demonstrated better gingival health, decreased plaque accumulation, and reasonable survival rates longitudinally, especially for molars. Further research on various brands of Zirconia crowns and methods for longer retention of crowns in anterior teeth should be planned.Based on the findings of this study, we conclude that Zirconia pediatric crowns preserve and maintain gingival health. They also prevent microbial plaque accumulation similar to the natural tooth. These pediatric crowns have long-term survival rates with good retention and marginal integrity, indirectly preventing secondary caries. Furthermore, these crowns do not intervene with the path of eruption of infra-occluded traumatized teeth. Hence, Zirconia pediatric crowns are a well-suited restoration of choice for primary teeth rehabilitation.


Evaluation of the different causes listed in literature for the rate of success of Zirconia based restorations.With the help of PRISMA guidelines , this Systematic review was carried out. For a time span of 18 years that is from 2003 to 2020, articles were searched using three electronic data bases which are PubMed , Cochrane Library and Sciencedirect. The selected 27 articles which included the in vivo as well as the in vitro studies presented the performance of zirconia-based prosthetic restorations. The studies also stated the commonest reason for failure which ultimately depicted the rate of success of the fixed dental prosthesis. Due to heterogeneity of gathered information , meta analysis could not be carried out.Failure of bond between veneer material and zirconia sub-structure could be related to the cause of fracture of veneering porcelain hypothetically.Mechanical connection and building up of compressive strength due to thermal contraction at the time of cooling after sintering process is the reason for the bond developed amongst the two materials. Key words:Zirconia based restorations, zirconia failure cause. Esthetics satisfying the contemporary consideration for attractiveness is treated with the help of prosthodontic treatment by traditionally restoring the lost function of speech, chewing and deglutition. Social burden and welfare of the profession maximizes the conditioned necessity of esthetics .The materials of choice for the cases in which esthetics is the key expectation are ‘Ceramics’ in recent times of which ‘yttria-stabilized tetragonal zirconia polycrystals’ (Y-TZP) is the most advanced core ceramic . This paEven if zirconia-based ceramics being a prime material for fabricating FPDs, its high resistance to fracture could also endure high occlusal loads adding a major advantage.Nonetheless, cohesive fractures of the veneering ceramic is a ‘weak link’ of the restoration seen as the short-term clinical letdowns of zirconia-based restorations .-Review QuestionIn vivo as well as in vitro studies performed with zirconia based restorationsPopulation – Intervention – Studies with success rate of anterior and posterior zirconia based restorations as FPDs or single crowns.Outcome –Overall success rate of the zirconia based restorations-Literature searchFrom 2003 to 2020, articles were searched using three electronic data bases which are PubMed , Cochrane Library and Sciencedirect. Articles with full texts that contended the criteria for inclusion were attained. To include all relevant articles and for improving the electronic search, a final manual search was carried out amongst the selected articles to get cross references and citations.PubMed provided 114 articles and Science direct provided 8 articles and citation search provided 17 articles after the electronic and manual search was done. So far , no systematic review has been published on the current topic. Total 80 articles were excluded and 59 articles were screened. These 59 articles were completely analyzed by the title and abstract leading to selection of only 27 relevant articles which served the criteria for inclusion considered for the systematic review.-Results of data extractionBy gathering all the data after excluding the duplicates, full text of these 27 articles was attained lead by thorough screening of the remaining 59 articles. Therefore, for this systematic review 27 articles was the final sample size.-Results of included studiesNo inference has yet touched regarding the attempt to substitute the metal in metal ceramic restorations having ceramics of greater resistance. Its discussion began at the end of the 20th century. In current situations, Zirconium oxide the foremost target of research and trials held clinically. Chemical along with dimensional stability, mechanical resistance, hardness, and modulus of elasticity of the similar demand that of stainless steel are the primary characteristics supporting its usage as a biomaterial.Chipping of veneers often goes overlooked by the patient and is simply corrected by intraoral polishing or repair inferring that it is an esthetic defect of slight status. This is the reason which leads the rate of survival of zirconia-based fixed dental prostheses and metal ceramic restorations equivalent upto 97 to 99% over a period of 5 years.The greatest numbers of problems due to the usage of zirconium oxide in prosthetic conducts occur with fixed partial prostheses or fixed bridges. Various studies clinically showed cohesive type of fracture of the veneer material as a major and utmost liability. However, there is a debate as to the rate of occurrence of this mechanical letdown because of variations in the variables evaluated in various studies and the success rate of the prostheses have been calculated. They have been summarized in In vivo and In vitro studies.The studies were divided into groups - In vivo performance in studies-Fixed prostheses with zirconia substructure – The authors Pospiech (follow-up of 24 months) , BornemaVult von Steyern in his study with 24 months of follow up period , Peláez Finally, diversity in the studies carried out by Raigrodski (with 31months follow-up) , Sailer In vitro performance studies-Fixed prostheses with zirconia substructure – Concerning the mechanical performance of fixed prosthetic restorations, resisting the force of chewing deprived of getting fractured is the vital need. The first molar is exposed to a 300-800N force, although the anterior region is exposed to 60-200N of mastication force. Forces can exceed upto 1000N in few parafunctional cases : Fracture without disturbing the interface of ceramic core.Adhesive: Fracture occuring at the ceramic to core bond.in vitro studies that when there is fracture in the samples, a cohesive fracture pattern is suffered in the occlusal zone which is adjacent to antagonist’s contact point.It is seen in various Tsalouchou evaluated to static resistance loading of zirconia crowns which were 50 in quantity, analysing of the transversal plane by SEM analysis is done and also display that the most recurring type of fracture resulted as cohesive fracture.Similarl-SummaryThe rate of success was significantly predisposed by site, with crowns seated in the molar area displaying further biological and technical difficulties than anterior crowns. Fractures of ceramic material were also knowingly influenced by site, with molar crowns showing knowingly more risk for these fractures than anterior crowns.In vitro full-coverage restorations studies have seen a greater occurence of cohesive type of fracture for zirconia restorations. The higher incidence of chipping is explained in a study by Martin Rosentritt (2009) that assayed zirconia restoration fracture resistance, finding that all samples suffered cohesive fractures due to inadequate performance of the veneer material .The relationship between chipping phenomenon and risk factors occurring clinically, chiefly occlusal aspects, ought to be taken into consideration in upcoming prospective studies. Speciﬁc attention should be given by dental practitioners to clinical constraints when performing zirconia based restorations till an answer is found to enhance the mechanical resistance of the materials.


The use of novel ingredients is increasing in popularity within the pet food industry. Products utilizing both grain-free and more traditional ingredient profiles are widely available. Unfortunately, gastrointestinal impacts associated with changes in starch content and gelatinization from different grain or carbohydrate sources are unknown. The purpose of this work was to measure starch content and gelatinization in commercially manufactured dry dog food products and assess differences related to ingredient profile. Gelatinized starch was proportionately lower in products manufactured with traditional ingredients . Diets manufactured with non-traditional ingredients including alligator, buffalo, venison, kangaroo, squid, quail, rabbit, and salmon along with plant-based ingredients including tapioca, chickpeas, lentils, potato, and pumpkin had higher proportions of starch gelatinized. The degree of gelatinization can impact digestibility in the canine digestive tract and is an important area requiring further investigation.n = 10) utilizing protein sources including chicken, chicken by-product meal, meat and bone meal and plant-based ingredients including rice, barley, oats, and corn were examined in comparison with non-traditional diets (n = 10) utilizing protein sources including alligator, buffalo, venison, kangaroo, squid, quail, rabbit, and salmon along with plant-based ingredients including tapioca, chickpeas, lentils, potato, and pumpkin. Total starch and gelatinized starch  were measured with variation due to ingredient type assessed using Student’s t-test in SAS 9.4. Significance was set at p < 0.05. Total starch (as a percent of diet) was higher in traditional diets compared to non-traditional diets formulated for maintenance (p < 0.0032) or all life stages (p < 0.0128). However, starch gelatinization as a proportion of total starch was lower in traditional diets formulated for maintenance (p < 0.0165) and all life stages (p < 0.0220). Total starch and gelatinized starch had a strong negative correlation  in diets utilizing traditional ingredients. These novel data reveal important differences between starch content and gelatinization and may impact selection of various ingredient types by pet food manufacturers.Starch gelatinization in pet food may be affected by moisture, retention time, and ingredients used. Starch gelatinization has been associated with changes in digestibility but is not well studied using non-traditional ingredients in canine diets. The objective of this research was to examine differences in starch content and gelatinization associated with changes in ingredient profile  and nutrient content requirements associated with differing life stages. Traditional diets ( Non-traditional dietary formulations utilizing grain-free ingredient profiles for canines have become increasingly popular in recent years. These diets exclude traditional cereal grains but frequently contain other carbohydrates such as legumes or tubers. Legumes are frequently high in lysine, often a first-limiting amino acid, and as such are commonly utilized as complementary proteins to balance amino acid content ,2. PriorUnfortunately, digestibility studies are not plentiful in canine diets formulated with non-traditional ingredients. Although some data have reported digestibility potential for individual protein sources ,5, more Prior characterization studies have examined total starch, resistant starch, and starch gelatinization in commercially available pet foods . Howevern = 20) commercially available, dry extruded dog foods were purchased from two pet food suppliers (www.chewy.com and www.amazon.com) (accessed on 4 March 2019) during March 2019. Diets were selected based on ingredient profile and were classified as traditional (T), (n = 10) or non-traditional (NT), (n = 10) formulations. Traditional diets utilized animal-based ingredients including chicken, chicken by-product meal, meat and bone meal and plant-based ingredients including rice, barley, oats, and corn. Non-traditional formulations utilized meat-based ingredients including alligator, buffalo, venison, kangaroo, squid, quail, rabbit, and salmon along with plant-based ingredients including tapioca, chickpeas, lentils, potato, and pumpkin. Diets were further stratified within ingredient profile by life stage such that traditional diets included formulations for all life stages (n = 5) and for adult maintenance (n = 5). Similarly, non-traditional diets included formulations for all life stages (n = 5) and adult maintenance (n = 5). All diets selected were manufactured to meet or exceed NRC recommendations  were collected and shipped to a commercial analytical laboratory . All diets were analyzed for three variables of interest. First, total starch  including all hydrolysable carbohydrates, was analyzed following a method adapted from AOAC 996.11 and YSI Application Note 319. Second, gelatinized starch  was measured using a method adapted from AOAC 996.11 . Finallyt-test was used to analyze total starch, starch gelatinization, and total gelatinized starch in non-traditional and traditional ingredient formulations for maintenance and all life stage diets. Linear regression and correlation were used to determine the relationship between total starch content and total gelatinized starch in traditional and non-traditional diets. Values are expressed as mean (±SD). Significance was set at p < 0.05.Data were analyzed using SAS Studio . Student’s When ingredients were examined for the two diet formulation types, twenty-two animal and thirty-five plant sources were reported on product packaging see . Non-trap = 0.0032)  compared to non-traditional diets (22.3 ± 8.0%) formulated for adult maintenance ( 0.0032) .p = 0.0128).Similarly, traditional ingredient profile also had increased total starch for diets manufactured for dogs of all life stages (36.0 ± 7.8%) compared to non-traditional formulations (19.9 ± 3.4%) (p = 0.0165). Similarly, greater gelatinized starch was found in diets formulated for all life stages with traditional diets (30.9 ± 5.9%) compared to non-traditional diets (19.4 ± 3.4%) (p = 0.0220).When gelatinized starch  was examined across both ingredient profiles, traditional diets were higher than non-traditional diets  formulated for maintenance . Non-traditional diets had proportionately greater total starch gelatinization at 95.4 ± 2.1% and 97.6 ± 2.3% for maintenance and all life stages, respectively.r = −0.78; p < 0.01) was demonstrated in traditional diets, as shown in r = 0.28; p = 0.43) between total and gelatinized starch was measured in non-traditional diets . It is especially interesting to note that the negative correlation observed in traditional diets between gelatinization and total starch content was absent in non-traditional diets. Reasons for this difference are unclear but may include differences in amylose:amypectin ratio, degree of starch resistant to digestion, and other processing considerations.In 2008, extruded dog foods were estimated to make up approximately 95% of dry pet foods ,9. By alAlthough improved digestibility is commonly assumed with increased starch gelatinization, starch source can impact digestibility . Prior wOther work comparing grain-free and ancient grain diets  demonstrated full digestion of starch even with lower starch gelatinization for the ancient grain diet ,18. The The popularity of novel ingredients that is currently driving formulations including these ingredients must be approached systematically. Variation in components has significant impacts on gastrointestinal microbiota. Changes in microbial taxa have been reported for dogs offered raw meat diets with varying starch sources . It is pThe diets analyzed in our study differed in carbohydrate source. Grain-free diets have previously been found to have lower crude fiber, higher total dietary fiber (TDF), higher starch content, and a lower degree of processing . These fFuture studies should include in vitro digestibility trials to determine the relationship between starch content, starch gelatinization, and digestibility for a wide variety of commonly utilized pet food ingredients, including those that are from non-traditional sources. Ultimately, comprehensive digestibility trials utilizing AAFCO approved standards should be completed in order to identify impacts to canine health associated with ingredient variation and interactions .Although non-traditional dry extruded canine diets had less starch proportionally than traditional diets, the amount of starch gelatinized as a percentage of total starch was markedly higher in diets formulated with non-traditional ingredient profiles. Potential impacts of these differences are unknown and require future investigations. In conclusion, despite differences in total and gelatinized starch for traditional vs. non-traditional diets, no difference was measured based on life stage  despite markedly different nutrient requirements.