GRN), microtubule-associated protein tau (MAPT), and the chromosome 9 open reading frame 72 (C9orf72) repeat expansion, which could cause neurodegenerative pathological changes years before symptom onset. Noninvasive quantitative molecular imaging with PET or single-photon emission CT (SPECT) allows for selective visualization of the molecular targets in vivo to investigate brain metabolism, perfusion, neuroinflammation, and pathophysiological changes. There was increasing evidence that several molecular imaging biomarkers tend to serve as biomarkers to reveal the early brain abnormalities in familial FTLD. Tau-PET with 18F-flortaucipir and 11C-PBB3 demonstrated the elevated tau position in patients with FTLD and also showed the ability to differentiate patterns among the different subtypes of the mutations in familial FTLD. Furthermore, dopamine transporter imaging with the 11C-DOPA and 11C-CFT in PET and the 123I-FP-CIT in SPECT revealed the loss of dopaminergic neurons in the asymptomatic and symptomatic patients of familial FTLD. In addition, PET imaging with the 11C-MP4A has demonstrated reduced acetylcholinesterase (AChE) activity in patients with FTLD, while PET with the 11C-DAA1106 and 11C-PK11195 revealed an increased level of microglial activation associated with neuroinflammation even before the onset of symptoms in familial FTLD. 18F-fluorodeoxyglucose (FDG)-PET indicated hypometabolism in FTLD with different mutations preceded the atrophy on MRI. Identifying molecular imaging biomarkers for familial FTLD is important for the in-vivo assessment of underlying pathophysiological changes with disease progression and future disease-modifying therapy. We review the recent progress of molecular imaging in familial FTLD with focused on the possible implication of these techniques and their prospects in specific mutation types.Familial frontotemporal lobar degeneration (FTLD) is a pathologically heterogeneous group of neurodegenerative diseases with diverse genotypes and clinical phenotypes. Three major mutations were reported in patients with familial FTLD, namely, progranulin ( MAPT), progranulin (GRN), and the repeat expansions in the chromosome 9 open reading frame 72 (C9orf72). These mutations could lead to neurodegenerative pathological changes years before symptom onset  encompasses a set of clinical syndromes characterized by progressive abnormalities in behavior, executive function, language, or motor function. Patients with FTLD may present clinical syndromes with the behavioral variant of frontotemporal dementia (bvFTD), the nonfluent variant of a primary progressive aphasia (nfvPPA), a semantic variant of PPA (svPPA), and some patients also have amyotrophic lateral sclerosis (ALS), corticobasal syndrome (CBS) or progressive supranuclear palsy (PSP) . Approxiom onset .MAPT gene located on chromosome 17q21 first reported in 1998  and intron 10  tend to form four tandem microtubule-binding domain repeat (4R-tau) pathology, while mutations outside exon 10  tend to form mixed 3R/4R tauopathy similar to the tauopathy in Alzheimer's disease  repeat expansions of the  in 2011 , 13. TDPpansions . The moscarriers , 15.in vivo to investigate the brain topographic and pathophysiological changes. The former included metabolism, perfusion, neuroinflammation, synaptic function, and neurotransmitters' activity, and the latter comprised Tau and Aβ aggregation. There was increasing evidence that several molecular imaging biomarkers tend to serve as biomarkers to reveal the early brain abnormalities in familial FTLD. Identifying molecular imaging biomarkers for familial FTLD is important for the in-vivo assessment of underlying pathophysiological changes with disease progression and future disease-modifying therapy. Thus, we review the recent progress of molecular imaging in familial FTLD with a focus on the possible implication of these techniques and their prospects in specific mutation types.Currently, the visual inspection of MRI was demonstrated to easily increase the diagnostic confidence of underlying FTLD , 17. TheMAPT,” “progranulin” or “GRN,” and “chromosome 9 open reading frame 72” or “C9orf72” for genes; “positron emission tomography” or “PET,” “single-photon emission CT” or “SPECT,” and “dopamine transporter imaging” for molecular imaging biomarkers). The retrieval deadline was 1 December 2021. All the relevant articles were retrieved, placing restrictions on #elds (free-text terms searched exclusively in the title or abstract of the articles) and publication type .We performed electronic searches of Medline, PubMed, and Embase databases using the combination of a number of medical subject headings, Emtree subject headings, and free-text terms   , 21. In 11C-PBB3 , 18F-MK6F-MK6240 , and 18Fd 4R tau , 26 comptaucipir . For 18Fd design –31. 18F-MAPT mutations is regarded as tauopathy  and intron 10  had 4R tau pathology, while with MAPT mutations outside exon 10  had 3R/4R paired helical filament tau pathology  had little to no uptake, but the other MAPT R406W mutation carrier had a signal in the AD range   , 70, 76,rbetapir , 18F-florbetaben , 18F-fluafuranol , 18F-fluemetamol , 79, etcMAPT mutation indicated negative results with 11C-PiB or 18F-florbetapir PET -PET is a technique for measuring glucose metabolism in vivo , 18F-fluoropropyl-DTBZ, etc.], transporter density , or postsynaptic terminals  , etc.] . DopaminMAPT mutation carriers, particularly individuals with MAPT N279K mutation. Tracers such as 11C-DOPA and 2b-carbomethoxy-3b-(4-trmethylstannylphenyl) tropane (11C-CFT) were used to reveal dopaminergic function. The 11C-CFT uptake in the putamen was mildly low in asymptomatic MAPT N279K mutation carriers  and technetium-99m-ethylcysteinate dimer (99mTc-ECD), both which are distributed proportionally to rCBF  to assess brain function . The tra to rCBF . Perfusi to rCBF , includiMAPT mutation carriers, including eight in P301L, two in G272V, and one in G389R, significant hypoperfusion detected by 99mTc-HMPAO SPECT was found in the asymmetric frontotemporal lobes  have been developed to target TSPO to visualize neuroinflammation in vivo  mutation carriers or sporadic FTLD  is a transmembrane protein ubiquitously expressed in secretory vesicles of synapsis in all the brain areas . It is c1C-UCB-J . Reducedom onset . There i11C-MP4A PET could reflect acetylcholinesterase (AChE) activity in vivo. A study showed reduced AchE activity in the temporoparietal cortex in one of three asymptomatic MAPT N279K mutation carriers , Science and Technology Innovation 2030 Major Projects (2022ZD0213600), and Natural Science Foundation of Sichuan (2022NSFSC1325).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Klinefelter Syndrome (KS) is the commonest disorder that affects the sex chromosomes , occurriKS is a multisystem disease that does not just impact sperm production. Many issues surrounding hypogonadism, psychosexual morbidity, gynaecomastia and risk of developing further disease such as diabetes, osteoporosis, venous thromboembolism and cancer need to be discussed and require more research . The carEspehana et al., and describes a holistic, patient-centered approach that encompasses the key specialties and support required for MDT care in these patients—endocrine, urology and fertility, genetics, psychology and psychosexual medicine with expert radiology, pharmacy and specialty nursing support. A key feature is a patient liaison from a Klinefelter patient group, such as the KSA (Klinefelter Association) in UK. The success of the MDT approach has been quantifiable in terms of faster access to a wider set of clinicians, greater awareness of KS issues, greater patient and primary care satisfaction, and a cohesive management plan that acts as a roadmap for managing the condition (The framework for the development of a multidisciplinary team (MDT) based clinic is described in the paper by ondition .Potter and Prezzi, describes the characteristics of these imaged lesions often found in KS, and the challenges they present. Most have appearances of benign Leydig Cell tumors, which can be safely followed up or biopsied at the time of microsurgical sperm retrieval. Multiparametric ultrasound shows potential for improved lesion characterization and may assist in their management in future. With prompt discussion within an MDT setting, unnecessary surgery or prolonged imaging follow-up could be avoided in the presence of lesions with benign sonographic features; with prompt organ sparing surgery directed to lesions with malignant features.Another key feature of the MDT approach is the ability to discuss abnormal findings detected during investigation of patients, such as indeterminate lesions of the testis. These are common sonographic findings in infertile men, including those affected by KS. A commentary by Kailash et al.'s review of surgical sperm retrieval (SSR) in KS, and Pook et al.'s analysis of fertility preservation in the pediatric KS setting. Both papers highlight the need for a holistic multidisciplinary approach in order to best optimize patients for fertility harvesting. In adults, evidence is presented that suggests that the single most significant determinant for successful SSR is the age of the patient . Although these results appeared to be encouraging, limited sample sizes and the wide range of SSR rates is likely due to variation in testosterone pre-treatment, mosaicism/non-mosaicism, and level of testicular dysfunction. As such, no current guidelines exist on optimal timing for FP in KS, although from available evidence an age range of 16–30 is potentially optimal. The ethical considerations of fertility preservation in the pediatric age group are also discussed.Fertility topics are explored in two papers,  patient –12. The  patient , 13. Sev patient . A protoetrieval . In a reetrieval  Bryson eRaheem et al. reviewed the impact and management of excessive breast tissue (gynaecomastia) in KS, which affects 80% of KS patients (The final paper in the series looks at another often missed manifestation of KS that can have immense psychological impact. patients . In KS, patients  and its patients  has rendThe wide range of specialist topics covered in this comprehensive review reveals the clinical issues important to KS including and beyond fertility that would ideally benefit from a multi- disciplinary team approach. This approach has now been recommended in recent guidelines , releaseThe author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


CSMD1) is a novel candidate tumor-suppressor gene. We investigated CSMD1 in esophageal squamous cell carcinoma (ESCC) by performing an integrated analysis of somatic DNA alterations  with RNA expressions (mRNA and target miRNAs) on specimens from the same ESCC patients, using data from SNP, miRNA, and RT-PCR arrays. Our results indicate that the CSMD1 gene may play a role in the development of ESCC through complex patterns involving somatic alterations and mRNA expression. Furthermore, somatic copy number alterations in SNPs located in non-coding regions of CSMD1 appear to influence expression of both this gene and its target miRNAs.Human Cub and Sushi Multiple Domains 1  is a novel candidate tumor-suppressor gene that codes for multiple domains, including complement regulatory and adhesion proteins, and has recently been shown to have alterations in multiple cancers. We investigated CSMD1 in esophageal squamous cell carcinoma (ESCC) by performing an integrated analysis on somatic copy number alterations (CNAs), including copy-number gain or loss, allelic imbalance (AI), loss of heterozygosity (LOH), and the expressions of mRNA and its target miRNAs on specimens from the same patients with ESCC. Results: (i) Two-thirds of ESCC patients had all three types of alterations studied—somatic DNA alterations in 70%, and abnormal expressions of CSMD1 RNA in 69% and in target miRNAs in 66%; patterns among these alterations were complex. (ii) In total, 97% of 888 CSMD1 SNPs studied showed somatic DNA alterations, with most located near exons 4–11, 24–25, 39–40, 55–56, and 69–70. (iii) In total, 68% of SNPs with a CNA were correlated with expression of CSMD1. (iv) A total of 33 correlations between non-coding SNPs and expression of CSMD1 target miRs were found. Conclusions: Our results indicate that the  CSMD1) gene is thought to be a novel candidate tumor-suppressor gene that codes for multiple domains, including complement regulatory adhesion proteins and a membrane protein with an extracellular region, a single transmembrane domain, and a short cytoplasmic domain, although its function in human tumors is currently unknown  of 20 paired samples from Affymetrix 500k and 36 paired samples from Affymetrix 5.0 were loaded by using R/bioconductor and aroma.affymetrix. The aroma.affymetrix package can process nearly all Affymetrix Copy Number and SNP arrays and can provide estimates of total copy number for all loci and allele B fractions for all  SNPs. For more details, go to http://hgdownload.soe.ucsc.edu/goldenPath/hg19/database/ (accessed on 22 June 2022)). Transcripts with accessions starting with “NM_” were used. A total of 888 CSMD1 SNPS were found to have the necessary information  to be included in our analysis, and all are located in intronic regions of CSMD1.For identifying physical location (exon or intron), an in-house python script was developed to retrieve the gene name and intron location for each SNP in our list from hg19 Refseq database from the UCSC genome browser  The copy number (CN) was defined by using the Log R ratio (LRR) as follows: CN high gain ≥ 1.0, CN gain > 0.2, CN big loss ≤ −1.0, and CN loss < −0.2. (iii) AI and LOH were identified by using B Allele Frequency (BAF) as follows: Allelic imbalance was identified when BAF was 0.2–0.45 or 0.55–0.80, and LOH was identified when BAF was <0.2 or >0.8.The criteria for DNA segment alterations are as follows: (i) There was a significance threshold of 5.0 × 10−ΔΔCT algorithm. Details of the 2−ΔΔCT method have previously been described [http://docs.appliedbiosystems.com/search (accessed on 22 June 2022); [−ΔΔCT method was used to calculate relative changes in gene expression determined from real-time quantitative PCR experiments. In the present study, the data are presented as the fold change in each exon of CSMD1 expression in tumors normalized to the internal control gene (GAPDH) and relative to the normal control . The results of the real-time PCR data were represented as CT values, where CT was defined as the threshold cycle number of PCR at which the amplified product was first detected. There is an inverse correlation between CT and amount of target: lower amounts of target correspond to a higher CT value, and higher amounts of target have lower CT values. The average CT was calculated for both the target genes and GAPDH, and the ΔCT was determined as  minus . The ΔΔCT represented the difference between the paired tissue samples, as calculated by the formula ΔΔCT = . The N-fold differential expression in the target gene of a tumor sample compared to the normal counterpart was expressed as 2−ΔΔCT [http://docs.appliedbiosystems.com/search (accessed on 22 June 2022)) [Gene expression was analyzed by using the 2escribed  (http://e 2022); ). Briefls 2−ΔΔCT  (http://e 2022)) . In the http://www.lifetechnologies.com/about-life-technologies.html (accessed on 22 June 2022)). Signals for miRNA present either in tumor only or normal only were dropped from our analysis. The fold change was calculated by using the 2−ΔΔCT method, as described previously [http://www.targetscan.org/)  and Sanger miRBase (http://www.mirbase.org/) to identify conserved miRNAs in the 3′ UTR for CSMD1. A total of 22 miRNAs were analyzed in the present study.RQ Manager integrated in software from ABI was used to normalize the entire signal generated. The expression level (as fold change) was calculated when both tumor and normal samples had signals in the assays, using DataAssist software v2.0  for each exon of CSMD1 with each of its target miRNAs. All analyses were conducted by using R software. If there were less than five cases showing signals on a miRNA or a probe of an exon of CSMD1, the miRNA or probes for an mRNA were excluded from the analysis. All analyses considered comparisons with p-values < 0.05 to be significant .To determine whether SNPs in the non-coding region affected mRNA expression or not, we performed Spearman correlation analyses between the signal value of each SNP with a somatic DNA alteration (Log RR for CNA) and the value (fold change) of each exon of CSMD1 by using the Kaplan–Meier Estimator in the survival analysis package in R software, which also included hazard ratio estimations from proportional hazards models. The patients were divided into two groups according to fold change: >1 fold change (FC) and ≤1 FC. A significant difference in survival status was detected by using the log-rank test and p-value < 0.05.We performed a survival analysis for the mRNA expression of each miRNA target and each exon of CSMD1 as CNAs, AI, and LOH and determined that 70% of cases (39/56) had such DNA alterations  and showed that overexpression was the predominant category   in the 56 ESCCs. Finally, we examined the relations among the three alterations  in erations . Second,category a. Indivicategory b. Third,ression) a, and alression) b. Table erations a,b, whic in ESCC .CSMD1 were examined in the present study, all located in non-coding regions. In brief, 97% of these SNPs (864/888 SNPs) showed somatic CNAs, with most alterations around exons 4–11, 24–25, 39–40, 55–56, and 69–70  of CSMD1. These somatic CNAs showed complex patterns, suggesting ESCC heterogeneity. For example, case E0844 and case E1293 showed the CN gain on the entire gene, while case E1210 and case E1242 showed CN loss throughout CSMD1. The distribution of DNA alterations in each of the 56 ESCC cases is summarized in CSMD1 in the 56 ESCC patients. Overall, 70% of patients (39/56) showed somatic alterations, including CN loss/LOH/AI (36%), CN gain/AI (16%), and only AI (11%). ESCC patients could be divided into two groups based on the somatic alterations: 70% of patients (39/56) had somatic CNAs, while 30% (17/56) had no CNAs on CSMD1  and 24 overexpressed (average FC ≥ 2). We also noted that patients with and without somatic DNA alterations showed similar percentages with abnormal expression of CSMD1 70% (26/37) and 65% (11/17), respectively.We selected specific exons in  regions  were useCSMD1 gene expression. (iii) The status for cases with mixed CN gain/loss and LOH and their mRNA expression on CSMD1 was also complex. For example, case E0410 had CN gain from exons 24 to 70 and LOH in the same region  A subset of cases (8/26 = 31%) with CNAs/AI/LOH followed the same direction as their mRNA expression, while other cases showed opposite patterns . (ii) Wee region , while me region .http://www.targetscan.org/)  and Sanger miRBase (http://www.mirbase.org/) to identify 22 conserved miRNAs in the 3′ UTR for CSMD1, which are thought to be preferentially conserved. We used TargetScan  and underexpressed in just 1 (≤0.5 FC). Similar abnormal expression rates of CSMD1 target miRNAs were observed both for patients with (67%) and without DNA alterations (65%).Sixty-six percent of cases (37/56) had abnormal expression of −5), 179 were nominally significant (p < 0.05)  with rho values ranging from 0.64 to 1.00, and 42% that were negatively (inversely) correlated, with rho ranging from −1.00 to −0.64  correlated in oppositive directions at different regions . These results suggest that some SNPs with somatic CNAs located in introns are capable of influencing gene expression, either directly or indirectly, although the mechanisms are currently unclear.Among the 888 total SNPs examined, correlations were evaluated between each of the 123 SNPs with somatic CNAs in non-coding regions and the 11 miR probes. Although none of these comparisons reached the significance threshold after adjustment for multiple comparisons (3.69 × 10 < 0.05) . These 1to −0.64 a. Among CSMD1 targeted miRNA were nominally significant (p < 0.05). These included 27 positively correlated SNPs with rho values from 0.79 to 1.0, and 6 inversely correlated SNPs with rho values from −0.86 to −1.0 . Thirty-three correlations between 30 SNPs with CNAs and expressions of 10  to −1.0 b. Of not to −1.0 .CSMD1 target miRNAs were significantly positively associated with expressions of exons 5–6, 8–9, 9–10, 10–11, 24–25, 55–56, and 69–70  (The expressions of seven to 0.64) c. Three to 0.64) .CSMD1 expressions as well as selected miRNA expressions , especially at exons 4–5  and exons 8–9  , miR-135b , miR-449a , and miR-449b ). In contrast, four miRNAs were expressed at lower levels in patients with DNA alterations than those without )  a. A tota lower)) b. These p = 0.026). Frequencies of somatic DNA alterations were similar between patients with (0.56) and without (0.53) lymph node metastasis.The study unexpectedly showed that patients without a family history of upper gastrointestinal tract cancer had a significantly higher frequency of somatic alterations than those with a positive family history . Only 2 of the 22 targeted miRNAs had expressions that were significantly associated with survival based on patients with expression (FC < 1 or FC ≥ 1).Finally, we examined expression levels of p = 0.014; median survival 30.1 vs. 21.3 months; p = 0.001; median survival 50.2 vs. 23.1 months; Patients with expression level < 1 FC had longer survival than those with ≥1 FC for miR19b ), 2.5 (1.17–5.2), CSMD1 somatic DNA in conjunction with both mRNA and targeted miRNA expression in the same patients and samples. Our results show complex patterns among alterations of DNA and expression of RNA and target miRNAs related to this gene. We identified CSMD1 DNA alterations in two-thirds of ESCC patients. When we divided patients by the presence/absence of these alterations to further examine relations with mRNA and miRNA expression, our results indicated that CSMD1 exhibited substantial heterogeneity in tumor nucleic acid alterations and expression levels, even though the cases all arose from the same high-risk geographic area and might be expected to show more molecular homogeneity. Recently, scientists have shown that investigating tumor heterogeneity may help us to understand the biology of tumors [The present study is the first analysis in ESCC patients to simultaneously examine alterations of f tumors .CSMD1 or, alternatively, that the diverse methods used across these studies  may have influenced these variations [CSMD1 were in introns adjacent to exons 4–11, 24–25, 39–40, 55–56, and 69–70, an observation that may facilitate detecting potentially meaningful SNPs to examine in related genes. Related previous studies showed that CSMD1 had biallelic instability and somatic mutations in exons in Chinese ESCC patients [CSMD1 plays a role in the development of ESCC, although the patterns of somatic CNA, AI, LOH, and mutations appear complex and highly heterogeneous.Somatic DNA alterations were identified in 70% of ESCC cases in this study, a higher prevalence than has been reported in other cancers, such as breast 55%) or lung cancer 46%) [5% or lun% [5% or patients ,18,19. CCSMD1, we had a similar question, namely, did the non-coding SNPs with DNA alterations we identified influence gene expression in CSMD1? To answer this question, we performed qRT-PCR with 11 probes, covering 16 coding regions and splice sites to capture expression in the DNA-alteration regions. In brief, we observed three phenomena: First, we found that 69% of ESCC patients with somatic DNA alterations had abnormal expression of the CSMD1 gene. Second, we found consistency between expression of mRNA and somatic DNA alterations in 31% of cases , but not in others ; case E1862, CN gain but underexpression (0.24 FC)). This phenomenon may result from several factors, such as mutations, structural variants, or methylation. Third, we found that CSMD1 expression levels varied across different coding regions. For example, in case E0387 (with CN loss and AI), two probes were underexpressed (FCs 0.32 for exons 7–8 and 0.35 for exons 6–7), two probes (for exons 10–11 and 24–25) showed average expression, and six probes were overexpressed (2.20–9.16 FC) for other exons. Overall, expression in this case averaged 2.77 FC, but it ranged from 0.32 to 9.16 FC. There are three genes at the 5′ end of CSMD1, including two pseudogenes , and one uncharacterized LOC107986907 (an RNA gene affiliated with the ncRNA class). It is unknown whether these genes influence CSMD1 expression. Richter et al. reported that aberrant splicing can cause the alteration of CSMD1 expression in cancer cell lines and may change CSMD1 via formation of a nonsense codon through deletion of exons [CSMD1 gene expression in tumors. Our results indicated that SNPs with CNAs in non-coding regions could affect gene expression, particularly since methylation may influence expression in these regions. The first exon of CSMD1 contains a CpG island that is thought to cause underexpression of mRNA [CSMD1 and so did not select probes to cover exon 1. Thus, we are unable to evaluate methylation on exon 1. Further, larger studies are needed to better understand individual gene expression and to determine whether splicing variants differentially influence function or expression. Other technologies, such as RNA-seq, might also help to better understand splicing variants of unknown significance.A major challenge in human genetics is pinpointing which non-coding genetic variants affect gene expression and influence disease risk. Since all the SNPs that showed somatic alterations in our study were located in non-coding regions of  of mRNA . HoweverGenome-wide association studies (GWASs) have emerged as powerful and successful tools to identify common SNPs associated with the risk of human diseases, including cancer. One approach to investigate the link between GWAS-identified variants and function is to look for variants that influence phenotype, for example, by comparing GWAS variants for differences in gene expression as determined by examination of expression quantitative trait loci (eQTL). Studies on genome-wide eQTLs in humans can help us to prioritize likely causal variants among the multiple SNPs within the regions identified by GWAS. However, most significant SNPs identified by GWAS are in non-coding regions of genes or in intergenic regions far from genes; thus, there remain challenges to explain the exact roles that “risk” SNPs identified by eQTL play in human diseases. One of our previous studies showed that SNPs in non-coding regions not only appeared to be potential risk biomarkers by classic eQTL, but also carried somatic CNAs that may influence gene expression in ESCC . The preCSMD1 expression based on their genome-wide profiling of miRNAs expressed in glioblastoma stem cells and normal neural stem cells [CSMD1 in ESCC. For example, miR-130a and miR-130b were positively correlated with expression in multiple exons in CSMD1. The abnormal expression of miR-130b has also been reported in other cancers  [CSMD1 target miRNAs in ESCC . Finally, seven SNPs with somatic alterations had a correlated expression of both CSMD1 and its target miRs, although the number of SNPs was small. Altogether, our results suggest that somatic alterations in non-coding SNPs may affect the expression of both mRNA and miRs in ESCC.Gene-expression status is also influenced by target miRNAs. Lang et al. reported that both miR10a and miR-10b could repress em cells . Our stuCSMD1. In addition, information on other types of somatic alterations  was not available for these cases. Future studies are also needed to help determine whether this integrated approach may lead to the identification of biological markers that may be used to determine clinical relevance or other mechanisms of action between CSMD1 and ESCC.There are several limitations in the present study, including the relatively small sample size. Thus, some of the significant associations reported here likely occurred by chance, and these findings need to be replicated and validated in future larger studies. Furthermore, the SNP arrays used here did not include SNPs in the exons of CSMD1 identified somatic DNA alterations and abnormal expressions of mRNA and its target miRNAs in two-thirds of ESCC patients. We also found that alterations in SNPs in non-coding regions of CSMD1 influenced the expression of both CSMD1 and its target miRs.In summary, our evaluation of


The analysis of the recorded SERS spectra based on the assignments of Raman bands from the density-functional theory (DFT) calculations and based on the SERS surface selection rules reveals that when molecules of dopamine are adsorbed from an aqueous solution to which no electrolytes have been added, they adopt a flat orientation versus the silver surface; whereas, the molecules of dopamine co-adsorbed with various ions interact with the silver surface, mainly via phenolic groups, and they adopt a perpendicular orientation versus the metal surface. An addition of electrolytes also significantly influences the intensity of the recorded SERS spectrum; for example, an addition of Na2SO4 to a final concentration of 1 M induces an increase in the intensity of the measured SERS spectrum by a factor of ca. 40. This means that the addition of electrolytes to the analyzed solution can reduce the limit of detection of dopamine by SERS spectroscopy. The abovementioned findings may facilitate the construction of dopamine SERS sensors.The abnormal metabolism or imbalance of dopamine may lead to some neurological disorders. Therefore, the facile and fast detection of this neurotransmitter is essential in the early diagnosis of some diseases. One of the methods that can be used for the detection and determination of dopamine is the surface-enhanced Raman scattering (SERS). In this contribution, we report a very strong influence of some salts (we used salts containing Na Dopamine  is one of the most important neurotransmitters in the human body, and it belongs to the catecholamine family (the structure of the dopamine molecule is shown in −7 M [−7 M [−9 M [−10 M) was achieved using SERS spectroscopy [Many methods for detecting dopamine have been reported, including high-performance liquid chromatography (HPLC) ,12, elec−7 M  and 3 ×  M [−7 M , respect M [−9 M . Howevertroscopy . Moreovetroscopy ,20. In ttroscopy .In summary, many various methods can be used for the detection and determination of dopamine. All developed methods have some advantages and disadvantages, but SERS spectroscopy, due to its low limit of detection, was considered to be a very promising technique for carrying out such analyses. Moreover, in SERS measurements, it is possible to record a good quality spectrum even from a single molecule ,22; therThe determination of the orientation of the molecules of dopamine versus the silver surface was carried out using SERS surface selection rules and assignments of the Raman bands of dopamine, which were previously obtained in density-functional theory (DFT) simulations ,27. DFT Sodium fluoride, sodium chloride, sodium bromide, sodium iodide, sodium sulfate(VI), and dopamine hydrochloride were purchased from Sigma-Aldrich Poland . Potassium chloride and ethanol were acquired from Chempur . All of the reagents mentioned above were used without further purification. The water was purified using a Millipore Milli-Q system . The silver plates were acquired from Mennica Polska .The nanostructured silver substrates for the SERS experiments were prepared by electrochemical roughening of silver plates. The silver electrodes were obtained by cutting 10 × 5 mm rectangles from the silver sheet. Immediately before carrying out the electrochemical nanostructuring, the surface of the silver electrodes was sanded with sandpaper to remove the outer oxidized layer, and the electrodes were then thoroughly washed with ethanol and water. When placed in an electrochemical cell, only the fragment of the electrode with dimensions of 7 × 5 mm was immersed in the electrolyte. The electrochemical roughening was carried out in a three-electrode system . The primary electrolyte used was a 0.1 M KCl solution. First, three successful positive–negative–positive cycles were performed from –0.3 V to 0.3 V to –0.3 V. The sweep rate was 5 mV/s. Then, the silver electrode was kept at –0.4 V for 5 min to reduce all of the remaining AgCl.The Microlab 350  microscope was used for the scanning electron microscopic (SEM) morphological characterization of the surface of the nanostructured silver.The electrochemical roughening of the silver electrodes was performed in a conventional three-electrode cell using a PGSTAT204 potentiostat/galvanostat .All Raman measurements were performed using a Horiba Jobin-Yvon Labram HR80  spectrometer equipped with a Peltier cooled charge-coupled device detector (1024 × 256 pixels), a 600 groove/mm holographic grating, and an Olympus BX40 microscope  with a long-distance 50× objective. A Nd:YAG laser provided the excitation radiation, with a wavelength of 532 nm; the power of the laser beam on the laser head was 70 mW.The SERS measurements were carried out for the SERS silver substrates immersed in a small vessel into which 1 mL of an appropriate solution  was introduced. The dopamine solutions  were obtained by the appropriate diluting of a dopamine hydrochloride solution with a concentration of 0.1 M. In the case of the measurements in the electrolyte solution, an appropriate amount of salt was introduced into 5 mL of dopamine solution or into 5 mL of water.The SERS measurements were made on the surface of a macroscopic nanostructured silver electrode  have been assigned to various C–H out-of-plane bending; whereas, the two other bands (at 1291 cm−1 and 1630 cm−1) are due to relatively complex ring vibrations [The experiments were started by recording the SERS spectra of dopamine  a local increase in the intensity of the electric field in close proximity to the illuminated plasmonic nanostructures due to the excitation of the localized surface plasmons, where the increase in the efficiency of the generation of the Raman spectrum is roughly proportional to the fourth power of such field enhancement ,33, and −5 M dopamine aqueous solution to which various amounts of Na2SO4 and NaF have been added, respectively . As it can be seen from the comparison of the spectra presented in change in the spectral patters in the recorded SERS spectra;change in the intensity of the recorded SERS spectra.Dopamine is usually administered as a drug in the form of a solution in saline (0.9% or 0.15 M aqueous solution of NaCl). Therefore, we decided to check how the presence of some electrolytes in the solution from which dopamine is adsorbed on the SERS substrates affects the intensity of the measured SERS spectrum. −1 due to a catechol ring breathing contributed mainly by the stretching of a carbon–carbon bond to which oxygens are attached  as a function of the concentration of the introduced halides. As it can be seen from As it can be seen from 2SO4 to a final concentration of 1 M induces an increase in the intensity of the measured SERS spectrum by a factor of ca. 40. The abovementioned dependences should be taken into account during the interpretation of the results obtained with the dopamine SERS sensors .Dopamine is one of the most important neurotransmitters in the human body, and disturbed concentration levels of dopamine have been connected to various neurological disorders, such as Parkinson’s disease, Alzheimer’s disease, or schizophrenia. Therefore, the sensitive detection of this neurotransmitter is necessary, for instance, in the diagnostics of diseases or in the control of medical treatments. One of the most sensitive techniques for the detection and determination of dopamine is SERS spectroscopy . In this


Using visual bibliometric analysis, the application and development of artificial intelligence in clinical esophageal cancer are summarized, and the research progress, hotspots, and emerging trends of artificial intelligence are elucidated.http://bibliometric.com/) and an online browser plugin  were used.On April 7th, 2022, articles and reviews regarding the application of AI in esophageal cancer, published between 2000 and 2022 were chosen from the Web of Science Core Collection. To conduct co-authorship, co-citation, and co-occurrence analysis of countries, institutions, authors, references, and keywords in this field, VOSviewer (version 1.6.18), CiteSpace (version 5.8.R3), Microsoft Excel 2019, R 4.2, an online bibliometric platform . In terms of the H-index (45) and citations (9925), the United States topped the list. The journal “New England Journal of Medicine” of Medicine, General & Internal (IF = 91.25) published the most studies on this topic. The University of Amsterdam had the largest number of publications among all institutions. The past 22 years of research can be broadly divided into two periods. The 2000 to 2016 research period focused on the classification, identification and comparison of esophageal cancer. Recently (2017-2022), the application of artificial intelligence lies in endoscopy, diagnosis, and precision therapy, which have become the frontiers of this field. It is expected that closely esophageal cancer clinical measures based on big data analysis and related to precision will become the research hotspot in the future.An increasing number of scholars are devoted to artificial intelligence-related esophageal cancer research. The research field of artificial intelligence in esophageal cancer has entered a new stage. In the future, there is a need to continue to strengthen cooperation between countries and institutions. Improving the diagnostic accuracy of esophageal imaging, big data-based treatment and prognosis prediction through deep learning technology will be the continuing focus of research. The application of AI in esophageal cancer still has many challenges to overcome before it can be utilized. Atypical hyperplasia and infiltration of the esophageal squid epithelium or glandular epithelium leads to the emergence of esophageal carcinoma (EC), which is mainly divided into two types, esophageal adenocarcinoma (EAC) and esophageal squamous cell carcinoma (ESCC), according to the proliferation of different epithelia. Globally, ESCC accounts for 84% of esophageal cancer cases, while EAC accounts for 15% . EC is tArtificial intelligence (AI) has been established and has played a considerable role in the medical industry, owing to the emergence of deep-learning algorithms, computer hardware developments, and the exponential rise of data that is generated and used for clinical decision-making , 11. AI While there are many reasons to be optimistic, there are several obstacles to the successful integration of AI in clinical esophageal cancer. In terms of EC detection of premalignant and malignant lesions, while histopathologic examination is the gold standard for establishing the diagnosis of EC and determining the presence of dysplasia, endoscopists must collect targeted biopsies from particular areas that host the real lesion. AI can assist clinicians in performing directed biopsies rather than depending on random samples by detecting locations that may harbor Barrett’s esophagus with or without dysplasia; this AI assisted method of biopsy has been presented as a potential solution to the aforementioned problem , 16. SomKeeping up with the rapidly changing corpus of literature is vital not just because new discoveries occur from a wide range of fields, but also because new results may profoundly alter the collective knowledge of everyone researching AI . As inteHere, we determine the countries, institutions, authors, or journals with the highest citations/publications of AI in the field of EC by collecting literature data in the database, and therein describe the challenges faced in the EC clinical translation of AI. The aim of this study is to characterize the application and progress of AI in EC from 2000 to 2022 utilizing bibliometric analysis and to identify the current research progress, hotspots, and emerging trends of AI in EC, which may assist new researchers comprehending future research and identifying areas of interest for further research.The data source was The Science Citation Index Expanded (SCI-Expanded) of Clarivate Analytics’ Web of Science Core Collection (WoSCC). Web of Science applies a strict screening process. Bradford’s Law in bibliometrics states that only prominent academic publications from many areas are included. SCIE, as a journal citation subdatabase of WoSCC, is a multidisciplinary comprehensive database covering the field of natural science, with over 8,600 global authoritative journals encompassing 176 topic categories.To guarantee that no data updates were made, two researchers from our organization examined the information of papers concerning AI in the field of EC simultaneously and finished the search in one day. The articles’ titles, keywords, abstracts, authors, institutions, and reference data were obtained and stored in plain text format. The following was the search formula:1#: esophag* (Topic) or oesophag* (Topic) or gullet (Topic) and Article OR Review (Document Type) and English (Language) 2#: cancer* (Topic) or tumor* (Topic) or tumor* (Topic) or neoplas* (Topic) or onco* (Topic) or carcinoma* (Topic) and Article OR Review (Document Type) and English (Language) .3#: 1# AND 2# 4#: artificial intelligent* (Topic) or computational NEAR/5 intelligence (Topic) or expert* system* (Topic) or intelligent learning (Topic) or feature* extraction (Topic) or feature* mining (Topic) or feature* learning (Topic) or machine learning (Topic) or feature* selection (Topic) or unsupervised clustering (Topic) or image* segmentation (Topic) or supervised learning (Topic) or semantic segmentation (Topic) or deep network* (Topic) or bayes* network (Topic) or deep learning (Topic) or neural network* (Topic) or neural learning (Topic) or neural nets model (Topic) or artificial neural network (Topic) or data mining (Topic) or graph mining (Topic) or data clustering (Topic) or big data (Topic) or knowledge graph (Topic) or AI (Topic) and Article OR Review (Document Type) and English (Language) .5#: 3# AND 4# .Two researchers independently analyzed the data to ensure the accuracy of the data and the repeatability of the research. Microsoft Excel 2019 was used to analyze and export the files of top-cited or productive authors, countries/regions, publications, journals, and institutions. H-index was a hybrid index proposed by Hirsch that can be used to evaluate academic achievements . AltmetrCiteSpace is a popular information visualization method in the field of knowledge graphs . This rehttp://bibliometric.com/) and the bibliometrix R package for bibliometric analysis were used. The calculation of the almetrics is performed as “Almetric Attention Scores” through a free browser plugin provided by Almetric .The VOSviewer is a scientific knowledge graph application that can depict the structure, progression, coordination, and other aspects of knowledge fields by constructing linkages and visually analyzing literary knowledge items . In thisThe data used in this study were acquired from an open source and do not require approval by any ethical committee.As per the study strategy flowchart, we eventually gathered 918 publications from the Web of Science (SCI-E) panning the previous 22 years, comprising 769 articles and 149 reviews Figure 1In this category, 53 countries/regions have published relevant publications. According to the global map in All of the papers were published in a total of 638 journals, with 378 of the journals publishing at least 10 articles. More than 118 universities played an active role in AI application research at the European Commission, with the top three TLSs being the University of Amsterdam (TLS = 72), Catharina Hospital (TLS = 64), and Eindhoven University of Technology (TLS = 53). In addition, the Co-institutions knowledge mapping was constructed, with N = 530 and E = 745. (Density was 0.053). The research includes 5 979 authors and 39,962 co-cited writers. Mani Menon, Ficarra V, and Tewari Ashutosh K had the most citations in a co-cited author network study. Tewari A’s and Kattan MW’s BC were as high as 0.4 and 0.35, respectively, showing that their contributions had a significant impact in this sector. The modularity Q value was used to assess the network’s clustering impact. The higher the value is, the better the network’s clustering performance. The silhouette value, which was used to quantify network homogeneity, was another indication. The modular Q value was 0.7218, and the mean silhouette S value was as high as 0.9248, as shown in This analysis included a total of 918 publications, 80 of which were cited at least ten times. Horie Y et al.  had the Keyword frequency analysis clarifies the research patterns in this study. As seen in Clusters are characterized along a horizontal timeline in a timeline visualization. Generally, citation bursts indicates the evolution of the study issue over time, just as shown in Artificial intelligence (AI) research has accelerated in the previous 22 years, with clinical applications being examined for the majority of medical professions. The discipline of EC, which is highly dependent on imaging studies, is no exception. Bibliometric analysis, as opposed to systematic review, uses visual tools to completely examine the current literature to intuitively comprehend the development pattern of research and identify future research hotspots. This is the first studies to use bibliometric analysis to summarize the contemporary use of AI in EC and intuitively illustrate the development trend and future research hotspots by applying two commonly used literature measurement software tools: VOSviewer and CiteSpace.The trajectory of the average citations of each article every year over the last 22 years allows us to see changes in the volume and quality of AI research in the field of esophageal cancer. The instability of the average citations per paper per year in the early days has developed into a relatively stable fluctuation range in the past 6 years, which suggests that the development of the entire discipline is maturing. China contributed the most to total publishing volume of any country , the creation of objective scoring systems for risk stratification, forecasting disease prognosis, or therapeutic response. Co-citation analysis is frequently used to assess an author’s academic influence. As shown in The changes in research fronts can be seen through the timeline view of clustering results of co-cited references . We may The study of the frequency of keywords may reflect the development tendency of research hotspots from another point of view, which further confirms the findings of this study. As shown in Comparing the application in esophageal cancer with other fields, modern research on the prognosis, survival and risk factors for esophageal cancer is bound to become a hot spot in the future. In particular, the word “database” appeared for the first time in 2018. Obviously, with the emergence of big data, the processing and application of large amounts of data has become an important research method. Through big data, we can apply artificial intelligence to conduct comprehensive analysis and extensive research on clinical esophageal cancer. However, at the same time, data require many human and financial resources, making data collection very difficult and valuable, which may be one of the reasons for the lack of cooperation in most studies.Looking at the entire study, the application of artificial intelligence in esophageal cancer has gone through two significant stages. The early emphasis was on esophageal oncogenes such as p53, classification/identification, and comparison of esophageal cancer. Meanwhile, the risk factors and prognosis of esophageal cancer were intermingled. Recently, the database was coupled with deep learning, convolutional neural networks, and machine learning. These areas are considered is hotspots which are the recent frontier in the examination, diagnosis, and therapy combination choices for esophageal cancer. The subject of artificial intelligence research for esophageal cancer is now approaching a new stage that will lead to the term “precision”. As a result, it will undoubtedly influence preoperative and postoperative nursing and clinical procedures for patients with esophageal cancer.AI currently appears to have indisputable potential, and in laboratory settings it has shown good enough performance and high enough precision to enhance the care of cancer patients and impact the cancer field more broadly. With the further development of artificial intelligence, the overall development of esophageal cancer toward precise inspection, diagnosis and treatment appears promising. The challenges of applying AI to esophageal cancer in the future may mainly lie in individualized data collection of esophageal cancer , data quality (such as ethnic differences in data differences), and data processing specifications . Inconsistency), AI code reproduction , and auxiliary diagnosis credibility decision-making . Finally, in terms of retrieval time, it may result in the loss of research hotspots in 2022. Only records before April were included this year. Last but not least, our study in WoSCC only contains English literature, which may result in the absence of essential literature in other languages. In addition, future research, databases such as Scopus and Google Scholar might be incorporated and compared for more thorough results.via deep learning technology, therapy and prognosis prediction based on big data. The difficulties of AI application in esophageal cancer may be mostly found in personalized data collection, data quality, data processing requirements, AI code reproduction, and helped diagnosis decision-making dependability.In conclusion, artificial intelligence is steadily taking over esophageal cancer research. Although China has the most published articles in this discipline, the United States, the Netherlands, and the United Kingdom have a greater influence and involvement in this field. The frequency of national research collaboration must be increased, particularly for emerging nations. Nations should work hard to retain strong ties with industrialized countries such as the United States. The use of AI in the field of esophageal cancer is generally behind, and the focus of this area will shift to increasing diagnosis accuracy The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The study was created by LW and X-QZ. J-XT gathered the data and prepared the paper. The data were examined by X-TL, H-QY, S-LY, L-FD, and R-LZ. The manuscript was revised and reviewed by LW, X-QZ, and J-XT. The essay was written by all of the writers, and the final version was approved by all of them.This study was supported by the Economic Evaluation Project of Early Diagnosis and Early Treatment of Urban Cancer in Jiangxi Province (No. JXTC2020040486), the National Natural Science Foundation of China , and the Nanchang University Students’ innovation and entrepreneurship training program (2022).The authors would like to express their gratitude to Dr. Jie KUANG and Dr. Yong LIU for their assistance in polishing the English content of this publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Pyrroloquinoline quinone (PQQ), present in breast milk and various foods, is highly recommended as an antioxidant, anti-inflammatory agent, and a cofactor in redox reactions in several biomedical fields. Moreover, PQQ has neuroprotective effects on nervous system disorders and immunosuppressive effects on different diseases. Herein, we report on the optimum fabrication of electrospun CS/PVA coaxial, core/shell, and uniaxial nanofibers. The morphological, elemental, and chemical structure of the fabricated nanofibers were investigated and discussed. PQQ, as a drug, was loaded on the uniaxial nanofibers and in the core of the coaxial nanofibers and the sustained and controlled release of PQQ was compared and discussed. The results revealed the privilege of the coaxial over the uniaxial nanofibers in the sustained release and reduction of the initial burst of PQQ. Remarkably, the results revealed a higher degree of swelling for CS/PVA hollow nanofibers compared to that of the uniaxial and the coaxial nanofibers. The coaxial nanofibers showed a lower release rate than the uniaxial nanofibers. Moreover, the CS/PVA coaxial nanofibers loaded with PQQ were found to enhance cell viability and proliferation. Therefore, the CS/PVA coaxial nanofibers loaded with PQQ assembly is considered a superior drug delivery system for PQQ release. Pyrroloquinoline quinone (PQQ), present in breast milk and various foods, is highly recommended as an antioxidant, anti-inflammatory agent, and a cofactor in redox reactions in several biomedical fields. The obtained nanofibers, from the aforementioned technique, have diameters that range from a few nanometers to several micrometers.2 The applied external voltage causes repulsive forces within the polymer's solution to exceed the surface tension of a solution droplet at the tip of a needle. Consequently, it creates deformation upon the polymer solution's spherical droplet to elongate forming a Taylor cone, which in turn creates the strands of nanofibers. In fact, the created nanofibers still have to travel a sufficient distance from the needle to the collector to allow them to dry out from any remaining solvent, producing dry, well-separated solid nanofibers.3 There are numerous factors that can affect the fibers' morphology and diameter including the polymer viscosity, humidity, flow rate, tip-to-collector distance (TCD), and applied voltage.4–7 Moreover, the accumulation of high surface area and high porosity nanofibers to form a membrane is a perfect candidate to be used in a wide range of medical applications, including tissue engineering and drug delivery.8The process of forming nanofibers using an applied external field has been reported for the first time by Formhals in 1934.9–11 In comparison with uniaxial nanofibers, coaxial fibers have many advantages where the coaxial technique enables the spinning of the material, which is not easily spun via uniaxial electrospinning.12,13 Moreover, in medical applications, sensitive bioactive substances such as antibiotics, drugs, DNA, and proteins, which are spun as the core solution, can be safely isolated from the harsh environment by the shell layer of nanofibers. Subsequently, controlling the drug release is possible via tuning the thickness of the core and shell.13,14 In addition, similar work has been done using novel biomaterial coaxial nanofibers for bone and cartilage regeneration.15,16Coaxial electrospinning is an improvement of the uniaxial electrospinning process in which two blunt-tip needles are connected to a high voltage with the same or different flow rates. Coaxial electrospinning provides nanofibers with a different and separate polymer solution for the core and shell, a tool used to fabricate hollow nanofibers or fibers with a varying core–shell.N-acetyl glucosamine and glucosamine linked by glycosidic bonds.17 Many previous studies reported that the electrospinning of pure CS is not easy due to high viscosity, limited solubility and the strong molecular interaction of CS.18,19 Therefore, to facilitate the synthesis of CS nanofibers, chitosan/other polymer blends such as PVA have been used.19,20 CS/PVA nanofibers have been used in many biomedical applications such as tissue engineering, drug delivery and wound healing due to their antibacterial, biocompatible and biodegradable properties. Our study selects coaxial CS/PVA nanofibers loaded with the natural component pyrroloquinoline quinone (PQQ) as a promising material for biomedical applications such as a drug delivery system for many medical applications.In the present study, chitosan (CS) and polyvinyl alcohol (PVA) were selected as core and shell polymers, respectively. CS is a natural biopolymer with a repeated polysaccharide unit, which consists of 21,22 PQQ was initially identified as a redox enzyme cofactor and later proved to be an essential nutrient for animal growth. It has been reported that the removal of PQQ from the diet causes growth impairment in animals.23,24 As an essential nutrient, PQQ has been classified as a new B vitamin with many other beneficial effects, such as anti-inflammatory,25 hepatoprotective,26 cardioprotective,27 and antioxidative properties.28,29 Furthermore, much evidence shows that PQQ is a key player in the central and peripheral nervous systems. For example, PQQ shields the brain against reversible middle cerebral artery occlusion,28 and increases the stimulus-response and the nerve growth factor levels in astroglial cells.30 In addition, it enhances the locomotor function after spinal cord injury in rats. This is achieved by lessening the secondary neuronal damage following primary mechanical injury31 and it further improves the regeneration of rat sciatic nerves.32 Consequently, PQQ is considered a perfect candidate for many medical applications and a key solution for complications, especially in the nervous system.PQQ is an anionic, water-soluble compound that has been discovered in several fruits, vegetables, breast milk, and tissues of mammals.Herein, we demonstrate the fabrication of CS/PVA uniaxial and coaxial nanofibers and compare their efficiency for the sustained release of pyrroloquinoline quinone (PQQ). The fabricated materials were fully characterized and compared. Also, the cell viability and proliferation of the cells on both classes of nanofibers were evaluated for use in biomedical applications.Mw: 190.000–310.000 Da), PVA , and acetic acid  were purchased from Merck, USA. PQQ was purchased from Sigma (USA).CS  3 g of CS (3% w/v) were dissolved in 100 ml of AA/distilled water solution (2% v/v) and heated in a water bath at 90 °C with vigorous stirring for 2 h until complete dissolution, and used as a stock.(2) 20 g of PVA (20% w/v) were dissolved in 100 ml of distilled water and heated in a water bath at 90 °C with vigorous for 3 h until complete dissolution, and used as a stock.34(3) 70 ml of PVA/distilled water solution were mixed with 30 ml of CS/AA/distilled water solution in volume ratios of 30/70 of CS/PVA, with vigorous stirring till reaching a homogenous solution.33,34 The pH and viscosity of various prepared solutions were adjusted using a pH meter and a viscometer.(4) After complete homogenization, 40 ml of distilled water were added to 60 ml of CS/PVA homogenous mixture and were allowed to vigorously stir for 1 h at 60 °C to maintain the viscosity of the solution.(5) For drug loading, PQQ (15% w/v) was added to the mixture, where PQQ solution was prepared by dissolving in distilled water. The CS/PVA solution loaded with PQQ was homogenized using a homogenizer for 1 h at 3000 rpm. Finally, the solutions (CS/PVA and CS/PVA loaded with PQQ) were left on the bench overnight to get rid of air bubbles before the electrospinning process.−1 and a TCD of 15 cm. For the hollow CS/PVA nanofibers, CS/PVA solution was loaded in the syringe of the outer needle only while the inner needle was filled with air. The applied voltage, flow rate, and TCD were 27 kV, 0.4 ml h−1, and 15 cm, respectively. The temperature and humidity were controlled at 25 °C and 30–40%. However, for the fabrication of CS/PVA uniaxial nanofibers and CS/PVA/PQQ coaxial nanofibers, CS/PVA and CS/PVA/PQQ solutions were loaded in two separate syringes. The CS/PVA solution was loaded as the outer needle (shell) and the CS/PVA/PQQ solution was loaded in another syringe as the inner needle (core). The applied voltage and TCD were 27 kV and 15 cm, respectively. The temperature and humidity were controlled at 25 °C and 30–40%. The flow rate of the outer needle's syringe was 0.4 ml h−1, while the flow rate of the inner needle's syringe was 0.3 ml h−1.An electrospinning apparatus  was used to fabricate nanofibers with hollow and core/shell structures. The blunt-tip coaxial needle was composed of a 23 G  inner needle concentrically mounted on an 18 G  outer needle where each of them has a separate syringe pump. In order to fabricate uniaxial CS/PVA/PQQ nanofibers, CS/PVA/PQQ solution was loaded in the syringe of the uniaxial set with an applied voltage of 27 kV, a feed rate of 0.4 ml hThe elemental and morphological analysis of the fabricated nanofibers were performed using a field emission scanning microscope  and a transmission electron microscope . To confirm the chemical bonding, Fourier Transform Infrared spectroscopy  was used to characterize the chemical structure of the uniaxial CS/PVA/PQQ, CS/PVA hollow and CS/PVA/PQQ coaxial nanofibers.Wi) was recorded. For the swelling test, the pieces of nanofibers were immersed in phosphate buffer solution  at 37 °C for 1, 3, and 7 days, and the samples were weighed again immediately after removing them from PBS and recorded as (W). Subsequently, the excess amount of PBS absorbed with filter paper and the weight of the nanofiber sample (Wx) were recorded. The degree of swelling of the nanofibers was calculated using W is the weight of swollen nanofiber samples by immersion in PBS, pH = 7.4 at 37 °C for 1, 2 and 3 days. Wd is the weight of the dried nanofiber sample.CS/PVA uniaxial, hollow and core/shell membranes were cut into small pieces of 3 × 3 cm and the initial weight of nanofibers (Wd) was recorded. The degradation index (%) was calculated using Wi is the initial weight of the nanofibers.Regarding the degradation test, the nanofibers were immersed in phosphate buffer solution  at 37 °C for 1, 3, and 7 days, and the nanofibers were dried in a vacuum oven for 24 h and the weight of the dried nanofiber sample . The nanofibers were placed in the donor compartment, while PBS with pH 7.4 was placed in the receptor compartment with adjusting temperature at 37 °C and the stirring rate at 200 rpm. Samples (2 ml) were taken at 24, 72, 120 and 168 hours and diluted to measure the PQQ concentration by using a UV-visible spectrophotometer at a λmax of 280 nm.Using a Franz diffusion cell apparatus  CS/PVA/PQQ uniaxial nanofibers, and CS/PVA and CS/PVA/PQQ coaxial nanofibers were prepared to test drug release. The area of Franz diffusion cells was 1.76 cm35 Briefly, 100 μl of the test bacteria were grown in 10 ml of fresh media until they reached a count of approximately 108 cells per ml for bacteria.36 100 μl of microbial suspension was spread onto agar plates corresponding to the broth in which they were maintained. Isolated colonies of each organism that might be playing a pathogenic role should be selected from primary agar plates and tested for susceptibility by the disc diffusion method.37,38 Gram(+) bacteria such as Staphylococcus aureus and Bacillus subtilis and Gram(−) bacteria such as Escherichia coli and Pseudomonas aeruginosa were incubated at 35–37 °C for 24–48. The diameters of the inhibition zones were measured in millimeters.35 Standard discs of ampicillin  served as positive controls for antimicrobial activity but filter discs impregnated with 10 μl of solvent  were used as a negative control. Blank paper disks  with a diameter of 8.0 mm were impregnated with 10 μl of tested concentration of the stock solutions. When a filter paper disc saturated with a tested chemical is placed on agar the chemical will diffuse from the disc into the agar. This diffusion will place the chemical in the agar only around the disc. The size of the area of chemical infiltration around the disc is determined via the solubility of the chemical and its molecular size. If an organism is placed on the agar it will not grow in the area around the disc if it is susceptible to the chemical. This area of no growth around the disc is known as a “Zone of inhibition” or “Clear zone”. For the disc diffusion, the calibers of the National Committee for Clinical Laboratory Standards38 were used to quantitatively measure the inhibition zone diameters. The agar-based methods including tests such as Etest and disk diffusion can be good substitutes for broth-based methods due to their simplicity, higher accuracy, and the less time taken for the test.39,40The antimicrobial activity of the nanofiber samples was determined using a modified Kirby–Bauer disc diffusion method.in vitro), in a Fibrolife® serum-free medium, human dermal fibroblasts, adult (HDFa)  cell cultures were cultivated at 37 °C and 5% CO2 in T75 culture flasks  for 24–27 hours. In order to determine the effect of CS/PVA hollow and CS/PVA/PQQ core/shell nanofibers on the proliferation of HDFa via 3--2,5-diphenyl-2H-tetrazolium bromide (MTT) assay the percentage viable cell was used41 as follows:Regarding the evaluation of human fibroblast cell proliferation (4 cell per well in Corning® 96-well tissue culture plates.(1) CS/PVA uniaxial, hollow and core/shell nanofibers were suspended (three replicates for each sample) with HDFa in a Fibrolife® serum free medium at a concentration of 5 × 10(2) Then the plates were incubated for 72 hours.(3) Vehicle control with DMSO was run for each 96 well plate (untreated cells).42,43(4) After incubating for 72 h, the number of viable cells was then determined by adding 10 μl of 12 mM MTT stock solution  into each well, including also the untreated control.2 for 4 hours.(5) The 96-well plates were then incubated at 37 °C and 5% CO(6) The cells were periodically viewed under an inverted microscope  to detect the presence of intracellular punctuate purple precipitation. The optical density was measured at 490 nm with a microplate reader  to determine the number of viable cells.t is the mean optical density of the wells treated with the tested samples (drug or the selected formulation), while ODc is the mean optical density of the untreated cells.(7) The percentage of viability was calculated using the following −1. On the other hand, the core was made hollow by filling the syringe with air using the same flow rate of the shell solution (0.4 ml h−1). The coaxial electrospinning technique was used to fabricate CS/PVA hollow nanofibers using acetic acid/distilled water solution as a solvent. On one hand, CS/PVA nanofibers, as a shell, were synthesized under constant processing conditions where the concentration of CS/PVA mixtures was 50%, with a voltage of 27 kV, TCD of 15 cm, humidity 30–40%, and flow rate of 0.4 ml h−1 for core solution and 0.4 ml h−1 for shell solution). Note that the coaxial nanofibers exhibited uniform fibers with a shell/core structure. The presence of core fibers with a smaller diameter within the shell fiber is evident. To further investigate the presence of the core/shell structure within the nanofibers, the core/shell as well as the hollow nanofibers were scanned by TEM. The TEM images shown in The morphology of the uniaxial CS/PVA/PQQ electrospun nanofibers was observed by FESEM. −1 confirms the presence of a saccharide structure as it is a major characteristic peak of the C–H bending in chitosan. Moreover, the 1096 cm−1 peak can be attributed to the C–O stretching vibrational mode in chitosan. Also, the peaks at 1375 and 1415 cm−1 are characteristic of the CH3 symmetrical deformation mode. The peak at 1640 cm−1 can be ascribed to the amide bond in chitosan. The vibrational band between 2790 and 2980 cm−1 can be ascribed to the C–H stretching of the alkyl groups. Finally, the O–H stretching shown in the broad band from 2980–3660 cm−1 is caused by the intramolecular and intermolecular hydrogen bonding in PVA. The PQQ spectra show 4 peaks located between 1505 and 1723 cm−1 that are characteristic of the C PBM data was replaced with SVG by xgml2pxml:<glyph-data id="z.dbd" format="PBM" resolution="300" x-size="8" y-size="10" xml:space="preserve">00000000000000000000000000000000111111110000000011111111000000000000000000000000</glyph-data><svg xmlns="http://www.w3.org/2000/svg" version="1.0" width="13.200000pt" height="16.000000pt" viewBox="0 0 13.200000 16.000000" preserveAspectRatio="xMidYMid meet"><metadata>Created by potrace 1.16, written by Peter Selinger 2001-2019</metadata><g transform="translate scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg>O stretching vibrations of the bonding between quinone and carboxyl groups. As is lucid from the CS/PVA/PQQ spectra, the reaction between PQQ and CS/PVA caused the minimization/loss of most of the peaks except for the minor two peaks located at 1632 and 2363 cm−1 and the minor broad band at 3430 cm−1. They were indicated on the figure due to their low transmission.The FTIR spectra were collected to elucidate the chemical bonding for CS/PVA, PQQ, and CS/PVA/PQQ as shown in To assess the physical properties of the fabricated nanofibers for biomedical applications, the degree of swelling and degradation are considered critical parameters that need to be quantitatively evaluated. The results of the degree of swelling are represented in −1.44 During the first hour of the test, 20% of PQQ from the uniaxial nanofibers was released. However, only 10% of PQQ was released from the coaxial nanofibers without initial burst release. Moreover, the PQQ release from the core of the coaxial nanofibers reached 57.2% after 168 hours of the experiment. In contrast, in the same time interval of the test, a cumulative release of 74% of PQQ occurred from the uniaxial nanofibers. Note that the coaxial nanofibers have a lower release rate than the uniaxial nanofibers. Since PQQ exists in the core of coaxial nanofibers, the CS/PVA shell acted as a barrier to slow down the rate of release of PQQ. In comparison, the uniaxial nanofibers, in which PQQ was loaded on their surface, have shown a rapid release. The difference in the release profile shown in 44In order to verify the previous results and better compare the uniaxial and coaxial nanofibers, a release study of the PQQ was carried out by loading PQQ in the core of the CS/PVA//CS/PVA coaxial nanofibers as well as by loading the surface of CS/PVA uniaxial nanofibers, see 45 Secondly, PQQ is naturally used in bacteria to change glucose into gluconic acid, which has a pH of 5–5.5.46 Since PQQ is added in excess to the bacteria (loaded on the nanofibers), more glucose is changed into gluconic acid, which consumes high energy. The bacteria used in the test work were at neutral to slightly alkaline pH. Thus, the accumulation of gluconic acid causes slight impairment of the bacterial behavior, motility, and growth.47 The aforementioned concluded modes of action can be confirmed by the less inhibition zone diameter of the hollow CS/PVA nanofiber due to the absence of PQQ. There is not much difference between the inhibition of Gram positive and Gram negative bacteria, which could be ascribed to the broad spectrum of the investigated coaxial/uniaxial nanofibers. In addition, it could also be attributed to the antibacterial mechanism that does not interact with the cell membrane.p < 0.05)  . The difIn conclusion, electrospun CS/PVA coaxial, core/shell, and uniaxial nanofibers were successfully fabricated. FESEM imaging revealed the good uniformity of the fabricated nanofibers with a fine structure that is free from beads. Moreover, their TEM images proved the hollow and core/shell structures. The elemental composition and the chemical bonding for CS/PVA, PQQ, and CS/PVA/PQQ were elucidated using EDX and FTIR analyses. The results proved the privilege of CS/PVA coaxial nanofibers in comparison with uniaxial nanofibers when loaded with PQQ for biomedical applications. The release study of PQQ showed that CS/PVA coaxial nanofibers have a lower release rate than uniaxial nanofibers with less initial release burst in comparison with uniaxial nanofibers. In addition, coaxial nanofibers have lower degradation and swelling rates than uniaxial nanofibers. Furthermore, it enhances the proliferation of HDFa cells. The obtained results strongly suggest the fabricated CS/PVA coaxial nanofibers loaded with PQQ in the core for use in many applications as an alternative to many complicated and expensive drug systems.There are no conflicts to declare.


Significant progress in parasite disease control has been made recently by applying novel tools and methodologies developed in genomic, epigenomic, proteomics, and metabolomics research. The research topic aimed to introduce some of the key developments and challenges of the field, focusing on parasite-induced immune response, genetic and epigenetic regulation mechanisms, and potential therapeutics. This Research Topic comprises 41 papers, including three reviews and 38 original research articles, spans topics that cover parasite biology, host pathology, epidemiology, and prevention and control programs.Liu et al. successfully obtain a high-quality genome assembly of Moniezia expansa (M. expansa) by applying the next-generation sequencing techniques   on different coverage depth datasets of Piroplasm  biosynthesis are differently expressed and correlated significantly with immune response in the placenta and colorectal cancer, indicating a potential biomarker or immune therapy targets for CRC.Whole-genome sequencing (WGS) provides the most comprehensive characterization of the genome of a given organism. It allows for discovering previously unknown genes or variants associated with a unique parasite phenotype. BioNano) . De novo species . An origiroplasm . The autch topic . It has ch topic . The parch topic . HoweverFeng et al. carried out the first comprehensive comparative epigenomic analysis in Trichinella species ; Toxocara canis—dog ; Coccidia microbiota—host ; Cysticercus cellulosae—host ; Schistosome japonicum—human host  and Trichinella spiralis—mice . For example, Yao et al. found that T. gondii virulence factor ROP18 of the type I RH strain (TgROP18I) interacted with human TRIM21. This E3 ligase plays a vital role in anti-infection responses against intracellular pathogens for its immune escape. The supporting evidence includes 1) the interaction of TgROP18I with human TRIM21, 2) the phosphorylation of TRIM21 promoted by gROP18I, 3) TgROP18I promoted TRIM21 degradation, and 4) TRIM21 restricted T. gondii replication through NF-κB activation. This study revealed the possible mechanisms of the different disease outcomes caused by type I and type III T. gondii. In the review article by Lu et al., Coccidia microbiota interactions and their effects on the host were discussed. The direct and indirect interactions between Coccidia and gut microbiota were highlighted. The role of mechanical mucosal damage, chemical mucosal damage, disruption of the immune system, and probiotics on Coccidia development were discussed.Host-parasite interactions play a central role in the evolution and dramatically influence their biology. Identification of the critical determinants of the host-parasite interactions could provide novel molecular targets for developing tools against parasitic diseases. This research topic covered various interactions between parasites and their host , such asTogether, this research topic brings important new data and provides a broad and diverse overview of cellular and molecular basis in parasitic diseases control. The collected articles are timely and address important questions in the field with overwhelmingly positive feedback . We hope this research topic will promote further investigations and drive the field toward better control for parasitic diseases.


Three structurally related gelators, each containing octadecyl chains, an α-diketo group at the 9,10 positions, and each with a different N-amide group—isobutyl (DIBA), isopentyl (DIPA) or N-(2-(dimethylamino)ethyl) (DMEA)—have been synthesized. Their neat structures as well as the thermal mechanical, and photophysical properties in their gel states with various liquids have been investigated. The gelator networks of DIBA and DIPA in octane, hexylbenzene and silicone oil consist of bundles of fibers. These gels are partially thixotropic and mechanically, thermally (to above their melting or silicone oil gelation temperatures), and photophysically stable. They are mechanically and thermally stronger than the gels formed with DMEA, the gelator with a tertiary amine group. The lone pair of electrons of the tertiary amine group leads to an intra-molecular or inter-molecular charge-transfer interaction, depending on whether the sample is a solution, sol, or gel. Neat, solid DMEA does not undergo the charge-transfer process because its amino and diketo groups are separated spatially by a large distance in the crystalline state and cannot diffuse into proximity. However, the solution of DIPA upon the addition of triethylamine becomes unstable over time at room temperature in the dark or (more rapidly) when irradiated, which initiates the aforementioned charge-transfer processes. The eventual reaction of the gelators in the presence of a tertiary amine group is ascribed to electron transfer from the lone-pair on nitrogen to an α-diketo group, followed by proton transfer to an oxygen atom on the anion radical of the α-diketo group from a methyl or methylene group attached to the nitrogen atom of the cation radical. Finally, the formation of an α-diketyl radical leads to irreversible electronic and structural changes that are observed over time. Gelators are molecules that can self-assemble into 3-dimensional (3-D) networks in the presence of a liquid through covalent or non-covalent processes and forces, such as hydrogen-bonding, van der Waals interactions, dipole–dipole interactions, or London dispersion interactions . The liqAggregation-induced emission enhancement (AIEE) in LMOGs containing a luminogen within their molecular structure is another area of growing interest . Such syRelated to these observations, bimolecular photoinitiator systems comprised of an α-diketone and a tertiary amine have been used as dental restorative materials ,18. A meIn this work, we report the aggregation, viscoelastic, and photophysical properties of three alkyl amide derivatives of DODA: N-isobutyl-9,10-dioxooctadecanamide (DIBA), N-isopentyl-9,10-dioxooctadecanamide (DIPA), N-(2-(dimethylamino)ethyl)-9,10-dioxooctadecanamide (DMEA) . All conOf special interest here is the nature of the interactions between the tertiary amine and α-diketo groups in DMEA . If the The research presented here focuses on three aspects of these topics: (1) synthesizing, characterizing and applying new classes of gelator molecules with different n-alkyl, tertiary carbon and tertiary amine groups at the terminus of the amide chains to study their gelating abilities in organic solvents; (2) gaining a deeper insight into the thermal, mechanical, and bulk properties of the organogels formed from these gelators; and (3) understanding the photophysical properties of these gelators both in solution states and in aggregates , especially contrasting when they do (as in DMEA) and do not (as in DIPA) contain a tertiary amino center. In addition, the effects on gelation of the DIPA homologue, DIBA, are explored. Detailed descriptions of the syntheses and physical properties of DIBA, DIPA and DMEA are included in the d , benzophenone , isostearyl alcohol , anhydrous magnesium sulfate , molecular sieves , p-xylene , sodium lumps , cyclohexane , dimethyl formamide , sodium metabisulfite , sodium bisulfite , and TEM copper grids with carbon/formvar coating . Ethyl oleate , acetic anhydride , potassium permanganate , sulfuric acid , sodium bisulfate , sodium bicarbonate , sodium chloride , sodium sulfate , methanol , hexane , ethyl acetate , tetrahydrofuran , triethylamine , ethyl chloroformate , acetic acid , Isobutylamine , isopentylamine , N,N-dimethyl ethylenediamine , dimethyl sulfoxide , silicone oil DC 550 , carbon tetrachloride , octane , decane , 1-butanol , acetonitrile , benzene , toluene , chlorobenzene , nitrobenzene , 1-hexylbenzene , acetone , chloroform-All reagents were used as received except tetrahydrofuran, which, before use, was dried over 4 Å molecular sieves, refluxed with sodium metal and benzophenone and distilled, to remove water .1H NMR and 13C NMR spectra were obtained on a Varian 400 MHz spectrometer operating at 400 MHz (100 MHz for 13C NMR) with tetramethylsilane (TMS) as the internal standard. The IR spectra of neat gelators were obtained on a Perkin–Elmer Spectrum One FT-IR spectrometer using an attenuated total reflection accessory (ATR) in a range of 4000–400 cm−1 as the average of 8 scans with a resolution of 2 cm−1.Melting points were obtained at a heating rate of ~1 °C/min on a Leitz 585 SM-LUX-POL microscope equipped with crossed polars, a Leitz 350 heating stage and an Omega HH503 microprocessor thermometer attached to a J-K-T thermocouple. Polarized optical micrographs were obtained on a Leitz 585 SM-LUX-POL microscope and images were captured on a Photometrics CCD camera interfaced to a computer using the flame-sealed flattened capillary tubes (Vitro Dynamics). Elemental analyses of neat gelators were performed using a Perkin–Elmer 2400 CHN elemental analyzer with acetanilide as the calibration standard. Differential scanning calorimetry (DSC) scans were obtained on a Q200 differential scanning calorimeter (TA Instruments) in TZero hermetically sealed aluminum pans with a nitrogen flow rate of 50 mL/min. Thermogravimetric analyses (TGA) for DIBA and DIPA were performed on a TA 2910 differential scanning calorimeter interfaced to a TA Thermal Analyst 3100 controller in open aluminum pans under a slow stream of nitrogen. Samples were equilibrated initially at 30 °C and the heating rate was 5 °C/min. TGA for DMEA was performed on a TA Instrument Q5000IR TGA controller under a slow stream of nitrogen in a 100 μL open platinum pan. Samples were equilibrated initially at 23 °C, and the heating rate was 3 °C/min. Transmission electron microscopy (TEM) experiments were performed using JEOL JEM-2100 Plus Electron Microscope with an accelerating voltage of 200 kV. Images were analyzed using TEM center for JEM-2100 Plus Software by JEOL. The width of the fibers was obtained from ImageJ software  . Rheology experiments were performed on an Anton Paar-Physica MCR 302 strain-controlled rheometer attached to a Peltier temperature-controlled base plate using a parallel plate geometry (diameter 25 mm) and a 0.5 mm gap between the plates. Data were analyzed using Rheoplus/32 Service V3.62 Software by Anton Paar. UV-vis absorption spectra were obtained at room temperature using a Cary 5000 UV-vis-NIR spectrophotometer from Agilent Technologies in quartz cuvettes 1 × 1 cm for solutions or 2 mm capillaries for gels. Steady-state excitations and emissions spectra were recorded using a SYS 2459 Photon Technology International Fluorimeter, with a 75 W Xenon lamp, a Quantum Northwest Peltier temperature controller, an Omega temperature probe and a PTI 814 photomultiplier detector. Felix 32 software by PTI was used for data analyses.d was passed through basic alumina before recording the 1H NMR spectrum of DMEA to remove traces of DCl/HCl, which is formed over time and can protonate a tertiary amine and alter proton shifts in the NMR spectrum [Chloroform-spectrum . Gels were prepared by placing weighed amounts of DIBA, DIPA or DMEA and a solvent in a 5 mm (id) borosilicate glass tube that was flame-sealed, heated to the boiling point of the solvent (or to 90 °C) and then placed directly into an ice-water bath (~0–2 °C) for 10–15 min. The samples were then kept at room temperature for 1 h (fast-cooled method). The slow-cooled gels were heated as above and then allowed to return to room temperature over 3–4 h while remaining in the slowly cooling water bath. Samples were considered gels if they did not fall over periods of ~1–2 min when the tubes were inverted and if their appearance did not indicate the formation of macro-crystalline aggregates .Samples, prepared by the fast-cooling method, were placed in sealed capillaries, inverted and submerged in a stirred water bath, and heated at a rate of 1–2 °C/min to determine the melting ranges reported in the tables. CGCs were taken as the lowest concentrations at which a sample did not fall under the force of gravity at room temperature. Stabilities of fast-cooled gels are the periods of time at room temperature before unperturbed samples fell or phase-separated visibly under the influence of gravity.Aliquots (5–10 mg) of gels from 3 wt% DIBA, DIPA or DMEA in silicone oil, made by the fast-cooling method, were placed in TZero hermetically sealed aluminum pans, equilibrated at 0 °C for 5 min, heated to 110 °C (or 90 °C for DMEA gels), and kept at 110 °C (or 90 °C for DMEA gels) for 5 min. The cooling cycles involved cooling from 110 °C (or 90 °C for DMEA gels) to 0 °C and keeping them at 0 °C for 5 min. The heating and cooling cycles were repeated 3 more times. For neat DIBA, DIPA or DMEA, heating cycles started at 40 °C for 1 min, before heating samples to 120 °C (or 95 °C for DMEA) and keeping them at 120 °C (or 95 °C for DMEA) for 10 min. The cooling cycle involved cooling from 120 °C (or 95 °C for DMEA) to 40 °C and keeping the samples at 40 °C for 30 min. The cycles were reproduced 3 more times. Heating and cooling rates were 5 °C/min for all samples.The methodologies for the different aspects of the TEM experiments are described below .Sample preparation method 1: One drop of a sol of 0.05% DIBA in octane was placed on a 400-mesh copper TEM grid supported with carbon/formvar coating and dried in air for 2–3 days (leading to a noticeable solid on the grid). The procedure was repeated 4 more times to deposit more DIBA on the grid. One drop of a sol of 0.5 wt% DIBA in hexylbenzene was placed on a 400-mesh copper TEM grid supported with carbon/formvar coating and dried in air for 2–3 days .Sample preparation method 2: Small amounts of gel samples of 2 wt% DIBA and DIPA in octane were placed on a glass plate. The same type of TEM grid was brought into contact with the glass plate containing the gel sample momentarily and then the TEM grid was dried in the air for 2–3 days .Gel samples were prepared by placing hot sols of 7 wt% DIBA or DIPA in silicone oil in round, special glass capillaries  with 1 mm interior thicknesses. The samples were kept at ~4–6 °C for 15–20 min and kept at room temperature for ~1 h. A gel sample of 7 wt% DMEA in silicone oil was prepared by the fast-cooling method and a polyimide (Kapton) tube with an inner diameter of 0.81 mm was dabbed over the gel to fill the tube with the gel. Powder X-ray data were not collected until/unless the sample was evenly distributed within the tube and without any air bubbles apparent by viewing through a microscope. Samples of neat powders of DIBA, DIPA, and DMEA for powder XRD were prepared by dabbing a Kapton tube over crystals that had been ground into a fine powder. Solidified melts of DIBA, DIPA and DMEA for XRD measurements were prepared by heating them to slightly above their melting temperatures for 10 min and incubating them at room temperature for 1 h before dabbing a Kapton tube over them. 3 wt% DIBA, DIPA or DMEA gels in silicone oil, prepared by the fast-cooling method, were placed on the lower rheometer plate with a gap of 0.5 mm set between the parallel plates. The linear viscoelastic region (LVR) for strain sweeps was selected as 1 Hz based on examination of the frequency sweeps. Frequency sweep measurements were performed from 0.01 to 100 Hz at a constant strain of 0.01%  with a Teflon cap after bubbling nitrogen through the solutions for 5–10 min. All measurements were made within an hour of sample preparation unless mentioned otherwise. For photophysical studies on an oxygen-free solution of 6 × 10−3 M DMEA in acetonitrile, the sample was prepared by the same method mentioned above, transferred to a 5 mm  pyrex capillary and flame-sealed after three freeze-pump-thaw cycles at 20 mTorr. The measurements were made at room temperature. Photophysical studies on solutions of DIBA, DIPA and DMEA in acetonitrile were conducted at 6 × 10−3 M DIPA in acetonitrile  that had been kept at room temperature for 14 days, and then the mixture was sonicated for 2 h and used to record excitation and emission spectra. In one experiment, triethylamine  was added to a solution of 6 × 10Photophysical studies on sols and gels of DIPA and DMEA in silicone oil were conducted at 3 wt% concentrations. Gels were prepared by the fast-cooling method in 2 mm  pyrex capillaries and sealed with a rubber cap. All excitation and emission spectra were recorded at increments of 1 nm in 2 s intervals. The flattened pyrex capillaries were placed inside a 1 cm × 1 cm quartz cuvette containing silicone oil and the Peltier temperature controller attached to the fluorometer was used to heat and cool the samples. The temperature was calibrated by placing a thermometer inside the cuvette containing the silicone oil. It showed less than ±2 °C differences from the Peltier settings.The lengths of conformationally extended single molecules of DIBA, DIPA and DMEA were calculated by the B3LYP/6-31G DFT method using the Gaussian 16 package ,30,31,32Experiments performed using DIBA, DIPA and DMEA with several low-polarity solvents resulted in gelation . PreviouFurthermore, the samples in 6 mm (i.d.) tubes were inverted and tapped twice gently to determine qualitatively the strength of the gels formed using DIBA, DIPA and DMEA. If a gel started to flow upon being tapped, it was considered weak; if it remained unchanged, it was considered a strong gel. DIBA and DIPA formed strong gels with octane, silicone oil and hexylbenzene; of these, a strong gel formed only with silicone oil as the liquid when DMEA was the gelator. DMEA formed weak gels with most of the liquids in The stabilities of the DMSO, hexylbenzene, octane and silicone oil gels were observed over time at room temperature . The gelDifferential scanning calorimetry (DSC) thermograms of neat DIBA and DIPA show one endotherm (melting point) and one exotherm and, thus, the formation of crystalline networks upon cooling a,c. The DSC curves for 3 wt% DIBA and DIPA gels in silicone oil b,d exhibSuccessive heating and cooling cycles of neat DMEA were not reproducible e. AccordThe gel-to-sol transition temperatures and the melting temperatures of the gelators determined by optical microscopy  are compThe morphologies of these gels on the nanoscale were obtained from transmission electron microscopy (TEM) images . The widd-spacings from Bragg’s law (Equation (1)) for neat gelators, gels, and solidified melts of gelators are tabulated in The molecular packing arrangements of the gels in silicone oil and their respective gelators were investigated by powder X-ray diffraction (PXRD). Diffraction from neat silicone oil was subtracted empirically from the diffraction patterns of the silicone oil gels with 7 wt% DIBA, DIPA or DMEA . The 2θ Cu = 1.54 Å), d is the spacing of the crystal layers, θ is the incident angle, and n is an integer.λ is the wavelength of the x-ray radiation (λd-spacing (as well as 2θ) values for the solidified melts of gelators and their respective neat gelators are comparable, indicating similar packing arrangements upon melting the gelators and cooling them to room temperature  for neat DIBA (31.7 Å), DIPA (33.4 Å) and DMEA (34.2 Å) are near to their extended lengths, which were calculated by DFT to be 28.8 Å, 29.5 Å and 30.9 Å, respectively  follow a 1:d-spacing of 33.4: 15.7: 11.1: 7.4: 4.9 Å corresponds to a 1:d-spacing values of 34.2: 15.8: 11.1: 9.1: 5.6 Å correspond to a 1:The lowest angle 2θ peaks  domains (L) was calculated using the Scherrer equation (Equation (2)) , where KBased on the L values in The mechanical properties and stabilities  of 3 wt%The cross-over points  at tan . The cro4 Pa) indicate the presence of strong interactions within these gel networks. The magnitude of G’ in the DIPA gel was slightly higher than that of the DIBA gel, which we tentatively attribute to conformational changes at the amide part caused by the different number of carbon atoms in the alkyl chains attached at the amide nitrogen. On the other hand, the frequency sweep for the corresponding DMEA gel  decreased in each successive cycle, destroying the network incrementally and irreversibly.Some of the gels regained part of their viscoelasticity upon cessation of the application of destructive strain (DS) ; they arThe 3 wt% DIPA in silicone oil gel at 600% DS recovered less than 2% of its initial G’ value upon cessation of DS during four cycles . Thus, e−3 M), its excitation spectrum showed one peak at 419 nm and a shoulder, which over a period of hours to days, developed another peak at 364 nm (λem = 479 nm). Thus, the process responsible for the transformation of DMEA must involve the ability of the tertiary amine and α-diketo groups to interact over short distances. As a neat powder, stored under a nitrogen atmosphere at 4–6 °C, DMEA underwent no discernible changes in its photophysical or structural properties over a period of one year. As mentioned, neat, solid DMEA packs in a lamellar arrangement in which there is a large separation between tertiary amine and α-diketo groups both intermolecularly and intramolecularly. When DMEA is freshly dissolved in acetonitrile  of DIPA solution displayed a maximum at 419 nm with a shoulder at ~396 nm. However, as mentioned above, the DMEA solution developed another shoulder at ~364 nm along with peaks at 419 nm and ~396 nm  were recorded at various times after the preparation of a of 6 × 10−3 M DMEA solution in acetonitrile (−3 M DIPA solution in acetonitrile were unchanged for as long as 13 days after sample preparation (em = 479 nm) showed the new peak at ~364 nm and a broad peak centered at ~419 nm 4 days after sample preparation (The excitation spectra (λ ~396 nm . The sho ~396 nm . To probonitrile . Two dayparation . Even afparation . vide infra). No photophysical changes were observed in DIPA, which contains an α-diketo group but lacks a tertiary amine group. To determine whether the spectral changes in DMEA are a result of specific interactions between the α-diketo and tertiary amine groups, a 4-molar excess of triethylamine (TEA) was added to a 6 × 10−3 M DIPA in acetonitrile solution and the photophysical changes were tracked over time (em = 479 nm) and emission spectra (λex = 419 nm) were observed during the first 3 days after the addition of TEA when the sample was kept in the dark. However, after 8 days, the excitation spectrum at λem = 479 nm showed a decrease in the intensity of the excitation peak at 419 nm and the emergence of a new peak centered at ~364 nm. No changes were observed in the emission spectrum at λex = 419 nm. These photophysical changes resembled those observed in DMEA after 2 days  for interactions between TEA and α-diketo groups of  in acetonitrile upon excitation at 355 nm [Malval et al. reported a high rate constant (~10t 355 nm . Excitatt 355 nm . In the t 355 nm . Subsequt 355 nm . The corem = 457 nm) for the DIPA gel showed a shoulder at ~381 nm with two maxima at ~400 and ~420 nm (em = 454 nm) of the DMEA gel showed a broad emission maximum centered at ~364 nm with two other peaks of comparable intensity at ~395 nm and ~420 nm, and a shoulder at ~381 nm (em = 454 nm) displayed a peak at ~364 nm which was already more intense than the peaks at 395 nm and 419 nm. On day 15 after sample preparation, only one peak, centered at ~364 nm, was observed, and the peaks at 395 nm and 419 nm had disappeared  is a part of the gelator network. However, a small fraction of the DMEA molecules, corresponding to approximately the CGC, remains free to migrate as it would in the sol or solution phase . The CGCTo determine whether the electron transfer process can be reversed thermally, a freshly prepared 3 wt% DMEA in silicone oil gel was converted to a sol by heating to 75 °C , it was found to undergo unimolecular and aggregate electron transfer processes (depending on its concentration and phase) that lead to its reactivity in solution/sol and gel states. Results from powder X-ray diffraction explain why neat DMEA in its crystalline state is both thermally and photochemically stable—the tertiary amine and diketo sites are not near each other and cannot diffuse into proximity. By contrast, the structural analog of DMEA, with an isopentyl group (DIPA), lacks a tertiary amine group (and a lone pair of electrons!) along the amide chain and is more stable to heat and light in its solution/sol and gel phases. However, when a tertiary amine, triethylamine, was added, DIPA became unstable (such as DMEA) due to