My Life as a Molecular Evolutionist” from the Institute of Genomics and Evolutionary Medicine, Temple University Philadelphia, PA. The purpose of this memoir is to discuss the origin and development of the discipline of molecular evolution from the point of view of a cofounder of MBE. He was born in Japan in 1931 and immigrated to the United States in 1969 to do the work on molecular evolution. Since then, most of his professional work was done in the United States. In this memoir, he discusses how he grew up in Japan and how he ended up with working on molecular evolution, including some of enjoyment and grief in conducting the work. He emphasizes the importance of working with collaborators and indicates that many of his studies were done in these collaborations. The memoir is about 200 pages long and consists of 9 chapters and additional sections (see Box 1).Molecular Biology and Evolution (MBE) is pleased to announcethat Masatoshi Nei recently published a memoir entitled “Box 1.Contents of Nei memoirPreface  IntroductionChapter 1.      Chronological TimelineMy ChildhoodChapter 2.  University EducationChapter 3.        University of Miyazaki      Kyoto UniversityStudies in the USAChapter 4.        University of California at Davis      North Carolina State UniversityNational Institute of Radiological Sciences in Chiba, JapanChapter 5.        Marriage and Children      Kimura’s Neutral Theory of Molecular Evolution      Use of Computers in Genetics      International Congress of Genetics in Tokyo and KyotoBrown University in the USAChapter 6.        Adapting to the American Way of Life      Theory of Genetic DistanceUniversity of Texas Health Science Center at HoustonChapter 7.  vs. The Theory of Mutation-Driven Evolution      Neutral Theory of Molecular Evolution       Gene Genealogy, Pseudogenes, and Neutral theory      Human Evolution      Meetings in Texas      Visit to India: A Mysterious Country      Moscow and Other European Cities      China and Sweden      Titisee Meeting in Germany      Natural Selection at the Major Histocompatibility (MHC) LociMolecular Population Genetics and Evolution      Book on       Neighbor-Joining Method of constructing Phylogenetic Trees      Gordon Research ConferencesMolecular Biology and Evolution (MBE)”      Founding a New Journal called “      International and Domestic MeetingsPennsylvania State UniversityChapter 8.        Work on Committees      Molecular Evolution and Phylogenetic Trees      Australia: a Continent of Isolation      Origins of the Japanese Populationthe Society for Molecular Biology and Evolution (SMBE)     Official Start of       MEGA: Molecular Evolutionary Genetics Analysis Software      Molecular Evolution and Phylogenetic Trees      Australia: a Continent of Isolation      Origins of the Japanese Population      Trip to Northern Greece and Hungary      Sabbatical leave in Japan, 1997     Penn State—SMBE Symposium on Molecular and Genomic Evolution      Another trip to China      Meeting with the emperor and other recognitionsTemple UniversityChapter 9.        A Mild Form of Stroke and Temple University      Symposium on Molecular Evolution at Temple University      My Work at Temple UniversityEpilogue and AcknowledgementsReferencesIndexpress.arctangent.com.Masatoshi Nei’s studies of evolutionary problems started from an investigation on the origin of genetic variation in populations, but as genetics has grown up into molecular biology, he has also become a molecular evolutionist. Working alone or with his students and collaborators, he continuously developed various new concepts of evolution and new statistical methods. Early on in these studies, he understood the importance of using computers in biology, bioinformatics, and genomics. Because of these researches, his work is widely appreciated. For those who are interested, the memoir is now available for purchase at the following website: https://


The purpose of this study was to prepare poly (D-L) lactide-co-glycolide (PLGA) and poly ε-caprolactone (PCL) nanofibers containing metronidazole and amoxicillin using an electrospinning process as intrapocket sustained-release drug delivery systems for the treatment of periodontal diseases. Scanning electron microscopy showed that the drug containing PLGA and PCL nanofibers produced from the electrospinning process was uniform and bead-free in morphology. The obtained nanofibers had a strong structure and resisted external tension according to the tensiometry results. The cytotoxicity results indicated acceptable cell viability (>80%). Quantification by high-performance liquid chromatography showed almost complete in vitro drug release between 7 and 9 days, whereas 14 days were required for complete drug release in vivo. No significant signs of irritation or inflammatory reaction were detected after three weeks of subcutaneous implantation of nanofibers in the animal models, thus indicating suitable compatibility. The results therefore suggest that the designed nanofibers can be used as potential commercial formulations in the treatment of periodontitis as controlled-release intrapocket drug delivery systems that can increase patient compliance. This is due to their ability to reduce the frequency of administration from three times daily in a systemic manner to once weekly as local delivery. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Fusobacter nucleatum are common species that can invade the tissue of the periodontium [Periodontitis refers to any inflammatory and infectious condition affecting one or more parts of the periodontium. The main pathogenesis of periodontal diseases is subgingival plaque formation, which can be caused by up to 400 species of bacteria. odontium . There aodontium ,3. Treatodontium . Generalodontium .Nonsurgical protocols are usually recommended for first-line therapy in mild to moderate conditions, in which scaling and root planning (SRP) is a common method of treatment. In this method, all of the possible factors involved in inflammation and infection are removed from the surface of the root and periodontal space around a tooth .Antibiotic therapy is necessary for the eradication of microbial plaque because of its important role in the progression of periodontitis . One of Surgical therapies are the most effective method available to treat severe types of periodontal diseases. In this method, dentists focus on reducing gingival depth or removing residual parts that can cause recurrences. Surgery can also be used to repair any possible losses due to long-term inflammation .Local drug delivery systems reduce systemic adverse effects as well as providing appropriate therapeutic concentrations. They have no first-pass effect or rapid onset and can also prevent microbial resistance against certain bacterial species . Their l®, Actisite®, Arestin®, Periostat®, and PerioChip® [The ability to act as a natural reservoir with access to blood circulation and the capacity to enable solid forms to reach all parts around an infected area make intrapocket delivery systems desirable . CommercrioChip® . PolymerrioChip® ,18,19. PrioChip®  and peririoChip®  deliveryPLGA is a favorable synthetic polyester that has high biocompatibility and good mechanical properties. Its disadvantages include hydrophobicity and limited bioactivity. These properties make it a potential polymer for use in commercial formulations to achieve better treatment goals ,23,24. PNanofibers are modified-release formulations suitable for use as an intrapocket drug delivery system. Electrospinning is the most commonly used and efficient method for the preparation of nanofibers, and it has been applied at industrial and laboratory scales. It generally consists of a syringe pumping a polymer solution, a high voltage electric reservoir, and a conductive rotating collector ,26,27,28In this study, PLGA and PCL nanofibers containing amoxicillin and metronidazole were designed and evaluated for the treatment of periodontitis as intrapocket drug delivery systems. These formulations were then characterized for their physicochemical properties, safety and toxicity, and in vitro/in vivo evaluation. It was expected that these systems would achieve controlled release of both drugs.Poly-(D-L-lactide-co-glycolide) (PLGA), poly(ε-caprolactone) (PCL), amoxicillin, metronidazole, dichloromethane (DCM), ethyl acetate (EA), ether, ketamine, xylazine, and ethanol were obtained from Merk Company . Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum, trypsin, sabouraud dextrose agar (SDA), sabouraud dextrose broth (SDB), soybean casein digest broth (SCDB), tryptic soy agar (TSA), thioglycolate culture medium, 3--2,5-diphenyltetrazolium bromide (MTT), and phosphate-buffered saline (PBS) were obtained from Bioidea Company . Rats and rabbits were obtained from an internal animal laboratory in Kermanshah University of Medical Sciences. Human gingival fibroblasts (HGFs) were obtained from the Pasteur Institute . The pH 7.4 used in the in vitro studies was prepared by using dihydrogen phosphate dodecahydrate and orthophosphoric acid, both purchased from Merck .w/v of PLGA in a mixture of DCM/EA (9:1 v/v ratio) and the other by dissolving PCL at 10% w/v concentration in a similar solvent mixture. The solutions were stirred magnetically (300 rpm) at room temperature (25 °C) for 5 h to obtain clear and uniform solutions. Each polymeric solution was separated into two portions; metronidazole and amoxicillin were individually dissolved in each portion at 15% w/w of drug to polymers and stirred at 300 rpm for 2 more hours to obtain a clear solution. Finally, for each formulation, two electrospinning solutions were developed, one loaded with metronidazole and the other with amoxicillin. Blank nanofibers were prepared by dissolving polymers in the solvent system without a drug.Two electrospinning solutions were prepared separately; one by dissolving 13% Through two individual electrospinning processes, PLGA and PCL nanofibers were prepared using a double-nozzle electrospinner. Each nozzle was filled with 10 mL of PCL/metronidazole and PCL/amoxicillin for the electrospinning of the PCL nanofibers. The same procedure was used in the case of the PLGA formulations (10 mL of PLGA/metronidazole and PLGA/amoxicillin). The temperature was kept between 25 to 30 °C for the PCL formulation and between 16 to 20 °C for the PLGA formulation using a temperature regulator. Other factors were kept the same for both samples. In the electrospinning process, a maximum voltage of 17 kV and a drum rotation rate of 250 rpm were applied. The needle was swept between distances of 110 to 190 mm from the baseline. A distance of 100 mm between the needle and the drum was also applied. The injection took place at a rate of 1 mL/h. Nanofibers were collected on the drum and stored in a refrigerator (2–8 °C) till required.SEM  was used as a method to observe the surface morphology, uniformity, and structure of the nanofibers. Small pieces of the nanofibers were separated and sputter-coated with a thin layer of gold. The surface characteristics were examined under a vacuum and at 30 kV accelerating voltage. Various magnifications were used to ascertain the different morphologies.−1.FTIR spectroscopy  was used to determine any interactions or changes at the molecular level of the starting materials used in the preparation of the nanofibers and also for the nanofibers themselves. A comparison of the nanofibers’ spectra can provide information about possible interactions. A small amount of the appropriate sample  was mixed with KBr and then compressed by an FTIR manual press under a pressure of 10 tons for 2–3 min. The samples were scanned between 400–4000 cm2 pieces and placed in the tensiometer apparatus for strength measurements. The samples were pulled at a 1 mm/min stretching rate.The tensile strength properties of the nanofibers were determined using a tensiometer . This method indicates the resistance of the nanofibers against an external force when placed between two clamps. The prepared nanofibers were cut into 2 × 2 cmThe sterility of nanofibers was studied by placing 20 mg of blank nanofibers into SDB, SCDB, and thioglycolate culture medium designed for fungal and aerobic and anaerobic bacterial growth. After 7, 14, and 21 days of incubation at 25 °C for the fungi and 37 °C for bacteria, the culture media were evaluated for possible occlusion due to microorganism growth compared to positive (cultured microorganisms) and negative (pure culture media) control samples.Escherichia coli and Staphylococcus aureus were selected for this study as anaerobic and aerobic microorganisms, respectively. They were activated in TSA culture media for 24 h and incubated at 37 °C. The antibacterial activity of the nanofibers was assessed using the agar diffusion test. Disks of 5 mg of the formulation were measured and sterilized for 30 min under UV light at room temperature (25 °C) and placed on the cultured media. This was then incubated for another 24 h at 37 °C and the diameter of the inhibition zone around the nanofiber disks was measured.In vitro drug release was evaluated using the Franz diffusion cell method. A specific amount of nanofibers  were cut and put into a pocket made of the diffusible cellulose dialysis membrane . The membrane was then kept in a release medium containing 25 mL of phosphate buffer (pH = 7.4) at 37 °C stored in a shaker incubator with a rotation rate of 100 rpm. At certain predetermined time intervals, 5 mL samples were collected from the release medium for drug content analysis using HPLC see . The samThe drug release data were fit to different kinetics models, including zero-order, first-order, Higuchi, and Korsmeyer–Peppas models. The model with the highest correlation coefficient was chosen as the best-fitted kinetic model. The equations used were as follows:Zero-order release:First-order release:Higuchi model:Korsmeyer–Peppas:0 denote the remaining drug in the formulation (unreleased) and the initial amount of drug in the formulation respectively. K0, K1, KH, and Kp are constant drug releases and n in Equation (4) is the release exponent and indicates the mechanism of drug release.In these equations, Qt is the amount or percentage of drug released at time t and Qr and Qv/v ratio; the pH of the mixture was adjusted to 4) at a 1 mL/min injection rate with the detection wavelength set at 220 nm. The HPLC analytical column (5 µm) Perfectsil ODS-2  was utilized. A twenty microliter sample loop was applied with retention times of 1.733 and 2.317 min for amoxicillin and metronidazole, respectively. Data were analyzed using the Excel program (Microsoft Office 2016). Regression equations were constructed based on the standard solutions to calculate the concentrations of drugs in the samples withdrawn from the Franz diffusion cell.Standard solutions with defined concentrations were prepared by dissolving drugs (amoxicillin and metronidazole) in distilled water at a stirring speed of 300 rpm at room temperature for 2 h to complete dissolution. Standard solutions (20 µL) were injected into the HPLC apparatus, where the mobile phase contained acetonitrile:phosphate buffer  were incubated in 1 mL of DMEM in sterile conditions for 48 h. The supernatant was then separated and used as an extract for direct cell contact.HGFs were cultured in sterile plates in DMEM medium and incubated for 24 h at 37 °C in an incubator. The supernatant from the DMEM, which was initially incubated with the formulated nanofibers, was added to cultured cells and incubated for another 24 h in standard conditions for further evaluations.The cultured cells were seeded into 96-well plates. Cell viability of human gingival fibroblasts was evaluated with the MTT test. The culture medium was discarded and plates were washed with PBS. Trypsin was added to free cell attachments and 10 µL of MTT solution (5 mg/mL) was then added to the wells. Control samples were prepared by adding DMSO to cultured cells for full cell inhibition. Plates were incubated for 4 h in an incubator at 37 °C and the absorbance of these samples was measured by UV spectrophotometry at a wavelength of 570 nm. The cell viability percentage was calculated by the following equation, in which C is the absorbance of the test samples and A is the absorbance of the control samples:Due to the small sizes of the oral cavity in animal models such as rat and rabbit models, which tend to be the most popular models for in vivo studies of intrapocket formulations, a replacement method was adopted . This mev/w ketamine. They were shaved on the dorsal part and two subcutaneous pockets of 1 × 1 cm2 were cut. Each pocket was used for the implantation of one formulation. The nanofibers cut into tiny pieces were implanted and pockets were closed by 000 size sutures. After 24, 48, 72, 96, 120, 144, 168, 216, 264, and 312 h, nanofiber samples were taken out from the subcutaneous pockets and prepared for HPLC method analysis.To sterilize the nanofibers for in vivo testing, the formulated nanofibers were cut into 25 mg pieces  and were subjected to UV light for 30 min before use. Twelve adult male rats (200–250 g weight) were chosen and treated in standard feeding and nursing conditions. They were anaesthetized using an injection of 10% v/v ratio)) and characterized using the HPLC method, as previously described in The implanted nanofibers were explanted from the rats’ bodies after the rats were euthanatized by ether. The nanofiber samples were then washed with distilled water and dried in an oven at 60 °C for 30 min to obtain the primary weight of the nanofibers. The nanofibers were then dissolved in the least amount (~1 mL) of solvent mixture  for 15 min each  (n = 3). Sections were paraffined and cut into 6 µm slides. These were then colored using hematoxylin and eosin (H&E). Slides were observed with a light microscope and evaluated for the existence of inflammatory cell lines [In this experiment, the nanofibers were subcutaneously implanted in two animal models (rats and rabbits) with the procedure described in ll lines .v/v used [The nanofibers obtained using the electrospinning method had a white to yellowish color with smooth surface structures, as shown in v/v used .−1 was assigned to the hydroxyl and amino groups of amoxicillin and metronidazole. The absorption bands at 2933 cm−1 and 2866 cm−1 were related to the symmetric and asymmetric C-H stretching vibrations of metronidazole and PCL, respectively. The 1732 cm−1 peak was attributed to the stretching vibrations of the carbonyl (C=O) groups of PCL and amoxicillin, whilst the peak at 1721 cm−1 corresponded to the carbonyl group of the beta-lactam ring. The peak at 1685 cm−1 merged with a wide 1721 cm−1 peak was indicative of the C=O group adjacent to the NH2. The C=C stretching vibration of metronidazole appeared at 1537 cm−1. The C-N stretching of metronidazole and amoxicillin was also present at 1473 cm−1. The stretching of the N=O group in the metronidazole structure was detectable as a peak at 1369 cm−1. The peak at 1186 cm−1 was related to the C-O-C group of PCL.A comparison of the nanofiber spectra for both formulations with the individual excipients used in the preparation of the nanofibers showed no interactions . The cha−1 was assigned to the -OH and NH2 groups. Symmetrical and asymmetrical stretching vibrations of C-H appeared as two characteristic peaks at 2939 cm−1 and 2862 cm−1. The carbonyl stretching vibrations of the PLGA and the amoxicillin beta-lactam ring appeared at 1721 cm−1. The peak at 1685 cm−1 was a result of the C=O groups adjacent to the NH2 and was merged with the side peak. The peak at 1469 cm−1 was assigned to the C-N stretching of both drugs. The characteristic N=O stretching peak was detectable at 1365 cm−1 in the peak of the PLGA nanofibers. The C-O-C stretching appeared at 1184 cm−1.In the case of the PLGA nanofibers, a wide peak at 3435 cmIn conclusion, there were no significant changes, such as decreases in the intensity or elimination of characteristic peaks. It can therefore be concluded that no significant interactions occurred between the drug and the polymers.The tensile strength results showed that the PLGA nanofibers were more resistant to tension than the PCL nanofibers when external stress was applied. This is due to the high inherent stiffness of PLGA . Chou etThe sterility of the blank nanofibers (without drug) was evaluated by placing the sterilized nanofibers into the selected culture media to observe possible microorganism growth. After incubation time , the test samples were just as clear as the negative controls in comparison to turbid positive controls. No microorganism growth was detected in all the culture media, indicating the sterility of the prepared nanofibers. The sterility was thus achieved by performing the whole preparation process under aseptic conditions and storing the samples in a sterile environment.E. coli and 23 ± 2 mm for the PCL nanofibers and 12 ± 1 mm for the PLGA nanofibers on S. aureus. It can thus be concluded that these nanofibers have antimicrobial efficacy against both Gram-positive and Gram-negative bacteria. Incubated bacterial cultures containing sterilized nanofibers were detected for the growth inhibition zone, which represents an inhibition effect of the formulated nanofibers on the studied microorganisms. The diameter of the growth inhibition zones was measured to be 11 ± 1 mm for the PCL nanofibers and 12 ± 1 mm for the PLGA nanofibers on Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli).Haider et al. prepared PLGA/CuO hybrid nanofiber scaffolds to determine their inhibitory effect on microorganisms involved in oral pathogenesis . The autThe amount of drug release was quantified by HPLC. Using the drug release data, the release profiles of the drugs from the nanofiber formulations were plotted and are shown in All formulations released most of the drugs within 180 h. These release profiles showed two phases of drug release. Almost 50% of the drug was released within the first day followed by a gradual release over 6–8 days. For example, the PLGA formulation released 45.28 ± 1.39% of the amoxicillin and 43.30 ± 4.16% of the metronidazole within the first 24 h whereas the PCL nanofibers released 55.34 ± 1.37% of the metronidazole and 51.32 ± 1.41% of the amoxicillin over the same period. It seems that, in the second phase, the release of drugs from the PLGA nanofibers was faster than the drug release from the PCL nanofibers. For instance, almost 80% of both the amoxicillin and metronidazole were released within 78 h, whereas in the case of the PCL nanofibers the time taken to release 80% of the amoxicillin and metronidazole was 145 h and 97 h, respectively.The more gradual and controlled release of drugs from the PCL nanofibers (196 h for the release of the metronidazole and 216 h for the release of the amoxicillin) compared to the PLGA nanofibers (168 h for the release of both drugs) may have been due to the higher hydrophobicity of PCL compared to PLGA . This hiTseng et.al prepared PLGA nanofibers containing lidocaine for topical delivery which showed a controlled release pattern for 14 days in the laboratory setting . Dias etH for metronidazole release from the PLGA and PCL formulations was determined to be 0.1019 and 0.1069, respectively, whilst the KH for amoxicillin release from the PLGA and PCL formulations was 0.1079 and 0.0916, respectively.The Higuchi release constants were calculated for both formulations and the results indicated that their values were very close to each other. The KThe percentages of cell viability based on concentration as determined by the MTT method are shown in It is important to carry out in vivo release studies, as these can provide more realistic data on the drug release from the nanofibers. As previously mentioned in the methodology section, the nanofibers were implanted surgically in subcutaneous pockets after sterilization. The surgical procedure in the rat model is depicted in The cumulative percentages of drugs released are shown in Nanofibers were surgically implanted into subcutaneous pockets in rat and rabbit models for biocompatibility assessments. The comparison of the first- and third-week samples with the control slides in the rat and rabbit models for the presence of neutrophils, macrophages, and giant cells as inflammatory markers was undertaken . Figure Nanofibers are one of the most promising formulations for use in modified release systems for direct drug delivery that are made of polymers that are compatible with body tissues. In this study, PLGA and PCL nanofibers containing metronidazole and amoxicillin were successfully manufactured and characterized. These nanofibers were able to release their drug contents in a controlled manner and keep the concentration of the drug at the right level in the local area to treat the infection. SEM studies of the nanofibers showed that they had smooth surfaces and could be used in an intrapocket drug delivery system. Solid-state analysis ruled out any interactions between the drugs and the polymers used in the nanofibers’ manufacture. Nanofibers were also studied for cytotoxicity and biocompatibility in both cell culture and animal model experiments. The promising results suggest the possibility of using these new formulations as an effective therapeutic method for the treatment of periodontitis. In vitro results showed good controlled drug release for the treatment goals. Drugs were released at a suitable concentration over a period of 7 to 10 days in an animal model. The prolonged-release profiles indicate that these systems could increase patient compliance by reducing the administration frequency. It can therefore be concluded that these systems have the potential to enter clinical studies and to be used as routine effective treatment modalities against periodontitis.


NSR, the research team led by Jiang Liu of Beijing Institute of Genomics, publishes a paper [In this issue of  a paper  on the eFor geneticists, especially the modern brand to whom the detailed molecular mechanism is the only reason to study biology, major evolutionary transitions would be the testing ground of that conviction. This is where vertebrates come in. Deuterostome appears to get the anterior–posterior axis backward (as far as protostomes see it). Étienne Geoffroy Saint-Hilaire forwarded the curious hypothesis that vertebrates are upside-down creatures compared with invertebrates. The endo- vs. exo-skeletal system is another distinguishing feature between them; we lump in the backbone even though it is unique in its own way. Finally, for a good measure, there is the adaptive immunity acquired at this evolutionary transition.Is it then possible to understand, in molecular terms, anything about vertebrates that sets them apart from invertebrates? For specific features, there are answers. Saint-Hilaire's hypothesis has in fact acquired some support from modern molecular genetic studies ,3. For aet al. show that DNA-methylation reprogramming is associated with the development, reproduction and adaptive immunity in vertebrates, but not in invertebrates. No less interesting, they show that the HOX cluster, which has been speculated to be the genic key to this transition, shows dynamic DNA-methylation patterns in vertebrates, but not in invertebrates.If the conundrum of the vertebrate–invertebrate transition is to be cracked in a general way, where might be the molecular entry point? Xu et al. at least offer an alternative, which may play a partial role in orchestrating the transition from invertebrates to vertebrates. Unlike other descriptions of the vertebrate–invertebrate transition, methylation is a regulatory mechanism of a rather complex kind in itself.Although this study does not provide specific molecular mechanistic answers, it is also dubious that the wide gaps in major evolutionary transitions can be bridged this way. Xu


AGAMOUS-like 15 (AGL15) is a member of the MADS domain family of transcription factors (TFs) that can directly induce and repress target gene expression, and for which promotion of somatic embryogenesis (SE) is positively correlated with accumulation. An ethylene-responsive element binding factor-associated amphiphilic repression (EAR) motif of form LxLxL within the carboxyl-terminal domain of AGL15 was shown to be involved in repression of gene expression. Here, we examine whether AGL15′s ability to repress gene expression is needed to promote SE. While a form of AGL15 where the LxLxL is changed to AxAxA can still promote SE, another form with a strong transcriptional activator at the carboxy-terminal end, does not promote SE and, in fact, is detrimental to SE development. Select target genes were examined for response to the different forms of AGL15. AGAMOUS-LIKE 15 (AGL15) is a MADS-domain transcription factor (TF) that accumulates to the highest levels during embryogenesis ,3. PreviArabidopsis thaliana SIN3 associated protein P18, At2g45640) that is part of a SWI-INDEPENDENT/HISTONE DEACETYLASE (SIN3/HDAC) complex. HDACs remove acetyl groups from histones, resulting in more compact chromatin and typically a decrease in gene expression of associated genes . Specifiew, see ,11. An eiew, see . AGL15 hiew, see . Likewisiew, see .LEAFY COTYLEDON2 (LEC2), FUSCA3 (FUS3), and ABSCISIC ACID INSENSITIVE3 (ABI3) [LEC2 is well known to enhance SE when overexpressed, perhaps AGL15′s ability to enhance SE is mainly (or entirely) due to its ability to induce gene expression, including that of LEC2. Thus, we asked if we changed AGL15 such that its repressive ability is lost and it only acts as an inducer of gene expression, would it still be able to promote SAM SE? We made two modified forms of AGL15 and report here that a form where the leucines within the EAR domain are changed to alanines can still promote SAM SE, but a form that includes a strong transcriptional activation domain reduces the ability to form SAM SE.Several transcription factors key for embryogenesis were found to be AGL15-direct up-regulated targets including 3 (ABI3) . Because35S:AGL15-AAA). This would be predicted to eliminate interaction with SAP18 and TPL/TPR, and recruitment of HDACs through these proteins [Two different forms of AGL15 that are predicted to eliminate the repressive function were generated A. In oneproteins ,11,12. Tproteins . Althougproteins , this foproteins .AGL15 transcript accumulation. As shown in 35S:AGL15-AAA was able to promote SAM SE when transcript accumulated to higher levels  did not show a significant increase in SAM SE over all biological replicates (35S:AGL15-AAA-1 compared to wild type (wt). This was the highest expressing line we obtained and transcript accumulation was comparable to 35S:AGL15. The form of AGL15 with the VP16 domain produced significantly reduced SAM SE , there appears to be cosuppression. As expected for reduced AGL15 expression, there is a significant decrease in SAM SE for this line.Transgenic lines were assessed for ability to produce SAM SE and for  line 1, C. Lines plicates B,C. Howeplicates C. SAM SEd SAM SE B, even w5S:AGL15 C. IntereAGL15 gene expression. Potentially there would be increased AGL15 transcript from the endogenous gene causing increased SAM SE. To examine this, we generated oligonucleotides that could differentiate between transcript from the endogenous gene (LxLxL) and transcript from the 35S:AGL15-AAA1 transgene (AxAxA oligonucleotide) . Thus AGL15-AAA is not promoting SAM SE by upregulation of the endogenous gene; in fact, this form of AGL15 is still able to repress expression from the endogenous gene.We previously reported that AGL15 represses its own expression [leotide) . As showAGL15 overexpression promotes SE, including SAM SE, is that it may up-regulate genes such as LEC2, FUS3 and LEAFY COTYLEDON1 (LEC1) that have been shown to promote SE or SE programs post-embryonically [ABI3 is a direct target of AGL15 [LAFL. If the promotion of expression of these key embryo TFs was all that was needed for SE, and repression of other genes did not factor into ability to promote SE, we would expect the 35S:AGL15-VP16 lines to have abundant SE development because LEC2 and FUS3 are direct targets, albeit LEC2 is a weaker target with a peak at the 3′ end of the gene that is below cutoffs used in CisGenome to call peaks . AGL18 th AGL15 ,24,25. Ath AGL15 . Therefoo Col wt . There i were assessed for response to different forms of AGL15 using qRT-PCR. All showed significant reduction in transcript accumulation compared to Col wt when in the 35S:AGL15 background, agreeing with the high-throughput data. We found a significant upregulation of AP3, SHP2 and SEP3 gene in 35S:VP16-46 line compared to the Col wt control, but not of AGL18 or SVP. SHP1 did exhibit increased transcript accumulation, but this was not significant. 35S:AGL15-AAA showed reduced transcript accumulation from SHP2 and SVP  but not others , while AGL15-VP16 can activate some genes , but not others .A number of other MADS-domain proteins besides AGL18 have been reported to interact with AGL15 , and man and SVP . Therefo35S:AGL15-AAA transgene was expressed at sufficient levels . This modification should eliminate recruitment of proteins needed for histone deacetylation. Histone deacetylation is associated with more compact chromatin leading to repression of gene expression. We had hypothesized that repression of expression of some genes as measured by mRNA accumulation may be important for SE [35S:AGL15 is able to upregulate LEC2, which encodes a gene product that is sufficient to induce SE on seedlings post-germination [LEC2, and FUS3 mRNA, and, for LLL and AAA forms, LEC1. FUS3 is a direct expressed AGL15 target and can promote embryo programs after completion of germination, although not to the extent observed for LEC2 and LEC1 [LEC2 and FUS3, LEC1 is an indirect target, so we would not expect association with AGL15-VP16 and induction of expression, unless this was a result of indirect regulation. While the overall means corresponded with the ability to promote SAM SE, numbers varied between biological replications and results across all experiments were not significant, with the exception of LEC2 in 35S:AGL15 and 35S:AGL15-AAA. Thus, this could at least in part explain why AGL15-AAA can promote SAM SE, but does not explain why AGL15-VP16 shows reduced SE as these key regulators are not reduced compared to Col wt in this background.Interestingly, elimination of the LxLxL/EAR domain by site directed mutagenesis to replace codons for leucine with codons for alanine  did promote SAM SE when the t for SE , so thismination ,20. Thusmination . Indeed,and LEC1 ,16,17. I35S:AGL15-AAA besides line 1 showed in some experiments a significant increase in SAM SE, this was not significant over all replicates of the experiment. However, no other lines showed as high an accumulation of AGL15-AAA transcript as did line 1, which was comparable to 35S:AGL15. Thus, it appears that the AAA transcript can support SAM SE development when expressed at sufficient levels, but not reproducibly in lines with lower levels of accumulation. No lines of 35S:AGL15-VP16 had levels of transcript accumulation comparable to 35S:AGL15; line 46 was one of the highest expressing lines. However, even at this level of transcript accumulation, a significant decrease in SAM SE was observed. Similarly, 35S:AGL15-VP16, line 32 showed a reduction in SAM SE, indicating that strong transcriptional activation of direct targets is detrimental to promotion of SAM SE. Another independent transgenic line (line 47) showed decreased AGL15 transcript accumulation . We used primers that were specific to the AAA and the LLL forms and found that the AAA form did not lead to overaccumulation of AGL15 transcript from the endogenous gene, but rather significantly decreased transcript accumulation compared to wild type  causes delays in the transition to reproductive development. In addition, the agl15 agl18 mutant flowers early in short days [AGL24 did not show significant responses to the different forms of AGL15, SVP did. Therefore, we were curious as to the trend for the other MADS investigated. As shown in AGL18 and SVP show decreases in transcript accumulation as the SAM transitions through the plant life cycle. Meanwhile AP3 and SEP3 increase in transcript abundance as the SAM progresses through the plant life cycle.The ability of the SAM in the SAM SE system to produce SE can be interpreted in several ways. The initial observation of SAM SE production involved mutants with unusually large meristems and induction of SE with 2,4-D could promote SE from these meristems . Howeverort days ,24. SVP ort days . While AFinally, AGL15 has been proposed to influence dedifferentiation, perhaps by regulating stress responsive genes including transcription factors associated with dedifferentiation ,37. TherPI is expressed in embryo contexts [Summarizing these observations, it is interesting that the MADS-box genes that respond to AGL15-VP16 tend to 1, not have detectable mRNA in a somatic embryo culture tissue  and/or lcontexts ,35. Evencontexts  and has contexts . Thus, aAGL15, SVP, SHP2) still able to be repressed by the AAA form but others  are not? All of these targets are potentially directly regulated  but not others (AGL18 and SVP)? More work needs to be performed, but again, perhaps particular interactions at different loci may lead to these different responses. While addition of VP16 clearly activates some targets, presumably by overriding repressive domains, others have reported that the EAR domain within other proteins can override the transcriptional activation of the VP16 domain. It is intriguing that the genes that are activated by AGL15-VP16 appear to have one binding site associated, whereas AGL18 shows association with AGL15 at the 5′ and the 3′ ends of the gene, while SVP has multiple regions at the 5′ end of the gene associated with AGL15  and the transgenic lines were surface sterilized as done by . Seeds wTUA3. Significance was determined using a Student’s t-test.Total RNA from the frozen samples was isolated using the RNeasy Plant Mini Kit (Qiagen) with 1% polyethylene glycol added to the RLC buffer for seeds/seedlings ,46. One


A was introduced. The formation of nanofiber-wrapped structural yarns was discussed using three values of A. The starting point of each twist was the same position when A = 0 with a constant corner angle β. However, the oriented nanofiber broke or was pulled out from the gripping point when it was twisted, and it appeared disordered. The forming process of electrospun nanofiber-wrapped yarn displayed some unique phenomena, including the emission of directional nanofibers during collection, fiber non-continuity, and twist angle non-uniformity. The conclusions of this research have theoretical and practical value to guide the industrial preparation of nanofiber yarns and their wrapped yarns.To analyze the feasibility of electrospinning nanofiber yarn using a wrapping yarn forming device, electrospun nanofiber-wrapped yarns and multiscale yarns were prepared by self-made equipment. The relationship between the surface morphology and properties of yarn and its preparation process was studied. The process parameters were adjusted, and it was found that some nanofibers formed Z-twisted yarns, while others showed exposed cores. To analyze the forming mechanism of electrospun nanofiber-wrapped yarn, the concept of winding displacement difference in the twisted yarn core  Electrospun nanofiber materials have shown great application prospects in the field of functional textiles due to their ultra-fine fiber size, flexible material selectivity, structural controllability, and easy surface functionalization ; howeverFiber and yarn are the main materials used in the traditional textile industry, but only fibers with a diameter larger than several microns can be processed using existing textile processing techniques . ElectroThere are various methods for preparing nanofiber bundles with specific orientations. One is to use an auxiliary electrode method, including a spacer conductive plate collection device and a disk collection device to collect oriented nanofibers between parallel grounded double disks. This method is simple and effective, and the fiber orientation degree is high. The other method involves using the mutual attraction of oppositely charged fibers. Negative and positive spinning nozzles emit fibers with opposite charges to form fiber bundles at certain locations because of their charge attraction, which produces continuous oriented nanofibers; however, these fiber bundles are relatively fine . ThirdlyIn contrast to electrospun pure nanofiber yarn, multiscale composite yarn is composed of traditional microfibers and electrospun nanofibers according to some structure rules. A nanofiber-wrapped core yarn or yarn wrapped on a core yarn may be a disordered or oriented nanofiber, and the oriented nanofiber is generally wrapped on the core yarn in a twisting manner. Zhou used electrospinning to collect nanofibers on parallel-aligned monofilaments and then aggregated and twisted monofilaments with nanofibers on the surface . The nanMultiscale yarn is made of a variety of fibers with different scales, which can fully integrate the structural characteristics and performance advantages of traditional microfibers and electrospun nanofibers. This allows them to make up for each other’s disadvantages while giving full play to the advantages of the textile yarn structure . The weaN, N-dimethylformamide (DMF) was purchased from Guangdong Jinhua Chemical Reagent Co., Ltd.  and was analytically pure. 75D/36F twistless polyester multifilament was purchased from Fujian Jinjiang Technology Co., Ltd. . High voltage DC power DW-P503 was purchased from Dongwen High Voltage Power  Co., Ltd. An LSP-10-18 micro-injection pump was purchased from Baoding Co., Ltd. .This process involves collecting nanofibers into a certain orientation, followed by twisting the fiber bundles to improve the yarn’s strength and collective properties. Polyacrylonitrile (PAN) was purchased from Taicang Kelda Plastic Materials Co., Ltd. , with a molecular weight of 150,000. The electrospinning nanofiber yarn forming device is shown in PAN powder with a mass fraction of 12% was added to DMF solvent and stirred at a constant temperature of 40 °C overnight. An electrospinning nanofiber yarn forming device was arranged in accordance with The surface morphology of the electrospun nanofiber yarn and its wrapped yarn was studied by a JSM-6390 scanning electron microscope . The voltage was 15 kV, and the magnification was 200, 300, and 5000 times. The SEM images were analyzed using Adobe Acrobat 9 Pro software to calculate the average diameter of yarns and fibers. The prepared yarn was subjected to a tensile testing standard ISO 2062:1993 using an Instron 3365 electronic strength tester , and the average was taken from 10 repeated measurements for each sample.To theoretically verify the feasibility of electrospinning nanofiber-wrapped structures using a yarn-making device, Maxwell analysis software was used to analyze the electrostatic field to determine the scalar potential distribution of electric fields generated by charge distribution or an applied potential. Because the movement of nanofibers in an electrostatic field is very complex, we simplified the electric field model; therefore, the electric field distribution formed between the nozzle, the receiving ring, and the receiving disk is the main factor to consider in this simulation.The three-dimensional space model established according to the experimental device is shown in As shown in When the distance from the ring to the disk was set to ncreased . At a coThe ring-to-disk distance The ring-to-disk distance The ring-to-disk distance It can also be seen from The mechanical tests return the breaking strength, which must be converted into the fracture strength as follows σ = Fb/So. The cross-sectional area of the sample was measured from the SEM images, and the diameter is converted according to the area formula.The distance The ring-to-disk distance The ring-to-disk distance The ring-to-disk distance The formation of electrospun nanofiber-wrapped structural yarns occurs via the formation of many oriented nanofibers between the disk and the ring. The rotation of the disk drives the oriented nanofibers to coat the core yarn at an angle. Its schematic diagram is shown in 0 (mm), and the length of the oriented nanofibers is v (mm/s). The twist angle is β (°), the disk rotation speed is n (r/min), the core yarn is assumed to be a cylinder with a diameter d (mm), the ring diameter is D1 (mm), the disk diameter is D2 (mm), and the disk aperture is D0 (mm), and the twisting time is t (s).It is assumed that the ring aperture is exactly the same as the diameter of the core yarn, and the inner and outer diameters of the ring are the same. After this simplification, according to the electric field nephogram in β, i.e., the angle between the nanofiber surface and the yarn axis , d is the core yarn diameter (mm), and L is the nanofiber displacement along the core yarn axis when twisted at a fixed angle, β. At this time,In the formula, In the formula, λ.The distance the core yarn moves in the axial direction in a twist, i.e., the distance the core yarn is wound during a twist is v is the core yarn winding speed, and n is the disk rotation speed.In the formula, A of the core yarn wound in one twist turn is the difference between the core yarn winding distance λ in one twist turn and the displacement L of the nanofiber moving along the axial direction of the core yarn in one twist turn.The displacement difference A can adopt one of three different values: A = 0, A > 0, and A < 0.A = 0,(1) The winding speed is equal to the displacement of nanofibers moving along the axial direction of the core yarn in a given twisting time, i.e., the starting point of each twist is the same. At this time,A schematic diagram of the disk twisting is shown in β is constant, which can be applied to the traditional twist coefficient Formula (7).In this case, the twist angle The twisting of the nanofiber yarns on a cylindrical core yarn is somewhat similar to winding the yarns on a cylindrical bobbin at a constant winding angle. The difference between the two is that winding yarns is continuous because the yarns can be continuously supplied, while oriented nanofibers twisted on a core yarn are discontinuous. Oriented nanofibers are formed between the ring and the disk. During twisting, the oriented nanofibers are wrapped on the core yarn, while the two ends of the oriented nanofibers remain between the ring and the disk. The twist angle of the same oriented nanofiber remains constant when twisted. The greater the twist angle β of the yarns with the same twist, the greater the twist of the fibers and the thicker the yarns. Some researchers have used the edge of a disk to collect oriented nanofibers and then used the disk to twist the nanofibers to form yarns. By increasing the rotational speed of the disk, the twist angle increased accordingly. This study also verified Formulas (5) and (7) .L0 > L, the oriented nanofibers are stretched during twisting. It is assumed that the elongation at break of the nanofibers is ε, the value of which depends on the nature of the polymer used in the electrospinning solution and the electrospinning process parameters.The length of the wrapping of each twisted nanofibers is given by:Lq, then:When the oriented nanofibers are twisted, the number of twists is m, and the length of the oriented nanofibers is β. The nanofibers will wrap on the core yarn with the oriented nanofibers irregularly formed behind them during twisting. Therefore, only some of the oriented nanofibers that are twisted on the core yarn form a stable twist angle β, and most irregularly wrap on the core yarn. Some nanofibers are directly deposited on the core yarn, and they are also irregular. These inferences are consistent with the experimental phenomena in this paper. There are many oriented nanofibers on the same layer that are irregularly wrapped on the core yarn. The phenomenon in which some nanofibers are twisted on the yarns in an irregular form in At this time, the oriented nanofibers will break, and they will no longer be twisted on the core yarn by a twist angle A > 0(2) Here, there are two situations. The first is A = 0.When When A > 0, it is assumed that the twist angle is fixed. The actual situation is that the twist angle changes, and the twist angle β’ has the following characteristics:In the above two situations where β’ changes continuously over time, and the twist angle β’ constantly increases. The maximum twist angle β0’ has the following formula:Formula (10) shows that the twist angle A < 0(3) During a twist,At this time, according to the disk twisting diagram in In the formula, x is the xth twist.β” is a variable, and as the number of twists increases, the twist angle becomes larger. During a twist, L in Formula (12) becomes smaller as the number of twists is increased.Formula (12) shows that Formula (8) shows that the wrapping length formula of each twisted nanofiber is as follows.Lq, thenWhen oriented nanofibers are twisted, the twisting number is m, and the length of the oriented nanofibers is β′. The nanofibers will wrap on the core yarn with the oriented nanofibers irregularly formed behind them during twisting. Therefore, only some of the oriented nanofibers that are twisted on the core yarn will form a stable twist angle β, and most will wrap on the core yarn in an irregular form. Some nanofibers will directly deposit on the core yarn, and this part of the nanofibers is also irregular. However, when the fiber head end is twisted to a certain degree, the nanofibers will be pulled out from the grip of the disk and twisted into the yarn body by the next layer of nanofibers in an irregular shape.At this time, the oriented nanofibers will break, and the nanofibers are no longer twisted on the core yarn by a twist angle, 1, and the distance between the end of the disk and the center of the core yarn is x2. The twisting of oriented nanofibers is shown in All the above inferences and conclusions are based on the assumption that the ring aperture and the core yarn diameter are identical, as are the inner and outer diameters of the ring. The actual situation is that the ring aperture and the core yarn diameter are different, and the inner diameters and the outer diameter of the ring are also different. According to the electric field vector diagram in β. The position at which the twisting begins is shown by p0 in When the oriented nanofiber is twisted, it will always contact the core yarn, and when it is twisted further, it will wrap the core yarn at a twist angle In the The above inferences and conclusions are based on the assumption that oriented nanofibers are formed at a fixed position between the ring and the disk, i.e., the environment in which oriented nanofibers is fixed. The actual situation is relatively complicated. One of the two ends of an oriented nanofiber may be located at any point between the inner and outer diameters of the ring, while the other end can be at any point on the non-central hole of the disk. The twisting process mainly involves disk twisting, so only the position of the nanofibers on the disk affects the twist angle.xi between the nanofibers at the disk and the center of the core yarn is a random variable, which can be a finite number or infinity and can be listed one by one, and its twist angle is one or several finite or infinite intervals. This random variable xi is p(xi), and the sum of the products is the mathematical expectation of the discrete random variable xi, denoted as E(x).The distance x is:The value of the discrete random variable x corresponds to the value, then:0 in In other words, the mathematical expectation 0 in The above inference is also applicable in the case of a coreless yarn, i.e., in the case where the core yarn diameter d = 0 . At thisFormula (6) becomes Electrospun nanofibers move spirally in an electric field, and their trajectory continually changes. The collection of oriented nanofibers between the ring and the disk is guided by the electric field of the grounding ring and the disk, and oriented nanofibers are deposited between the ring and the disk. These oriented nanofibers are only some of the fibers formed by electrospinning . Some nat), and the angle Formula (18) shows that oriented nanofibers have intermittent formation, but also have a certain angular relationship with each other during mechanical twisting. The discontinuity of fiber length direction is caused not only by the intermittent formation of oriented nanofibers, but also by nanofiber breakage described by the formula:The mechanical twisting and wrapping process is obviously different from the wrapped structure, in which the outer wrapping yarn is generally continuously wrapped around the core yarn under the action of machinery and fluid. However, the length of the nanofibers during mechanical twisting wrapping herein is discontinuous.A > 0, A = 0, the nanofibers are not broken and are fixed in a certain layer in the yarns. The oriented nanofibers are formed at a fixed position between the disk and the ring. This constant twist angle under only certain conditions is identical to the traditional spinning process. Nanofibers have different twist angles in different yarn layers, which is consistent with the characteristics of traditional spinning.The analysis in A > 0  can be introduced to represent the average twist angle.According to A of the core yarn within one twist. When A = 0, the starting point of each twist was the same. When the oriented nanofiber was twisted to a certain degree, it broke, or the fiber tip was pulled out and was no longer held. The nanofibers no longer twisted on the core yarn at a twist angle β, and they irregularly wrapped the core yarn by the latter layer. Compared with traditional spinning processes, the electrospinning nanofiber wrapping process has unique properties, such as the uncertainty of the oriented nanofiber collection process, the discontinuity of the fiber lengthwise direction during twisting, and the non-uniform twist angle. The research conclusion of this paper has guiding significance for accurately controlling the mass production of electrospun micro- and nanofibers.In this paper, a self-made device was used to form oriented nanofibers between a disk and ring. By rotating the disk, the oriented nanofibers were twisted into yarns with a specific angle, and electrospun nanofiber yarns and its multiscale wrapping yarns were prepared. The electric field simulation analysis of the electrospinning nanofiber yarn forming was in agreement with experimental observations. During the preparation of electrospun nanofiber yarns, the average twist angle and diameter of the nanofiber yarns was correlated with changes in the ring-to-disk distance, the tip and the center of the disk, and the disk rotation speed. When preparing electrospun nanofiber-wrapped yarns, oriented nanofibers were arranged in a specific direction. This paper also discussed the mechanism of the nanofiber-wrapped yarn in three cases, according to the displacement difference CV% were not analyzed because of software limitations. This paper only prepared micro/nanocore spun yarn using ordinary yarn as the inner layer and nanofiber as the outer layer, and it did not study the fabric prepared by this micro/nano-structured yarn or the functionality of this fabric. The research team will carry out further research concerning these limitations. Interestingly, we are studying the electrospun nanofiber-core spun yarn wrapped with a layer of ordinary yarn. This sandwich yarn will protect the nanofibers in the middle layer, which can be processed by traditional weaving technology; thus, they will have broader potential applications in areas such as supercapacitors and functional textiles.In this paper, the 24-h output of pure electrospun yarn was about 672 cm, and the output of electrospun micro/nanocore spun yarn was about 2016 cm. It also has certain industrial application value for high value-added tissue engineering, sensors, and other fields. If the single needle is replaced by multiple needled or needle-free electrospinning technology, the output will be much higher than that of traditional single-needle electrospinning. The diameter distribution and yarn strength with the


Management of endometrial cancer, an adenocarcinoma of the endometrium which occupies most uterine corpus neoplasms, including uterine sarcomas, has been more relevant due to its increasing incidence. Extensive research on tumorigenesis molecular mechanisms and molecular characterization across cancers has brought paradigm shifts in the treatment of various malignant tumors. Endometrial cancer treatment has been traditionally guided according to the disease extent or histology types, while recent studies on molecular features have led to the introduction of targeted agents into clinical use, along with conventional chemotherapeutic agents in patients with recurrent or metastatic disease. Considering the proven efficacy and relatively tolerable toxicities of targeted therapies across malignant tumors, improvement of treatment outcomes is also expected in endometrial cancer by adopting an individualized therapy depending on the specific molecular features. Efficacy assessment of new biological agents is still ongoing based on previous preclinical data on endometrial cancer molecular features. Here, endometrial cancer molecular characterization will be reviewed, and then, we will introduce preclinical data, directing the adoption of new biological agents. Endometrial cancer (EC), arising from the epithelium of the uterine corpus, is one of the leading causes of cancer mortality among women in developed countries , 2. EC hAlong with tremendous efforts on clarifying the molecular mechanisms of tumorigenesis across cancers, therapies targeting molecules involved in carcinogenesis have been dramatically developed for several decades. Having been fueled by successful treatment outcomes of targeted therapies, guiding treatment of malignant tumors according to molecular aberrations is now generally regarded as one of treatment options for promising outcomes. On the other hand, EC has been relatively distant from benefiting from targeted therapies compared to other malignant tumors, but recent efforts to understand the disease biology have released results of preclinical studies, leading to the development of clinical trials to test the potential of novel biological agents in EC treatment. In 2016, the U.S. National Cancer Institute (NCI) organized a Uterine Clinical Trial Planning Meeting to promote the design of clinical trials for the advance of molecular targeted therapies in EC  in patients with advanced or recurrent HER2-positive serous EC [HER2 also showed clinical responses when treated in combination with chemotherapy [Although amplification/overexpression of HER2 has been reported to be frequently found in ECs, particularly in serous ECs , 86, theoutcomes . Lapatinoutcomes . Howevererous EC . An earlotherapy , giving otherapy . CombinaARID1A in EC [ARID1A deficiency, resulting in impairment of HR DNA repair , has pro1A in EC  led to tp = 0.43) was shown by adding bevacizumab in chemotherapy [Bevacizumab, a monoclonal antibody against vascular endothelial growth factor-A (VEGF-A), has shown activity across cancers. Efficacy of the single-agent bevacizumab was examined in recurrent or persistent ECs in a phase II clinical trial. Results in this study, an RR of 13.5% and a median PFS of 4.2 months, encouraged further investigation on its efficacy in EC patients . The effotherapy . No beneotherapy . HoweverResults of clinical trials for targeted agents other than hormonal agents or immunotherapy are summarized in Tailored therapy according to the molecular characterization has been expected to be one of the promising treatment modalities to improve the outcomes across solid cancers, despite various challenging interpretations of genetic aberrations to directly guide treatments. Efforts to clarify the molecular features of EC have established distinct molecular classifications. Despite the extensive amounts of data on molecular features of ECs, clinical application of targeted agents has been limited. Currently, systemic chemotherapy is the only standard treatment strategy for advanced and recurrent ECs. Targeted therapies based on molecular features might be one of the solutions for unmet needs for the treatment of ECs. However, several limitations to be solved exist to establish targeted therapies as a standard therapy in EC treatment. First, due to the fact that considerable genetic aberrations are context-dependent according to tumors, thorough preclinical studies should be performed to address exact function of the genetic aberrations. Second, exploration of appropriate biomarkers for targeted agents should be continued to improve the treatment outcomes. Third, the clinical benefit of targeted agents should be demonstrated in large prospective clinical trials. Efforts to develop molecular targeted agents are expected to improve treatment outcomes, especially high-grade histology subtypes, by extending clinically applicable treatment modalities.


Hevea brasiliensis. However, this traditional process is facing a huge economic and environmental challenge. Compared with the acid coagulation, spontaneous or microorganism coagulation is an ecofriendly way to obtain NR with excellent performance. To clarify the influence of different sugars on NR quality, several sugars were used to assist the coagulation process. Influence of different sugars on microstructure and cold crystallization were examined by 1H NMR, DSC, etc. The results indicated that sugars exhibit different biological activity on terminal components of fresh field latex and can influence the resultant molecular structure and basic properties. Brown sugar exhibits higher metabolic activity and is inclined to decompose the protein and phospholipids crosslinking compared with other sugars. The larger molecular weight of sugar molecule is beneficial for the formation of the stable α lamellae phase and higher overall degree of crystallization.Natural rubber (NR) as a renewable biopolymer is often produced by acid coagulation of fresh natural latex collected from  Hevea brasiliensis, has been used in many applications for its outstanding properties [Natural rubber (NR), an important renewable biopolymer material which is often produced via coagulation of the fresh natural latex collected from the tree operties . Compareoperties ,5,6,7,8.Nowadays, technological processing of fresh natural latex coagulated by some types of acid is popular in the Chinese tropical area. However, this traditional process is facing the huge challenge of adapting to the rising severe environmental demands since the process will cost a large amount of ammonia and acid, which makes the traditional process neither economical nor eco-friendly. Additional treatment is needed to avoid pollution of the acidic effluent ,10. MoreIn recent years, the spontaneous or biological coagulation to obtain raw material natural rubber has attracted increasing attention due to not only their eco-friendly and economical nature, but also the excellent performance of NR, e.g., good mechanical properties, high cure rate, thermal stability, good dynamic mechanical properties ,12,13,14Moreover, in recent research of natural rubber, crystallization has attracted a considerable attention worldwide from both academic and industrial points of view due to its significant role in many mechanical performances and in some practical applications such as sealing ,24,25,26To our best knowledge, few studies have been focused on comparing the effect of sugars on the microstructure and crystallization behavior of NR. Information on the difference of the properties of raw natural rubber prepared by different kinds of commercial sugars in an assisted spontaneous coagulation process is not available in the literature so far. Hence, we studied the microstructure parameters and the cold crystallization behavior of raw NR with a view not only towards further insight of influence of sugars on basic properties but also the regulation and control of natural rubber quality. In order to clarify the difference of biological conversion and the influence of sugars on NR quality, several type of commercial sugars were used to assist the coagulation process of fresh field latex. Based on the difference of molecular degree of polymerization and purity of sugars, glucose, white sugar, brown sugar, trehalose, and cassava starch were used in this work. Metabolic activity of those sugars on terminal components, branching degree of natural rubber molecules, Mooney viscosity, rheological property were examined and the difference of cold crystallization behavior were also investigated.Fresh natural Latex was collected from Xida Farm located in Chengmai County, Hainan Province, China. Toluene (A.R.), glucose (A.R.) was purchased from Guangzhou chemical reagent factory, Guangzhou, China. Trehalose (99%) was manufactured by Hayashbara Co., Ltd., Okayama, Japan. White sugar (food-grade) and brown sugar (food-grade) were purchased from local supermarket. Cassava starch (food-grade) was from Wuxi Shenglunte international trade co. Ltd., Wuxi, China.Hevea brasiliensis was determined by Microwave set  according to the method provided in the instrument manual. Then, a quantitative volume of fresh natural latex containing 150 g dry rubber was diluted with deionized water to 20 wt% and then added 1 wt% aqueous sugar substance with slow stirring for 30 min. As for cassava starch, it needs to be gelatinized at 90 °C before being added into the latex. The above mixture was eventually adjusted to the 18 wt% of the dry rubber content to coagulate spontaneously in the stainless steel square plate with a glass cover at the room temperature until the wet gum formed completely. The coagulation and maturation were carried out at room temperature for around 60 h under typical tropical circumstances. Afterwards, the wet gum was washed, creped out, dried in an electro-thermostatic blast oven  for about 72 h to obtain the raw material natural rubber. The control sample was coagulated by acetic acid according to the traditional acid coagulation procedure.Raw NR was prepared via a spontaneous coagulation process assisted by the sugars. The typical process is as follows: First, the dry rubber content of the fresh natural rubber latex collected from the tree Molecular weight of NR samples were determined by viscometric method. The intrinsic viscosity, Huggins’ constant and Kramer’s constant were measured with a single bulb Ubbelohde viscometer (diameter 0.54 mm) at 30 ± 0.1 °C in toluene. All of the solvents and rubber solutions were filtered through a Bacteria funnel (30~50 micrometer) before measurement. The measurement was repeated until three consecutive readings differed by ±0.2 s. The intrinsic viscosity  of natural rubber sample was performed using MV-2000 Mooney viscometer . The sample is preheated at 100 ± 1 °C for 1 min before starting the motor and further continuously measured for 4 min with small size rotor. The measurement was repeated 3 times for each sample.Mooney viscosity  at 100 °C, with the strain of 14% and sweeping frequency range of 0~30 Hz. The cylindrical test pieces (45 mm diameter and 4 mm thick) were prepared with a volumetric sample cutter after the homogenization of raw material rubber with a two-roll mill.2. The mold was then cooled down to room temperature under pressure over 12 min. The resulting thin NR sheets (2 mm thick) were then die cut with type C dumbbell die. Determination of green strength was carried out using a Gotech AI-3000 model based on GB/T 528-92. The testing crosshead speed of 500 mm/min was applied with load cell of 100 N.The raw natural rubber were first compression molded at 100 °C for 10 min under the pressure of 200 Kg/cmc = −25 °C) for 8 h. After the end of crystallization, the sample was cooled to −40 °C and heated to 90 °C at the rate of 10 °C/min. According to the previous work, the degree of crystallization (cX) can be calculated by the following Equation (3).y is the weight ratio of filler, . About 10 mg natural rubber sample was sealed in standard Aluminum crucible to remove the thermal history by heating at 90 °C for 5 min and quickly cooled to crystallization temperature Tc = − °C for 867.3 J/g .6 to 1.49 × 106. As a control sample, the measured molecular weight of raw natural rubber coagulated by acid is 1.22 × 106 which is only higher than that of sample from coagulation using brown sugar. Among those sugars used in the experiment, brown sugar resulted in the lowest molecular weight of raw natural rubber while white sugar got the highest molecular weight of raw natural rubber. Other sugars including glucose, trehalose and cassava starch resulted in medium molecular weight. This may be attributed to molecular activity of different sugars due to their different molecular structure and purity. Although brown sugar and white sugar both belong to the disaccharide, they have different activity to promote biological action of microorganism in the latex system due to their nuanced component and chemical structure. Brown sugar usually contains some other nutritious constituents such as vitamins and trace mineral which are beneficial to biological action of microorganism. Thereafter, brown sugar is inclined to decompose the protein and phospholipids crosslinking and obtain the lower molecular weight. In other words, the effect of sugars on the resulted molecular weight can be ascribed to their different ability of decomposing the initial crosslinking points between the natural rubber molecular chains through prompting the metabolism of microorganism.It is well-known that factual molecular size of NR is related to the branching extent between single molecular chains. Our previous results demonstrated glucose exerted an obvious effect to adjust the number of ω-terminal and α-terminal of natural rubber molecular chains via the metabolism of microorganisms . In orde1H NMR measurement was carried out on the NR sample coagulated by traditional acid and sugars, respectively. Since the above molecular weight data indicated that the brown sugar has a higher ability of this kind, brown sugar was chosen as the typical representation of sugars to compare the traditional acid. 1H NMR spectra for NR samples from brown sugar and from acetic acid (see 3), methylene proton (CH2) and methine proton (CH) of cis-1, 4-isoprene unit of NR molecular chain, respectively. Compared with spectrum (b), spectrum (d) appeared additional signals at 1.60 ppm, 1.51 ppm, 1.43 ppm which indicated that brown sugar has the ability to decompose the proteins and phospholipids. This was consistent with the previous reference [To further confirm the ability of sugar to decompose the protein and phospholipids, acid see . Three meference  which teThe possible decomposition of crosslinking points between the terminals of the natural rubber molecular chains was shown in k′) and the sum value of Huggins’ constant and Kramer’s constant (β). k′ and β can be used to describe the branching degree of natural rubber molecules [k′ is the qualitative index of long chain branching, which goes up with the increment of the number of branching while it is independent of molecular weight and molecular weight distribution. If k′ ranges from 0.45 to 0.65, it shows that macromolecules are branched; if k′ is about 0.3, the macromolecules are linear [k′ and β equals 0.5, However, the value of the sum of k′ and β has apparent deviation from 0.5.olecules . k′ is te linear . In regak′ value and the sum of k′ and β is very close to the 0.5, which indicating that the corresponding NR sample from auxiliary coagulation by brown sugar and glucose may have a lower branching degree and the rubber chains are close to linear molecules. While white sugar gained a higher k′ value and the sum of k′ and β deviated from 0.5. Therefore, the sample of natural rubber solidified by white sugar had a higher branching degree. Trehalose and cassava starch got a middle k′ value. In addition, the sum of k′ and β had a similar tendency to k′ with respect to the all sugars used in the work, which indicated the parameter of k′ agreed well with the parameter of the sum of k′ and β in assessing of the branching degree of the natural rubber molecular chains.From the Mooney viscosity which indicates the flow properties of NR is an important assessment parameter for assessing the raw material natural rubber quality and processability . The MooRubber Processing Analyzer (RPA) was widely used to determine the rheological properties of rubber melts ,36. The Moreover, the loss factor (tanδ) which is defined as the ratio of viscous modulus to elastic modulus reflects the recovery characteristics of the melts after deformation and a larger tanδ value indicates a more liquid-like character . Figure Sugars exist different biological action with non-rubber constituent and have different decomposition effect on the branching points of the natural rubber molecular chains which will consequently affect the stress-strain behaviors of the unvulcanized rubber that are important to certain processing operations in the rubber industry . The greβ lamellae and the higher melting peak could be produced by thermodynamically more stable α lamellae. Chenal [β lamellae, and the higher temperature peak for the phase of α lamellae. Therefore, the influence of sugar type on the formation of α lamellae and β lamellae was determined by DSC (see α lamellae and β lamellae, respectively. It is commonly accepted that peak position of melting peak is related to lamellae thickness and crystal perfection [β peak shifted slightly toward lower temperature with increasing molecular weight of sugar roughly, while α peak shifted toward higher temperature very slightly. However, the shift of α peak was much smaller than that of the β peak. This indicated that α lamellae phase was thicker and or/more perfect when lower molecular weight of sugar used in the spontaneous coagulation of fresh natural latex. The sequence of β peak position towards the higher temperature was glucose, brown sugar, trehalose and Cassava starch, and white sugar.For the cold crystallized natural rubber c natural rubber, there are always two different peaks detected in the DSC curves. According to Philips , the low. Chenal  showed t DSC see . From thrfection ,41. The α and β lamellae were calculated (see α lamellae and β lamellae. The fraction of α lamellae and β lamellae of control sample was also determined and was 7.37% and 6.03%, respectively, which indicates that the fraction of β lamellae of rubber samples from coagulation with these sugars was lower than that from control sample. It is obvious that glucose as a monosaccharide resulted in the higher content of less stable β lamellae than stable α lamellae among all sugars used in the experiment. Except for the sample from the assisted coagulation by glucose, the fraction of α lamellae was higher than that of β lamellae in the samples from the assisted coagulation by other disaccharides and polysaccharide. It is noteworthy that the ratio of α lamellae to β lamellae was increasing with the molecular weight of sugar. That is to say, the ratio of α lamellae to β lamellae of rubber samples would be higher from coagulation with monosaccharide to disaccharide and then polysaccharide. Additionally, polysaccharide was in favor of formation stable lamellae α and higher total degree of crystallization.Based on the soft-ware built in the Mettler-Toledo DSC system, the melting enthalpy and the crystallinity of both ated see . Figure α lamellae to β lamellae and the overall crystallinity of rubber sample coagulated with different kinds of sugars. The fraction of α lamellae and β lamellae of control sample was also determined and was 7.37% and 6.03%, respectively, which indicated that the fraction of β lamellae of rubber samples from coagulation with these sugars was lower than that from control sample. It is obvious that the overall crystallinity in the natural rubber sample increased with the increase of molecular weight of sugars used in the experiment. Moreover, the effect of sugars on the ratio of α to β in crystallographic phase was roughly similar to the overall crystallinity. Among the three kinds of disaccharides, trehalose resulted in higher ratio of α to β.α lamellae phase and higher overall degree of crystallization. The results may provide an alternative way for the conversion of field coagula into suitable forms for long-term storage and marketing.A green way to obtain raw material natural rubber was completed via assisted coagulation of natural latex by sugar substance. In this process, microorganisms exhibit different biological activity with different sugars and the given terminal components of original fresh field latex as nutrition and further influence the resultant molecular structure and basic properties including molecular weight, branching degree, Mooney viscosity, rheological performance and green strength. Cold crystallization behavior as an important performance for the natural rubber is also influenced by different sugars. The saccharide molecules of larger molecular weight are beneficial to the formation of the stable


Communicating about antimicrobial resistance (AMR) and antimicrobial stewardship (AMS) requires technical knowledge, consideration of audience values and appropriate identification of communication strategies for multiple audiences. Within the context of animal agriculture, communicating about AMR represents an important and complex endeavour for veterinarians, governmental agencies, producers and the industry to convey policy and practice information regarding the use of antimicrobials in food animals.To assess the science communication challenges related to AMR by identifying the motivations, goals and struggles of animal agriculture stakeholders when communicating about AMR and AMS.N = 80) completed a workshop on science communication, including small group meetings with oral/written comments collected. Participants included veterinarians, government agency representatives, industry stakeholders and producers.Participants attending a meeting on AMR communication in animal agriculture (Results indicated participants believed providing more accurate information would resolve misunderstanding and concern about AMR to other stakeholders, counter to recommendations of science communicators. Other participants noted beliefs about the utility of stories in trying to explain how AMS is normative and consistent with the values of all parties interested in animal agriculture. Participants noted the importance of public engagement, even if the participants’ perceived target audiences did not include the public.Communicating about AMR and AMS in animal agriculture contexts provide unique challenges. Few evidence-based recommendations are available for science communicators in these contexts and more research is needed to improve the quality of communication about AMR and AMS in animal agriculture. Farmers and producers, those who manage and run operations where animals may be treated with antibiotics, may perceive their stakeholders to be retailers and consumers who buy their product, veterinarians who treat their animals and government officials who might draft regulations producers must follow. Each of these stakeholder groups has unique obligations and limitations related to AMR. For example, veterinarians must adhere to regulations from government agencies, discuss treatment plans for clients and serve as a public-facing group given the visibility of their profession to mainstream audiences. Similar complexities exist for other animal agriculture stakeholders whose diverse obligations, audiences and communication goals require varied, and effective, science communication strategies. At present, few studies examine how the many stakeholder groups in animal agriculture manage and address communication challenges relevant to AMR.The current study reports the outcomes of an interactive, in-person workshop focused on improving science communication about AMR among animal agriculture stakeholders. The objective of the workshop evaluation reported here was to increase understanding about the communicative challenges facing animal agriculture stakeholders and provide a foundation for improving AMR communication efforts in the future.N = 80) responded to prompts about (i) why communication about AMR is important to their job; (ii) the audience with which they wanted to better communicate about AMR; (iii) a challenging AMR communication situation faced in the past; (iv) the cause of said communication challenge; and (v) how to improve communication efforts in the future.Participants in this study were science communication workshop attendees at a national antibiotic symposium hosted by the National Institute of Animal Agriculture. The workshop included participants representing five stakeholder groups  by the public across multiple  stakeholder groups.All stakeholder groups indicated the need to improve consumer/public understanding of AMR and AMS at varying levels of priority. Policymakers  were the only other audience segment of interest mentioned by most groups.Most participants, across groups, indicated a lack of knowledge (of some audience) hindering communication attempts. Unfamiliarity with animal agriculture practices and policies was another frequently cited source of communication challenges.Participants noted inconsistent messaging, the complexities of AMR policies and concerns, and lack of consumer knowledge as drivers of communication challenges. Other reasons for challenges included beliefs about media messages influencing myths and misperceptions and that animal agriculture messaging often focuses more on precision/accuracy than clarity.ineffective in science communication.Many participants believed using stories and analogies focusing on shared values  and emotions might be a useful approach. Other participants advocated for a deficit model approach, relying on the provision of information to the target audience to correct a perceived knowledge deficit. While this strategy remains a popular and oft-cited approach for public outreach, it has been found to be consistently The current study suggests that agricultural stakeholders are aware of the considerable challenges in communicating about AMR to audiences of interest. Many responses for improving communication aligned with strategies that science communicators advocate . However, there were still participants who favoured simply filling the perceived gap in knowledge related to AMR and AMS, which science communication research has shown repeatedly to be unproductive. Findings from this study suggest a need for more engagement between animal agriculture stakeholder groups and social scientists who could provide recommendations/training about communication and public engagement theory and practice that move beyond the deficit model. Moreover, the breadth of opinions and issues addressed by these stakeholders highlights the need for more nuanced approaches to communication/outreach with a diversity of stakeholder groups with animal agriculture interests. Without a larger evidence base, AMR communication will continue to be a challenge, hindering progress toward widespread adoption of AMS behaviours. Evidence-based recommendations will improve communication and facilitate behaviour change that contributes to AMS efforts across animal health contexts.


In this theoretical study, we report on the molecular electrostatic potential (MEP) of titled molecules confined by repulsive potentials of cylindrical symmetry mimicking a topology. Our calculations show that the spatial restriction significantly changes the picture of the MEP of molecules in a quantitative and qualitative sense. In particular, the drastic changes in the MEP as a function of the strength of spatial confinement are observed for the BrCN molecule. This preliminary study is the first step in the investigation of the behavior of the MEP of molecular systems under orbital compression. AZ denotes the charge on nucleus A, located at a distance AR, and ρ(r′) is the molecule’s electronic density. In contrast to ρ(r), which represents solely the electronic density at the point r, MEP involves contributions from each nucleus and electrons in the entire molecule, thus providing a total electrostatic picture. The first term of Equation (1) brings in the positive contribution of the nuclei, while the second one represents the negative effect of the electrons. Hence, the sign of V(r) in any particular region depends on whether the nuclear or the electronic term is dominant there. It is worth mentioning that MEP belongs to the quantities that are observable physically, and thus, it can be derived directly from the wave function or experimental electron densities available from, e.g., X-ray diffraction [Incorporation of the molecular electrostatic potential (MEP) concept into the chemistry and molecular physics community is owed to the pioneering works of Scrocco and Tomasi in the early 1970s ,3,4,5. Ifraction . Typicalfraction ,9,10, jufraction . An impofraction ,13,14,15fraction .Recent years have seen a significant strengthening of studies concerning the description of noncovalent complexes exposed to high pressure or embedded in confining environments  of atomic and molecular systems  in the molecular complexes ,25,26. Aω = 0 and ω = 0.6) are presented in The results of the calculations of MEP  for the investigated molecules  and minimum  values of MEP. It should be noticed that s,maxV and s,minV correspond to the positive and negative of the MEP of the investigated molecules, respectively. In s,maxV of the confined HF molecule significantly decreases in comparison with the situation without spatial restriction. This finding becomes clear after analyzing the results presented in s,maxV , s,minV is located on the extension of the covalent bond H-F  is already observed for ω ≈ 0.3. It is an important finding that may have very serious consequences in the theoretical description of molecular complexes, where the σ-hole idea is applied (see Introduction). Because the σ-hole, in the classical meaning, disappears under strong spatial confinement (in this condition (ω = 0.6), the value of MEP on the extension of the covalent bond Br-CN is negative and equal to −2.6 s,minV. The location of s,minV for BrCN is not changed but their value is more negative, i.e., More drastic changes are observed for the BrCN molecule. The significant reorganization of the MEP is observed in the presence of spatial restriction in comparison with the free molecule . In particular, the semi-empirical, range-separated hybrid functional ωB97X as an approximation of the exchange–correlation energy functional in DFT  combinedous work . It is wIn summary, the present study focuses on the investigation of behavior of the MEP as a function of the strength of spatial restriction . In order to model this effect, the cylindrical symmetry of the repulsive potential  was applied. Our preliminary studies showed that that the spatial restriction may significantly change the picture of the MEP of molecules in a quantitative and qualitative sense. This finding suggests that the many processes that occur in such conditions (in which electrostatic interactions play important role) may show different faces in comparison with the ambient conditions. It would be interesting to propose experimental measurements for the confirmation our findings. It possible to imagine that the electron densities (rather than MEP) can be derived from the X-ray diffraction experiment, as the molecular crystals contain investigated molecules. However, the measurements should be carried out as a function of the external pressure. The model confining potential often correctly described high pressure effects. It should be noted that the two-dimensional harmonic oscillator model is a hypothetical approximation. It is mainly connected with the fact that the presented model describes only repulsive forces (this type of interaction is a result of the Pauli exclusion principle and increases rapidly when the wave functions of guest and host molecules start to overlap) but neglects the van der Waals force of attraction from the neighboring molecules. Thus, this simple model of the spatial confinement corresponds to a non-polarizable, electronically inert environment. This important restriction should of course be taken into account. However, it should be underlined that this is the first study analyzing the effect of spatial confinement on MEP of molecular systems. The authors would like to express their hope that more studies will follow so that it will be possible to generalize the conclusions presented in this work.