The best alpha-synuclein threshold was characterized by an Area Under the Curve = 0.85, Sensitivity = 82.0% and Specificity = 76.0%. CSF alpha-synuclein was associated with CSF AT(N) biomarkers positivity (p < 0.01) but not with hippocampal atrophy or white matter lesions. Conclusion: CSF Alpha-synuclein evaluation could help to early differentiate patients with DLB and AD in association with existing biomarkers.Background: Alpha-synuclein, abnormally aggregated in Dementia with Lewy Bodies (DLB), could represent a potential biomarker to improve the differentiation between DLB and Alzheimer’s disease (AD). Our main objective was to compare Cerebrospinal Fluid (CSF) alpha-synuclein levels between patients with DLB, AD and Neurological Control (NC) individuals. Methods: In a monocentric retrospective study, we assessed CSF alpha-synuclein concentration with a validated ELISA kit (ADx EUROIMMUN) in patients with DLB, AD and NC from a tertiary memory clinic. Between-group comparisons were performed, and Receiver Operating Characteristic analysis was used to identify the best CSF alpha-synuclein threshold. We examined the associations between CSF alpha-synuclein, other core AD CSF biomarkers and brain MRI characteristics. Results: We included 127 participants . CSF alpha-synuclein levels were significantly lower in DLB than in AD (1.28 ± 0.52 ng/mL vs. 2.26 ± 0.91 ng/mL, respectively,  Over the past ten years, Cerebrospinal Fluid (CSF) biomarkers (β-amyloid (Aβ), total Tau (t-Tau), phosphorylated Tau181 (pTau)) have dramatically improved the accuracy of the molecular diagnosis of Alzheimer’s Disease (AD) in clinical practice. However, several differential diagnoses remain challenging due to the poor specificity of clinical symptoms linked to neurocognitive disorders and sometimes because of the overlap between neurodegenerative disorders, vascular cognitive impairment and brain aging. The diagnosis of Dementia with Lewy Bodies (DLB), the second cause of neurodegenerative cognitive impairment after AD . These cDLB is neuropathologically characterized by an accumulation in the brain of abnormally aggregated and phosphorylated alpha-synuclein (α-syn), a 140 amino-acids protein, which is mainly expressed in presynaptic terminals . This acOur primary objective was to compare the CSF α-syn levels, measured by a validated ELISA kit, in extensively-phenotyped AD, DLB patients or neurological control individuals from a clinical practice cohort of patients recruited in a tertiary memory clinic. The secondary objectives were to study the relationships between the concentration of CSF α-syn and (i) CSF amyloid and Tau biomarkers, (ii) the main neuroimaging characteristics of these individuals, (iii) the Mild Cognitive Impairment (MCI) or dementia stages, in patients with DLB or AD. p < 0.001). Patients with DLB reported hallucinations in 28.9%, fluctuations in 22.2%, parkinsonism (or positive DAT-SCAN imaging) in 60.0% and RBD in 33.3%. There was no difference regarding hippocampal atrophy between individuals with AD and those with DLB (p = 0.80).We retrospectively selected 85 DLB patients and 105 NC, among whom 45 DLB patients and 32 NC individuals met all the inclusion criteria See ; we alsop < 0.001). There was no significant difference between DLB and NC. In addition, α-syn levels were higher in individuals with AD than in NC (2.26 (0.91) ng/mL vs. 1.56 (0.60) ng/mL, p < 0.001) (The mean CSF α-syn levels were significantly lower in the DLB group than in the AD group: 1.28 (0.52) ng/mL vs. 2.26 (0.91) ng/mL, respectively (< 0.001) .p < 0.001). Further analysis did not show any significant differences between early (MCI) and late (dementia) stages in both patients with AD (p = 0.62) and in those with DLB (p = 0.30)  . The best threshold of α-syn to discriminate patients with DLB from patients with AD was 1.50 ng/mL. With this value, we obtained a sensitivity (Se) = 82.0%, a specificity (Spe) = 76.0%, and hence, an area under the curve (AUC) = 0.85 .The mean concentration of total CSF α-syn was significantly increased in patients with brain amyloïdopathy (A+), tau pathology (T+) and neurodegenerescence (N+) . There wIn this study, we observed significantly higher concentrations of CSF α-syn in AD patients than in those diagnosed with DLB or in NC individuals. However, there was no significant difference in CSF α-syn concentrations between the DLB and NC groups. CSF α-syn was neither associated with the stage of hippocampal atrophy nor with white matter lesions on brain MRI. Among the DLB or AD groups, there was no difference in CSF α-syn concentrations regarding the stage of cognitive impairment (MCI vs. dementia). 2O2, associated with delayed cell death, in comparison with control cell lines [α-syn has been identified as a toxic protein to human neurons and a trigger to neuronal death . More prll lines . Therefoll lines ,30,31. Tll lines . As in tll lines , this reAs a result, both aggregates of α-syn in DLB and overexpression of wild-type α-syn in AD favor synaptotoxicity. Nevertheless, previous evidence suggested that the destruction of the synapse was more important in AD than in other neurodegenerative diseases . PatientSo far, several studies have examined CSF α-syn in DLB in comparison with AD, other neurodegenerative disorders or NC  Paris Nord  France. This department has expertise in the diagnosis and care of patients with cognitive disorders and neurodegenerative diseases.BioCogBank is a monocentric clinical practice cohort of 1500+ patients with cognitive complaints prospectively recruited in the CNC since 2008. All patients included in BioCogBank underwent neurological and neuropsychological assessments, neuroimaging and CSF AD biomarkers analysis for cognitive complaints. Plasma and CSF samples have been biobanked for most of these patients. CSF results were classified according to the AT(N) classification . A conseWe retrospectively identified, from participants included in the cohort between 2008 and 2021, patients fulfilling criteria for DLB, taking into account the information recorded during the clinical follow-up  and confThe following cardiovascular risk factors were collected: smoking status (including active smoking or having quit < 3 years ago), hypertension (systolic blood pressure ≥ 140 mmHg and/or diastolic blood pressure ≥ 90 mmHg), diabetes (defined by two measures of glycemia after fasting ≥ 1.26 g/L ) and dysThe following clinical symptoms associated with DLB were recorded: visual hallucinations (reported by the patient or a relative/caregiver), parkinsonism or dopamine denervation , fluctuating cognition, Rapid eye movement sleep Behavior Disorders (RBD), reported by patients and relatives and/or using a polysomnography when performed. Global cognitive function was assessed by the Mini Mental State Examination (MMSE) score . APOE ε4 allele. Several characteristics of brain MRI were collected for the present analysis: hippocampal atrophy using the visual Scheltens score, defined by the mean values between the right and left sides and periventricular white matter hyperintensities (Fazekas score). DNA was extracted from peripheral blood samples for ApoE genotyping, and participants were classified as carrier or non-carrier of at least one g at 4 °C for 10 min and aliquoted in polypropylene tubes of 1.5 mL and stored at −80 °C until further analysis, according to international guidelines [®; Fujirebio, Ghent, Belgium) until July 2018, according to the manufacturer’s instruction. Then, the concentration of these biomarkers was measured by the analyzer Cobas 601 with the electrochemiluminescent reagents Elecsys® . The CSF profiles of patients were classified in A+T+N+/A-T-N-/or intermediate according to cut-offs determined by the laboratory.CSF samples were obtained by lumbar puncture, immediately centrifugated at 1000× idelines . AD CSF idelines . Aβ42, ACSF levels of α-syn were measured using an ELISA kit  available via EUROIMMUN, Lübeck, Germany), validated for research for the quantification of human total α-syn in CSF . Comité d’évaluation et d’Ethique pour la recherche Paris Nord) and the Commission Nationale Informatique et Libertés.All the participants provided oral and written information about the opportunity to collect additional blood and CSF samples and perform genetic analyses for further research analyses, according to the BioCogBank© protocol. Written informed consents were obtained for all participants except for two subjects with dementia, under guardianship. Accordingly, consent was obtained from their legal guardian. This study was approved by the local Ethics Committees ) and categorical variables were presented as percentages (number of subjects). Analysis of variance (ANOVA) was carried out to compare the CSF concentration of biomarkers between the diagnostic groups, taking into account the stage of cognitive impairment (MCI or dementia). For variables with non-Gaussian distribution, comparisons were made using a Kruskall–Wallis test. Regarding categorical variables, intergroup comparisons were performed using Chi-squared or exact Fisher tests.We compared the CSF α-syn concentrations with regards to the diagnostic groups and the stage of cognitive impairment, then with regards to their CSF amyloid (A), p-Tau (T) and t-Tau (N) status. Finally, we compared the CSF α-syn concentration given the presence or absence of brain MRI abnormalities. The analysis of Receiver Operating Characteristic (ROC) curve, Se, Spe and AUC were carried out to determine the best α-syn threshold for discriminating subjects with DLB from those with AD or NC. Statistical analyses were performed using R-studio (v1.4.1106).Our study demonstrated that α-syn could represent another promising biomarker for improving the discrimination between AD and DLB patients, even at the prodromal stage of the disease. The higher CSF α-syn levels in AD than in DLB may reflect both aggregation of misfolded α-syn in DLB as well as inflammation and faster neurodegeneration in AD. The implementation of CSF α-syn measurement to AT(N) CSF biomarkers could be useful as part of a biomarker panel to support DLB diagnosis. This novel approach could enable an earlier and more accurate diagnosis as well as a better selection of patients for clinical trials of future disease-modifying treatments for DLB. α-syn may also provide additional insights into the prognosis of patients with DLB. Further studies are warranted to precisely determine the role of this biomarker in clinical practice and in therapeutic research.


Isoelectric focusing followed by immunoblotting is a method routinely used in human medicine to assess the presence of oligoclonal bands (OCBs) in cerebrospinal fluid (CSF) and serum. The detection of OCBs is a valuable diagnostic test, especially important in patients with the suspicion of multiple sclerosis (MS), in which at least two OCBs are found in the CSF not present in paired serum samples in up to 95% of patients. So far, presence of OCBs in CSF and serum of dogs has only been investigated in a small cohort of dogs diagnosed with degenerative myelopathy and healthy dogs. The main objective of the current study was to describe the method used for OCB detection and compare two different canine anti-IgG antibodies: a canine rabbit-anti-IgG antibody (Jackson ImmunoResearch) vs. a canine goat-anti-IgG antibody (Bio-Rad). The method was performed according to the instructions of the commercial kit used. The canine goat-anti-IgG antibody showed a better performance than the canine rabbit-anti-IgG antibody. The availability of the technique of OCB detection in the dog paves the way for further studies, especially in the field of inflammatory diseases of the canine central nervous system, and comparison between specific human and canine diseases. The analysis of cerebrospinal fluid (CSF) is an important diagnostic test for various neurological diseases in both human and veterinary patients. Besides the standard macroscopic, biochemical, and cytological analyses, the assessment of IgG oligoclonal bands (OCBs) is routinely performed in human medicine. B cells and plasma cells involved in an inflammatory process are responsible for the production of oligoclonal immunoglobulin within the CNS; the term oligoclonal means the immunoglobulin is derived from few antibody clones . OCBs thThe purpose of the present study was to describe a modified method for OCB detection in dogs as well as to compare two different canine anti-IgG antibodies for detection of OCBs in CSF and serum samples of dogs. For this purpose, isoelectric focusing followed by immunoblotting used in human medicine was investigated in a clinical laboratory specialized in human CSF diagnostics.The experimental procedures used in this study were approved by the Ethical Committee of the Veterinary Service, Cantone of Bern (BE121/2020).n = 8, idiopathic epilepsy (IE) n = 2, intracranial neoplasia (IN) n = 1, intervertebral disc herniation (IVDH) n = 2, steroid-responsive meningitis-arteritis (SRMA) n = 3, eosinophilic meningoencephalitis n = 1], as well as three dogs that did not receive a final diagnosis. The medical records including signalment of all dogs were available. Moreover, all dogs included received a complete diagnostic work-up with physical and neurological examination, as well as CSF analysis. The further diagnostic work-up differed according to the suspected underlying disease and included one or more of the following examinations: hematology, biochemistry, infectious disease testing, urine analysis, thorax and/or abdominal radiographs, MRI of the brain and/or spinal cord. The samples used for this brief research report were investigated to describe the method and compare two different canine anti-IgG antibodies. The description of CSF-specific OCBs in neurological and non-neurological diseases and a statistical comparison among different groups are not objects of this communication.Dogs included in the study were presented for diagnostic purposes to the Small Animal Clinic, Division of Clinical Neurology, Vetsuisse Faculty Bern, as well as to the Neurology Department, Clinic of Small Animal Surgery, Vetsuisse Faculty Zurich and were diagnosed with different neurological diseases [meningoencephalitis of unknown origin (MUO) g, at 20°C) and CSF and serum samples frozen at −80°C within 1 h after collection. Samples were thawed prior to the measurement of the IgG concentration and further processing for OCB detection.Paired CSF and serum samples were collected from all dogs after the MRI examination of the brain and/or spinal cord for the underlying disease in general anesthesia. CSF was obtained by atlanto-occipital puncture in lateral recumbency, harvested into plain polypropylene tubes, and clinical standard examinations were performed within 30 min. Standard clinical CSF analysis included total protein, white blood cell count and cell differentiation. Blood samples mostly were taken at the same time or within 24 h by puncture of a peripheral vein . Serum samples were centrifuged  was used according to the manufacturer‘s protocol. A pilot study was performed to determine suitable serum and CSF dilutions to be used in the ELISA, using samples from two dogs. Ten-fold serum dilutions ranging from 1:1,000 to 1:1,000,000 were made and CSF was tested undiluted and in serial dilutions up to 1:10,000. Values were within the standard curve using a serum dilution of 1:100,000 and CSF dilution of 1:1,000. These dilutions were used for all following IgG measurements in CSF and serum. Moreover, serum and CSF dilutions of the two dogs initially used to establish the ideal dilution were used as controls for all further ELISA plates measured. The median IgG concentration in CSF and serum was 137.66 mg/L and 23.32 g/L with a mean of 352.52 mg/L and 24.46 g/L, respectively.IEF and subsequent immunoblot were performed according to the instructions of the human kit provided .Aqua ad iniectabilia to reach equal IgG concentrations of 25 mg/mL. The diluted serum and CSF samples were vortexed and 7 μL per sample were pipetted into the wells of the agarose gel (1%). In case the original IgG concentration was <25 mg/mL, the total amount was increased accordingly to reach an equal amount of IgG. As a standard canine positive control was not available due to restricted CSF and serum sample volumes in canine patients, a human CSF and serum sample were used as controls , quickly dried between two soft paper tissues and then placed onto the gel in order to remove excess proteins and fluid. The lower right corner of a second blotting membrane was removed, and the membrane placed into a plastic container with 20 mL Aqua destillata and 1.6 mL methanol 100% (helps to transfer the proteins onto the membrane). The membrane was then placed onto the gel, carefully smoothed with gloves until no air bubbles or irregularities were visible and covered with the smooth surface of a blotter G paper. A second aluminum plate was placed on top of the membrane and two weights (each 1 kg) applied on top. The membrane was blotted for 30 min.After completion of the IEF, the gel was removed from the frame and placed onto a soft paper tissue. Both contact strips were removed and the gel placed centrally on an aluminum plate. One blotting membrane (nitrocellulose membrane) was immerged in a plastic container with Aqua destillata for 3 and 2 min, respectively. The “final reaction solution” was prepared by adding 4 mL of the CSF Substrate  to the NBT CSF (Nitroblue tetrazolium CSF) and shaking it for 10 min. The blotting membrane was then transferred into a small plastic container and the whole solution was placed on top of the membrane. The membrane was incubated on a plate shaker for 5–7 min; depending on the time necessary until the bands of the control samples were nicely visible. Following the incubation, the membrane was washed again twice for 5 min first with 0.9% saline solution and then Aqua destillata and dried using soft tissues and a weight.After blotting, the membrane was placed with the blotting side upwards into a plastic container with 20 mL of Binding agent 1 and incubated for 30 min on a shaker. The blotting membrane was then washed twice in 0.9% saline solution for 2 min and placed into a plastic container with 20 mL of Binding agent 2 (=dilution of Binding agent 1: 1:10). Two canine anti-IgG antibodies were used for the measurements. The first one was a canine rabbit-anti-IgG antibody , of which 80 μL in a concentration of 0.6 mg/mL were used. This antibody was chosen based on the material used in the only clinical study on canine OCBs  working in the human laboratory specialized for OCB detection and using the method on a daily basis for more than 5 years. The presence of the positive control, overall correct running of the samples on the gel, and correct blotting were evaluated by all examiners. The result was classified as sufficient for interpretation, if the OCB pattern of the human control sample assessed with the human anti-IgG antibody was reflected by the result of the samples assessed with the canine anti-IgG antibody. Additionally, at least two out of three examiners needed to be able to read and evaluate the samples of a patient. The final evaluation of the OCB result was performed using an adapted scheme of the current guidelines in human medicine describing five patterns: pattern 1 = no OCB in CSF and serum; pattern 2 = CSF-specific OCBs, i.e. presence of OCBs in CSF, not in serum; pattern 3 = CSF-specific OCBs with additional, identical bands in CSF and serum; pattern 4 = identical OCBs in CSF and serum; and pattern 5 = monoclonal bands in CSF and serum . CSF-speCSF and serum sample of 20 dogs were examined for IgG concentration and presence of OCBs. In those 20 paired samples, the performance of two different canine anti-IgG antibodies was evaluated. Results of 16/20 (80%) dogs examined using the canine rabbit-anti-IgG antibody compared to 20/20 (100%) of dogs assessed with the canine goat-anti- IgG antibody were sufficient for evaluation of the different patterns described above. Overall, there was an excellent agreement of pattern recognition between all three examiners (CSF-specific OCBs yes or no). On contrary, there was a disagreement concerning the number of visible OCBs in 8/16 cases (50%) of dogs evaluated with the canine rabbit-anti-IgG antibody and in 12/20 cases (40%) evaluated with the canine goat-anti-IgG antibody. In these cases, the blot was evaluated again by the most experienced examiner (AL) in order to reach a consensus. Both canine anti- IgG antibodies showed cross-reactivity with human serum and CSF samples; a feature that was used to include a human sample with clear visible CSF-specific OCBs as positive control. Overall, when comparing the performance of the canine goat-anti-IgG antibody and the canine rabbit-anti-IgG antibody, both consistently lead to the same conclusion concerning the presence or absence of CSF-specific OCBs in 16/16 (100%) cases. In comparison with the canine rabbit-anti-IgG antibody, the canine goat-anti IgG antibody caused significantly less pinkish background, which facilitated detection of faint bands  and madeThe assessment of OCBs using a commercially available isoelectric focusing protocol followed by an immunoblot is routinely used in human medicine, especially during the diagnostic work-up of a presumed MS. In contrast, the diagnostic value of assessing OCBs in various diseases in dogs has not been investigated. Therefore, the present study paves the way to use the method of OCB detection in a larger clinical trial in dogs for the future. For the assessment, dogs with a variety of different clinical diagnoses were included; inclusion of healthy dogs was limited by ethical restrictions. Kamishina et al. assessed the prevalence of OCBs in CSF and serum obtained post mortem in six clinically healthy dogs; none of those presented CSF-specific OCBs . Two difThe current study presents some limitations. Firstly, only a small amount of canine samples has been evaluated and results might differ in a larger cohort. Secondly, the three examiners did not agree on the exact number of OCBs found in each sample in 50 and 40% of the dogs, when evaluated with the rabbit canine anti-IgG antibody or the goat canine anti-IgG antibody, respectively. It is important to remember that the assessment of OCBs in serum and CSF is a qualitative, not quantitative assessment. Therefore, the main conclusion is the presence or absence of CSF-specific OCBs, per definition two or more OCBs uniquely present in the CSF sample, independently from the exact number of OCBs. A third limitation is linked to the intrinsic nature of the method, being the evaluation of OCBs a subjective assessment, which can lead to both false-positive and false-negative results. Finally, the antibodies used recognize both heavy and light chains of the immunoglobulin, therefore we cannot fully exclude that also additional immunoglobulin classes other than IgG have been detected.In summary, both antibodies enabled the interpretation for the presence or absence of CSF-specific OCBs in the canine samples; overall, the canine goat-anti-IgG antibody showed a better performance than the canine rabbit-anti-IgG antibody. Besides MS, OCBs have been detected in many different diseases in humans such as other inflammatory conditions, infectious or neoplastic diseases . In contThe availability of the methodology for OCB detection in dogs paves the way for further insights concerning the pathophysiology of inflammatory diseases and could support clinical diagnostics in these diseases.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The animal study was reviewed and approved by Ethical Committee of the Veterinary Service, Cantone of Bern (BE121/2020). Written informed consent was obtained from the owners for the participation of their animals in this study.JP: conceptualization, laboratory analyses, writing—original draft preparation, and writing—review and editing. VS: conceptualization, writing and editing, supervision, and resources. MZ: introduction and helped with laboratory work, methodology, evaluation, and interpretation of laboratory results. AL and IJ: conceptualization, methodology, evaluation, and interpretation of laboratory results. FS and TB: conceptualization and writing—review and editing. AM: conceptualization, writing and editing, and supervision. All authors read and approved the final version of the manuscript.This work was supported by the Board of Specialization of the Vetsuisse Faculty of Veterinary Medicine, Bern. The funding source was not involved in study design, laboratory work, writing of the article, or decision to submit the article for publication. Open access funding was provided by the University of Bern.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.


The use of gamete donation is expanding worldwide. At the same time, the practices, and perceptions of how donor families are established are changing.While some countries still mandate anonymous donation to ensure the privacy of the donor and protection of the family unit , a number of countries have introduced new legislation according to which open‐identity donation is the sole option .Given the availability of low‐cost DNA testing and commercial international DNA ancestry sites, anonymity can no longer be guaranteed for future and past donors.How do technologies, such as genetic testing and online fora, impact anonymity in donor conception? What challenges or possibilities emerge in relation to donor conception as new technologies are used? Are there ways to organize donor conception more responsibly in the future?The consequences of these technological developments are as yet unresolved. To address this issue, the ‘Reconfiguring Donor Conception’ network, consisting of social scientists from Japan and Denmark, organized a stakeholder workshop in Copenhagen, in August 2022. Participants included representatives from public and private fertility clinics, gamete donors, parents to donor‐conceived children, donor‐conceived people, private sperm and egg banks, and researchers within the field. Here, we describe the main perceptions and ideas that emerged from this workshop by discussing the following questions: Empirical evidence suggests that what is central to the well‐being of offspring is not whether the donor is anonymous but whether parents disclose the donor conception to their child early in life. Positive associations between early disclosure and feelings about donor conception have been found.Presently, both legislations and expectations regarding donor disclosure and anonymity are rapidly changing. For example, in Denmark, it was only in 2012 that legislation enabled the use of open donors. Furthermore, while medical staff today will support a recipient's decision to disclose how their child has been conceived, heterosexual couples were guided to keep donor conception a secret up until the 2000s.DNA surprise, an unexpected finding arising from a curiosity about one's ancestry and/or genetic make‐up. This is not only the case for donor‐conceived people. Donors' relatives may also face DNA surprises if the donor kept their donor status a secret. Often in such cases, those involved in the donation, including the donor's relatives, do not necessarily have anywhere to go for needed support.The changing morality and status of the donor often place donor families in challenging situations, in which regrets and contradictory perceptions of the donation may emerge together with conflicting perceptions of whose rights and interests should be safeguarded. This is especially likely to come about when a donor‐conceived person is told of their conception late in life, when a donor‐conceived person feels a strong need to identify the donor, or when parents regret the choice of anonymous donation, but at the time of conception had only this option. The changing norms surrounding donor conception have been further accelerated by direct‐to‐consumer genetic testing, as donor‐conceived people may learn of their conception as a A number of online fora and registers have been established because many parents, donor‐conceived people, and donors are faced with questions about how to be a donor family and wish to learn more about it or potentially connect with genetic relatives. However, not everyone feels safe sharing personal information on Facebook or unofficial, unregulated registries.At the workshop stakeholders did not agree on questions such as anonymity, or whether the number of donor offspring should be limited and to what extent. Instead, a consensus emerged around the idea of developing a more comprehensive support system to help donor families, including donor relatives to receive help as challenges emerge. A number of solutions were suggested.Firstly, support could be provided through an independent website. Such a website could provide information and stories regarding the lived experiences of donor‐conceived families, including the types of challenges that may emerge and how they can be overcome. Moreover, such a platform could provide information on current legal rights and policies related to donor conception.Stakeholders also strongly emphasized the need for professional counseling services catering to the various needs and perceptions and complex situations that unfold over time within donor‐conceived families. Some women may for example regret that they had transnational treatment in a country where they used an anonymous donor, living in a country where only open donation is available. In that case neither she, nor the child, would have any rights to donor information. The only way to potentially obtain information would be (as many do) to breach anonymity by using donor registers and DNA‐tests. This might result in finding genetic relatives which can prove to be a positive experience. However, for some the number of genetic relatives can also be overwhelming. Others fear the risk of consanguinity, or they question what types of relations can be established with a donor, and with his or her other genetic relatives. Central to these considerations is the open question: What is the role of genes in building relations, social networks, and families? Relatedly, what type of relations can parents, donors, and donor‐conceived people expect to establish based on shared DNA?These questions also serve as a call for an independent international and flexible registry that could enable different levels of contact between genetic relatives, including donors who wish to lift their anonymity.Overall, the workshop revealed that gamete donation can and must be organized in a way that supports those involved in donation over time as new moralities and challenges emerge. The need to rethink how this is done has become ever more emergent with the increasing availability of low‐cost DNA tests and online fora. At the same time, the workshop raised the questions of who should be responsible for providing and developing an independent support website; who should provide counseling programs to the parties involved in sperm donation; and who should develop, update, and keep a donor registry for gametes that are marketed and sold on the international market.The increased availability and use of third‐party reproduction and consumer genetic testing speak to transformations that are inherently global in nature. Therefore, solutions to address the accompanying ethical, regulatory, and practical challenges and unexpected consequences must yield cross‐country answers. While particular supranational, state‐level, and public and private actor responsibilities are not easily delegated, it is evident that the provision of lifelong support is necessary to those families involved in gamete donation. This demands a collective effort, including funding for further international research to understand how to develop better support for donor families, as well as setting up a comprehensive support system globally. The aforementioned questions represent a first step toward conducting donor conception more responsibly in the future, arising from collective stakeholder deliberation on experienced challenges. Considering the number of families involved in donor conception, it is about time that we start rethinking what donation means for families and donors and how it shapes families today.


We found no changes in CSF amyloid Aβ1–42 concentrations, regardless of etiology. We showed an increase in tau and phosphorylated tau concentrations in purulent CNS infections of the brain, compared to other etiologies. Moreover, the total concentrations of tau in the CSF correlated with the CSF absolute number of neutrophils. These findings and the Aβ 42/40 concentration quotient discrepancies in CFS between meningitis and encephalitis suggest that infections may affect the metabolism of AD biomarkers.Alzheimer’s disease (AD) has emerged as a growing threat to human health. It is a multifactorial disorder, in which abnormal amyloid beta metabolism and neuroinflammation have been demonstrated to play a key role. Intrathecal inflammation can be triggered by infections and precede brain damage for years. We analyzed the influence of infections of the central nervous system on biomarkers that are crucially involved in AD pathology. Analyses of the cerebrospinal fluid (CSF) levels of Aβ Extracellular Aβ clearance is accomplished by glial phagocytosis, involving microglia, astrocytes, and protease activity from neurons and astrocytes [Alzheimer’s disease (AD) is the most common neurodegenerative disease, and causes memory loss, cognitive decline, and crucially interferes with patients’ everyday lives. The hallmarks of AD include extracellular plaques, due to the deposition of amyloid-beta (Aβ) peptides, and intracellular neurofibrillary tangles, resulting from tau phosphorylation. Understanding Aβ and tau accumulation and clearance is crucial, in order to recognize AD pathways and develop future therapies. Cerebral amyloid angiopathy is another condition linked to Aβ deposition in the brain. Both diseases are driven by impaired Aβ clearance and share pathogenic implications . Aβ is nHerpesviridae family, such as herpes simplex virus-1 and -2 , varicella zoster virus (VZV), cytomegalovirus (CMV), and Epstein–Barr virus (EBV), are capable of reaching the central nervous system (CNS). Herpesviruses were first proposed to contribute to AD pathology [Numerous factors have been designated to promoting cerebral amyloid accumulation. Among them, aging and several genetic risk factors are well recognized . Currentathology . Herpes athology , stayingathology . Controlathology . An incrathology . Human iathology .Chlamydia pneumoniae antigens was observed both intra- and extracellularly in AD brains [Chlamydia pneumoniae [Borrelia spirochetes [The role of bacteria in AD pathways is uncertain. The immunoreactivity of D brains . Experimeumoniae . Morpholrochetes .The pathways leading to Alzheimer’s disease begin many decades before the diagnosis of AD dementia . It was Neisseria meningitidis (n = 2), Streptococcus pneumoniae (n = 2), Streptococcus agalactiae (n = 2), Haemophilus influenzae (n = 1), and Escherichia coli (n = 1). The bacterial aseptic group comprised 11 children diagnosed with Lyme neuroborreliosis caused by Borrelia burgdorferi.A total of 53 children were included in the study. All the patients in the study and in the control group were referred to the hospital with acute febrile illness and did not have any underlying chronic disorders. In twenty-six children, a viral cause of the infection was detected. This included 12 children with an infection caused by enteroviruses, 5 children with an infection caused by the varicella zoster virus (VZV), and 9 children with an infection caused by the tick-borne encephalitis virus (TBEV). The control group comprised 8 febrile children suspected of having meningitis, in whom the diagnosis was ruled out and who were eventually diagnosed with a non-specific viral infection. To broaden the analysis of the impact of CNS infection on biomarkers of AD, we also included 19 children with two types of bacterial infections of the CNS—purulent and aseptic. In the purulent group, there were eight children with infections caused by In the study, meningitis was defined as symptoms consistent with meningitis and a leucocyte count in the cerebrospinal fluid above 5 cells per µL. Encephalitis was defined, according to the International Encephalitis Consortium, as an altered mental status  for over 24 h, with no alternative cause identified and two of the following: seizures, focal neurologic findings, abnormalities in electroencephalography or magnetic resonance imaging suggestive of encephalitis, cerebrospinal fluid (CSF) pleocytosis, and fever. Enteroviruses and VZV were detected in the CSF by polymerase chain reactions. The diagnosis of TBE and LNB was confirmed with serological assays. Purulent infections were diagnosed based on positive CSF culture.1–42, Aβ1–40, tau, and pTau181, in the CSF were performed in the Department of Neurodegeneration Diagnostics, Medical University of Bialystok, Poland. The concentrations of AD biomarkers were assessed with a commercially available quantitative enzyme-linked immunosorbent assay, using IBL kits  for Aβ1–42 and Aβ1–40, and Fujirebio kits  for tau and pTau181 proteins. The standards and samples were run in duplicates, with a coefficient of variance (CV) < 20%. The assays were performed following the manufacturer’s instructions, and CSF samples were diluted 1:20 for Aβ1–42 and Aβ1–40, and were undiluted for tau and ptau.The biochemical measurements of neurochemical dementia diagnostic (NDD) biomarkers, such as Aβp-value was less than 0.05. The statistical analysis was performed with the use of TIBCO Software Inc. (2017) Statistica, version 13 .The summary statistics for continuous variables are presented as a median with interquartile range (IQR); categorical variables are presented as frequencies. The differences between groups were analyzed with the one-way analysis of variance (ANOVA), followed by Newman–Keuls post hoc test. Non-parametric tests were used for other analyses. The results were considered statistically significant when the The characteristics of the study groups are presented in 1–42 to Aβ1–40 in the CSF, regardless of etiology .The comparison of purulent and aseptic  CNS infections yielded similar results . In purufections . The SpeIn the comparison between viral and bacterial (including both purulent infections and Lyme neuroborreliosis) CNS infections, no significant differences were found in the levels of the biomarkers. 42/40 ratio . No significant correlations were found between amyloid isoforms, amyloid ratio, and the CSF parameters. In addition, the Spearman’s R analysis found no significant correlation between the Aβ42/40 ratio, amyloid isoforms, and all of the CSF parameters.In the search for possible causes of the reduced AβIn our study, we analyzed the CSF concentration of Alzheimer’s disease biomarkers in CSF in children with central nervous system infections of different etiologies. The distribution of etiologies in the study group reflects the epidemiology of CNS infections in our region. Northeastern Poland is an endemic area for tick-borne diseases, including tick-borne encephalitis ,17 and L1–42 in CSF is one of the best-established biomarkers of AD [1–42 levels may also be observed in infections of the CNS, where the abundance of plaques is low, suggesting downregulation of amyloid expression and/or processing in these diseases. Studies have found decreased Aβ1–42 levels in bacterial meningitis, multiple sclerosis, and HIV with cerebral engagement [1–42 were associated with worse outcomes of CNS infections [1–42 levels in diseases of the CNS. Presently, the cause of altered levels of amyloid isoforms in the CSF is not clear. In this study, we found no changes in the CSF concentrations of Aβ1–42 and Aβ1–40, regardless of etiology. The lack of statistical importance could be, to some extent, a result of the rather limited number of children included in each of the study subgroups. This is a result of the heterogeneity of infections observed during the study period. There was a trend towards lower concentrations of Aβ1–42 and Aβ1–40 in all the study groups; however, the differences did not reach statistical significance in all, but one, of the comparisons. In a further analysis, we found that the concentrations of Aβ1–40 were significantly lower in children with purulent meningitis, compared to the control group. In the search for a cause of altered amyloid isoform levels, we found no associations between the amyloids and the CSF parameters.A reduced concentration of Aβrs of AD . Key regrs of AD . It has rs of AD . Howevergagement ,23,24. Sgagement ,25,26. Afections . Thus, m1–42 was reported in tuberculous meningitis [Currently, the knowledge of inflammation-induced changes in Aβ peptide metabolism is mainly based on studies of acute infections. In a small case series study, a very low level of Aβningitis . The conningitis . The chaningitis . Thus, t42/40 ratio in CSF was found to be more accurate in the diagnosis of AD than Aβ1–42 alone [1–42 and an increase in Aβ1–40 CSF concentrations [42/40 ratio in children with viral meningitis, compared to children with viral encephalitis. However, the underlying cause was different than in AD, as we did not find an increase in any of the amyloid isoform levels. When compared to the controls, the median Aβ1–40 concentrations were reduced more in encephalitis than in meningitis patients, although the differences did not reach statistical significance. This might explain the observed variability in the relative concentration of amyloid isoforms observed in our study. It is noteworthy that most, if not all, of the research on Alzheimer biomarkers in various CNS infections is conducted in adult patients. The results of these studies are not the best reference for our research in children. Recently, a reduced Aβ42 alone ,31,32. Itrations . We repoTau is an important part in the stability and regulation of the axonal cytoskeleton. Increased tau levels in the CSF reflect neuronal cell death, with the release of tau-related proteins into the CSF. Hyperphosphorylation of tau causes the proteins to detach from the microtubules and destabilizes them . ImportaOne of the limitations of this study is the relatively small sample size in each of the study groups. In addition, the concentration of AD biomarkers displayed high variation between individuals, which might have affected the statistical analysis. Nevertheless, to the best of our knowledge, this is the first study evaluating the biomarkers of AD in children with infections of the CNS, caused by different viral and bacterial pathogens. Our results confirm the notion that infections may, in fact, impact amyloid metabolism, but further investigation is needed, in order to gain a more complete understanding of this finding.


Timely conversion of antimicrobials from intravenous (IV) to oral (PO) route has been shown to decrease costs and length of stay (LOS) without compromising safety and efficacy of therapy. Use of oral antimicrobials may additionally prevent complications related to IV catheters, such as infection, emboli, and patient discomfort. An approved IV to PO policy allowed pharmacists to convert orders for fourteen included antimicrobials and eligible patients at time of order verification.This single-center, retrospective, comparative study was conducted at AdventHealth Orlando, a 1,368-bed community teaching hospital in central Florida. In November 2020, six clinical pharmacist teams began receiving monthly feedback on IV to PO conversion rates in the form of a RePOrt Card, containing IV to PO conversion rates compared to other clinical teams, individual antimicrobial conversion rates, and comparison to prior team progress . RePOrt Cards were provided through October 2021. This study compared pre-intervention days of therapy (DOTs) of antimicrobials from November 2019-October 2020 to post-intervention DOTs from November 2020 to March 2022. The primary objective of this study was to assess the impact of monthly, team-based feedback on percentage of antimicrobials administered orally during a pharmacist-driven IV to PO stewardship initiative.Significantly more DOTs were administered orally in the post intervention group . Positive change in oral conversion rates was observed for all agents except linezolid, minocycline, and voriconazole (Table 1). The largest increase in percentage of DOT administered orally was observed for azithromycin (20%), rifampin (14%), and metronidazole (10%). Estimated monthly and total cost differences are available in Table 2. Minocycline represents the largest opportunity missed; while oral conversion rates remained the same, an increase in overall drug use creates opportunity to continue to prioritize oral conversion due to high cost.Monthly, team-based feedback positively impacted IV to PO conversion rates. Opportunities remain to optimize cost benefits in high-cost agents such as linezolid, minocycline, and voriconazole.Amy L. Carr, PharmD, BCIDP, Shionogi: Advisory Board.


The star copolymer also exhibited good absorption properties in the ultraviolet-visible (UV-Vis) and the near infrared (NIR) spectral regions, which give rise to an optical energy bandgap value as low as 1.43 eV. A HOMO energy level at −5.53 eV, which is below the air-oxidation threshold, was obtained by cyclic voltammetry (CV). Electrochemical impedance spectroscopy (EIS) ascertained the semiconducting properties of the macromolecule, which is characterized by a charge transfer resistance, ctR, value of 3.57 kΩ and a Bode plot-phase angle value of 75°. The combination of the EIS properties of G1PPT-co-P3HT and its highly electron-donating capability in bulk heterojunction (BHJ) active layer containing a perylene derivative, as demonstrated by photoluminescence quenching coupled to the observed Förster Resonance charge transfer, suggests its suitability as an electron-donor material for optoelectronic and photovoltaic devices.In this study, electron-donating semicrystalline generation 1 poly(propylene thiophenoimine)-co-poly(3-hexylthiophene) star copolymer, G1PPT-co-P3HT was chemically prepared for the first time. Copolymerization was achieved with high molecular weight via facile green oxidative reaction.  The past recent years have seen an increasing number of studies centered around the development of new organic materials for optoelectronic and photovoltaic applications ,2,3 resu2 stream for 48 h that produces generation 1 poly(propylene thiophenoimine), G1PPT  in CDCl3. The chemical shifts were compared to those found in the starting materials, generation 1 poly(propyleneimine) tetramine, G1PPI and 2-thiophene carbaldehyde  tetramine dendrimer  (0.3863 ), G1PPT  was confaldehyde . G1PPI waldehyde . On the aldehyde . G1PPT saldehyde .3 were put in a 2-neck round bottom flask containing chloroform (CHCl3). 3-hexylthiophene (81 µL) was added dropwise to the magnetically stirred suspension and the reaction mixture was stirred for 48 h in an N2 atmosphere. 100 mL of methanol was added to quench the copolymerization. The prepared material was successively extracted with methanol (100 mL), acetone (100 mL), and CHCl3 (100 mL) by column chromatography and filtration, and G1PPT-co-P3HT copolymer was collected from CHCl3. Number of average molecular weight, nM, and dispersity, PDI values were estimated from size exclusion chromatography, performed using HPLC grade dimethylformide (DMF) at a flow rate of 0.8 mL/min. Mn was found to be 16,239 g/mol and the average molecular weight, Mw 30,315 g/mol. The dispersity, PDI = Mw/Mn is calculated to be 1.87.Copolymerization of G1PPT to 3-HT followed the procedure reported by Liu et al.  for the Preparation of PDI-co-Carbazole was achieved by copolymerization of 1,7-dibromoperyle-ne-3,4,9,10-tetracarboxylic diimide) (PDI-2Br) to 9-(Heptadecan-9-yl)-2,7-bis-9H-carbazole (Carbazole), according to the synthetic route reported by Zhou et al. . Detaile1H NMR) spectroscopy was performed using Bruker Avance III HD 400 MHz Nanobay NMR spectrometer equipped with a 5 mm BBO using tetramethylsilane as internal standard, and Fourier-Transform Infra-Red (FTIR) spectroscopy was investigated on PerkinElmer model Spectrum 100 series equipment . Both techniques were used for structural characterization.For the effective investigation, characterization, and application of the prepared macromolecule, various analytical techniques were used. Gel permeation chromatography in the form of size exclusion chromatography (GPC/SEC) was performed using an Agilent 1260 Quad Pump, using HPLC grade DMF at a flow rate of 0.8 mL/min and DMAC (0.05% BHT + 0.03% LiCl) as eluent. This technique was used for determination of average molecular weight and number of average molecular weight. Single proton Nuclear Magnetic Resonance  spectroscopy measurements were performed in a quartz cuvette using a Nicolet Evolution 100 UV–Visible spectrometer from ThermoElectron Corporation  and photoluminescence (PL) spectroscopy studies were carried out on Horiba NanoLog™–TRIAX , with double grating excitation and emission monochromators with a slit width of 5 nm, and on Ocean Optics equipment. Microscopic characterization was achieved using high resolution transmission electron microscopy (HRTEM) using Tecnai G2F2O X-Twin MAT 200 kV field emission transmission electron microscope from FEI-ThermoFischer Scientific , scanning electron microscopy (SEM) using ZEISS ULTRA scanning electron microscope equipped with an energy dispersive spectrometer and a Tecnai G2F2O X-Twin MAT 200 kV field emission transmission electron microscope from FEI . Small-angle X-ray scattering (SAXS) analysis for particle size determination was carried out on an Anton Paar SAXSpace system , using copper Kα radiation (0.154 nm) equipped with a 1 D Mythen 2 position sensitive detector and a beam stop alignment. The sample was scattered at a thickness of 315 mm and the sample chamber was evacuated to below 5 mbar to avoid further air scattering. X-ray diffraction patterns for the material were obtained using a D8 advance diffractometer (BRUKER-AXS) employing copper Ka radiation with a wavelength of 0.154 nm, operating at a voltage of 40 kV and current of 40 mA.+ as reference electrode, was used. Additionally, for frontier orbital determination, cyclic voltammetric measurements were carried out with reference to ferrocene.Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) studies were conducted on CH instruments . Electrochemical measurements of G1PPT-co-P3HT drop-coated film on a working electrode were performed at ambient air in a solution of acetonitrile containing 0.1 M tetrabutylammonium hexafluorophosphate (Bu4NPF6). A three-electrode system with a gold (Au) working electrode, a Pt wire as the counter electrode and Ag/AgCl or Ag/Ag2. Prior to fabrication, the patterned ITO-coated glass substrates with resistance 20 Ω/m2 were cleaned. The cleaning procedure was as follows: the substrates were sonicated in Hellmanex solution for 15 min and rinsed twice in boiling water; then, they were sonicated for 15 min in N, N-isopropanol, and rinsed again twice in boiling water. Afterwards, the substrates were dried under an argon (Ar) gas stream. Subsequent to the cleaning process, the hole transporting layer, PEDOT:PSS (Heraeus Clevios PH-1000), was deposited on the ITO surface by spin coating at 5000 rpm for 30 s; then dried on a hot plate at 150 °C for 5 min. PEDOT-PSS was used to reduce the roughness of the ITO layer and acts as electron blocking layer (EBL) between the ITO and the active layer. PEDOT-PSS was gently wiped away from the etched part of the ITO using de-ionized water with a cotton bud to enhance contact with the electrode. The active layer consists of an intermixed donor: acceptor bulk heterojunction blends. The prepared donor:acceptor solutions in chloroform (50 µL) were spin-coated on the PEDOT:PSS at 900 rpm for 105 s, dried on a hot plate at 80 °C for 10 min and the edges were cleaned using chloroform. Finally, the top electrode Aluminum layer (103 nm) deposition was carried out by thermal evaporation on the glass/ITO/PEDOT-PSS/BHJ active layer substrate to complete the device fabrication. A drop of encapsulation epoxy was deposited on a glass substrate and used to seal the device that was then dried under UV lamp to avoid oxidation from air contact. Current-voltage (I–V) curves of the devices were recorded in the dark and under illumination using a solar simulator. All characteristic measurements were carried out at ambient air and recorded under simulated solar illumination of AM 1.5 with an incident power density of 100 mW/cm2 from −0.2 to 1 V using a Keithley output reader. Although the main device of interest is based on G1PPT-co-P3HT/PDI-co-Carbazole (2:1) bulk heterojunction blend, three other devices based on G1PPT-co-P3HT: PC61BM (1:2), P3HT: PDI-co-Carbazole (2:1), and P3HT:PC61BM (2:1) were also fabricated to investigate the photovoltaic characteristics of each prepared copolymer with respect to well-known widely used P3HT and PCBM as donor and acceptor, respectively.Devices were fabricated according to the following procedure, where the device configuration used was glass/indium tin oxide (ITO)/poly:poly(styrene sulphonate), (PEDOT:PSS)/active layer/Al with a pixel area of ~0.0256 cm1H NMR  spectroscopy was used as the key technique to ascertain the structure and purity of star copolymer poly(propylene thiophenoimine)-co-poly(3-hexylthiophene) (G1PPT-co-P3HT). New signals were observed with respect to G1PPT  and G1PPT, is found in −1. Bands in the spectral region 2954–2854 cm−1 are characteristics of aliphatic −C−H stretching signals of the hexyl group attached to the thiophene rings [−1 are the bands resulting from the molecular vibrations of C=N−H and C=C, respectively; the same vibrations were already observed in G1PPT at 1673 and 1632 cm1 with high intensity [−1, respectively, are found at 1473 cm−1 in G1PPT-co-P3HT. At 802 cm−1, the C−S stretching is within the thiophene rings that was also observed in 3-HT and G1PPT [−1 in G1PPT-co-P3HT, already present in G1PPT at 703 cm−1, is as a result of the alkyl groups -C-H bending vibrations of the proton at α-position with respect to the non-alkylated thiophene ring and next to the imine bond [to G1PPT . In geneR CDCl3, 0 MHz speR CDCl3, 0 MHz spene rings . At 1731ntensity . The C−Hnd G1PPT . Finallyine bond ,46. Mole1H NMR spectroscopy, G1PPT-co-P3HT HRTEM images show various regions with well-defined lattice fringes. The crystalline regions were confirmed by the SAED (Selected Area Electron Diffraction) image  image C in whicn) image  and 3-HTn) image , it is tSmall-angle X-ray scattering (SAXS) was used to obtain the particle size distribution of G1PPT-co-P3HT. The free-model pair distance distribution function of the star copolymer depicted in D is the mean crystallite size, K is the shape factor constant (0.89 rad), λ is the wavelength of the X-rays, β is the full width at half maximum (FWHM) of the (220) Bragg’s peak, and ϴ is the angle of reflection of the (220) peak. The calculated grain size was 12.6 nm which agrees with the results obtained from SAXS analysis.X-ray diffractogram of G1PPT-co-P3HT, −1.cm−1. This was characterized by two strong absorbance bands in the ultraviolet (UV) region with absorption maximum at 251 nm; and two new bands in the visible region. The two intense absorption peaks at higher frequency are due to the n-π* electronic transitions in the C=C within the thiophene moiety [Absorption properties of 725.5 µM G1PPT-co-P3HT investigated in solution from chloroform as well as on spin-coated thin films are found in e moiety , wherease moiety . We sugge moiety . At highe moiety ,54. On te moiety . Additioe moiety . It has e moiety . This cae moiety . The pree moiety .The photoluminescence spectrum of G1PPT-co-P3HT when excited at a wavelength of 365 nm is shown in a1 and ipa2 are found at 1.38 and 1.73 V, respectively, while the two reduction peaks, ipc1 and ipc2, are found at 0.97 and 1.21 V, respectively. Anodic peaks of P3HT have been reported to be found at a less positive, below 1 V. This shows that copolymerization to G1PPT has resulted in the oxidation processes to occur at higher positive potentials. Reports also demonstrated that one, two, and sometimes three anodic peaks could be detected in the cyclic voltammograms of polys, PATs depending on the extent of regioregularity of the conjugated chain [G1PPT-co-P3HT electrochemical behavior was studied using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Typical voltammetric behavior of G1PPT-co-P3HT, swept in a potential window between 0 and 2.5 V at a scan rate of 50 mV/s, is depicted in ed chain . While oed chain . While t+ in the presence of ferrocene . The calculations were made using Bredas et al., [ferrE = −0.03 V is the value for ferrocene vs the Ag/Ag+ electrode. Further cyclic voltammetric analyses of G1PPT-co-P3HT on gold-modified electrode versus Ag/Agerrocene B was conergy, EA . HOMO an et al.,  equationEIP and EA were calculated to be 5.53 and 3.6 eV, respectively, giving rise to an electrochemical band gap OCV) value in an organic photovoltaic cell [First oxidation and reduction onsets were found to be −0.63 and −2.2 V, respectively. From which energies . The HOMenergies . In addi4NPF6 (acetonitrile) at a scan rate of 50 mV/s using a three-electrode system with three different applied potentials of −1.162, −1.172, and −1.182 V. The Nyquist plots consist of two distinct parts: a small semi-circle in the high frequency region which describes the electron transfer-limiting processes and an inclined line in the medium to low frequency region which provides information about the diffusion-controlled processes [Z’ axis corresponds to the Ohmic/solution resistance (Rs), while the diameter of the semicircle along the Z’ axis relates to the charge transfer resistance (ctR), which was obtained after fitting the data using the equivalent circuit. The inclined line is attributed to the diffusion of tetrabutylammonium ions in the bulk of the extended conjugation known as the Warburg diffusion (WZ). The resulting parameters obtained from fitting simulation are recorded in cRt decreased with increasing applied potential, i.e., the lowest ctR was exhibited by the highest applied formal potential at −1.182 V. This indicates a faster charge transfer kinetic process comparatively to the other two applied potentials. For a pure capacitor, a vertical line should be exhibited at low frequency, i.e., phase angle at 90°. Deviation from the vertical line is attributed to the inner diffusion resistance for electrolyte ions that reduces phase angle degree, and which is usually strongly dependent on the applied formal potential [CPE) which describes the double-layer capacitance of the electrode-solution interface, a parallel parameter to ctR and ZW, is found not to be a function of the applied potential. Electrochemical impedance spectroscopy (EIS) measurements were used to investigate the electrical resistance and diffusive processes occurring at the electrochemical interface of the electrode. rocesses . The intotential . Microscotential . G1PPT-cmaxλ for the broad band between 470–680 nm corresponding to the intermolecular charge transfer (ICT) [Optical absorbance properties of G1PPT-co-P3HT:PDI-co-Carbazole bulk heterojunction blends were studied in chloroform within the UV-Vis-NIR spectral region as depicted in er (ICT)  shifted er (ICT) , but furer (ICT) .G1PPT-co-P3HT/PDI-co-Carbazole different bulk heterojunction blended photo-physical properties were also investigated using photoluminescence the spectroscopic technique. The nanocomposite fluorescence properties were studied. Fluorescence of pristine G1PPT-co-P3HT and PDI-co-Carbazole are presented in 61BM (used as a reference) that achieved a power conversion efficiency of 1.5% with OCV = 510 mV, short-circuit current density, SCJ = 7.7 mA/cm2, fill factor, FF = 0.38. G1PPT-co-P3HT:PDI-co-Carbazole (2:1) exhibited the lowest characteristics with OCV = 50 mV, SCJ = 2.65 µA/cm2, fill factor, FF = 0.11 which resulted in a power conversion far below 1%. Parameters of the device based on G1PPT-co-P3HT with the fullerene derivative PC61BM in a 1:2 ratio were slightly higher than in the case of the all-polymer device. However, the efficiency is still found to be far below 1%. It is commonly known that an ‘ideal’ exciton-splitting energy, the difference between the LUMO of electron-donating material and LUMO of electron-accepting material, 0.3 eV, is required to ensure proper charge-carrier separation. Although the exciton-splitting energy of G1PPT-co- P3HT:PDI-co-Carbazole (2:1) is calculated to be lower ((−3.6–(−3.87) eV) = 0.27 eV) compared to that of the device fabricated with G1PPT-co-P3HT: PC61BM (1:2) i.e., 0.5 eV where, PC61BM LUMO energy level is −4.1 eV [OCV to 180 mV, the SCJ to 62 µA/cm2 and FF = 0.26. The efficiency was thus increased by one order of magnitude.Preliminary investigation of the application of these BHJ layers into solar cells was conducted. Typical I–V curve characteristics of the BHJ devices are depicted in  −4.1 eV , the obt −4.1 eV . It is t −4.1 eV . AnotherVarious studies have demonstrated that organic photovoltaic device performances are strongly dependent on the BHJ blend morphology ,75. It i3 in chloroform was successfully achieved as confirmed by 1H NMR and FTIR spectroscopic analyses of the prepared dendritic star copolymer, G1PPT- co-P3HT. The analyses demonstrated several characteristic chemical shifts and vibrational bands associated with the features of poly(3-hexylthiophene) and the functionalized dendrimer. Cyclic voltammetric, microscopic, and optical spectral data were used to confirm the degree of regioregularity and type of morphologies, while prepared with high molecular weight, Mw 30,315 g/mol, the star copolymer exhibited an optical band gap energy as low as 1.43 eV, which falls within the optical band gap window of an ideal donating material in OPVs. The two shoulders, at 525 and 567 nm observed in G1PPT-co-P3HT solid-state optical thin film and associated with vibronic side-band and vibronic 0-0 structure, respectively, attested to the existence of different morphologies within the material. This was further confirmed when the G1PPT-co-P3HT-modified Au electrode exhibited two oxidation peaks in the positive potential region during voltammetric studies. These two anodic peaks ascertained the existence of regioregularity between 80–90% caused by amorphous polymer zones of shortly conjugated chains and more ordered regions with longer average conjugation length. The calculated HOMO frontier orbital of −5.53 eV, which confers an air-stability property, also suggested that this dendritic material could exhibit good open-circuit voltage, OCV, a key parameter for optimal OPVs performance. Further electrochemical investigation through EIS analysis, proved that G1PPT-co-P3HT has good semiconducting properties with phase angle above 70°. The semi-crystalline nature of the prepared dendritic star copolymer coupled to the exhibited electronic properties therefore qualified G1PPT-co-P3HT as a good electron-donating candidate for optoelectronic devices. Preliminary studies of I-V characteristics of fabricated devices based on G1PPT-co-P3HT and PDI-co-Carbazole with observable OCV suggest that if the morphologies and ratio are improved, and they could exhibit outstanding PCE; it also shows the applicability of these bulk heterojunction layers for organic light-emitting diodes.For the first time, oxidative copolymerization of functionalized dendrimer, G1PPT to poly(3-hexylthiophene) in the presence of FeCl


Selenoprotein P, a selenium-transporter protein, has been hypothesized to play a role in the etiology of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and Alzheimer’s dementia (AD). However, data in humans are scarce and largely confined to autoptic samples. In this case–control study, we determined selenoprotein P concentrations in both the cerebrospinal fluid (CSF) and the serum of 50 individuals diagnosed with ALS, 30 with AD, 54 with mild cognitive impairment (MCI) and of 30 controls, using sandwich enzyme-linked immunosorbent assay (ELISA) methods. We found a positive and generally linear association between CSF and serum selenoprotein P concentrations in all groups. CSF selenoprotein P and biomarkers of neurodegeneration were positively associated in AD, while for MCI, we found an inverted-U-shaped relation. CSF selenoprotein P concentrations were higher in AD and MCI than in ALS and controls, while in serum, the highest concentrations were found in MCI and ALS. Logistic and cubic spline regression analyses showed an inverse association between CSF selenoprotein P levels and ALS risk, and a positive association for AD risk, while an inverted-U-shaped relation with MCI risk emerged. Conversely, serum selenoprotein P concentrations were positively associated with risk of all conditions but only in their lower range. Overall, these findings indicate some abnormalities of selenoprotein P concentrations in both the central nervous system and blood associated with ALS and neurocognitive disorders, though in different directions. These alterations may reflect either phenomena of etiologic relevance or disease-induced alterations of nutritional and metabolic status. Amyotrophic lateral sclerosis (ALS) is an extremely severe neurodegenerative disease characterized by the progressive degeneration of motor neurons in both the brain and the spinal cord . ALS casAmong the environmental factors hypothesized to be involved in the etiology of neurodegenerative disease, possibly by interacting with genetic factors, are heavy metals and metalloids, pesticides and other chemicals ,10,11,12SELENOP and mainly produced in the liver [In addition to the uncertainties concerning the actual effects of overall selenium exposure on the nervous system, little is known about the single selenium compounds directly involved in the risk of neurological disorders, as well as the selenoproteins whose deficit and excess may be specifically implicated in such a relationship, including selenoprotein deficiencies due to gene mutations ,22,23. Vhe liver . Moreovehe liver . In recehe liver ,38,39,40he liver ,43,44,45he liver .In this study, we aimed at assessing for the first time CSF selenoprotein P content in vivo in patients with newly diagnosed ALS, AD and MCI as well as in controls and at characterizing the possible involvement of CSF and blood selenoprotein P concentrations in the etiology and the progression of these neurodegenerative diseases.Based on CSF sample availability, we eventually recruited 164 subjects (87 men and 77 women). Of these, 30 were diagnosed with AD and 50 with ALS; 54 individuals with MCI and 30 controls were also included. For all ALS patients and for controls, a serum sample was also available, while a serum sample was not available for four AD patients and seven individuals with MCI. In controls, CSF selenoprotein P levels were 22.1 ng/mL (interquartile range (IQR) 14.9–29.9 ng/mL), with higher values in males (28.7 ng/mL) compared to females (19.1 ng/mL) and in participants aged ≥65 years (29.5 vs. 20.1 ng/mL). Selenoprotein P levels in the serum amounted to 5615.9 ng/mL (IQR 4683.6–6235.7 ng/mL), with higher values in females (5694.0 ng/mL) than in males (5310.8 ng/mL). When considering the overall population, similarly, males exhibited higher CSF and serum levels of selenoprotein P compared to females .Selenoprotein P median and IQR concentrations in serum and CSF according to diagnosis are reported in The highest CSF selenoprotein P levels were found in the individuals with MCI and AD and the lowest in patients with ALS, while for serum, the highest levels emerged in the MCI subgroup and the lowest in controls. Individuals with MCI exhibited slightly higher selenoprotein-P-bound selenium levels compared to AD patients. Concerning neurodegeneration and amyloidosis biomarkers, individuals with MCI had higher average Aβ1-42 levels and lower t-tau and p-tau181 concentrations, compared with AD patients.In the spline regression analyses between CSF neurodegeneration and amyloidosis biomarkers and CSF selenoprotein P levels, Aβ1-42 levels slightly and positively correlated with selenoprotein P in the AD group. This association tended to also be positive in MCI individuals until 800 ng/L of selenoprotein P concentrations, before the curve flattened .Patterns for p-tau181 and t-tau levels in relation with selenoprotein P were substantially similar in the AD group, while in the MCI one, at high levels of both neurodegeneration biomarkers, we found an inverted-U-shaped pattern with turning points at around 80 and 500 ng/L of p-tau181 and t-tau, respectively.With reference to the association between selenoprotein-P-bound selenium and selenoprotein P concentrations in CSF , as showIn the logistic regression analysis , we founConversely, the association between CSF selenoprotein P levels and both AD and MCI risk was positive, while the corresponding risk estimates for serum selenoprotein P levels were much attenuated and close to unity. In the spline regression analysis , the relThe spline analyses for serum selenoprotein P concentrations yielded considerably different results, with a positive association in the lowest range and no or an inverse association at higher concentrations for the ALS-MCI and AD groups, respectively.This study about selenoprotein P concentrations in individuals affected by neurodegenerative disease is apparently the first ever to have been carried out in vivo and using a biomarker within the central nervous system, in addition to the serum levels of this protein. We found that relatively high CSF selenoprotein P concentrations characterized both prevalent AD and the milder degree of cognitive impairment represented by MCI, while in spline and logistic regression analysis, there was a stronger and monotonic association with AD risk compared with the inconsistent relation with MCI risk. This suggests that the occurrence of cognitive impairment and particularly of dementia is associated with alterations of selenoprotein P concentrations, which could be either a consequence of or a risk factor for these conditions. The considerably more inconsistent and variable pattern for MCI could be due to the fact that this condition is extremely heterogeneous, since it includes individuals not only with Alzheimer’s pathology who will likely progress to AD but also with other diseases causing cognitive impairment, and even subjects who may subsequently revert to “normal” cognitive status .Conversely, ALS was associated with considerably lower CSF selenoprotein P levels compared to controls and clearly even more to AD and MCI. This suggests either an effect of prevalent disease in selenoprotein P expression or a phenomenon of etiologic interest, possibly associated with an inadequate supply of selenium to the brain for selenoprotein synthesis, and therefore, a decreased capacity to counteract the oxidative stress anticipating and characterizing this disease ,48.However, interpreting associations between selenoprotein P concentrations in patients and disease risk as causal could be misleading. This is so both in the central nervous system (with reference to CSF concentrations) and in blood (as indicated by serum levels) for patients affected by a major neurodegenerative disease such as AD and ALS, due to an issue of reverse causation linked to altered nutritional status and metabolism. Higher CSF and serum selenoprotein P concentrations in patients could be due, for instance, to a higher degree of oxidative stress compared with controls, with a higher need for selenium availability for antioxidant selenoprotein synthesis and, therefore, of selenoprotein P as the main selenium transporter. However, since in ALS patients, such an increase was found for blood selenoprotein P concentrations but not for its CSF concentrations, mechanisms influencing selenoprotein P status even within the same patients could considerably differ, as well as the relation of this protein with disease risk.Contrary to the aforementioned findings for CSF selenoprotein P content, the relation between selenoprotein P concentrations in the serum and the risk of neurodegenerative disease systematically suggested a positive association at the lower range of protein levels, and no association (or for AD risk even an inverse relation) above an approximate threshold of 6000 ng/mL. These findings are difficult to interpret with reference to the etiology of ALS, AD and MCI, limiting the utility of determining circulating selenoprotein P levels for the diagnosis and follow up of neurodegenerative disorders. They also suggest an uncoupling of the blood and CSF selenoprotein P concentrations, possibly due to altered selenium availability and metabolism and disturbances of the brain–blood barrier. Therefore, despite some partial correlation between blood and CSF selenoprotein P content ,50, onlyThe positive association between selenoprotein P levels and selenoprotein-P-bound selenium in the CSF was expected, comparable in AD and MCI participants and substantially linear. However, it also showed some statistical imprecision and variability around its basic, monotonic trend. This could reflect an uneven distribution of selenoprotein P selenium contents across participants, due to a degree of variability in the number of selenium atoms incorporated in the protein ,54,55,56We found different patterns of association between CSF selenoprotein P levels and biomarkers of neurodegeneration, which defy simplistic interpretation. The positive associations detected in AD patients could reflect a reactive phenomenon linked to an increased need for selenium and synthesis of this key transporter of the trace element, to generate a compensatory response to counteract oxidative stress ,57. AlteThe paired analysis of selenoprotein P concentrations in both CSF and serum showed some positive association in controls and in subjects with MCI and ALS but not in patients with AD, possibly due to metabolic reasons or alterations of the blood–brain barrier. However, even in the former groups, the association between CSF and serum selenoprotein P concentration was neither strong nor statistically precise, as largely expected on the basis of the little evidence available ,50. OverIn conclusion, results of our case–control study indicate that concentrations of selenoprotein P in the CSF are decreased in ALS and increased in AD and MCI, thus suggesting alterations of the overall selenium availability in the brain and/or deleterious effects of abnormal selenoprotein P concentrations. Our findings also show that CSF selenoprotein P concentrations tend to linearly correlate with biomarkers of neurodegeneration in AD but not in MCI and have a considerably more linear, monotonic pattern of association with both ALS and AD risk as compared with the blood concentrations of this protein. To reliably assess the causes and the effects of abnormalities in selenoprotein P concentrations in patients with neurodegenerative diseases, studies with a prospective design are clearly needed.For this study, we selected all subjects diagnosed with AD and ALS as well as individuals with MCI at the Neurology Clinic of Policlinico University Hospital in Modena, Northern Italy. The study period spanned 2007–2020. Diagnosis of the aforementioned conditions was performed used clinical criteria reported in detail elsewhere ,58. Brieg for 10 min at controlled room temperature. After centrifugation, samples were aliquoted into polypropylene sterile tubes and stored at −80 °C awaiting testing. CSF amyloid beta 1-42 (Aβ1-42), phosphorylated (p-tau181) and total tau protein (t-tau) were determined with the enzyme-linked immunosorbent assay (ELISA) method (INNOTEST® β-AMYLOID(1-42), INNOTEST® hTAU-Ag and INNOTEST® PHOSPHO-TAU(181P), Innogenetics, Ghent, Belgium) following the manufacturers’ instructions. For the purpose of selenoprotein P determination, the remaining aliquots of each sample were then transported deep frozen in dry ice by air courier to the Molecular Biology and Metabolism Laboratory of Tohoku University .Each sample, CSF or serum, was collected in the morning in fasting subjects who underwent lumbar puncture and venipuncture according to standard clinical and operating procedures. Each sample was received by the adjacent Neurology Laboratory within 30 min from collection and was centrifuged at 2500× For the measurement of full-length selenoprotein P concentrations in CSF and serum, we sandwiched the ELISA system with an AA3 method. Clone AA3 antibody recognizing the selenoprotein P C-terminal region was, therefore, used as a capture antibody. In addition, clone AH5 recognizing the N-terminal region of selenoprotein P was labeled with horseradish peroxidase (HRP) and used as a detection antibody. These antibodies were prepared as previously described , and humNinety-six-well microtiter plates were coated for 18 h at 4 °C with rat anti-human selenoprotein P monoclonal antibody AA3 (5 μg/mL) in 0.05 M sodium bicarbonate buffer, pH 9.6, filtered before use. The wells were washed four times with PBS containing 0.05% Tween 20 (200 µL) and incubated at 37 °C with PBS containing Block Ace  for 1 h after washing the wells four times, selenoprotein P standard or CSF or serum sample  was added to each well and incubated at 37 °C for 1 h after washing the wells four times, HRP-conjugated rat anti-human selenoprotein P monoclonal antibody AH5 (20 μg/mL) was added and incubated at 37 °C for 1 h. Finally, TMB Peroxidase Substrate  was added to each well, and the protein–substrate reaction was allowed to proceed for 10 min. The reactions were stopped with the addition of 1 M sulfuric acid, and the absorbances were read at 450 nm in a SpectraMax iD5 . We determined at least four points in each CSF or serum sample, and the average value was used for analysis. This assay was shown to be highly reproducible when identical samples were compared intra-assay or inter-assay, with the relative standard deviations within determinations under 5.0% and 10.0%, respectively. Overall, the laboratory personnel performed selenoprotein P determinations while blinded to patient diagnosis.For all individuals diagnosed with AD and MCI subjects, we could also rely on CSF selenoprotein-P-bound selenium levels from a recent cohort study assessing whether the central nervous system selenium status could influence MCI progression to dementia . In thatWe used spline regression analysis to assess the correlation of full-length selenoprotein P concentrations in CSF with its serum levels and with biomarkers of neurodegeneration  and amyloidosis (Aβ1-42) in CSF, as well as selenoprotein-P-bound selenium. In more detail, we used a restricted cubic spline model with three knots at fixed percentiles , adjusting for age at sample collection, sex and education. We also used linear regression analysis to assess the linear trend of some associations, by computing its β coefficient and the related 95% confidence interval (CI). We computed the crude and adjusted odds ratio (OR) for AD, MCI and ALS, by increasing 10-unit selenoprotein P concentrations through crude and multivariable unconditional logistic regression adjusting for sex and age as continuous variables. We also modeled the relation between selenoprotein P and disease risk using the restricted cubic spline model with three knots at fixed percentiles , adding sex and age in the multivariable model. We used Stata 17.0 SE statistical package  for all statistical analyses.


SWEET) gene family, LoSWEET14, from Oriental hybrid lily ‘Sorbonne’ was identified. LoSWEET14 encodes a protein of 278 amino acids and is capable of transporting sucrose and some types of hexoses. The transcript level of the LoSWEET14 gene was significantly increased under various stress conditions including drought, cold, salt stresses, and abscisic acid (ABA) treatment. Overexpression of LoSWEET14 in tobacco (Nicotiana tabacum) showed that the transgenic lines had larger leaves, accumulated more soluble sugars, and were more resistant to drought, cold, and salt stresses, while becoming more sensitive to ABA compared with wild-type lines. Promoter analysis revealed that multiple stress-related cis-acting elements were found in the promoter of LoSWEET14. According to the distribution of cis-acting elements, different lengths of 5′-deletion fragments were constructed and the LoSWEET14-pro3(-540 bp) was found to be able to drive GUS gene expression in response to abiotic stresses and ABA treatment. Furthermore, a yeast one hybrid (Y1H) assay proved that the AREB/ABF (ABRE-binding protein/ABRE-binding factor) from lilies (LoABF2) could bind to the promoter of LoSWEET14. These findings indicated that LoSWEET14 is induced by LoABF2 to participate in the ABA signaling pathway to promote soluble sugar accumulation in response to multiple abiotic stresses.Sugar transport and distribution plays an important role in lily bulb development and resistance to abiotic stresses. In this study, a member of the Sugar Will Eventually be Exported Transporters ( Plants are exposed to multiple abiotic stresses during growth and development, including drought, extreme temperatures, and high salinity . In respIn higher plants, sugars, especially sucrose, act as the final product of photosynthesis, transported from mature leaves (source) to non-photosynthetic cells (sink) through phloem sieve elements (SE). There are two pathways for loading and unloading sucrose in the phloem, one is the symplastic pathway, mediated by plasmodesmata, and the other one is the apoplastic pathway, which is linked by specific sugar transporters (STs) ,8. After+-dependent sugar transporter, which needs to be coupled with protons to generate dynamical potential and thus drive the transport of sucrose, whereas SWEET does not require energy to complete the bidirectional transport of sucrose [Sucrose transporters (SUTs) and SWEETs are major transporters for sucrose transported through the apoplastic pathway ,8,11. SU sucrose ,13. SWEE sucrose ,15. Base sucrose ,17.VvSWEET10 in grape (Vitis vinifera) callus and tomato (Solanum lycopersium) significantly increased the content of glucose, fructose, and total sugar [SlSWEET15 function via (CRISPRs)/CRISPR editing significantly reduced fruit size and weight in tomato [SlSWEET7a or SlSWEET14 instead led to an increase in sugar content in ripe fruits [Zea mays L.) ZmSWEET4c and its rice (Oryza sativa) homolog, OsSWEET4, were defective in seed filling [SlSWEET15 resulted in severe defects in seed filling and embryo development [Glycine max L.), GmSWEET15 could mediate the export of sucrose from the endosperm to the embryo in early seed development [SWEETs have been shown to play roles in improving fruit quality by altering sugar accumulation in sink organs. Overexpression of al sugar . Eliminan tomato , while se fruits . SWEETs  filling , while eelopment . In soybelopment . SWEET9 elopment .Arabidopsis thaliana, disruption of AtSWEET11 and AtSWEET12 affected plant resistance to cold and frost [AtSWEET16 was downregulated and the AtSWEET16 overexpression lines could not accumulate fructose normally [AtSWEET15/SAG29 was also induced by low temperature, high salt, and drought stress [MdSWEET17 transgenic lines had higher tolerance to drought and accumulated more soluble sugars, especially fructose [DsSWEET17 and DsSWEET12 from Dianthus spiculifolius in Arabidopsis showed higher tolerance to osmotic and oxidative stresses [SWEET genes may be able to receive abiotic stress signals to improve tolerance to adversity by regulating sugar transport and distribution, but the exact signals that induce SWEET have not been fully studied.SWEET genes have also been identified to participate in resistance to abiotic stresses. In nd frost . At low normally . AtSWEETt stress . Under dfructose . Overexpstresses ,28. ThesABF, CBL9, and CIPK3, have been identified in enhancing plant tolerance to abiotic stress [Arabidopsis ABI3 (ABA Insensitive3) mutant was not sensitive to sugar during germination and showed sugar insensitivity (sis) and sucrose uncoupling (sun) phenotypes [Malus domestica), promoted soluble sugar accumulation by activating the expression of MdSUT2, and overexpression lines of MdAREB2 also accumulated more soluble sugar [The phytohormone abscisic acid (ABA) is known as a key signal for plants to respond to abiotic stress ,30. Abioc stress . The basc stress . On the c stress . The Araenotypes . In riceenotypes . Consideenotypes . MdAREB2le sugar . These rLoSWEET14, was cloned from oriental Lilium ‘Sorbonne’, and was found to be induced by drought, cold, salt treatments, and exogenous ABA. In LoSWEET14 transgenic tobacco (Nicotiana tabacum), more starch, sucrose, and soluble sugar were accumulated, and the transgenic plants showed higher tolerance to low temperature, drought, and high salt stresses and were more sensitive to ABA. In addition, the ABA-binding factor of lilies (LoABF2) could bind directly to ABREs on the LoSWEET14 promoter. These findings suggest that LoSWEET14 may be involved in the ABA signaling pathway to regulate sugar accumulation under abiotic stresses, which provides a new idea for improving the quality of lily bulbs to respond to abiotic stresses including cold vernalization.Lily (Lilium spp.) is one of the most important bulbous flowers worldwide. Bulb vernalization and development plays a crucial role in the development of the lily industry and has been proved to be closely related to carbohydrate metabolism ,39. In oLoSWEET14 was found based on the previous transcriptome data of our laboratory [LoSWEET14 was 837 bp (the full-length is shown in pI) of 8.49. The conserved domain sequence (CDS) analysis of LoSWEET14 on NCBI revealed that it contained two MtN3 domains , and was specifically induced again after 12 h. After 24 h, the expression level under cold and drought treatment began to return to a normal level  were significantly decreased, on the other hand, when the light switched on, expression levels of LoSWEET14 were relatively higher, indicating that LoSWEET14 might be regulated by photoperiod. Despite the influence of light/dark transition, the expression levels of LoSWEET14 were also induced by cold, mannitol, NaCl, and ABA apparently. These findings indicate that LoSWEET14 plays a role in abiotic stresses and ABA signaling. In addition, tissue-specific expression analysis revealed that LoSWEET14 was significantly higher expressed in the stem, followed by the flower, while being relatively lower in the bulb and leaf  with relatively higher expression were selected for subsequent study , three 5′-truncated fragments, pro2 (-807 bp), pro3 (−540 bp), and pro4 (−357 bp), were constructed, based on the position of the ABRE elements and distribution of other stress-related cis-acting elements , so the LoSWEET14 promoter fragments pro-LoSWEET14-ABRE1 (−1103 bp to −1000 bp) containing one ABRE element and pro-LoSWEET14-ABRE2 (−499 bp to −334 bp) containing two ABRE elements (LoABF2 gene was fused to the activation domain (AD) of pGADT7 vector. Yeast strains co-transformed pGADT7-LoABF2 with pAbAi-ABRE, pAbAi-LoSWEET14-ABRE1, or pAbAi-LoSWEET14-ABRE2 could grow well on SD/−Leu plates containing 50 ng/mL, 200 ng/mL, and 100 ng/mL AbA, respectively, while no growth was observed in strains co-transformed pGADT7-LoABF2 with the negative control bait pAbAi-ABRE (M) (LoABF2 transcription factor is involved in the regulation of LoSWEET14.When the 1106 bp fragment of the elements  were isoelements . The LoAABRE (M) . These rLoSWEET14 in our research , which also belonged to clade III and is closely related to AtSWEET10, could only transport sucrose [In higher plants, assimilates produced by photosynthesis in mature leaves are transported to sink organs to maintain plant growth and development, mainly in the form of sucrose, which requires the participation of sugar transporters ,9. In thresearch . In Arab sucrose . However sucrose , while t sucrose . Such diLoSWEET14 was significantly correlated with starch content in bulbs, and with soluble sugar content in leaves, indicating that LoSWEET14 might play a key role in sink–source relationship change in bulbs [LoSWEET14 transgenic tobacco accumulated more starch, sucrose, and soluble sugars compared with WT plants in leaves  [SWEET genes can change the external morphological characteristics of plants and affect fruit development by changing the accumulation and distribution process of sugar, which provides a new perspective to improve the quality of bulbs in lilies and fruit quality in production.Sugar not only provides essential energy for plant growth, but also has a profound effect on cell division and expansion . Solublein bulbs , but then leaves a, and thn leaves . SimilarWT lines . AtSWEETant size . Howeverstrains) . OverallLoSWEET14 were found to be significantly upregulated under drought, cold, and salt treatments  seedlings, it has been reported that low concentrations of glucose could enhance the activity of antioxidant enzymes, such as peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) [LoSWEET14 transgenic plants not only showed a significant increase in soluble sugar and sucrose content compared with WT lines, but also displayed an obvious upregulation in the transcript levels of antioxidant-related genes such as NtAPX1, NtCAT, and NtCAX3 by qRT-PCR analysis  . In our analysis . In addianalysis . These fOsSWEET13 and OsSWEET15 [AtSUC2, AtSUC4, and AtSUC9 of Arabidopsis were also upregulated by ABA treatment [LoSWEET14 was also found to be significantly increased under ABA treatment  was also found to be closely related to LoSWEET14 under ABA treatment , 2 h, 4 h, 6 h, 12 h, and 24 h immediately and all samples were frozen with liquid nitrogen quickly and stored at −80 °C.The oriental lily ‘Sorbonne’ was used as the experimental plant material in the present study and it was prepared as described in our previous study . Box plaNicotiana tabacum) ‘NC89′ was used for transgenic research of LoSWEET14. The tobacco plants were grown under conditions of 24 °C/18 °C, 16 h light/8 h dark with 1000lx light intensity, and 65% relative humidity.The wild type tobacco . The ProtParam tools  were used to predict molecular weight (MW) and theoretical isoelectric point (pI). The tertiary structure of the LoSWEET14 protein was predicted on SWISS-MODEL .The complete cDNA sequence  of LoSWE® Premix Ex TaqTM II  according to the protocol. Primers used in qRT-PCR were designed by Primer Premier 5.0, and primer sequences used in this study are shown in −ΔΔCt method with TIP1 as the reference gene in lily and Ntactin as the reference gene in tobacco.The total RNAs were extracted using an EASYspin Plus plant RNA rapid extraction kit . The degradation and contamination concentration of RNA were determined by 1% agarose gel. The obtained RNA was reversely transcribed into cDNA by the PC54-TRUEscript RT kit (+gDNA Eraser) . A Bio-Rad/CFX Connect™ Real-Time PCR Detection System  was used to perform qRT-PCR with TB GreenLoSWEET14 without a stop codon was inserted between the EcoRI and SpeI restriction sites of the pDR196 vector. The pDR196-LoSWEET14 fusion plasmid and the empty vector were transferred into the EBY.VW4000 yeast strain referring to the Quick Easy Yeast Transformation Mix Kit , respectively. Transformants were screened in an SD/-Ura plate added with 2% maltose and the transformed cells were cultured in an SD/-Ura liquid medium to OD = 0.5, then it was diluted and spot coated in SD/-Ura plates containing 2% maltose, 2% sucrose, 2% glucose, and 2% fructose, respectively. The growth of yeast cells was observed after three days incubation at 30 °C.The ORF of LoSWEET14 was cloned by referring to the instructions of the TAKARA Genome Walking Kit , and the cis-acting elements on the promoter were predicted on the web of PLACE . Details of the detected cis-acting elements are listed in pro1 (−1107 bp), pro2 (−807 bp), pro3 (−540 bp), pro4 (−357 bp), respectively , 2 µM ABA, 4 µM ABA, and 150 mM NaCl, respectively. The standard sucrose concentration of the MS medium was 3%. Phenotypic changes were observed, and germination rates were measured after 2 weeks. For drought stress, 5-week-old seedings were watered thoroughly, then withheld for 30 days followed by re-watering for 7 days. For low temperature stress, 3-week-old seedings were pretreated at 4 °C for 24 h and then treated at −4 °C for 6 h and −8 °C for 6 h to observe phenotypic changes.For salt and ABA stress, seeds of WT and Contents of soluble sugar, starch, and sucrose were determined according to the method described in a previous study . ContentLoSWEET14 promoter transgenic tobacco were planted on MS medium for 3 weeks and then were treated with 4 °C for cold treatment, and MS medium containing 50 µM ABA, 200 mM NaCl, and 200 mM mannitol, respectively, for 24 h. The treated leaves were sampled immediately and incubated in GUS reaction buffer  at 37 °C for 24 h in the dark. Then, chlorophyll of the samples was removed with 70% alcohol. The staining of samples was observed under white light of the Leica, Wetzlar, Germany, TL3000 Ergo microscope and photographed [GUS gene in LoSWEET14 promoter transgenic tobacco under different stresses were analyzed as described in T3 generation seeds of ographed . The expSacl and Xhol restriction sites (upstream of the AbA resistance gene) of the pAbAi vector (bait), respectively. The LoSWEET14 promoter fragments, pro-LoSWEET14-ABRE1 (−1103 bp to −1000 bp) containing one ABRE element and pro-LoSWEET14-ABRE2 (−499 bp to −334 bp) containing two ABRE elements (LoABF2 gene was inserted into the EcoRl and Xhol restriction sites of the pGADT7 vector (prey) and co-transformed into positive yeast strains transformed with the recombinant pAbAi vector. The obtained co-transformed yeast strain could grow in SD/-Leu plates containing the corresponding concentration of AbA indicating that LoABF2 is able to bind to this DNA sequence.The Y1H assay was performed referring to the Matchmaker Gold Yeast One-Hybrid System Kit . The ABRE (CACGT) and its mutation ABRE(M) (TGGAT) were inserted between the elements , were alp < 0.01 and * p < 0.05 compared with the corresponding controls using Student’s t test.In this study, each experiment contained a minimum of three plants per treatment and each value indicated mean ± SE (standard error) of three independent biological replicates. Asterisks indicate a significant difference ** LoSWEET14 (OP298002) and LoABF2 (OP298003) have been submitted to NCBI.The coding sequences for