Aspergillus is a filamentous fungus renowned for its extraordinary ability to produce active natural products of high therapeutic value and economic importance. This review is the first to focus on Aspergillus-derived alkaloids. Through an extensive literature review and data analysis, 263 alkaloids are categorized according to their structural features into those containing cytochalasans, diketopiperazine alkaloids, quinazoline alkaloids, quinoline alkaloids, indole alkaloids, pyrrolidine alkaloids, and others. These metabolites exhibited diverse biological activities, such as antibacterial activity, cytotoxicity, anti-inflammatory activity, and α-glucosidase, ACE, and DPPH inhibitory activities. The bioactivity, structural diversity, and occurrence of these alkaloids are reviewed in detail.Alkaloids represent a large family of natural products with diverse structures and bioactivities. These compounds and their derivatives have been widely used in clinics to treat various diseases. The endophytic  Aspergillus is one of the most widely studied filamentous fungi and renowned for its extraordinary productivity when it comes to active natural products with therapeutic values, making it of economic importance quinazoline-3,6-dione (163) and protuboxepin K (164). Compounds 153, 160–164 exhibited mosaic virus TMV inhibitory activity, with IC50 values of 34.8, 37.9, 32.2, 42.4, 39.5, and 35.2 μM, respectively -L-tryptophan methyl ester (236), N-[(2S)-2hydroxy-1-oxo-3-phenylpropyl]-L-tryptophan (237), acudioxomorpholine (238), and emindole SB (239) benzodiazepine-5,11-dione (241)  [The fungus ne 241) , Table 6 , Table de (242) .A. fumigatus was studied and yielded the known metabolites 14-norpseurotin (243) and pseurotin A (244). Compound 243 promoted the neurite outgrowth of PC12 cells at 10.0 µM [244 was also produced from Aspergillus sp. EJC08 associated with Bauhinia guianensis [A. fumigatus associated with Erythrophloeum fordii Oliv [A. fumigatus associated with Heteroscyphus tener (Steph.) Schiffn [A. fumigatus D associated with Edgeworthia chrysantha Lindl [Aspergillus sp. 87 [244 exhibited inhibition of S. aureus, B. subtilis, Pseudomonas aeruginosa, and E. coli, with MICs of 15.62, 31.25, 31.25, and 15.62 μg/mL, respectively [50 value of 5.20 µM [The endophyte  10.0 µM . Compounianensis , A. fumidii Oliv , A. fumi Schiffn , A. fumiha Lindl , and Asps sp. 87 . Compounectively , and inh 5.20 µM .A. fumigatus LN-4 obtained from Melia azedarach led to the discovery of 244 and pseurotin A1 (245), which demonstrated nontoxicity toward brine shrimps [A chemical study of  shrimps .246), and the known compound 11-O-methylpseurotin A (247) were collected from A. fumigatus Y0107 of Crocus sativus Linn (saffron). These compounds were inactive against Pantoea agglomerans, Agrobacterium tumefaciens, Erwinia sp, and Ralstonia solanacearum [A new alkaloid, 11-acetyl-pseurotin A2  and adenine (250). Compound 248 was also isolated from A. fumigatus LN-4 obtained from Melia azedarach. Compounds 248 and 250 suppressed melanin production in B16 melanoma cells with IC50 values of 144.7 ± 2.35 and 100.4 ± 3.05 µM, respectively [A chemical study on unds 248 , Table 7ectively ,38.251), obtained from A. fumigatus collected from Erythrophloeum fordii Oliv. (Leguminosae), did not inhibit NO production [The known alkaloid lumichrome (oduction .Aspergillus sp. TJ23 collected from leaves of Hypericum perforatum (St John’ Wort) yielded a new pyridone alkaloid, asperpyridone A (252), which improved glucose uptake in HepG2 cells at 50 μM [The endophyte at 50 μM .253) and 4-amino-2-hydroxymethylpyridin-5-ol (254) and the known compound 5-hydroxy-2-hydroxymethylpyridine-4(1H)-one (255) were obtained from A. flavus GZWMJZ-288 on Garcinia multiflora. None of them demonstrated any inhibitory activity against gram-positive S. aureus ATCC6538, S. aureus ATCC25923, MRSA, gram-negative Pseudomonas aeruginosa ATCC10145, or E. coli ATCC11775, nor against the pathogenic fungi C. albicans ATCC10231 or C. glabrata ATCC2001 at 100 μg/mL [New alkaloids 2-Hydroxymethyl-5-(3-oxobutan-2-yl)aminopyran-4(4H)-one , cyclopeptine (257), and trans-3-(3′-hydroxybenzylidene)-3,4-dihydro-4-methyl-1H-1,4-benzodiazepin2,5-dione (258). None of them showed any AChE inhibitory activity [A study on activity .259), together with known compounds penipanoid A (260), and fuscoatramide (261) were obtained from A. flavipes DZ-3 of Eucommia ulmoides Oliver. These compounds were inactive against antioxidant and α-glucosidase capacities [A new alkaloid, asperflaloid B (pacities .262), obtained from the fungus Aspergillus sp. 87, was devoid of antibacterial activity [A new alkaloid, aspergilamide A -1-(4-hydroxy-phenyl)-4-(4-methoxyphenyl)buta-1,3-diene-2,3-diyl)diformamide (263) was isolated from the A. fumigatus HQD24, but it did not display inhibition of splenic lymphocyte growth [N,N’-, followed by A. versicolor , A. flavipes , A. terreus , A. nidulans , other species  including A. aculeatus (1), A. amstelodami (1), A. cristatus (1), A. creber (1), A. micronesiensis (1), and A. ochraceus (1), and Aspergillus unknown species . Detailed analysis revealed that the discovery probability of known alkaloids is high (61.98%)  . The micrsicolor , 10.87%,82) displayed potent inhibitory activity against the human FLT3-ITD mutant AML cell line MV4-11, with an IC50 value of 0.22 μM [252) improved glucose uptake and is a potential hypoglycemic agent [The alkaloids summarized in this literature exhibited antibacterial activity; cytotoxicity; anti-inflammatory activity; and α-glucosidase, ACE, and DPPH inhibitory activities. Many of these metabolites demonstrated potent biological activity. For example, gartryprostatin C ( 0.22 μM . Asperpyic agent . HoweverAspergillus is a dominant community in natural products exploration. In this literature, the bioactivity, structural diversity, and biosources of alkaloids derived from plant endophytic Aspergillus species during January 2004 to May 2023 were described. Approximately 263 alkaloids isolated from 46 strains of Aspergillus species were reviewed according to their structural features, including cytochalasans, diketopiperazine alkaloids, quinazoline alkaloids, quinoline alkaloids, indole alkaloids, pyrrolidine alkaloids, and others. Among them, 149 alkaloids have significant physiological activities, such as antibacterial activity, cytotoxicity, anti-inflammatory activity, and α-glucosidase, ACE, and DPPH inhibitory activities. Therefore, these active alkaloids have tremendous potential as lead compounds for the exploitation of new drugs. The interdisciplinary research of chemistry, biology, and pharmacology for alkaloids derived from plant endophytic Aspergillus sp. has attributed to driving the application of alkaloids in the drug discovery and development.Plant endophytic fungi have provided abundant resources of natural products with unique structural features and diverse biological activities, which play a critical role for drug development. The plant endophytic


Limbic-predominant age-related TDP-43 encephalopathy neuropathologic change (LATE-NC)  frequentThe amygdala is also preferentially affected by argyrophilic grains (AGs) and age-related tau astrogliopathy (ARTAG). AGs are age-related lesions in which four-repeat tau is selectively accumulated . AGs areAt present, the pathogenic relationship between LATE-NC, AGs, and GFAs in the amygdala remains to be elucidated. For example, although a few previous studies supported the potential relationship between LATE-NC and AGs , 4, a reTo address these issues, first, we examined whether amygdala GFAs and AGs are independent risk factors of LATE-NC in a Japanese series with a low to moderate Braak neurofibrillary tangle (NFT) stage. Then, independent pathological risk factors of the formation of AGs in this series were also explored. Finally, whether LATE-NC, AGs, and amygdala GFAs have independent effects on severe loss of neurons in the amygdala was examined.From 1,180 autopsy cases who were registered in the database at the Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences as of the end of June 2023, we first selected 501 cases in which all of the following pathological data were available: Braak NFT stage, Thal phase, CERAD neuritic plaque score, LBD subtypes, Braak Parkinson’s disease stage, Saito AG stage , LATE-NCFrom 501 cases, we excluded any cases having following pathologies: NFTs with Braak stages V-VI, Lewy bodies in any regions including the olfactory bulb, NFTs which quantity fit the NINDS-PSP criteria, tufted astrocytes and astrocytic plaques in any region, amyotrophic lateral sclerosis or frontotemporal lobar degeneration with TDP-43-positive or FUS-positive inclusions, Pick’s disease, globular glial tauopathy, multiple system atrophy, trinucleotide repeat diseases, inflammatory diseases, leukodystrophies, and lysosomal storage diseases. Finally, 72 cases that were Braak NFT stages I-IV and Thal phases 0–4 but lacked any other neurodegenerative changes except for AGs, GFAs, LATE-NC, or HS which is closely associated with LATE-NC, were extracted .The mean age at death in 72 cases was 71.6 ± 11.6 years. Of 72 cases, 52 cases had GFAs in the amygdala (72.2%), and 26 had AGs (36.1%) Table : Fig. S2Four of 72 cases (5.6%) had HS Figs. E. All HSp < 0.001, Fig. Among 72 cases, five (6.9%) had severe neuronal loss in the amygdala (Table p = 0.0264) and amygdala GFA stage 4  were significant predictors of LATE-NC  demonstrated that the age at death  and amygdala GFA stage 4  were independent risk factors of AGs  revealed that the age at death  was an independent risk factor of severe neuronal loss with stage 3.In the multivariate binomial logistic regression regarding tissue degeneration in the amygdala , only Saito AG stages II-III  independently contribute to amygdala degeneration. Although LATE-NC was not a statistically significant factor in the present study, it might be explained by the small number of LATE-NC-positive cases in our series. Indeed, it was reported that LATE-NC was associated with the amygdala volume assessed using postmortem MRI . RegardiAdditional file 1: Fig. S1. Representative figures of neuronal loss stage in the amygdala. A–C Stage 0. Neuronal loss and glial proliferation is absent. D–F Stage 1. Mild neuronal loss with minimal gliosis is noted. G–I Stage 2. Moderate neuronal loss with gliosis is present, but tissue rarefaction is not evident. J–L Stage 3. Severe neuronal loss with remarkable glial proliferation is noted. Tissue rarefaction is also seen. Scale bars: A, D, G, J 200 μm, B, E, H, K 100 μm, C, F, I, L 50 μm.Additional file 2: Table S1. Demographic data in cases with Braak stages I-IV by LATE-NC status.Additional file 3: File S1. Supplementary file 1: Supplementary materials and methodsAdditional file 4: Fig. S2. TDP-43 pathology, granular fuzzy astrocytes (GFAs), argyrophilic grains, hippocampal sclerosis, and tissue degeneration in the amygdala in representative cases. A–F Pathological findings in a case with Braak NFT stage II, Thal phase 0, amygdala GFA stage 2, Saito AG stage III, and LATE-NC stage 2. A, B Phosphorylated TDP-43 accumulation in the amygdala A and dentate gyrus in the hippocampus B. pS409/410 immunohistochemistry. Scale bar: 25 μm. C A GFA in the amygdala. AT8 immunohistochemistry. Scale bar: 25 μm. D AGs in the amygdala. Gallyas method. Scale bar: 25 μm. E Hippocampal sclerosis. Hematoxylin-eosin stain. Scale bar: 100 μm. F Severe loss of neurons with gliosis in the amygdala. Hematoxylin-eosin stain. Scale bar: 25 μm. G–L Pathological findings in a case with Braak stage II, Thal phase 0, amygdala GFA stage 4, Saito AG stage III, and LATE-NC stage 0. G This case lacked phosphorylated TDP-43-positive lesion in any region. The amygdala. pS409/410 immunohistochemistry. Scale bar: 25 μm. H, I GFAs in the amygdala. AT8 immunohistochemistry. Scale bar: 25 μm. J AGs in the amygdala. Gallyas method. Scale bar: 25 μm. K Neither loss of pyramidal neurons nor gliosis is noted in the hippocampal CA1. Hematoxylin-eosin stain. Scale bar: 100 μm. L Severe neuronal loss with gliosis in the amygdala. Hematoxylin-eosin stain. Scale bar: 25 μm.


S-adenosylmethionine (SAM) and total cysteine (tCys) concentrations. Serum 5-MTHF concentrations demonstrated a stronger negative correlation with tHcy/tCys than with tHcy alone. The negative correlation between betaine and tHcy concentrations was stronger in the low 5-MTHF group than in the high 5-MTHF group. The 5-MTHF status could be linked to Hcy flux into the transsulfuration pathway via SAM. Therefore, the tHcy/tCys ratio may be a more sensitive indicator of the 5-MTHF status than tHcy alone. Furthermore, a low 5-MTHF status can enhance Hcy metabolism via betaine.Maintaining optimal one-carbon metabolism (OCM) is essential for health and pregnancy. In this cross-sectional study, folate status was assessed based on 5-methyltetrahydrofolate (5-MTHF) levels, and the association between 5-MTHF and OCM-related metabolites was investigated in 227 female Japanese university students aged 18–25 years. The participants were divided into high and low 5-MTHF groups based on their folate status. Serum samples of the participants were collected while they were fasting, and 18 OCM-related metabolites were measured using stable-isotope dilution liquid chromatography–electrospray tandem mass spectrometry. The association between serum 5-MTHF and OCM-related metabolite concentrations was assessed using Spearman’s rank correlation coefficient. Serum 5-MTHF concentrations were negatively correlated with total homocysteine (tHcy) concentrations and positively correlated with  S-adenosylmethionine (SAM)-dependent methyl transfer reactions [One-carbon metabolism (OCM) comprises a folate cycle and choline metabolic pathway linked to a methionine cycle; homocysteine (Hcy) in the methionine cycle is connected to the transsulfuration pathway . OCM is eactions , nucleiceactions ,2, and aeactions .2-dependent MTHF reductase  [S-methyltransferase , which is expressed primarily in the liver and kidneys and remethylates Hcy to produce methionine and dimethylglycine (DMG) [S-adenosylhomocysteine (SAH) [6-dependent enzyme cystathionine-β-synthase , which condenses serine to Hcy thiol [6-dependent enzyme cystathionine γ-lyase  to Cys [Folate mediates the transfer of one-carbon units in OCM . Folic a.5.1.20) . The maj.5.1.20) ,8. Methi.5.1.20) ,7,8. Cho.5.1.20) . Betainene (DMG) ,11. Methne (DMG) . SAM is ne (SAH) . Methylane (SAH) ,15. SAH ne (SAH) . Both thne (SAH) . Hcy is ne (SAH) ,19. Hcy cy thiol ,20. Then) to Cys ,21. Cys ) to Cys ,19.OCM metabolic markers have been linked to various diseases. Intracellular Hcy concentrations increase when OCM fails to function adequately ,22. The Previous studies have assessed folate concentrations in circulation using a microbiological assay ; howeverThe characteristics of the study population are summarized in 6, folate, and vitamin C intakes, but significantly lower sodium intake than the low 5-MTHF group.p = 0.059). However, tHcy and cystathionine concentrations and tCys/tHcy ratios were significantly lower in the high 5-MTHF group than in the low 5-MTHF group. Serum homocysteic acid, pyridoxamine, and pyridoxine concentrations were below the limit of quantification in all samples.The serum OCM-related metabolite concentrations in the high and low 5-MTHF groups and in the overall study sample are summarized in p < 0.001) than in the high 5-MTHF group . The positive correlation between DMG and tHcy concentrations was weaker in the low 5-MTHF group  than in the high 5-MTHF group . The correlation matrix between serum OCM-related metabolite concentrations and enzyme activity indices is summarized in p < 0.001) than in the high 5-MTHF group . The 5-MTHF concentrations and betaine/DMG ratios showed a significant positive correlation. The significant negative correlation between 5-MTHF concentrations and tHcy/tCys ratios  was stronger than the correlation between 5-MTHF and tHcy concentrations alone  than the low 5-MTHF group. On the other hand, the high 5-MTHF group exhibited higher energy and micronutrient intake than the low 5-MTHF group, suggesting a potential dietary influence on OCM. Because food nutrients are complex, it is likely that the intake of folate-rich foods, such as vegetables, altered the intake of other micronutrients. The intake of methionine, cystine, glycine, serine, zinc, riboflavin, and vitamin BHerein, the median serum 5-MTHF concentration in young Japanese women was 19.2 nmol/L, which was comparable to or slightly higher than that reported in previous studies involving healthy adults in countries where FA fortification of grains is not mandated, such as Japan ,64. HoweConcentrations of 5-MTHF exhibited a significant positive correlation with SAM and tCys concentrations and a significant negative correlation with tHcy concentrations. Similarly, a previous study involving older adults found a positive correlation between 5-MTHF and SAM concentrations, and FA intervention significantly increased plasma SAM concentrations in the intervention group compared with the control group . In addiThe negative correlation between serum 5-MTHF concentrations and tHcy/tCys ratios was stronger than that between 5-MTHF and tHcy concentrations alone . The tHcHerein, 5-MTHF concentrations exhibited significant positive correlations with betaine concentrations or betaine/DMG ratios . SimilarHerein, a negative association between serum 5-MTHF and cystathionine concentrations was observed . A negatSerum FA concentrations were not correlated with OCM-related metabolite concentrations associated with 5-MTHF concentrations. This suggests that the concentration of 5-MTHF, a bioactive folate molecule species, as a biomarker may be more useful than that of total folate, which was considered in previous studies .This study has several strengths. Early morning fasting blood samples were collected, and standardized blood collection protocols were used to minimize the following effects on OCM-related metabolites: changes in the blood concentration of the measured substance due to eating immediately before the blood sample is collected; errors related to human activities at the time of blood collection; and variation in the deterioration of the substance being measured. Furthermore, the serum concentrations of OCM-related metabolites were measured using the stable-isotope dilution mass spectrometry method with internal standards for all measured components, yielding high-quality quantitative results .12-dependent reaction, and vitamin B12 is an important determinant of Hcy [6 intake may have promoted transsulfuration pathway flux. Finally, single-nucleotide polymorphisms may have an impact on OCM [However, this study has several limitations. First, given the cross-sectional survey design, it is impossible to establish a causal association between OCM metabolic dynamics. Further intervention studies on FA or 5-MTHF are required. Second, serum OCM-related metabolites do not always directly reflect OCM dynamics in organ cells. Third, 5-MTHF-dependent Hcy remethylation is a vitamin Bt of Hcy ,107; howt of Hcy ; howevert of Hcy ,109. In t on OCM ; howeverThis study was conducted at Kagawa Nutrition University, Saitama, Japan, between October and December 2018. This was a cross-sectional study designed to investigate the association between diet and blood components such as biochemical test values, OCM-related metabolites, fatty acids, and antioxidants; we reported associations between serum OCM-related metabolites.Healthy female university students aged 18–25 years majoring in nutrition were included. The following students were excluded: (1) those with health conditions that may affect the biomarker concentration; (2) those with a history of, or who have serious hepatic, renal, cardiac, pulmonary, gastrointestinal (including gastrectomy), or organ disorders; diabetes; food allergies; or other serious diseases; (3) those receiving medicines that can affect lipid metabolism or FA metabolism or anti-inflammatory and antioxidant drugs that may affect the measured values in this study; (4) those pregnant and lactating or planning for pregnancy and lactation; (5) those with systolic blood pressure <90 mmHg; (6) those who had previously donated large amounts of blood; (7) those participating in other clinical trials or studies or within 4 weeks of the end of those studies; (8) those with BMI >25 because BMI may affect blood SAM and SAH levels ,111; andn = 9), lost contact (n = 6), health problems (n = 1), busyness (n = 4), and withdrawals (n = 11), leaving 227 participants. The participants completed a lifestyle questionnaire and underwent physical measurements, and their fasting blood samples were collected after at least 10 h of fasting.A total of 258 eligible participants consented to the study, including proxy consent for minors. After the study began, 31 participants dropped out owing to consent withdrawals (The study protocol was approved by the Ethics Committee of Kagawa Nutrition University (protocol code no. 204) and was conducted in accordance with the Helsinki Declaration. All participants provided written informed consent before participating in the study.g, and the serum was then separated within an hour and frozen at −80 °C. Under these storage conditions, the concentrations of OCM-related metabolites measured in this study have been reported to be generally stable [To minimize dietary effects ,67,112, y stable ,113,114.m/z 460.2→313.2 (5-MTHF), m/z 465.2→313.2 (5-MTHF−13C5), m/z 442.2→295.2 (FA), m/z 447.2→295.2 (FA−13C5), m/z 104.0→60.1 (choline), m/z 113.0→69.1 (choline-d9), m/z 118.1→58.0 (betaine), m/z 129.0→66.1 (betaine-d11), m/z 104.1→58.1 (DMG), m/z 110.1→64.0 (DMG-d6), m/z 150.1→104.0 (methionine), m/z 153.0→107.0 (methionine-d3), m/z 399.1→250.3 (SAM), m/z 402.2→250.2 (SAM-d3), m/z 385.2→134 (SAH), m/z 389.2→136.2 (SAH-d4), m/z 136.0→90.0 (Hcy), m/z 140.0→93.9 (Hcy-d4), m/z 184.1→138.3 (homocysteic acid), m/z 188.1→142.0 (homocysteic acid-d4), m/z 223.2→134.1 (cystathionine), m/z 227.1→138.1 (cystathionine-d4), m/z 122.1→59.1 (Cys), m/z 124.2→61.0 (Cys-d2), m/z 126.1→107.8 (taurine), m/z 128.1→110.2 (taurine−13C2), m/z 106.1→60.1 (serine), m/z 109.1→63.1 (serine-d3), m/z 76.0→30.0 (glycine), m/z 78.0→31.9 (glycine-d2), m/z 377.1→243.1 (riboflavin), m/z 383.2→249.1 (riboflavin−13C415N2), m/z 169.2→152.2 (pyridoxamine), m/z 172.1→155.1 (pyridoxamine-d3), m/z 170.1→134.1 (pyridoxine), and m/z 172.1→136.0 (pyridoxine-d2)) [We previously described the method for measuring OCM-related metabolites . After cine-d2)) . Hcy andine-d2)) . AnalystData regarding the participants’ ages were collected using a self-administered questionnaire.Dietary data were collected for 7 days before blood collection using a continuous, non-weighted dietary record in conjunction with digital images captured using a digital camera or smartphone according to a validated dietary survey method ,118,119.Blood pressure was measured using a P2000 Electronic Blood Pressure Monitor . The mean systolic and diastolic blood pressure was measured twice in the resting state according to the Japanese guidelines for managing hypertension .The participants’ anthropometric measurements were collected using standardized methods during laboratory visits for blood sampling. Body weight and body fat percentage were measured using TBF-110 .U test was used to compare these groups. The enzyme activity indices included the betaine/DMG ratio [p-value of <0.05. IBM Statistical Package for the Social Sciences Statistics, Version 28  was used for statistical analysis.The distribution of subject characteristics data, nutrient intake, and serum concentrations of OCM-related metabolites used in the analysis were mostly non-normal and continuous variables were expressed as medians . Since there are no thresholds established for serum 5-MTHF concentrations such as deficiency, insufficiency, sufficiency, or excess, based on a previous study , a mediaMG ratio  for BHMTMG ratio  for methMG ratio  for enzyPrevious studies assessed folate status without considering folate molecular species. Thus, we extended the findings of previous studies by comprehensively measuring OCM-related metabolites, including 5-MTHF, a major reduced folate. FA showed no association with important OCM-related metabolites, indicating that blood 5-MTHF levels could be useful for evaluating folate status. The 5-MTHF state could be linked to the Hcy flux into the transsulfuration pathway via SAM. Therefore, the tHcy/tCys ratio may be a more sensitive indicator of the 5-MTHF status than tHcy concentrations alone. A low 5-MTHF status may affect the choline metabolic pathway, particularly BHMT or CBS activity via betaine, implying that the folate cycle and choline metabolic pathway should be evaluated simultaneously in Hcy studies. Further intervention studies on FA or 5-MTHF are warranted to determine the causal association between the abovementioned findings.


Correction: Journal of Translational Medicine (2013) 11:29 10.1186/1479-5876-11-29Since the publication of our article , it has https://jeccr.biomedcentral.com/articles/10.1186/1756-9966-27-15. Copyright © 2008, Liao et al.; licensee BioMed Central Ltd.(A) The levels of CSE1L expression in B16-EV, B16-CSE1L, COLO-EV, and COLO-CSE1L cells were assayed by immunoblotting with anti-CSE1L antibody. The β-actin levels were assayed as a control. The invasive ability of the cells was analyzed by in vitro invasion assays using chemotaxis chambers, as described in “Materials and Methods”. The left panel is adapted with permission from Fig. 5C in Liao CF, Luo SF, Li LT, Lin CY, Chen YC, Jiang MC. CSE1L/CAS, the cellular apoptosis susceptibility protein, enhances invasion and metastasis but not proliferation of cancer cells. J Exp Clin Cancer Res. 2008; 27:15.


Infections after LT are common, but standard of care testing (SOCT) often fails to identify a cause. We prospectively compared longitudinal mcfDNA sequencing vs SOCT to determine the utility of mcfDNA sequencing for infection surveillance after LT.Single center prospective observational study of new LT patients (pts). We collected plasma on days 0, 1, 2, 3, 7, 14, and 30 post LT for mcfDNA sequencing; results were not available clinically. We reviewed outcomes and SOCT through 6 months (mo).Candida, oral or gastrointestinal commensals, Enterococci, or obligate pathogens ; most had polymicrobial mcfDNA sequencing results. In 36.67% (11/30), mcfDNA sequencing detected pathogens of unclear significance . In 8 pts with positive SOCT, mcfDNA sequencing did not detect at least 1 pathogen found on SOCT. In 13% (4/30), mcfDNA sequencing detected a pathogen before it was detected by SOCT (n=1 each): VRE surgical site infection (SSI), invasive candidiasis, Enterobacter pneumonia, and M. hominis SSI (a known donor-derived syndrome) with HHV-8/Kaposi Sarcoma .We enrolled 30 LT pts . 93.3% (28/30) received broad-spectrum antibiotics, although only 40.0% (12/30) had SOCT yielding pathogenic bacteria. 43.33% (13/30) had unexplained leukocytosis. Table 2 shows serial mcfDNA sequencing results, clinical course, and adjudication of mcfDNA sequencing. 96% (29/30) had at least 1 positive mcfDNA sequencing test, including 68.75% (11/16) with presumed infection or unexplained leukocytosis and negative SOCT, but diversity of organisms on mcfDNA sequencing, e.g., Table 1.Table 2 legend.Table 2.Plasma mcfDNA sequencing revealed potential occult explanations for unexplained leukocytosis and infectious syndromes with negative SOCT early post LT. mcfDNA sequencing also detected infection earlier than SOCT in 4 pts but did not detect known pathogens from 8 pts; > 1/3 had presumed FP mcfDNA sequencing results. Larger studies with longer follow-up are needed to determine the utility of mcfDNA sequencing in diagnosing infection, optimizing antibiotic use, and monitoring donor-derived infection post LT.Ghady Haidar, MD, Allovir: Grant/Research Support|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|Karius: Advisor/Consultant|Karius: Grant/Research Support|NIH: Grant/Research Support Timothy A. Blauwkamp, PhD, Karius: Board Member|Karius: Ownership Interest Georgios Kitsios, MD, PhD, Karius, Inc: Grant/Research Support


This study reports on the E. coli and 92 S. saprophyticus isolates were collected during 2022 from 45 medical centers located within the United States. Most isolates (77%) tested were cultured from urine specimens collected from patients seen in ambulatory, emergency, family practice, and outpatient medical services. Bacterial identifications were confirmed by MALDI-TOF MS. Isolates were tested for susceptibility by CLSI methods at a central laboratory (JMI Laboratories). MIC results for oral antibiotics licensed for the treatment of uUTI and drug-resistant subsets were interpreted per CLSI guidelines.A total of 1,001 50/90, 2/4 µg/mL) displayed good activity against 1,001 E. coli isolates as 98.8% of all observed gepotidacin MICs were ≤4 µg/mL (Table). Other oral agents tested against these isolates demonstrated the following rates of susceptibility (S): amoxicillin-clavulanate (85.2% S), ampicillin (55.6% S), ciprofloxacin (78.8%S), fosfomycin (99.3% S), mecillinam (93.5%S), nitrofurantoin (97.5% S), and trimethoprim-sulfamethoxazole (73.3% S). Gepotidacin maintained similar MIC50 values (ranging from 1 – 2 µg/mL) and MIC90 values (ranging from 2 – 4 µg/mL) against these drug-resistant subsets. Against S. saprophyticus isolates, gepotidacin  inhibited all isolates at ≤0.12 µg/mL. Most oral agents showed >90% S against S. saprophyticus isolates, except for penicillin (7.6% S).Gepotidacin (MICin vitro activity against contemporary E. coli and S. saprophyticus from US urine isolates. This activity remained unaffected by resistance to other oral standard-of-care antibiotics.Gepotidacin demonstrated S J Ryan Arends, PhD, Cipla: Grant/Research Support|GSK: Grant/Research Support Renuka Kapoor, PhD, GSK: Grant/Research Support Didem Torumkuney, PhD, GSK: Grant/Research Support Nicole E. Scangarella-Oman, MS, GSK: Employee and shareholder Rodrigo E. Mendes, PhD, AbbVie: Grant/Research Support|Basilea: Grant/Research Support|Cipla: Grant/Research Support|Entasis: Grant/Research Support|GSK: Grant/Research Support|Paratek: Grant/Research Support|Pfizer: Grant/Research Support|Shionogi: Grant/Research Support


Acinetobacter spp., including carbapenem-resistant Acinetobacter baumannii-calcoaceticus complex (ABC) organisms. In this study, the susceptibility testing of zosurabalpin was carried out against a panel of 150 randomly selected Acinetobacter spp. isolates (100 A. baumannii & 50 non-A. baumannii) representing 11 sites in China and a broad susceptibility profile (65% of which were multi-drug resistant [MDR]) collected in 2021.Zosurabalpin (RG6006) is the first representative of a novel class of tethered macrocyclic peptide antibiotics active against MICs were performed using the Clinical Laboratory Standards Institute (CLSI) broth dilution method cation-adjusted Mueller Hinton broth (CA-MHB) supplemented with either 20% of human serum (HS) or 20% of horse serum (HoS). For a fraction of isolates (10-25%), MIC determination for zosurabalpin is affected by aberrant readings  in CA-MHB. This effect complicates routine susceptibility testing. Supplementation of CAMHB with serum allows accurate MIC determinations without any major effects on the MIC distribution.Summary data from the study are shown in Table 1.Acinetobacter spp., with an MIC50/90 of 0.12/0.5 μg/mL and 0.25/1 μg/mL in CA-MHB supplemented with HoS and HS, respectively (MIC range of 0.015/0.03 to 8 μg/mL). Against ABC isolates (n=133), the MIC50/90 for zosurabalpin was 0.12/0.25 μg/mL and 0.25/0.5 μg/mL, in HoS and HS, respectively (MIC range of 0.015/0.03 to 1 μg/mL). A similar activity was observed against carbapenem-resistant ABC isolates.Zosurabalpin was active against all in vitro antibacterial activity against Acinetobacter clinical isolates circulating in China. These data support the continued clinical development of zosurabalpin for infections caused by ABC isolates, including difficult to treat carbapenem-resistant isolates.In addition to the potent activity observed against isolates from USA and Europe, zosurabalpin exhibited potent Stephen Hawser, PhD, Allecra: study funding|Innoviva Specialty Therapeutics, Inc.: Honoraria|Roche: Honoraria|Roche: This project has been funded by BARDA (HHSO100201600038C). Nimmi Kothari, PhD, Allecra: Allecra (study funding)|Innoviva Specialty Therapeutics, Inc.: Honoraria|Roche: Honoraria|Roche: This project has been funded by BARDA (HHSO100201600038C). Thomas Valmont, BS, Roche: Honoraria|Roche: This project has been funded by BARDA (HHSO100201600038C). Séverine Louvel, PhD, F. Hoffmann-La Roche Ltd: employee of the company Claudia Zampaloni, n/a, F. Hoffmann-La Roche Ltd.: Full time employee of Roche


Detection of SARS-CoV-2 in hospitalized patients is an important measure to prevent transmission to vulnerable individuals. Admission screening using nucleic acid amplification test such as PCR testing or rapid antigen testing (RAT) is proven to be beneficial in certain situations. However, the usefulness of SARS-CoV-2 antibody testing for COVID-19 screening in real-world setting is unclear.In this study, we aimed to investigate the effect of antibody testing on the results of SARS-CoV-2 screening tests. Anti-SARS-CoV-2 spike protein antibody (S-Ab) and anti-SARS-CoV-2 nucleocapsid protein antibody (N-Ab) were measured using an electrochemiluminescence immunoassay  in all patients who presented to the emergency department and subsequently administered to Nagasaki University Hospital. PCR testing and RAT were also performed for the detection of SARS-CoV-2. The diagnostic performance of each antibody testing to PCR and RAT was retrospectively evaluated by Fisher's exact test and odds ratio (OR).A total of 2,692 patients were tested for S-Ab, N-Ab, RT-PCR, and RAT from Jan 2022 to Feb 2023. Among them, S-Ab were positive ( >0.7 U/mL) for 2,195 patients (81.5%), N-Ab ( >1.0 U/mL) 307 patients (11.4%), PCR 215 patients (7.9%), RAT 174 patients (6.5%).P=0.35, OR 0.8397) and RAT 1.8% . For those with N-Ab positive (n=307), PCR positivity was 6.8%  and RAT 2.6% . Comparing the group of both S-Ab > 10,000 U/mL and N-Ab positive (n=128) with that of both negative (n=2564), PCR positivity was 4.7% vs. 8.1% , and RAT positivity was 1.6% vs. 6.7% .For those with S-Ab > 10,000 U/mL (n=625), PCR positivity was 7.0% (Antibody testing of S-Ab, N-Ab, or a combination of both showed performance in reducing the probability of positive PCR or RAT tests, indicating vaccine and infection-induced protective effect. However, the need for PCR and RAT tests for virus detection was suggested.Kosuke Kosai, M.D., Ph.D., FUJIFILM Toyama Chemical Co., Ltd.: Commissioned research|KYORIN Pharmaceutical Co.,Ltd.: Commissioned research Koichi Izumikawa, M.D., Ph.D., Asahi Kasei Pharma Corporation: Grant/Research Support|Asahi Kasei Pharma Corporation: Honoraria|Astellas Pharma Inc.: Honoraria|DAIICHI SANKYO COMPANY, LIMITED: Grant/Research Support|DAIICHI SANKYO COMPANY, LIMITED: Honoraria|KYORIN Pharmaceutical Co.,Ltd.: Honoraria|Merck & Co., Inc.: Honoraria|Pfizer Japan Inc.: Honoraria|Shionogi & Co., Ltd.: Grant/Research Support|Shionogi & Co., Ltd.: Honoraria|Sumitomo Pharma Co., Ltd.: Grant/Research Support|Sumitomo Pharma Co., Ltd.: Honoraria|TAIHO PHARMACEUTICAL CO., LTD.: Grant/Research Support Katsunori Yanagihara, MD, PhD, FUJIFILM Toyama Chemical Co., Ltd.: Commissioned research|KYORIN Pharmaceutical Co.,Ltd.: Commissioned research


Artemisia heptapotamica Poljak led to the isolation of ten known compounds, including four alkyl p-coumarates: octadecyl trans-p-coumarate (1), icosy trans-p-coumarate (2), docosyl trans-p-coumarate (3), and tetracosyl trans-p-coumarate (4), one sesquiterpene lactone: santonin (5), four flavonoids; axillarin (6), quercetin 3-O-methyl ether (7), luteolin (8), and quercetin (9), and one phenolic acid derivative: p-coumaric acid (10). The structures of the isolated compounds were identified by various spectroscopic analyses. Additionally, the antimicrobial activity of the total extract and different fractions was screened, and they exhibited no inhibition of the growth of Candida albicans, C. neoformans, Aspergillus fumigatus, methicillin-resistant Staphylococcus aureus (MRS), E. coli, Pseudomonas aeruginosa, Klebsiella pneumonia, and Vancomycin-resistant Enterococci (VRE) at the tested concentrations ranging from 8 to 200 μg/mL. The identification and tentative characterization of the secondary metabolites were conducted using LC-QToF analysis. This method helps in the putative characterization of sesquiterpene lactones, flavonoids, coumarate derivatives, and aliphatic compounds. The developed method identified 43 compounds, of which the majority were sesquiterpene lactones, such as eudesmanolides, germacranolides, and guaianolide derivatives, followed by flavonoids. The proposed LC-QToF method helps develop dereplication strategies and understand the major class of chemicals before proceeding with the isolation of compounds.Phytochemical investigation of the aerial parts of  Artemisia L. is the largest genus belonging to the family Asteraceae + and [M + Na]+ adduct precursor ions. The tentative characterization of compounds was processed based on the molecular features, such as accurate mass, fragment ions , and precursor ion molecular formula. The representative base peak chromatograms (BPC) in negative and positive modes, along with LC-DAD profiles at 210 nm, are shown in The secondary metabolites from aerial parts of Artemisia species. To perform characterization using LC-QToF, a few sesquiterpene lactones, e.g., santonin (compound 19), were isolated from aerial parts of A. heptapotamica in this study. Santonin mass fragmentation initiated with loss of water molecules, resulting in m/z 228.1219 [M + H-H2O]+. The following loss of -CO and sequential loss of -C2H4 and -CH4 resulted in m/z 201.1271, 173.0954, and 157.0648, respectively. Further loss of -CO ion resulted in the opening of the lactone moiety. Further loss of ketene moiety (-C2H2O) from the opened six-membered ring resulted in the formation of m/z 115.0542. Fragments with the least molecular weight (m/z 105.0698 and 91.0544) were useful in determining the backbone skeleton of the molecule. These characteristic fragments matched with the reported santonin fragmentation pathway − and fragment ions are shown in 25 showed m/z 509.1284 [M + H]+ with a C23H24O13 molecular formula. The corresponding fragment ions resulted in m/z 347.0762 [M + H-Glc]+, 331.0434, and 289.0329, respectively. The compound was characterized as 3,4,5,7-tetrahydroxy-3-methoxyflavone 7-O-β-D-glucopyranoside. Similarly, the flavone derivatives were characterized based on their fragmentation pathways. Along with hydroxy/methoxy flavone derivatives, simple aglycone flavonoids such as quercetin (compound 27) and luteolin (compound 26) were identified. Quercetin showed m/z 303.0491 [M + H]+ and m/z 301.0354[M-H]− with fragment ions at m/z 153 and 151, respectively. Furthermore, luteolin showed m/z 287.0548 [M + H]+ and 285.0406 [M-H]− in both positive and negative ionization modes. In addition, one quercetin derivative (compound 33) was identified with m/z 317.0658 [M + H]+ with fragment ions at m/z 301.0343 (such as quercetin aglycone), 274.0465, and 137.0233. The compound was characterized as quercetin 3-O-methyl ether.Using the isolated reference compounds and exact mass measurements, fourteen flavonoid compounds were identified. Most of the compounds are flavone derivatives. There are two quercetin derivatives, along with quercetin and luteolin. Flavonoids have established mass fragmentation patterns based on the literature previously reported ,23,24,25m/z 165.0546 [M + H]+ and 163.0398 [M-H]− with fragment ions at m/z 119.0502 [M-H-CO2]− in the negative mode of ionization. The compound was characterized as p-coumaric acid (compound 37). In addition, an aliphatic compound was identified with chemical formula C10H18O3. Compound 38 was tentatively characterized based on the exact mass at m/z 185.1186 [M-H]− and corresponding fragment ion at m/z 167.1077 [M-H-H2O]−. Compound 39 was identified as 3,4,5-tri-caffeoylquinic acid with a precursor ion at m/z 677.1524 [M-H]−. The corresponding fragment ions were at m/z 515.1198 [M-H-Glc]−, 353.0822 [M-H-Glc-Glc]−, and 191.0572 [quinic acid]  , respectArtemisia species.The isolated compounds, as well as chromatographic peaks with distinctive fragment ions, were confirmed based on the reported literature studies whose corresponding references are listed in 1H and 13C NMR spectra were recorded on a Bruker Avance 400 MHz instrument. HR-ESI-MS was taken on BrukerBioApex-FTMS with electron spray ionization. Solvents used in this work, e.g., n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), methanol (MeOH), and ethanol (EtOH), were purchased from Fisher Scientific, USA. Deuterated solvents purchased from Cambridge Isotope Laboratories, Inc., Tewksbury, MA, USA, including methanol-d4 (CD3OD), chloroform-d3 (CDCl3), and pyridine-d5 (C5H5N-d5), were used for nuclear magnetic resonance (NMR) spectroscopic analyses. Acetonitrile, methanol, and formic acid of HPLC-certified grade were used for LC-MS analysis, and water was purified using a Milli-Q system . Column chromatography (CC) was performed using silica gel 60 . Thin-layer chromatography (TLC) analyses were carried out using pre-coated silica G plates w/UV254 . An ultraviolet lamp  was used for visualization of spots on thin-layer chromatograms at 254 and/or 365 nm. Spots were visualized by spraying with 2% vanillin  in sulfuric acid–ethanol followed by heating at 110 °C.A. heptapotamica was collected in October 2021 from Kokpek village, Almaty, Kazakhstan. Identification and authentication were performed by Dr. Danilov Mikhail Petrovich. The sample was stored with the voucher number 0000723 in the Main Botanical Garden of the Institute of Botany and Phyto-introduction, Almaty, Kazakhstan. The same sample was assigned with the NCNPR number 25173 and stored in the National Center for Natural Product Research Botanical Repository, University of Mississippi, USA.The whole plant of A. heptapotamica were first dried in the shade and then crushed into small pieces. The dried plant material (1.65 kg) was extracted by maceration with 95% methanol three times at room temperature and was concentrated under reduced pressure to yield 253.35 g of the total extract. The total extract was mixed with a small amount of distilled water and successively fractionated with n-hexane and ethyl acetate. The fractions were concentrated under reduced pressure to produce n-hexane (29.1 g) and ethyl acetate fractions (58.74 g).The collected aerial parts of n-hexane and EtOAc  and finally washed with MeOH, affording 6 fractions. Fraction F-4 (395.9 mg) was subjected to silica gel CC , using DCM–MeOH gradient mixtures to increase the polarity gradually in 2% MeOH till 80 % MeOH and finally 100% MeOH were produced, yielding 37 subfractions. The seventeenth subfraction (21.1 mg) afforded a mixture of compounds 3 and 4. The eighteenth subfraction (12.9 mg) produced a mixture of compounds 2–4. The nineteenth subfraction (16.5 mg) produced a mixture of compounds 2–4. The twentieth subfraction (6.2 mg) furnished a mixture of compounds 1–4.The EtOAc (58.74 g) fraction was subjected to fractionation using VLC silica gel CC using 5 (45 mg). The eighteenth subfraction was precipitated to give compound 6 (27 mg). Subfraction F-5-21 (1.26 g) was purified on silica gel CC using DCM–MeOH gradient mixtures to gradually increase the polarity in 2% MeOH till 80% MeOH was produced, affording compound 7 (8.4 mg). Subfraction F-5-24 (342.8 mg) was rechromatographed over silica gel CC using DCM–MeOH gradient mixtures to gradually increase the polarity in 2% MeOH till 80% MeOH was reached, producing a mixture of compounds 7 and 8 (14.5 mg) and 9 (24.4 mg). Subfraction F-5-26 (450.1) was purified on silica gel CC using DCM–MeOH gradient mixtures to gradually increase the polarity in 2% MeOH till 80% MeOH was reached, producing compound 10 (8.2 mg).Fraction F-5 (14.3 g) was subjected to silica gel CC , using DCM–MeOH gradient mixtures to gradually increase the polarity in 2% MeOH till 80% and finally 100% MeOH were produced, affording 29 subfractions. The eleventh subfraction was crystalized to produce compound The antimicrobial activity of the total extract and different fractions was evaluated using the method reported by Samy et al. .About 25 mg of extract was sonicated in 1.0 mL of methanol for 5 min, followed by centrifugation for 15 min at 7000 rpm. The clear filtered supernatant solution was used for analysis.TM HSS C18 column . The mobile phase consisted of water with 0.1% formic acid (A) and acetonitrile with 0.1% formic acid (B) at a flow rate of 0.23 mL/min. Analysis was performed using the following gradient elution: 15% B to 40% B in 30 min, then to 100% B in the next 15 min. Each run was followed by a 5 min wash with 100% B and an equilibration period of 15 min with 85% A/15% B. Two microliters of the sample were injected. The column temperature was 40 °C.The liquid chromatographic system was an Agilent Series 1290, and separation was achieved on an Acquity UPLC2) flow rate, 13 L/min; drying gas temperature, 325 °C; nebulizer pressure, 30 psi; sheath gas temperature, 300 °C; sheath gas flow, 11 L/min; capillary voltage, 3500 V; nozzle voltage, 0 V; skimmer, 65 V; Oct RF V, 750 V; and fragmentor voltage, 125 V. All the operations, acquisition of data, and analysis of data were controlled using Agilent MassHunter Acquisition Software ver. A.10.1 and processed with MassHunter Qualitative Analysis software ver. B.07.00. Each sample was analyzed in positive and negative modes over the range of m/z 50–1700 and an extended dynamic range. Accurate mass measurements were obtained by employing ion correction techniques using reference masses at m/z 121.0509 (protonated purine) and 922.0098  in positive ion mode, while m/z 112.9856 (deprotonated trifluoroacetic acid-TFA) and 1033.9881 (TFA adducted HP-921) were used in negative ion mode. Samples were analyzed in all-ion MS–MS mode, where experiment 1 was carried out with a collision energy of zero and experiment 2 with a fixed collision energy of 45 eV.The mass spectrometric analysis was performed with a QToF-MS-MS  equipped with an ESI source with Jet Stream technology, using the following parameters: drying gas , which matched with the authentic samples of A. heptapotamica.Ten compounds were isolated from the ethyl acetate fraction of


Immune responses after COVID-19 vaccination should be evaluated in different populations around the world. This study compared antibody responses induced by ChAdOx1 nCoV-19, CoronaVac, and BNT162b2 vaccines. Blood samples from vaccinees were collected pre- and post-vaccinations with the second and third doses. The study enrolled 78 vaccinees, of whom 62.8% were women, with the following median ages: 26 years—ChAdOx1 nCoV-19; 40 years—CoronaVac; 30 years—BNT162b2. Serum samples were quantified for anti-RBD IgG and anti-RBD IgA and anti-spike IgG by ELISA. After two vaccine doses, BNT162b2 vaccinees produced higher levels of anti-RBD IgA and IgG, and anti-spike IgG compared to ChAdOx1 nCoV-19 and CoronaVac vaccinees. The third dose booster with BNT162b2 induced higher levels of anti-RBD IgA and IgG, and anti-spike IgG in CoronaVac vaccinees. Individuals who reported a SARS-CoV-2 infection before or during the study had higher anti-RBD IgA and IgG production. In conclusion, two doses of the studied vaccines induced detectable levels of anti-RBD IgA and IgG and anti-spike IgG in vaccinees. The heterologous booster with BNT162b2 increased anti-RBD IgA and IgG and anti-spike IgG levels in CoronaVac vaccinees and anti-RBD IgA levels in ChAdOx1 nCoV-19 vaccinees. Furthermore, SARS-CoV-2 infection induced higher anti-RBD IgA and IgG levels in CoronaVac vaccinees. Several preventive strategies have been developed and continue to be generated to control SARS-CoV-2 transmission, the most important being vaccines . CurrentMost vaccines developed against SARS-CoV-2 induce an immune response toward the receptor-binding domain (RBD), a portion of the spike protein present at the S1 region, which is essential for binding the virus to angiotensin-converting enzyme-2 (ACE2) on the cell surface and to establish cellular infection . Thus, aAfter COVID-19 vaccination, IgG and IgA are mostly involved in virus neutralization, especially IgA since this Ig can neutralize SARS-CoV-2 at the mucosal surface entrance . Some stThe anti-SARS-CoV-2 vaccines comprise platforms of inactivated viruses , proteinThe present study was evaluated and approved by the Research Ethics Committee of the Hospital das Clínicas, Universidade Federal de Goiás, under the number CAAE: 30804220.2.0000.5078 and is in accordance with Resolution 466/2012 of the National Health Council, which regulates research involving human beings. The collection of data and blood samples was conducted after the individuals selected for the sample had agreed to participate and had signed the informed consent form.Blood samples were collected from 78 volunteers before immunization (ChAdOx1 nCoV-19 and BNT162b2) and after the administration of the second doses of ChAdOx1 nCoV-19, CoronaVac, and BNT162b2 vaccines, and after the third dose (booster) with BNT162b2 vaccine. In addition, nasopharyngeal swab samples were collected from individuals belonging to the ChAdOx1 nCoV-19 and BNT162b2 vaccine groups at the pre-vaccine collection and 1 month after the second dose to assess whether these individuals could be infected with SARS-CoV-2 during the period.Participants were recruited by phone or in person while in the queue for vaccination, and during the vaccination of health professionals at the Instituto de Patologia Tropical e Saúde Pública (IPTSP), Universidade Federal de Goiás (UFG), Goiânia-GO, Brazil. The inclusion criteria were individuals at least 18 years old, vaccinated with two doses of CoronaVac, ChAdOx1 nCoV-19, or BNT162b2, and those who may have received the third dose, but not necessarily.Primary pre-vaccine samples were collected at the queues for vaccination, in a physical space adjacent to the line, right before the first dose was administered. Post-vaccination samples were collected in three different periods: 1 month and 4–6 months after the second dose, and 1 month after the third dose. Samples were collected 1 month after the second and third doses because the peak of the antibodies occurs around four weeks after vaccination ,30,31,32® Viral RNA Mini Kit  was used, following the manufacturer’s protocol. Samples were submitted to real-time polymerase chain reaction post reverse transcription (RT-qPCR) after the RNA extraction, using the Promega GoTaq® Probe 1-Step RT-qPCR System, according to the manufacturer’s protocol [For Ribonucleic acid (RNA) extraction, the commercial QIAampprotocol . The priprotocol  was usedg for 10 min at room temperature. All samples were processed on the day of collection. Serum was separated and stored in 2 mL polypropylene cryotubes in a −80 °C freezer for subsequent enzyme immunoassay ELISAs.After blood collection from vaccinated individuals, whole blood was centrifuged at 600× For the ELISAs, 96-well high-binding polystyrene half-area plates  were coated with 50 µL per well with the RBD protein, expressed according to Amanat et al. (2020) , at a co2 > 0.5. For all tests, p < 0.05 was considered significant.All analyses of individual samples were conducted using GraphPad Prism version 9 . The variables distribution patterns were evaluated via the Kolmogorov–Smirnov and Shapiro–Wilk tests. For paired comparisons between groups, the non-parametric Wilcoxon Matched-Pairs Signed Rank test was used. Paired analyses were assigned to samples from the same individuals at different collection times. Unpaired groups were analyzed with the non-parametric Mann–Whitney U test. The non-paired analyses were attributed to a total group. For frequency calculations, we used the Fisher exact test. Multiple group comparisons were analyzed by running non-parametric Kruskal–Wallis statistical tests and were corrected using Dunn’s and Dunnett’s methods. Spearman correlation test was used for association analyses. The existence of correlation was adopted for rn = 78) were divided into three groups according to the vaccines they had received: ChAdOx1 nCoV-19 (n = 33), BNT162b2 (n = 27), and CoronaVac (n = 18). The ChAdOx1 nCoV-19 group was composed of 33 individuals, 23 (69.7%) women and 10 (30.3%) men. The BNT162b2 group was composed of 27 individuals, 11 (40.7%) women and 16 (59.3%) men. Finally, the CoronaVac group was made up of 18 participants, 15 (83.3%) women and 3 (16.7%) men (Study participants (.7%) men . Therefo.7%) men . ParticiWhole blood collection periods comprise T1 (pre-vaccine), T2 (1 month post-second dose), T3 (4–6 months post-second dose or pre-third dose), and T4 (1 month post-third dose or booster dose). All participants who had their samples collected at T4 received the booster dose, mostly BNT162b2. Of 78 participants, 56 received the third dose. This information can be better observed in Only the pre-vaccine nasopharyngeal samples from the ChAdOx1 nCoV-19 group were tested via reverse transcription followed by the real-time polymerase chain reaction (RT-qPCR), and all samples were negative for SARS-CoV-2 RNA. In addition, a rapid test assay to detect IgM/IgG  was performed on all serum samples collected , and only a few samples were positive. Some of them belonged to individuals who reported SARS-CoV-2 infection during the application of the questionnaire. Therefore, we believe that the rapid test positivity was due to a previous SARS-CoV-2 infection.p = 0.0027, median—pre-vaccine: 0.6250, post-vaccine: 0.8310; anti-RBD IgG, p < 0.0001, median—pre-vaccine: 0.8990, post-vaccine: 10.16; anti-spike IgG, p < 0.0001, median—pre-vaccine: 1.549, post-vaccine: 10.75; BNT162b2: anti-RBD IgA, p < 0.0001, median—pre-vaccine: 0.6460, post-vaccine: 4.023; anti-RBD IgG, p < 0.0001, median—pre-vaccine: 0.9560, post-vaccine: 11.80; anti-spike IgG, p < 0.0001, median—pre-vaccine: 1.254, post-vaccine: 11.22   A,D,G.p < 0.0001, median—ChAdOx1 nCoV-19: 0.7890; BNT162b2: 3.731; IgG, p = 0.0268, median—ChAdOx1 nCoV-19: 9.991; BNT162b2: 10.87) and CoronaVac . Individuals immunized with CoronaVac exhibited lower anti-RBD IgG levels compared to those with ChAdOx1 nCoV-19   and CoronaVac   E. This w: 11.28) H.p < 0.0001, median—ChAdOx1 nCoV-19: 1.634; BNT162b2: 6.217) and IgG , and in comparison, to CoronaVac, for anti-RBD IgA . In addition, comparisons made in relation to the previous collection period (4–6 months after the second dose or before the third dose) showed differences between anti-RBD IgA and IgG levels, after the booster dose, for the ChAdOx1 nCoV-19    C and Corr: 1.925 C; anti-Rr: 11.07 F), with : 10.80) I.p < 0.0001). Individuals administered with BNT162b2 showed a higher frequency of anti-RBD IgA production compared to those administered with ChAdOx1 nCoV-19 (p < 0.0001) and CoronaVac (p < 0.0001) at 4–6 months after the second dose (p = 0.0030) (All study vaccinees (100%) produced detectable levels of anti-RBD and anti-spike IgG after each vaccine dose. The frequency of individuals who produced anti-RBD IgA at post-vaccination collection times is shown in ond dose B. At 1 m 0.0030) C.p < 0.0001; T1 vs. T3, p = 0.0028, median—T1:0.6250, T2: 0.8310, T3: 0.7890, and T4: 1.634)   (p < 0.0001) for anti-RBD IgA, and anti-spike IgG . The longitudinal analysis performed for BNT162b2 (p = 0.0313). For the CoronaVac group    , T2 (1 month post second dose vaccine), T3 (4–6 months post second dose vaccine), and T4 (1 month post third dose vaccine). The results of this evaluation for the ChAdOx1 nCoV-19 group A,D,G rev: 1.634) A and IgG: 10.59) D, and an: 10.76) G. The da: 10.76) A  C and IgG: 11.07) F and for: 10.80) I.p = 0.0165, median—men: 11.03; women: 10.63)  .p = 0.0089; IgG, p = 0.0146) and women also vaccinated with ChAdOx1 nCoV-19  (p = 0.0145)  ) (p = 0.0080) (median (BNT162b2 men: 11.49)) (ChAdOx1 nCoV-19 women: 10.46)). At 4–6 months after the second dose, the group of men vaccinated with BNT162b2 exhibited higher anti-RBD IgA levels compared to men (p = 0.0030) and women (p = 0.0003) immunized with ChAdOx1 nCoV-19 (p = 0.0273) and men (p < 0.0001) in the BNT162b2 group   ) (p = 0.0014) and men (p = 0.0024) in the BNT162b2 group and compared to men in the ChAdOx1 nCoV-19 group (p = 0.0448) (median (ChAdOx1 nCoV-19: men: 10.05)  (CoronaVac: women: 5.224)) (p = 0.0003) and women (p = 0.0182) vaccinated with BNT162b2  (CoronaVac: women: 9.895)) (p = 0.0021) and men (p = 0.0297) vaccinated with ChAdOx1 nCoV-19  (BNT162b2: men: 9.279)) (The comparison of sexes between vaccine groups at post-vaccination collection periods  indicate 0.0001) A,D. In a 11.90)) A. The ev 11.90)) G, reveal nCoV-19 B. At the0.6920)) B. Women  5.224)) E. Women  9.895)) H. At 1 m 9.279)) C. The an 9.279)) F,I.Furthermore, we stratified the data of vaccinees in three age groups—18–30 years, 31–50 years, and >50 years old—to evaluate the influence of age on antibody production.p = 0.0062) (median (ChAdOx1 nCoV-19: 0.9070) (BNT162b2: 4.135)). Individuals vaccinated with BNT162b2 also presented higher anti-RBD IgA levels (p = 0.0188) (median (BNT162b2: 18–30 years: 4.135) (ChAdOx1 nCoV-19: >50 years: 0.7880)) and anti-spike IgG (p = 0.0075) (median (BNT162b2: 18–30 years: 12.20) (ChAdOx1 nCoV-19: >50 years: 10.09)) than those older than 50 years vaccinated with ChAdOx1 nCoV-19 (p = 0.0038) and those older than 50 years (p = 0.0460) vaccinated with ChAdOx1 nCoV-19 (p = 0.0084) (p = 0.0379) in the CoronaVac group and compared to individuals aged 18–30 years in the ChAdOx1 nCoV-19 group (p = 0.0369)   (CoronaVac: 31–50 years: 0.7570)) (p = 0.0002) and ChAdOx1 nCoV-19 (p = 0.0405) compared to those aged 31–50 years in the CoronaVac group (p = 0.0218) (median (ChAdOx1 nCoV-19: 18–30 years: 9.872)  (CoronaVac: 31–50 years: 4.402)). Analysis of anti-spike IgG levels showed that individuals aged 18–30 years vaccinated with BNT162b2 presented higher levels of this antibody compared to those aged 31–50 years vaccinated with CoronaVac (p = 0.0030) (median (BNT162b2: 18–30 years: 12.12) (CoronaVac: 31–50 years: 9.761))  B. Those 0.7570)) B. Assessac group E. Furthe 9.761)) H.p = 0.0296), and those aged 18–30 years (p = 0.0142), both in the ChAdOx1 nCoV-19 group  (BNT162b2: 18–30 years: 7.252)) (p = 0.0259) (median (ChAdOx1 nCoV-19: 10.21) (BNT162b2: 12.08)) (p = 0.0347) (median (BNT162b2: 12.24) (ChAdOx1 nCoV-19: 10.30)) ) C. The as 12.08)) F. The an 10.30)) I.p = 0.0096, median—comorbidities: 10.67; no comorbidities: 12.06)  , none of: 12.06) . This wa: 12.06) .p = 0.0180, median—COVID-19+: 1.673; COVID-19−: 0.7415) and CoronaVac  groups compared to naïve individuals (uninfected)    (n = 3).We next determined serum antibody levels of vaccinees with a history of COVID-19, infected with SARS-CoV-2 at any moment, before or during the study. Of the 22 infected individuals, 13 were infected with SARS-CoV-2 before any vaccine dose was administered and nine were infected after one, two, or three vaccine doses. Due to the reduced number of individuals who had the infection, we were unable to analyze them according to the period in which they presented COVID-19. For anti-RBD IgA, individuals who were infected with SARS-CoV-2 at any moment showed higher antibody levels at 4–6 months after the second dose of ChAdOx1 nCoV-19 (nfected) A. The sa: 3.446) B. The ev: 3.446) C. We did: 3.446)  for anti: 10.86) . At 1 mo: 10.88) . However2 = 0.5979, p = 0.0013)  D, after  0.0013) E,F.In this study, we investigated the specific humoral immune response against RBD and spike proteins after COVID-19 vaccination with three vaccine formulations: ChAdOx1 nCoV-19, BNT162b2, and CoronaVac. The data showed that, after the primary vaccination regimen (two-dose regimen), all vaccinees showed an increase in anti-RBD IgG and IgA and anti-spike IgG levels and were able to maintain these antibody levels for 4–6 months after two doses. BNT162b2 induced greater production of anti-RBD IgG and IgA and anti-spike IgG compared to ChAdOx1 nCoV-19. These findings are similar to those of Zhang et al. (2022), who detected higher anti-spike and anti-RBD IgG levels induced by mRNA vaccines compared to adenovirus and recombinant protein vaccines . The booThe longevity of the immune response induced by vaccines may vary according to the vaccine platform used, among other factors. We found positive levels of anti-RBD IgG and IgA and anti-spike IgG antibodies up to 4–6 months after the second dose for the three vaccine formulations evaluated. Similarly, in other studies using sera from individuals immunized with mRNA vaccines such as mRNA-1273 (Moderna), neutralizing and binding antibody activity against SARS-CoV-2 variants was detected six months after the primary vaccination regimen ,42. GreaThe use of booster doses has already proven to be essential in improving the immune response, especially against SARS-CoV-2 variants . Many stTo assess whether host factors, such as sex, age, and comorbidities, could influence the response to the vaccines, these factors were evaluated to observe changes in anti-RBD IgG and IgA and anti-spike IgG production since some host characteristics can interfere with the development of immune responses against SARS-CoV-2 . No diffThe next evaluation conducted according to biological characteristics was the effect of age on antibody production. Aging has already been shown to be responsible for a decrease in cellular immune function due to the process known as immunosenescence ,53. It cAlthough the presence of comorbidities may hinder the induction of immune responses by different vaccines ,57,58, iOur research demonstrated higher production of anti-RBD IgG and IgA in individuals with a history of SARS-CoV-2 infection compared to naïve individuals, during the collection period 4–6 months after the second dose, suggesting that the infection could have potentiated antibody production, especially for individuals in the CoronaVac group. This hypothesis can be corroborated by a study conducted by Padoan et al. (2021), which showed increased anti-S-RBD IgG levels in individuals previously exposed to SARS-CoV-2 and immunized with two doses of BNT162b2 vaccine compared to naïve individuals . Chua etSome limitations of our study include the small size of the cohort and the differences between age and sex in each vaccine group. These limitations could have influenced the analysis of antibody production, but most of the results obtained in this work, such as the longevity of the antibody response and higher antibody levels induced by the heterologous regimen, are similar to those of other studies ,44,45,61In conclusion, two doses of the BNT162b2, ChAdOx1 nCoV-19, and CoronaVac vaccines were sufficient to induce detectable levels of anti-RBD IgG and IgA and anti-spike IgG antibodies. The heterologous booster dose with BNT162b2 increased the levels of anti-RBD IgA and IgG and anti-spike IgG for CoronaVac and anti-RBD IgA for ChAdOx1 nCoV-19 vaccinees. In addition, individuals who presented the SARS-CoV-2 infection at any time during the study had higher anti-RBD IgA and IgG levels at 4–6 months after the second dose, compared to uninfected individuals. Further studies involving larger cohorts should be conducted to investigate and find answers to unresolved questions.


Correction to: Lossy Image Compression in a Preclinical Multimodal Imaging Study https://doi.org/10.1007/s10278-023-00800-5The corresponding author for this paper should be Andreas Walter; details are below.Andreas WalterCentre of Optical Technologies, Aalen University, Aalen, Germanyandreas.walter@hs-aalen.deThe original article has been corrected.