Exploring microbial and moist-heat effects on Pu-erh tea volatiles and understanding the methoxybenzene formation mechanism using molecular sensory science.

Piling fermentation (PF) is crucial for Pu-erh tea aroma, yet its microbial and moist-heat impact on aroma quality is poorly understood. Solid-phase microextraction, solvent-assisted flavor evaporation, and gas chromatography-mass spectrometry were used to detected and analyses the samples of sun-green green tea, sterile PF and spontaneous PF. Microbiological action promotes the formation of stale aromas. Moist-heat action promotes the formation of plum-fragrance and sweet aroma. 20 microbial markers and 28 moist-heat markers were screened from 184 volatile components. Combining odor activity values and gas chromatography-olfactometry, 22 aroma-active compounds were screened (1,2,3-trimethoxybenzene, linalool, 1,2,4-trimethoxybenzene …), and analyzed during PF processing. Aroma omission and addition experiments verified its importance. Gallic acid addition experiments successfully verified that microorganisms are the main contributors to the synthesis of methoxybenzenes. Finally, Blastobotrys, Rasamsonia, and Thermomyces showed positive correlation with the synthesis of 1-ethyl-4-methoxybenzene, 1,2,4-trimethoxybenzene, 1,2,3-trimethoxybenzene, and 1,2-dimethoxybenzene. The formation mechanism of Pu-erh tea's aroma was clarified. Exploring microbial and moist-heat effects on Pu-erh tea volatiles and understanding the methoxybenzene formation mechanism using molecular sensory science.

Contribution of tea stems to large-leaf yellow tea aroma.

Large-leaf yellow tea (LYT) is processed from both leaves and stems, resulting in a distinctive rice crust-like aroma. Tea stems may contribute differently to the aroma of LYT than leaves. This study aimed to clarify the specific contribution of stems to LYT. The volatile compounds in different components of LYT were extracted and analyzed using a combination of headspace solid-phase microextraction and stir bar sorptive extraction coupled with gas chromatography-olfactory-mass spectrometry. The results revealed high concentrations of compounds with roasty attributes in stems such as 2-ethyl-3,5-dimethylpyrazine (OAV 153-208) and 2-ethyl-3,6-dimethylpyrazine (OAV 111-140). Aroma recombination and addition experiments confirmed that the roasty aroma provided by stems plays a pivotal role in the formation of the distinctive flavor of LYT. This study offers novel insights into the contribution of stems to the aroma of LYT, which can be used for processing and quality enhancement of roasted tea.

A bioactive calcium silicate nanowire-containing hydrogel for organoid formation and functionalization.

Organoids, which are 3D multicellular constructs, have garnered significant attention in recent years. Existing organoid culture methods predominantly utilize natural and synthetic polymeric hydrogels. This study explored the potential of a composite hydrogel mainly consisting of calcium silicate (CS) nanowires and methacrylated gelatin (GelMA) as a substrate for organoid formation and functionalization, specifically for intestinal and liver organoids. Furthermore, the research delved into the mechanisms by which CS nanowires promote the structure formation and development of organoids. It was discovered that CS nanowires can influence the stiffness of the hydrogel, thereby regulating the expression of the mechanosensory factor yes-associated protein (YAP). Additionally, the bioactive ions released by CS nanowires in the culture medium could accelerate Wnt/ß-catenin signaling, further stimulating organoid development. Moreover, bioactive ions were found to enhance the nutrient absorption and ATP metabolic activity of intestinal organoids. Overall, the CS/GelMA composite hydrogel proves to be a promising substrate for organoid formation and development. This research suggested that inorganic biomaterials hold significant potential in organoid research, offering bioactivities, biosafety, and cost-effectiveness.

Assessment of knowledge, attitudes, practices, and risk perceptions regarding COVID-19: a cross-sectional study from China.

Aim: To analyze the level of knowledge, attitudes, practices, and risk perception regarding COVID-19 among Chinese residents 1.5 years after the pandemic. Subject and methods: A cross-sectional study was carried out with both online and paper questionnaires. We included a variety of covariates that were characteristic-related factors such as age, gender, education level, and retirement status, as well as those closely associated with risk perception regarding COVID-19. Results: Participants (n = 3588), 53.49 ± 18.88 years old, from two provinces of China, of which 44.7% were male and 52.03% had a high school or greater level of education, answered the questions. More than 90% of participants had adequate background knowledge about COVID-19 and agreed or even strongly agreed with many attitude items related to the government's role in diagnosis, treatment, and dealing with COVID-19 infections. About three fifths of the participants reported fear of contracting COVID-19, but only a minority (18.63%) felt they were more susceptible than others. Respondents aged 45 years or younger were more likely to fear contracting the virus than those older than 45 years (adjusted OR = 1.464, 95% CI 1.196 to 1.794, P = 0.0002). High education level (adjusted OR = 1.503, 95% CI 1.187 to 1.904, P = 0.0007) and non-retired status (adjusted OR = 1.679, 95% CI 1.354 to 2.083, P < 0.0001) were associated with a higher perception of susceptibility to infection than others. Moreover, respondents who were not retired had a significantly reduced practice score (adjusted OR = 1.554, 95% CI 1.261 to 1.916, P < 0.0001). Age, retirement status, and education level were also associated with knowledge, attitude, and practice level. Conclusion: Our findings suggest that the public generally has trust in the COVID-19 vaccine and the government with regard to COVID-19 in China. We recommend that high-risk groups of communities, such as elders and patients with chronic diseases, be given greater consideration in the outbreaks. Health education campaigns combined with workplace preventive intervention should be aimed at improving COVID-19 knowledge and beliefs in order to encourage more optimistic attitudes and to maintain safe practices.

An integrated approach of high-performance liquid chromatography-mass spectrometry-based chemical profiling, network pharmacology, and molecular docking to reveal the potential mechanisms of Qishen Gubiao granules for treating coronavirus disease 2019.

Qishen Gubiao granules, a traditional Chinese medicine preparation composed of nine herbs, have been widely used to prevent and treat coronavirus disease 2019 with good clinical efficacy. In the present study, an integrated strategy based on chemical profiling followed by network pharmacology and molecular docking was employed, to explore the active components and potential molecular mechanisms of Qishen Gubiao granules in the therapy of coronavirus disease 2019. Using the ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry technique, a total of 186 ingredients corresponding to eight structure types in Qishen Gubiao preparation were identified or structurally annotated with the elucidation of the fragmentation pathways in the typical compounds. The network pharmacology analysis screened 28 key compounds including quercetin, apigenin, scutellarein, luteolin and naringenin acting on 31 key targets, which possibly modulated signal pathways associated with immune and inflammatory responses in the treatment of coronavirus disease 2019. The molecular docking results observed that the top 5 core compounds had a high affinity for angiotensin-converting enzyme 2 and 3-chymotrypsin-like protease. This study proposed a reliable and feasible approach for elucidating the multi-components, multi-targets, and multi-pathways intervention mechanism of Qishen Gubiao granules against coronavirus disease 2019, providing a scientific basis for its further quality evaluation and clinical application.

WITHDRAWN: Conjugation of Ruthenium Drugs to Nanocellulose using Boronic Ester

Since the authors are not responding to the editor's requests to fulfill the editorial requirement, therefore, the article has been withdrawn.Bentham Science apologizes to the readers of the journal for any inconvenience this may have caused.The Bentham Editorial Policy on Article Withdrawal can be found at https://benthamscience.com/editorial-policies-main.php. Bentham Science Disclaimer: It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneously submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submitting the article for publication the authors agree that the publishers have the legal right to take appropriate action against the authors, if plagiarism or fabricated information is discovered. By submitting a manuscript the authors agree that the copyright of their article is transferred to the publishers if and when the article is accepted for publication.

Dual drug delivery system of RAPTA-C and paclitaxel based on fructose coated nanoparticles for metastatic cancer treatment.

Ruthenium complexes have been widely studied as potential alternatives to platinum-type anticancer drugs due to their unique medical properties such as high selectivity, strong ability to inhibit solid tumour metastasis. However, non-specific biodistribution, and weak lethality of ruthenium to cancer cells limit its use in medical application. Drug delivery systems offer the ability to integrate multiple drugs in one system, which is particularly important to enhance the chemotherapeutic efficacy and to potentially achieve a synergistic effect of both drugs. Here, we report a dual drug nanocarrier that is based on a self-assembled biodegradable block copolymer, where the ruthenium complex (RAPTA-C) is chemically attached to the polymer chain, while another drug, paclitaxel (PTX), is entrapped in the core of the micelle. The dual drug delivery system was studied via in vitro tests using MDA-MB-231 breast cancer cells and it was observed that RAPTA-C in combination with PTX significantly enhanced anti-tumour and anti-metastasis activity.

Epidemiological survey and genetic characterization of type 3 vaccine-derived poliovirus isolated from a patient with four doses of inactivated polio vaccine in Henan Province, China.

BACKGROUND: Vaccine-derived poliovirus (VDPV) is a potential threat to polio eradication because they can reintroduce into the general population and cause paralytic polio outbreaks, a phenomenon that has recently emerged as a prominent public health concern at the end of global polio eradication. This study aimed to describe the epidemiology and genetic characteristics of the first VDPV identified from a patient with acute flaccid paralysis (AFP), with four doses of inactivated polio vaccine immunization in Henan Province, China in 2017. METHODS: The patient was diagnosed with type 3 VDPV. Subsequently, a series of epidemiological approaches was implemented, including a retrospective search of AFP cases, rate of vaccination assessment, study of contacts, and supplementary immunization activities. Fecal samples were collected, viral isolation was performed, and the viral isolates were characterized using full-length genomic sequencing and bioinformatic analysis. RESULTS: Phylogenetic analysis showed that the viral isolates from the patient were different from other reported genetic clusters of type 3 VDPV worldwide. They were identified as a Sabin 3/Sabin 1 recombinant VDPV with a crossover site in the P2 region. Nucleotide substitutions, including U → C (472) and C → U (2493), have been identified, both of which are frequently observed as reversion mutations in neurovirulent type 3 poliovirus. A unique aspect of this case is that the patient had been vaccinated with four doses of inactive polio vaccine, and the serum neutralizing antibody for Sabin types 1 and 3 were 1∶16 and 1∶512, respectively. Thus, the patient was speculated to have been infected with type 3 VDPV, and the virus continued to replicate and be excreted for at least 41 d. CONCLUSIONS: The existence of this kind of virus in human population is a serious risk and poses a severe challenge in maintaining a polio-free status in China. To the best of our knowledge, this is the first report of VDPV identified in the Henan province of China. Our results highlight the importance of maintaining a high-level vaccination rate and highly sensitive AFP case surveillance system in intercepting VDPV transmission.

Relative vaccine effectiveness against Delta and Omicron COVID-19 after homologous inactivated vaccine boosting: a retrospective cohort study.

ObjectiveTwo COVID-19 outbreaks occurred in Henan province in early 2022-one was a Delta variant outbreak and the other was an Omicron variant outbreak. COVID-19 vaccines used at the time of the outbreak were inactivated, 91.8%; protein subunit, 7.5%; and adenovirus5-vectored, 0.7% vaccines. The outbreaks provided an opportunity to evaluate variant-specific breakthrough infection rates and relative protective effectiveness of homologous inactivated COVID-19 vaccine booster doses against symptomatic infection and pneumonia. DESIGN: Retrospective cohort study METHODS: We evaluated relative vaccine effectiveness (rVE) with a retrospective cohort study of close contacts of infected individuals using a time-dependent Cox regression model. Demographic and epidemiologic data were obtained from the local Centers for Disease Control and Prevention; clinical and laboratory data were obtained from COVID-19-designated hospitals. Vaccination histories were obtained from the national COVID-19 vaccination dataset. All data were linked by national identification number. RESULTS: Among 784 SARS-CoV-2 infections, 379 (48.3%) were caused by Delta and 405 (51.7%) were caused by Omicron, with breakthrough rates of 9.9% and 17.8%, respectively. Breakthrough rates among boosted individuals were 8.1% and 4.9%. Compared with subjects who received primary vaccination series ≥180 days before infection, Cox regression modelling showed that homologous inactivated booster vaccination was statistically significantly associated with protection from symptomatic infection caused by Omicron (rVE 59%; 95% CI 13% to 80%) and pneumonia caused by Delta (rVE 62%; 95% CI 34% to 77%) and Omicron (rVE 87%; 95% CI 3% to 98%). CONCLUSIONS: COVID-19 vaccination in China provided good protection against symptomatic COVID-19 and COVID-19 pneumonia caused by Delta and Omicron variants. Protection declined 6 months after primary series vaccination but was restored by homologous inactivated booster doses given 6 months after the primary series.

Sulfonated polybenzothiazole cathode materials for Na-ion batteries.

A new flexible aromatic polymer sulfonated polybenzothiazole (sPBT-SE) with sulphone and ether units is reported as an advanced cathode material for storing Na+, which delivers a high discharge capacity of 103 mA h g-1 after 350 cycles at 30 mA g-1.

Phase Behavior of Ionic Liquid-Based Aqueous Two-Phase Systems.

As an environmentally friendly separation medium, the ionic liquid (IL)-based aqueous two-phase system (ATPS) is attracting long-term attention from a growing number of scientists and engineers. Phase equilibrium data of IL-based ATPSs are an important basis for the design and optimization of chemical reactions and separation processes involving ILs. This article provides the recent significant progress that has been made in the field and highlights the possible directions of future developments. The effects of each component (such as salting-out agents and ILs) on the phase behavior of IL-based ATPSs are summarized and discussed in detail. We mainly focus on the phase behavior of ATPSs by using ILs, expecting to provide meaningful and valuable information that may promote further research and application.

A case-case study on the effect of primary and booster immunization with China-produced COVID-19 vaccines on prevention of pneumonia and viral load among vaccinated persons infected by Delta and Omicron variants.

Using a three-prefecture, two-variant COVID-19 outbreak in Henan province in January 2022, we evaluated the associations of primary and booster immunization with China-produced COVID-19 vaccines and COVID-19 pneumonia and SARS-CoV-2 viral load among persons infected by Delta or Omicron variant. We obtained demographic, clinical, vaccination, and multiple Ct values of infections ≥3 years of age. Vaccination status was either primary series ≥180 days prior to infection; primary series <180 days prior to infection, or booster dose recipient. We used logistic regression to determine odds ratios (OR) of Delta and Omicron COVID-19 pneumonia by vaccination status. We analysed minimum Ct values by vaccination status, age, and variant. Of 826 eligible cases, 405 were Delta and 421 were Omicron cases; 48.9% of Delta and 19.0% of Omicron cases had COVID-19 pneumonia. Compared with full primary vaccination ≥180 days before infection, the aOR of pneumonia was 0.48 among those completing primary vaccination <180 days and 0.18 among booster recipients among these Delta infections. Among Omicron infections, the corresponding aOR was 0.34 among those completing primary vaccination <180 days. There were too few (ten) Omicron cases among booster dose recipients to calculate a reliable OR. There were no differences in minimum Ct values by vaccination status among the 356 Delta cases or 70 Omicron cases. COVID-19 pneumonia was less common among Omicron cases than Delta cases. Full primary vaccination reduced pneumonia effectively for 6 months; boosting six months after primary vaccination resulted in further reduction. We recommend accelerating the pace of booster dose administration.

Effectiveness of Inactivated COVID-19 Vaccines Against Symptomatic, Pneumonia, and Severe Disease Caused by the Delta Variant: Real World Study and Evidence - China, 2021.

WHAT IS ALREADY KNOWN ABOUT THIS TOPIC?: Effectiveness of China's 2 inactivated vaccines (BBIBP-CorV and CoronaVac) against pre-Delta severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) variants ranged from 47% to over 90%, depending on the clinical endpoint, and with greater effectiveness against more severe coronavirus disease 2019 (COVID-19). During an outbreak in Guangdong, inactivated vaccine effectiveness (VE) against the Delta variant was 70% for symptomatic infection and 100% for severe COVID-19. However, separate or combined VE estimates for the two inactivated vaccines against Delta are not available. WHAT IS ADDED BY THIS REPORT?: In an outbreak that started in a hospital, VEs of completed primary vaccination with inactivated COVID-19 vaccines against symptomatic COVID-19, COVID-19 pneumonia, and severe COVID-19 caused by the Delta variant were 51%, 61%, and 82%. Completed primary vaccination reduced the risk of progressing from mild to moderate or severe COVID-19 by 74%. VE estimates for BBIBP-CorV and CoronaVac or combined vaccination were similar, and partial vaccination was ineffective. WHAT ARE THE IMPLICATIONS FOR PUBLIC HEALTH PRACTICE?: Completed primary vaccination with either of the 2 inactivated COVID-19 vaccines reduces risk of symptomatic COVID-19, COVID-19 pneumonia, and severe COVID-19 caused by the Delta variant. Completion of the completed primary vaccination with two doses is necessary for protection from Delta.

Different epitopes of Ralstonia solanacearum effector RipAW are recognized by two Nicotiana species and trigger immune responses.

Diverse pathogen effectors convergently target conserved components in plant immunity guarded by intracellular nucleotide-binding domain leucine-rich repeat receptors (NLRs) and activate effector-triggered immunity (ETI), often causing cell death. Little is known of the differences underlying ETI in different plants triggered by the same effector. In this study, we demonstrated that effector RipAW triggers ETI on Nicotiana benthamiana and Nicotiana tabacum. Both the first 107 amino acids (N1-107 ) and RipAW E3-ligase activity are required but not sufficient for triggering ETI on N. benthamiana. However, on N. tabacum, the N1-107 fragment is essential and sufficient for inducing cell death. The first 60 amino acids of the protein are not essential for RipAW-triggered cell death on either N. benthamiana or N. tabacum. Furthermore, simultaneous mutation of both R75 and R78 disrupts RipAW-triggered ETI on N. tabacum, but not on N. benthamiana. In addition, N. tabacum recognizes more RipAW orthologs than N. benthamiana. These data showcase the commonalities and specificities of RipAW-activated ETI in two evolutionally related species, suggesting Nicotiana species have acquired different abilities to perceive RipAW and activate plant defences during plant-pathogen co-evolution.

Effects of carboxylated multi-walled carbon nanotubes on bioconcentration of pentachlorophenol and hepatic damages in goldfish.

Carboxylated multi-walled carbon nanotubes (MWCNT-COOH) exerts strong adsorption capacity for pentachlorophenol (PCP) and they inevitably co-occur in the environment, but few studies have characterized the effects of MWCNT-COOH on the bioavailability of PCP and its oxidative and tissue damages to fish. In this work, we assessed the PCP accumulation in different organs and the induced oxidative and tissue damages of goldfish following 50-d in vivo exposure to PCP alone or co-exposure with MWCNT-COOH. Our results indicated that PCP bioaccumulation in goldfish liver, gill, muscle, intestine and gut contents was inhibited after co-exposure with MWCNT-COOH in uptake phase. PCP exposure alone and co-exposure with MWCNT-COOH evoked severe oxidative and tissue damages in goldfish bodies, as indicated by significant inhibition of activities of antioxidant enzymes, remarkable decrease in glutathione level, simultaneous elevation of malondialdehyde content, and obvious histological damages to liver and gill. The decreased accumulation of PCP in the presence of MWCNT-COOH led to the reduction of PCP-induced toxicity to liver tissues, as confirmed by the alleviation of hepatic oxidative damages. However, co-exposure groups had higher concentrations of PCP in the tissues than PCP treatment alone (p < 0.05 each) in the depuration phase, revealing that MWCNT-COOH-bound pollutants might pose higher risk once desorbed from the nanoparticles. These results provided substantial information regarding the combined effects of PCP and MWCNT-COOH on aquatic species, which helps to deeply understand the potential ecological risks of the emerging pollutants.

Perfusion Cultivation of Artificial Liver Extracellular Matrix in Fibrous Polymer Sponges Biomimicking Scaffolds for Tissue Engineering.

A major challenge in tissue engineering and artificial scaffolding is to combine easily tunable scaffolds biomimicking the extracellular matrix of native organs with delivery-controlled cell culturing to create fully cellularized, large artificial 3D scaffolds. Aiming at bioartificial liver construction, we present our research using galactose-functionalized, ultraporous polylactide 3D nanofiber sponges fabricated out of electrospun fibers. Sponge biomodification by blend galactosylation and in-solution coating is performed, respectively, using a polylactide-galactose carrier-copolymer that promotes cell delivery and features a pronounced autofluorescence. It allows us to verify the galactosylation success, evaluate its quality, and record dye-free, high-resolution images of the sponge network using confocal laser scanning microscopy. The galactose carrier and its impact on scaffold cellularization is validated in benchmark to several reference systems. Verification of the human hepatic cell asialoglycoprotein receptor presence and galactose interaction in culture is performed by Cu2+ receptor-blocking experiments. The culture results are extensively investigated in and ex situ to trace and quantify the cell culture progress, cell activity, and viability at different culture stages. Bioreactor cultivation of sponges reveals that the galactose carrier does not only facilitate cell adhesion but also enhances cellular distribution throughout the scaffold. The promising 3D culture results allow us to move forward to create mature in vitro liver model research systems. The elaboration into ex vivo testing platforms could help judging native cell material interactions with drugs or therapeutics, without the need of direct human or animal testing.

The impact of universal live attenuated hepatitis A vaccines in Henan, China, 2005-2018.

OBJECTIVES: To analyze the effects of one dose of live attenuated hepatitis A vaccine in a developing country. METHODS: The reported cases of hepatitis A virus (HAV) infection from 2005 to 2018 in Henan province, China, were analyzed. Data of vaccinated children were assessed on the childhood immunization information management system. Questionnaire survey and blood sample collection were randomly conducted in six counties and districts of Henan province to analyze the prevalence of HAV lgG among the population aged 0-70 years. RESULTS: In 2008, Henan province began to expand its program on immunization, and children aged 18 months were given one dose of live attenuated hepatitis A vaccine (HepA-L). From 2005 to 2007, the HAV incidence remained steady at above 5000 cases per year and increased to 7489 in 2007. Since 2008, the HAV incidence decreased cumulatively from 4576 to 237 in 2018, indicating a 94.8% decrease, which was particularly pronounced among adolescents (98.2%). The proportion of hepatitis A cases in patients younger than 10 years continually decreased from 41.6% in 2012 to 3.8% in 2018. The reduction of reported cases older than 40 years was slower than that of children. In 2012, the proportion of hepatitis A cases older than 40 years was 27.6%, and continually increased to 69.2% (164/237) in 2018. The results of serological investigation showed that the 0-1.5-year age group had the lowest anti-HAV IgG prevalence (38.6%), which increased to 75.0% in the 4-6-year age group, covered by this immunization program. CONCLUSIONS: The data indicated a large decrease in HAV infections in Henan province from 2008 onward in response to the introduction of a planned immunization program of HepA-L.

Molecular interaction balanced one- and two-dimensional hybrid nanoarchitectures for high-performance supercapacitors.

Hybridisation of one-dimensional (1D) and two-dimensional (2D) materials to enhance charge storage has received considerable attention, but a fundamental understanding of the inherent ratio-dependent charge transfer mechanisms associated with the modulation of their molecular interactions is still within the community. Herein, we examined 1D surface oxidised carbon nanotubes (Ox-CNTs) and 2D reduced graphene oxide (rGO) to understand their ratio-dependent charge transfer and molecular interaction dynamics. We found that stepwise ultrasonication and the self-assembly process can control the thermodynamic molecular interactions, which result in rGO and Ox-CNT suspensions not only well dispersed in N,N-dimethylformamide but also self-organised into sandwiched nanoarchitectures. We reveal that the enhanced charge storage performance originated from the Ox-CNT-mediated low contact resistance between the active material and the current collector, and the incorporation of rGO leads to a significant ion diffusion coefficient and gives rise to numerous ion diffusion channels for high rate retention. Through a systematic electrochemical characterisation, we found that the GC5 : 5 hybrids (mass ratio of rGO to Ox-CNT) provide the best compromise-balance ratio between rGO and Ox-CNT for realising a champion energy density (9 W h kg-1) and power density (10 kW kg-1) beyond the state-of-the-art performance of the individual materials. Our results herald the advent of molecular level hybridisation of 1D-2D materials for high-performance electrochemical energy storage.

Correlation between Drug Loading Content and Biological Activity: The Complexity Demonstrated in Paclitaxel-Loaded Glycopolymer Micelle System.

Drug delivery carriers are now widely established because they can increase the therapeutic efficiency of drugs. In general, the aim in this field is to create effective carriers that have large amounts of drugs loaded to minimize drug carrier material that needs to be disposed of. However, there has been little attention so far in the literature on the effect of the amount of loaded drugs on the biological activity. In this paper, we are trying to answer the question of how the drug-loading content will affect the in vitro activity. We use two methods to load paclitaxel (PTX) into micelles based on the glycopolymer, poly(1- O-methacryloyl-ß-d-fructopyranose)- block-poly(methyl methacylate) (Poly(1- O-MAFru)35- b-PMMA145). In the one-step method, the drug is loaded into the particles during the self-assembly process. However, the size of nanoparticle increased with the PTX content from 26 to 50 nm, triggering enhanced cellular uptake by MCF-7 and MDA-MB-231, which was caused by changes in diameter size and not by changes in drug concentration. To keep the nanoparticle size constant, preformed micelles were loaded with PTX (two-step process). The increasing amount of loaded drug led to decreased cellular uptake and reduced cytotoxicity by the cancer cell lines. Small-angle neutron scattering and small-angle X-ray scattering, supported by transmission electron microscopy and dynamic light scattering, exposed the PTX location in the shell. This caused shrinkage of the shell and lower levels of shell hydration, resulting in lower cellular uptake and lower cytotoxicity. Upon the release of PTX, the shell regained its original level of hydration. We could show that because drug loading causes morphology changes, in either the shell or the size, it is impossible to separate the parameters that will influence the biological activity. Although the same phenomenon may not apply to every drug delivery system, it needs to be considered that except for the well-known parameters that affect cell uptake-size, shape, surface chemistry, type of nanoparticle, and presence of bioactive groups-the amount of loaded drugs might change the physicochemical parameters of the nanoparticle and thus the in vitro and potentially the in vivo outcomes.