Deciphering the toxic effects of polystyrene nanoparticles on erythropoiesis at single-cell resolution

Classified as probably not a substantial claim.

The pseudotime starting point was determined based on expression of myb, a known marker for HSCs in zebrafish erythropoiesis.

“The pseudotime starting point was determined based on the expression levels of the myb gene, a known marker for hematopoietic stem cells (HSCs) in zebrafish erythropoiesis (Zhang et al., 2021).”

• cites Zhang et al., 2021 sourceSupported

Classified as probably not a substantial claim.

Cell type annotation was performed by verifying expression of established marker genes from ZFIN.

“Cell type annotation was performed by verifying the expression of previously established marker genes using the Zebrafish Information Network (ZFIN, https://zfin.org/) (Bradford et al., 2022), resulting in the identification of 21 cell types:”

• cites Bradford et al., 2022 sourceSupported

May combine two claims.

The nanoscale size of plastics allows them to penetrate cellular membranes, disrupting growth, development, and metabolic processes in exposed organisms.

“The nanoscale size of these plastics allows them to easily penetrate cellular membranes, thereby disrupting growth, development, and metabolic processes in exposed organisms (Pedersen et al., 2020; Shen et al., 2019).”

• cites Pedersen et al., 2020 sourcePartly supported

  The Results (Transcriptomic analysis; page not provided) report altered developmental and metabolic pathways after 200 nm nanoplastic exposure in larval zebrafish. The paper does not show that nanoplastics easily penetrate cellular membranes or that membrane penetration causes those changes. Its Abstract also reports no effect on hatching or deformities, so the claim of disrupted growth and development is broader than the observed outcomes. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites Shen et al., 2019 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Extensive polystyrene microplastic contamination has been detected in river systems such as the Mulde and Elbe, primarily downstream of industrial sources.

“For example, extensive polystyrene microplastic contamination has been detected in river systems such as the Mulde and Elbe, primarily downstream of industrial sources (Laermanns et al., 2021).”

• cites Laermanns et al., 2021 sourcePartly supported

  The Results (section 3.2) identify many sampled spheres as PS or PS-DVB, and the Discussion (section 4.1) reports a remarkable amount downstream of the Elbe–Mulde confluence, most likely coming from the Mulde. But section 4.1 says the data cannot trace the particles to an industrial source, and section 4.2 describes overall microplastic concentrations as remarkably low; the claim overstates both the source attribution and the extent of contamination. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

Disruptions to erythropoiesis could lead to anemia-like symptoms, resulting in reduced physical performance, impaired feeding activity, and potential organ damage.

“These disruptions to erythropoiesis could lead to anemia-like symptoms, resulting in reduced physical performance, impaired feeding activity, and potential organ damage (Chiabrando et al., 2014).”

• cites Chiabrando et al., 2014 sourcePartly supported

  The sections on erythroid disorders of heme biosynthesis and erythropoietic protoporphyria report anemia associated with disrupted heme synthesis; the latter also describes tissue damage and possible liver failure in that specific disorder. The paper does not report reduced physical performance or impaired feeding activity, or establish that anemia itself causes the organ damage described. Page numbers are unavailable in our copy. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The definitive wave drives the formation of all blood cell types, establishing the complete hematopoietic system.

“The definitive wave drives the formation of all blood cell types, establishing the complete hematopoietic system (Hu & Jing, 2023).”

• cites Hu & Jing, 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

alas1 plays a critical role in the early stages of heme synthesis.

“including alas1, which plays a critical role in the early stages of heme synthesis (Chiabrando et al., 2014).”

• cites Chiabrando et al., 2014 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The primitive wave initiates the generation of blood cells to supply oxygen and establish the immune system.

“The primitive wave initiates the generation of blood cells to supply oxygen and establish the immune system (Davidson & Zon, 2004; Kulkeaw & Sugiyama, 2012).”

• cites Davidson & Zon, 2004 sourceNot checked — no full text of the source
• cites Kulkeaw & Sugiyama, 2012 sourcePartly supported

  Section 1 says the primitive wave generates erythrocytes and macrophages, and the Introduction explains that erythrocytes supply oxygen. The paper does not attribute establishment of the immune system as a whole to the primitive wave: Section 1 assigns production of lymphocytes and other blood cell types to the definitive wave. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The transient wave serves as an intermediate stage, producing some blood cells and leading to the emergence of temporary blood components.

“The transient wave serves as an intermediate stage, producing some blood cells and leading to the emergence of temporary blood components (Zhang et al., 2021).”

• cites Zhang et al., 2021 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Extensive research has investigated the toxic effects of polystyrene nanoparticles on aquatic organisms, primarily focusing on oxidative stress and inflammation.

“Extensive research has investigated the toxic effects of polystyrene nanoparticles on aquatic organisms, primarily focusing on oxidative stress and inflammation (Feng et al., 2022; Pei et al., 2022; Wu et al., 2022).”

• cites Feng et al., 2022 sourcePartly supported

  The Conclusions (page unavailable) report oxidative stress in zebrafish exposed to polystyrene nanoplastics and suggest that inflammation may contribute to DNA damage. The Introduction also discusses oxidative stress and inflammation, but largely in relation to microplastics. These passages do not establish that extensive research on polystyrene nanoparticles in aquatic organisms has primarily focused on both responses. (LLM, conf=0.82)

  Experimental — accuracy not yet measured
• cites Pei et al., 2022 sourceNot checked — no full text of the source
• cites Wu et al., 2022Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

Reduced RBC levels impair oxygen transport, leading to diminished physical capabilities, impaired feeding behaviors, increased organ damage, and compromised immune systems.

“Reduced RBC levels impair oxygen transport, leading to diminished physical capabilities, impaired feeding behaviors, increased organ damage, and compromised immune systems (Canny et al., 2023; Ekiz et al., 2005).”

• cites Canny et al., 2023 sourcePartly supported

  The Abstract says healthy RBC numbers are critical for tissue oxygenation, and the Overview of Red Blood Cell Homeostasis defines anemia as insufficient healthy RBCs. Our copy does not link reduced RBC levels to the claimed changes in physical capability, feeding behavior, organ damage, or immune function. No page numbers are provided for these passages. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites Ekiz et al., 2005 sourceNot checked — no full text of the source

May combine two claims.

HR-TEM analysis revealed well-defined, spherical polystyrene nanoparticles with diameters ranging from 10 to 20 nm (average 14.3 nm).

“HR-TEM analysis of the polystyrene nanoparticles revealed well-defined, spherical nanoparticles with diameters ranging from 10 to 20 nm (average size=14.3 nm) (Figure 1C), consistent with the values obtained from DLS measurements.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Cell-type annotation identified 20 distinct cell types.

“Cell-type annotation using established markers identified 20 distinct cell types (Supplementary Figure S1B–D).”

May combine two claims.

rps7 knockdown embryos exhibited facial malformations and pericardial edema.

“treated embryos exhibited facial malformations and pericardial edema (Figure 3G).”

May combine two claims.

Polystyrene nanoparticle exposure decreased mature erythrocytes and increased immature erythrocytes in zebrafish embryos.

“revealing that polystyrene nanoparticle exposure disrupted erythrocyte differentiation, as evidenced by the decrease in mature erythrocytes and concomitant increase in immature erythrocytes.”

• AssumptionRests on an assumption: The gata1:DsRed transgenic marker identifies undifferentiated erythrocytes.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The cell types representing over 5% of total cell population included mesenchymal cells (n=5,439), keratinocytes (n=2,790), epithelial cells (n=2,052), pharynx cells (n=1,619), RBCs (n=1,475), fin cells (n=1,389), and notochord cells (n=1,061).

“Subsequent analyses focused on cell types representing over 5% of the total cell population (Supplementary Figure S1E), which included mesenchymal cells (n=5 439), keratinocytes (n=2 790), epithelial cells (n=2 052), pharynx cells (n=1 619), RBCs (n=1 475), fin cells (n=1 389), and notochord cells (n=1 061) (Figure 3A)”

May combine two claims.

Trajectory inference identified distinct stages of RBC differentiation, with the RBC1 cluster representing early differentiation and the RBC7 cluster representing final differentiation.

“Trajectory inference identified distinct stages of RBC differentiation, with the RBC1 cluster representing early differentiation and the RBC7 cluster representing final differentiation (Supplementary Figure S3A).”

The quote only partly states this claim.

DEGs in the pharynx cluster were identified between the CTRL and PS20 nm groups.

“DEGs in the pharynx cluster were identified between the two groups (Supplementary Table S1).”

Classified as probably not a substantial claim. May combine two claims.

Quality control retained 20,159 cells, including 10,184 from CTRL and 9,975 from PS20 nm-exposed group.

“Quality control analysis of single-cell RNA sequencing data using the Seurat package retained 20 159 cells, including 10 184 from the CTRL group and 9 975 from the PS20 nm-exposed group (Supplementary Figure S1A).”

May combine two claims.

DLS analysis showed polystyrene nanoparticles had a uniform size distribution with an average particle size of 16.7 nm.

“DLS analysis demonstrated that the polystyrene nanoparticles exhibited a uniform size distribution, with an average particle size of 16.7 nm (Figure 1A).”

May combine two claims.

Zebrafish embryos exposed to polystyrene nanoparticles showed physiological alterations including increased eye distance and elevated heart rate, but no severe morphological changes.

“Although no severe morphological changes were observed, physiological alterations, including increased eye distance and elevated heart rate, were detected (Figure 2B–G).”

May combine two claims.

In PS20 nm-exposed zebrafish embryos, neither p53 nor TNF-α expression was elevated.

“Interestingly, in PS20 nm-exposed zebrafish embryos, neither p53 nor TNF-α expression was elevated (data not shown), which are hallmark mechanisms of DBA.”

May combine two claims.

Comparison of cell proportions between CTRL and PS20 nm-exposed groups revealed a substantial increase in CMP-like cells and a significant reduction in the proportion of mature erythrocytes in the PS20 nm-treated group.

“A comparison of cell proportions between the CTRL and PS20 nm-exposed groups revealed a substantial increase in CMP-like cells in the PS20 nm-treated group, accompanied by a significant reduction in the proportion of mature erythrocytes (Figure 4D; Supplementary Figure S3C).”

May combine two claims.

Genes involved in translation in the RBC cluster showed marked down-regulation, correlating with reduced expression of genes crucial for erythropoiesis, such as rps7.

“Specifically, genes involved in translation in the RBC cluster showed marked down-regulation, correlating with reduced expression of genes crucial for erythropoiesis, such as rps7 (Figure 3D–H).”

Probably not a substantial claim: may not pass S4 (based).

Polystyrene nanoparticles are likely to interact with red blood cells during circulation.

“Given that polystyrene nanoparticles are likely to interact with red blood cells (RBCs) during circulation, these cells may also be vulnerable to polystyrene-induced toxicity.”

Classified as probably not a substantial claim.

Quality control measures excluded low-quality cells based on specified criteria.

“Quality control measures were implemented to exclude low-quality cells for subsequent analyses. The criteria for exclusion were: (1) Genes expressed in fewer than 12 cells (~0.1% of the data) and cells with fewer than 300 detected genes; (2) Cells expressing more than 3 300 or fewer than 300 unique genes; and (3) Cells with mitochondrial gene content exceeding 10%.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Nanoplastics can infiltrate biological systems with great efficiency, accumulating in aquatic species such as fish, crustaceans, and shellfish.

“Notably, nanoplastics can infiltrate biological systems with great efficiency, accumulating in aquatic species such as fish, crustaceans, and shellfish.”

• cites Timilsina et al., 2023 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

All procedures followed IACUC guidelines from Pusan National University.

“All procedures followed the guidelines of the Institutional Animal Care and Use Committee (IACUC) from Pusan National University (PNU-2023-0359).”

Probably not a substantial claim: may not pass S3 (load-bearing).

All experiments were performed in triplicate.

“All experiments were performed in triplicate.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

In Nile tilapia, 0.1 μm polystyrene nanoparticles accumulate in the gills at levels 3.4-fold higher than 0.5 μm polystyrene nanoparticles, with approximately 0.8 μg per gram detected in the gills.

“For instance, in Nile tilapia (Oreochromis niloticus), 0.1 µm polystyrene nanoparticles accumulate in the gills at levels 3.4-fold higher than those of 0.5 µm polystyrene nanoparticles, with approximately 0.8 µg per gram detected in the gills (Wang et al., 2023).”

• cites Wang et al., 2023 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Polystyrene's aromatic ring structure renders it more toxic than polyethylene or polyethylene terephthalate.

“However, its aromatic ring structure renders it more toxic than polyethylene or polyethylene terephthalate, amplifying its potential ecological impact (Alimi et al., 2022).”

• cites Alimi et al., 2022Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S4 (based).

Red blood cells may be vulnerable to polystyrene-induced toxicity.

“Given that polystyrene nanoparticles are likely to interact with red blood cells (RBCs) during circulation, these cells may also be vulnerable to polystyrene-induced toxicity.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Smaller nanoparticles exhibit enhanced bioavailability and organ penetration compared to larger nanoparticles.

“Studies have indicated that nanoparticle size plays a critical role in their absorption and distribution within tissues, with smaller particles exhibiting enhanced bioavailability and organ penetration (Wang et al., 2023).”

• cites Wang et al., 2023 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

RT-qPCR results were normalized to β-actin as an endogenous control.

“The expression levels of the target genes were analyzed using the comparative threshold method, and the results were normalized to β‐actin as an endogenous control.”

May combine two claims.

This study revealed that exposure to polystyrene nanoparticles inhibits translation and heme synthesis, leading to the formation of immature erythrocytes.

“Nonetheless, this study revealed that exposure to polystyrene nanoparticles inhibits translation and heme synthesis, leading to the formation of immature erythrocytes.”

May combine two claims.

These findings provide compelling evidence that polystyrene nanoparticles inhibit heme biosynthesis, leading to impaired erythropoiesis.

“These findings provide compelling evidence that polystyrene nanoparticles inhibit heme biosynthesis, leading to impaired erythropoiesis.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Previous studies investigated mammalian models exposed to micro-/nanoplastics for 42 days, identifying erythrocyte reductions linked to changes in HSCs mediated by interactions with gut microbiota and cytokines.

“For example, (Jing et al., 2022) investigated mammalian models exposed to micro-/nanoplastics for 42 days, identifying erythrocyte reductions linked to changes in HSCs mediated by interactions with gut microbiota and cytokines.”

• cites Jing et al., 2022 sourceNot checked — no full text of the source

May combine two claims.

Polystyrene nanoparticles directly impacted translation-related pathways and RBC differentiation.

“Instead, polystyrene nanoparticles directly impacted translation-related pathways and RBC differentiation.”

May combine two claims.

Erythrocyte differentiation was arrested at the CMP stage in PS20 nm-exposed zebrafish embryos, resulting in the accumulation of immature erythrocytes and disrupted erythrocyte differentiation.

“Results revealed that erythrocyte differentiation was arrested at the CMP stage (Figure 4D–H), resulting in the accumulation of immature erythrocytes and disrupted erythrocyte differentiation.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Nanoparticles with diameters of approximately 20 nm are particularly suitable for toxicity studies due to their ability to cross biological membranes and penetrate cells, enabling accurate assessment of their toxicological effects.

“Nanoparticles with diameters of approximately 20 nm are particularly suitable for toxicity studies due to their ability to cross biological membranes and penetrate cells, enabling accurate assessment of their toxicological effects (Ding et al., 2024).”

• cites Ding et al., 2024Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

These findings provide insights into how polystyrene nanoparticles disrupt erythrocyte differentiation, leading to a reduction in erythrocytes.

“These findings provide insights into how polystyrene nanoparticles disrupt erythrocyte differentiation, leading to a reduction in erythrocytes, and emphasize the importance of examining the effects of nanoplastic exposure on erythrocyte differentiation under varied conditions.”

Classified as probably not a substantial claim.

Concentrations of microplastics and nanoplastics in natural water and marine environments typically range from 6.35×10−3 to 6.94×103 μg/L.

“The concentrations of microplastics and nanoplastics in natural water and marine environments are typically reported to range from 6.35×10−3 to 6.94×103 μg/L.”

• cites Wang et al., 2023 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Hematopoiesis in zebrafish embryos occurs in three distinct waves: primitive, transient, and definitive.

“Hematopoiesis in zebrafish embryos occurs in three distinct waves: primitive, transient, and definitive (Davidson & Zon, 2004).”

• cites Davidson & Zon, 2004 sourceNot checked — no full text of the source

May combine two claims.

Polystyrene nanoparticles induce physiological stress responses, even in the absence of overt morphological defects, pointing to potential sub-lethal toxicity.

“These findings suggest that polystyrene nanoparticles induce physiological stress responses, even in the absence of overt morphological defects, pointing to potential sub-lethal toxicity.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Prior research reported gut cell damage after prolonged exposure to 500 ng/mL polystyrene nanoparticles for 28 days.

“While prior research has reported gut cell damage after prolonged exposure to 500 ng/mL polystyrene nanoparticles for 28 days (Deng et al., 2023), our study demonstrated significant effects on pharynx cells and RBCs within just 48 h of exposure (Figure 3B).”

• cites Deng et al., 2023 sourceNot checked — no full text of the source

May combine two claims.

This study validated results from single-cell RNA sequencing through in vivo analysis, confirming disruptions at the mRNA level.

“This study validated results from single-cell RNA sequencing through in vivo analysis, confirming disruptions at the mRNA level.”

Classified as probably not a substantial claim.

Laboratory studies often employ higher concentrations (1-100 mg/L) to assess toxicological effects.

“However, laboratory studies often employ higher concentrations (1–100 mg/L) to assess the toxicological effects (Wang et al., 2023).”

• cites Wang et al., 2023 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Smaller nanoparticles exhibit a higher surface area-to-volume ratio, enhancing their chemical reactivity, biological interactions, cellular uptake, and tissue penetration.

“Smaller nanoparticles exhibit a higher surface area-to-volume ratio, enhancing their chemical reactivity, biological interactions, cellular uptake, and tissue penetration (Jin et al., 2019; Shin et al., 2023; Teng et al., 2022).”

• cites Jin et al., 2019 sourceNot checked — no full text of the source
• cites Shin et al., 2023 sourceNot checked — no full text of the source
• cites Teng et al., 2022 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

As a result, the hematological disruptions induced by polystyrene nanoparticles may have far-reaching consequences that extend beyond individual organisms, impacting the ecosystem as a whole.

“As a result, the hematological disruptions induced by polystyrene nanoparticles may have far-reaching consequences that extend beyond individual organisms, impacting the ecosystem as a whole.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

These physiological impairments may cascade into reduced reproduction and survival rates, disrupting population dynamics and ecosystem balance.

“These physiological impairments may cascade into reduced reproduction and survival rates, disrupting population dynamics and ecosystem balance.”