Journal: npj Viruses

Article Title: Experimental Usutu virus infection in Eurasian blackbirds ( Turdus merula )

doi: 10.1038/s44298-025-00133-w

Figure Lengend Snippet: a Weight loss is expressed in % of initial weight. The humane endpoint was set at 75% of initial weight (dashed line). Results averaged from three individuals infected with USUV Afr3 and Eu3 and two control individuals, bars represent SD. Statistically significant difference was observed between the Eu3 and ctrls group on 6 d.p.i. as indicated by the asterisk * p = 0.03. b Survival curves of blackbirds. No statistically significant difference was observed in survival among the three groups of blackbirds.

Article Snippet: USUV Afr3 (isolated in 2016 from Turdus merula , GenBank accession MH891847.1 ) and USUV Eu3 (isolated in 2017 from Turdus merula , GenBank accession MN122189.1 ) were grown and passaged three times on Vero cells (African green monkey kidney epithelial cells, ATCC CCL-81).

Techniques: Infection, Control

Journal: npj Viruses

Article Title: Experimental Usutu virus infection in Eurasian blackbirds ( Turdus merula )

doi: 10.1038/s44298-025-00133-w

Figure Lengend Snippet: GAMM fit on the individual blackbirds’ square-root transformed hourly-total moved distances (in meters) for each day. Due to the small sample size, we combined the Afr3 and Eu3 infected birds in one treatment group (USUV-infected), and compared these birds to the control group. For visualization purposes, the time of day is fixed to the average value within the total data set (whilst the model corrects for diurnal activity patterns). Intervals around the lines depict standard errors of the predicted daily values. The gray rectangle indicates the day on which the experimental infection took place.

Article Snippet: USUV Afr3 (isolated in 2016 from Turdus merula , GenBank accession MH891847.1 ) and USUV Eu3 (isolated in 2017 from Turdus merula , GenBank accession MN122189.1 ) were grown and passaged three times on Vero cells (African green monkey kidney epithelial cells, ATCC CCL-81).

Techniques: Transformation Assay, Infection, Control, Activity Assay

Journal: npj Viruses

Article Title: Experimental Usutu virus infection in Eurasian blackbirds ( Turdus merula )

doi: 10.1038/s44298-025-00133-w

Figure Lengend Snippet: Results averaged from three individuals infected with Afr3 and Eu3 viruses. a Levels of RNAemia expressed in Ct value. b Amount of virus shed through pharyngeal route expressed in Ct value, c Amount of virus shed through cloacal route expressed in Ct value. Error bars represent SD.

Article Snippet: USUV Afr3 (isolated in 2016 from Turdus merula , GenBank accession MH891847.1 ) and USUV Eu3 (isolated in 2017 from Turdus merula , GenBank accession MN122189.1 ) were grown and passaged three times on Vero cells (African green monkey kidney epithelial cells, ATCC CCL-81).

Techniques: Infection, Virus

Journal: npj Viruses

Article Title: Experimental Usutu virus infection in Eurasian blackbirds ( Turdus merula )

doi: 10.1038/s44298-025-00133-w

Figure Lengend Snippet: a The amount of infectious virus in several tissues collected postmortem from the infected blackbirds is expressed in TCID 50 /g of tissue. Results averaged from three individuals for USUV Afr3 and USUV Eu3 groups, bars represent SD. A statistically significant difference (* p = 0.02) was observed between USUV Afr3 titer (2,3 × 10 9 TCID 50 /g) and USUV Eu3 titer (1 × 10 8 TCID 50 /g). b The amount of viral RNA detected in the lung and pallium of infected blackbirds is expressed in Ct value. Results averaged from three individuals for Afr3 and Eu3 groups, bars represent SD. No statistically significant differences were observed in either of the two tissues (lung p = 0.1; pallium p = 0.8).

Article Snippet: USUV Afr3 (isolated in 2016 from Turdus merula , GenBank accession MH891847.1 ) and USUV Eu3 (isolated in 2017 from Turdus merula , GenBank accession MN122189.1 ) were grown and passaged three times on Vero cells (African green monkey kidney epithelial cells, ATCC CCL-81).

Techniques: Virus, Infection

Journal: npj Viruses

Article Title: Experimental Usutu virus infection in Eurasian blackbirds ( Turdus merula )

doi: 10.1038/s44298-025-00133-w

Figure Lengend Snippet: USUV-associated lesions (H&E) ( a , c , e , g ) and USUV tropism ( b , d , f , h ) in the main target organs of USUV Afr3-infected blackbirds. a , b Spleen of blackbird n.4: a small multifocal areas of necrosis in the spleen (arrows) and infiltration of plasma cells and mott cells (arrowheads). b USUV antigen in the cytoplasm of large mononucleated cells morphologically compatible with splenic macrophages ( c , d ) Liver of blackbird n.4: c diffuse vacuolization and pigment deposition in the cytoplasm of hepatocytes, multiple cells with pyknotic nuclei and hyper eosinophilic cytoplasm (single cell necrosis) (arrowheads). d USUV antigen in the cytoplasm of hepatocytes (arrowhead), endothelial lining of the sinusoids (arrow) and Kupffer’s cells (asterisks). e , f Brain, pallium of blackbird n.1: e multiple neurons undergoing acidophilic neuronal necrosis (arrowheads) and mild perivascular infiltration of mononucleated leukocytes (arrow). f USUV antigen in the cytoplasm of neurons (arrowheads). g , h Brain, cerebellum of blackbird n.1: g Multiple Purkinje cells in variable degree of degeneration from chromatolysis to acidophilic neuronal necrosis (arrowheads). h USUV antigen in the cytoplasm of Purkinje cells (arrowhead) and adjacent neurons of the molecular layer (arrow).

Article Snippet: USUV Afr3 (isolated in 2016 from Turdus merula , GenBank accession MH891847.1 ) and USUV Eu3 (isolated in 2017 from Turdus merula , GenBank accession MN122189.1 ) were grown and passaged three times on Vero cells (African green monkey kidney epithelial cells, ATCC CCL-81).

Techniques: Infection, Clinical Proteomics

Journal: npj Viruses

Article Title: Experimental Usutu virus infection in Eurasian blackbirds ( Turdus merula )

doi: 10.1038/s44298-025-00133-w

Figure Lengend Snippet: Eight Eurasian blackbirds were sampled on day -7 (blood, serum, pharynx and cloacal swabs and breast feathers); after sampling, the blackbirds were singularly housed in cages in isolators and left to acclimatize for 7 days and activity and health status were monitored. On day 0, blackbirds were either inoculated with USUV Afr3 ( n = 3), Eu3 ( n = 3), or sterile PBS ( n = 2). On 1, 3 and 5 days post inoculation (d.p.i.) blackbirds were sampled (blood, serum, pharynx and cloacal swabs and breast feathers). At the humane endpoint, blackbirds were euthanized, necropsied, and tissue samples were collected for histopathology, tissue titration and RT-qPCR. Created in BioRender, agreement licence number OO28BUTARJ.

Article Snippet: USUV Afr3 (isolated in 2016 from Turdus merula , GenBank accession MH891847.1 ) and USUV Eu3 (isolated in 2017 from Turdus merula , GenBank accession MN122189.1 ) were grown and passaged three times on Vero cells (African green monkey kidney epithelial cells, ATCC CCL-81).

Techniques: Sampling, Activity Assay, Sterility, Histopathology, Titration, Quantitative RT-PCR

Journal: Cell Death Discovery

Article Title: Bridging the marrow: a co-culture-platform of leukemia cells and MS5-derived stromal cells or adipocytes

doi: 10.1038/s41420-025-02631-5

Figure Lengend Snippet: A and B B-precursor ALL cell lines EU3, KOPN-8, RCH-ACV, Nalm6, Reh, and RS4;11 (each dot represents one cell line) were cultured alone or together with stromal cells or adipocytes derived from MS5 cells. Cell viability ( A ) and the number of viable cells per well ( B ) were determined after 7 days by FACS. C EU3 mono-culture was compared to co-culture with MS5-derived stromal cells or adipocytes cultured in DMEM/F12 medium without FCS, compared to medium with increasing concentrations of FCS as indicated. Viability and the number of viable cells per well were determined by FACS analysis after 7 days. D The B-precursor ALL cell line RS4;11 was labeled with CellTrace Violet (CT) and cultured alone or together with MS5-derived stromal cells or adipocytes, either directly or in transwell inserts. Viability and mean fluorescence intensity (MFI) of CellTrace Violet were determined on day 7 after seeding. E RS4;11 cells were co-cultured in transwell inserts over medium (mono-culture), MS5-derived stromal cells, or adipocytes for 72 h. Subsequently, ATP production was analyzed by the Seahorse Extracellular Flux Analyzer. F MS5-derived stromal cells (left panel) and adipocytes (right panel) were incubated with leukemia-conditioned media for 24 h and subsequently analyzed by the Seahorse Extracellular Flux Analyzer. ATP production rates are depicted. Data are presented as mean ± SEM ( A , B , D ) or as mean ± SD ( C , E , F ) of at least three independent experiments performed in triplicate ( A – C ) or duplicates ( D ) or in three to five technical replicates ( E , F ). RM one-way ANOVA with Tukey’s multiple comparisons test ( A , B ), two-way ANOVA with Dunnett’s ( C , E , F ) or Tukey’s ( D ) multiple comparisons test, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: B-precursor ALL cell lines Nalm6 (DSMZ no. ACC 128; Species: human), Reh (DSMZ no. ACC 22; Species: human), RS4;11 (DSMZ no. ACC 508; Species: human), RCH-ACV (DSMZ no. ACC 548; Species: human), EU3 (also known as cell line 697; DSMZ no. ACC 42; Species: human), and KOPN-8 (DSMZ no. ACC 552; Species: human) were maintained in RPMI (Gibco, Thermo Fisher Scientific) supplemented with 10–20% FCS and 2% L -glutamine.

Techniques: Cell Culture, Derivative Assay, Co-Culture Assay, Labeling, Fluorescence, Incubation

Journal: Cell Death Discovery

Article Title: Bridging the marrow: a co-culture-platform of leukemia cells and MS5-derived stromal cells or adipocytes

doi: 10.1038/s41420-025-02631-5

Figure Lengend Snippet: A – C PDX B-precursor ALL samples ( n = 24) were cultured alone or together with MS5-derived stromal cells or adipocytes. Viability ( A ), the number of viable cells per well ( B ), and mean fluorescence intensity (MFI) of CellTrace relative to day 0 ( C ) were determined after 7 days. D and E Three different human PDX samples (X07, X12, X14) were stimulated with the indicated concentrations of daunorubicin ( D ) or doxorubicin ( E ), and PI positivity was measured after 7 days. F B-precursor ALL cell lines EU3 and Nalm6 were stimulated with increasing concentrations of dexamethasone as indicated in co-culture with either MS5-derived stromal cells or adipocytes. Cell death was assessed by PI positivity at the FACS after 96 h. Data are presented as mean ± SD of individual PDX B-precursor ALL samples represented by single dots measured in triplicate ( A – E ) or of three independent experiments performed in triplicates ( F ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001; ordinary one-way ANOVA with Tukey’s multiple comparisons test ( A – C ), two-way ANOVA with Šídák’s multiple comparisons test ( D – F ).

Article Snippet: B-precursor ALL cell lines Nalm6 (DSMZ no. ACC 128; Species: human), Reh (DSMZ no. ACC 22; Species: human), RS4;11 (DSMZ no. ACC 508; Species: human), RCH-ACV (DSMZ no. ACC 548; Species: human), EU3 (also known as cell line 697; DSMZ no. ACC 42; Species: human), and KOPN-8 (DSMZ no. ACC 552; Species: human) were maintained in RPMI (Gibco, Thermo Fisher Scientific) supplemented with 10–20% FCS and 2% L -glutamine.

Techniques: Cell Culture, Derivative Assay, Fluorescence, Co-Culture Assay