Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A ) Timeline of chronic recordings. ( B ) Imaging and stimulation setups. Widefield and two-photon imaging was performed on the implanted hemisphere, and visual stimulation was applied to the contralateral eye. ( C ) Example widefield images of neural (Ca 2+ dF/F 0 ) and OEF hemodynamic responses induced by 25-Hz ICMS. Scale bar = 500 µm. ( D and E ) Time course (left) and distance profile (right; measured at 5 [red] and 20 [blue] seconds after stimulation onset) of induced neural (D) and hemodynamic (E) responses. Mean ± SD across four trials. Shaded bar indicates stimulation period (0-10 s); vertical dashed lines mark the time points of distance profiles.

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques: Imaging

Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A ) Coefficient of variation (CV, pixel-wise standard deviation normalized by mean over time) of widefield Ca 2+ (i) and hemodynamics (ii) during pre-stimulation periods at day 0 (top) and day 14 (bottom). ( B ) CV (top) and silencing index relative to 350 µm (SI 350 = [CV d -CV 350 ]/CV 350 , where d = 50, 150, 250, 350, or 450 μm) during pre-stimulation periods (bottom) across distances and imaging days. ( C ) CV map during visual stimulation. ( D ) CV and SI 350 during visual stimulation. * in SI 350 plots indicate significant differences from 0 (one-sample t-test with Bonferroni correction, p < 0.05; colors correspond to days after implantation).

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques: Standard Deviation, Imaging

Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A-C ) Example colormaps from a single mouse showing Ca 2+ responses during ( A ) 25-Hz ICMS, ( B ) 250-Hz ICMS, and ( C ) visual stimulation at (i and ii) day 0 and (iii and iv) day 21. Only increases from pre-stimulation baseline are shown in red. Corresponding time courses within 500 µm from the stimulation site are shown in (v) (mean ± SD across 4 trials). ( D-F ) Chronic comparisons of Ca 2+ activation magnitude (D), spatial spread (E), and duration (F) across days post-insertion. Mean ± SEM across mice. * next to line plot indicates a significant effect of day (LME model).

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A ) Example Ca 2+ response images at days 0 and 7 under (i) 25-Hz ICMS, (ii) 250-Hz ICMS, and (iii) visual stimulation. Corresponding time courses are shown in (iv; mean ± SEM across cells for soma, mean ± SEM across repeats for neuropil; scale bars = 5 s and 5%). ( B and C ) Chronic comparisons of somatic activation magnitudes (B) and durations (C). ( D and E ) Somatic depression magnitude (D) and duration (E) across days. ( F and G ) Neuropil activation magnitude (F) and duration (G) across days. ( H and I ) Neuropil depression magnitude (H) and duration (I) across days. Mean ± SEM across cells (B-E) or mice (F-I); “<“ and “>“ indicate significant differences compared with days 0 and 84, respectively (LME model followed by Welch’s t-test with Holm-Bonferroni correction).

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A-C ) Example colormaps of a single mouse showing Ca 2+ responses after ( A ) 25-Hz ICMS, ( B ) 250-Hz ICMS, and ( C ) visual stimulation at (i and ii) day 0 and (iii and iv) day 21. Only decreases from pre-stimulation baseline are shown in blue; darker blue indicates stronger depression. Corresponding time courses within 500 µm from the stimulation site are shown in (v) (mean ± SD across 4 trials). Traces during the stimulation period (0-10 s) are omitted for clarity. ( D-F ) Chronic comparisons of Ca 2+ depression magnitude (D), spatial spread (E), and duration (F) across days post-insertion. Mean ± SEM across mice.

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques:

Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A-C ) Example OEF response colormaps from a single mouse during and after ( A ) 25-Hz ICMS, ( B ) 250-Hz ICMS, and ( C ) visual stimulation at (i and ii) day 0 and (iii and iv) day 21. Corresponding time courses within 500 µm of the stimulation site are shown in (v; mean ± SD across 4 trials). OEF represents deoxyhemoglobin relative to total hemoglobin (see 2.4.4), where decreases from baseline (0.25) indicate relative increases in oxyhemoglobin, reflecting enhanced metabolic supply. ( D-F ) Chronic comparisons of OEF magnitude (D), spatial spread (E), and duration (F) across days post-implantation. Mean ± SEM across mice. * next to line plot indicates a significant effect of day (LME model).

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques:

Journal: bioRxiv

Article Title: Chronic alteration of Ca 2+ and hemodynamic signals induced by intracortical microstimulation in the visual cortex of awake mice

doi: 10.1101/2025.10.23.679242

Figure Lengend Snippet: ( A ) Example epileptiform Ca 2+ activity induced by 25-Hz ICMS, showing extremely strong activation spreading across the entire visual cortex and persisting >10 s after stimulation offset (i-v, maps; vi, time courses at each distance). ( B ) Overview of Ca 2+ activity induced by 25-Hz ICMS (i), 250-Hz ICMS (ii), and visual stimulation (iii). Each line represents one mouse. ( C ) Distributions of magnitude, duration, and spatial spread of Ca 2+ activation. Colors indicate clusters identified using agglomerative clustering (see Methods). Epileptiform activity was defined from the green cluster with magnitude >100%, duration >20 s, and distance >800 µm. ( D ) Number of mice exhibiting epileptiform activity at least once across all days, conditions, and repeats (Epi) versus not (Norm). ( E ) Probability of epileptiform activity across stimulation conditions for all days, mice, and repeats. ( F ) Probability of epileptiform activity over days for all ICMS and visual stimulation conditions. Mean ± SEM across animals.

Article Snippet: Custom stimulation microelectrodes (Q1×4-3mm-50-703-CM16LP, NeuroNexus Technologies, single 3-mm-long shank, 30-μm-diameter four channels, 50-μm site spacing, electrochemically activated iridium/iridium oxide sites ) were implanted at an angle of 20-30 degrees slanted towards the posterior region.

Techniques: Activity Assay, Activation Assay

Journal: Cell Death & Disease

Article Title: Mesenchymal stem cells infected with Mycoplasma arginini secrete complement C3 to regulate immunoglobulin production in b lymphocytes

doi: 10.1038/cddis.2014.147

Figure Lengend Snippet: MSCs inhibited IgE production in B cells. ( a ) Purified B cells secreted IgE in the presence of LPS (10 μ g/ml) and IL-4 (30 ng/ml). When MSCs were co-cultured, IgE secretion from B cells was significantly decreased. IgE downregulation was observed in the absence of cell–cell contact between MSCs and B cells in a transwell system. CM harvested from MSCs after 3-day cultivation also markedly inhibited IgE production. Extracellular protein levels of IgE were quantitatively measured by ELISA. ( b ) LPS-IL-4-mediated IgE induction was significantly suppressed by serial dilution of MSC-CM. NC, negative control with no stimulus; CC, co-culture; TW, transwell system; CM, MSC-CM

Article Snippet: B cells were stimulated with 10 μ g/ml LPS (Sigma) and 30 ng/ml mouse IL-4 (Prospec-TanyTechnogene, Rehovot, Israel).

Techniques: Purification, Cell Culture, Enzyme-linked Immunosorbent Assay, Serial Dilution, Negative Control, Co-Culture Assay

Journal: Cell Death & Disease

Article Title: Mesenchymal stem cells infected with Mycoplasma arginini secrete complement C3 to regulate immunoglobulin production in b lymphocytes

doi: 10.1038/cddis.2014.147

Figure Lengend Snippet: CM harvested from mycoplasma-free MSCs did not inhibit IgE production in B cells. ( a ) In addition to IgE, production of other Ig isotypes including IgM and IgG1 was also decreased by mycoplasma-contaminated MSC-CM in LPS/IL-4-stimulated B cells. However, Ig production (IgE, IgG1, and IgM) was not observed when mycoplasma-free MSC-CM was added. ( b ) Mycoplasma infection was determined by PCR. myco(+), CM from mycoplasma-infected MSCs; myco(−), CM from mycoplasma-free MSCs

Article Snippet: B cells were stimulated with 10 μ g/ml LPS (Sigma) and 30 ng/ml mouse IL-4 (Prospec-TanyTechnogene, Rehovot, Israel).

Techniques: Infection

Journal: Cell Death & Disease

Article Title: Mesenchymal stem cells infected with Mycoplasma arginini secrete complement C3 to regulate immunoglobulin production in b lymphocytes

doi: 10.1038/cddis.2014.147

Figure Lengend Snippet: M. arginini specifically downregulated IgE production in B cells. ( a ) To estimate the minimal numbers of infecting mycoplasma required to infect host cells, two cell types including MDF and MSC were infected with 10–80 cfu/ml of cultured M. arginini . MDF was included because it is a type of fibroblast without stem cell properties. M. arginini -specific PCR was performed for validity of infection. ( b ) IgE levels in LPS/IL-4-stimulated B cells were measured after incubation with CMs harvested from mycoplasma-free cells or from M. arginini -infected cells. In parallel, M. arginini infection to MDF or MSC was determined by PCR. ( c ) Several numbers of M. arginini (1, 2, 4, and 20 cfu/ml) were directly added to LPS/IL-4-stimulated B cells and then secreted IgE concentration was measured by ELISA. Significant existence of M. arginini in CM was determined by PCR. ( d ) After M. arginini (20 cfu/ml) was added to LPS/IL-4-stimulated B cells, IgG1 and IgM levels were determined by ELISA. myco(+), CM from mycoplasma-infected MDFs or MSCs; myco(−), CM from mycoplasma-free MDFs or MSCs, Ma, M. arginini

Article Snippet: B cells were stimulated with 10 μ g/ml LPS (Sigma) and 30 ng/ml mouse IL-4 (Prospec-TanyTechnogene, Rehovot, Israel).

Techniques: Infection, Cell Culture, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Mesenchymal stem cells infected with Mycoplasma arginini secrete complement C3 to regulate immunoglobulin production in b lymphocytes

doi: 10.1038/cddis.2014.147

Figure Lengend Snippet: MSC-CM lacking M. arginini still inhibited IgE production in B cells. ( a ) CM harvested from M. arginini -infected MSCs was serially diluted up to 25 600-fold and added to LPS/ IL-4-stimulated B cells. Each dilution in hexaplicates was subjected to IgE measurement. Relative IgE concentrations over 90% and under 40% to positive control (IgE in LPS/IL-4-stimulated B cells with no CM) were shown in white (○) and black (●) circles, respectively. Existence of M. arginini was determined by M. arginini -specific PCR reactions. ( b ) Mycoplasma-infected MSC-CM was filtrated through a 0.22- μ m syringe filter. The filtrate and non-filtered MSC-CM were added to LPS/IL-4-stimulated B cells and then IgE was measured by ELISA. M. arginini -specific PCR was performed. myco(+), CM from mycoplasma-infected MSCs

Article Snippet: B cells were stimulated with 10 μ g/ml LPS (Sigma) and 30 ng/ml mouse IL-4 (Prospec-TanyTechnogene, Rehovot, Israel).

Techniques: Infection, Positive Control, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Mesenchymal stem cells infected with Mycoplasma arginini secrete complement C3 to regulate immunoglobulin production in b lymphocytes

doi: 10.1038/cddis.2014.147

Figure Lengend Snippet: M. arginini -infected MSCs secreted soluble factors inhibiting IgE production in B cells. ( a ) Mycoplasma-infected MSC-CM was heat-inactivated by boiling for 5 min (boiled CM). The effect of boiled CM on IgE production in B cells was assessed by ELISA. ( b ) MSCs were treated with 2 μ M monensin (Mon) or vehicle for 24 h and then CMs were harvested. Each CM was added to LPS/IL-4-stimulated B cells followed by IgE measurement. ( c ) Mycoplasma-infected MSC-CM was size-fractionated and then added to LPS/IL-4-induced B cells. Secreted IgE levels were determined by ELISA. ( d ) FPLC was performed with 200-fold concentrated M. arginini -infected MSC-CM, MDF-CM, and medium control. ( e ) After FPLC fractions (from fraction numbers 8 to 25) were added to LPS/IL-4-stimulated B cells, the effect of each fraction on IgE downregulation was examined. ( f ) ELISA was conducted to assess the effects of M. arginini -infected MSC-CM fractions (11–14), MDF-CM fractions (11–14), and medium fractions on IgE downregulation

Article Snippet: B cells were stimulated with 10 μ g/ml LPS (Sigma) and 30 ng/ml mouse IL-4 (Prospec-TanyTechnogene, Rehovot, Israel).

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Mesenchymal stem cells infected with Mycoplasma arginini secrete complement C3 to regulate immunoglobulin production in b lymphocytes

doi: 10.1038/cddis.2014.147

Figure Lengend Snippet: C3 secreted from M. arginini -infected MSC-CM was responsible for Ig downregulation in B cells. ( a ) Secreted C3 level in M. arginini -infected MSC-CM was quantitatively measured by ELISA. C3 secretion was apparent only in mycoplasma-infeced MSCs. C3 was not detected in B-cell-CM (B-CM) regardless of mycoplasma infection. ( b ) When mouse C3 (10 ng/ml) was added to LPS/IL-4-stimulated B cells, it significantly inhibited IgE production as mycoplasma-infected MSC-CM. IgE downregulation was abrogated when boiled C3 was added to the culture. ( c ) Elevated production of IgG1 and IgM in LPS/IL-4-stimulated B cells was also reduced with C3 treatment (10 ng/ml). ( d ) Compstatin suppressed IgE downregulation by mycoplasma-infected MSC-CM in a dose-dependent manner. ( e ) IgE production inhibited by a FPLC fraction number 13 was restored by compstatin. ( f ) Compstatin treatment restored the reduced production of IgG1 and IgM by mycoplasma-infected MSC-CM. ( g ) Blimp-1 expression in LPS/IL-4-stimulated B cells was examined by semi-quantitative RT-PCR. In B cells, Blimp-1 expression induced by LPS/IL-4 stimulation was not observed by addition of mycoplasma-infected MSC-CM or by C3 treatment. In the presence of compstatin, Blimp-1 downregulation by the MSC-CM was restored. It is likely that boiled C3 treatment does not downregulate Blimp-1 expression. myco(+), CM from mycoplasma-infected MSCs or B cells; myco(−), CM from mycoplasma-free MSCs or B cells. Significance was * P <0.01, ** P <0.001, or *** P <0.01

Article Snippet: B cells were stimulated with 10 μ g/ml LPS (Sigma) and 30 ng/ml mouse IL-4 (Prospec-TanyTechnogene, Rehovot, Israel).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: International Journal of Nanomedicine

Article Title: Simultaneous Intramuscular And Intranasal Administration Of Chitosan Nanoparticles–Adjuvanted Chlamydia Vaccine Elicits Elevated Protective Responses In The Lung

doi: 10.2147/IJN.S218456

Figure Lengend Snippet: Ags-specific T-cell responses in spleen. Notes: Splenocytes from immunized mice were seeded into 24-well plates and incubated at 37°C in 5% CO 2 for 48 hrs and assayed for Ags-specific CD4 + T-cell response. ( A ) Ags-specific proliferative responses in splenocytes were induced by different immunization routes. ( B ) Frequency of CD4 + /IFN-γ cells was calculated according to IFCS. ( C ) Frequency of CD4 + /IL-4 cells was calculated according to IFCS. ( D ) Ags-specific CD4 + T-cell response was determined by intracellular IL-4 and IFN-γ staining. The FACS plots are representative of the mean percentages of 6 mice in each group. Each bar indicates the mean ± SD of 6 mice per group from three independent results. *P < 0.05; **P < 0.01. Abbreviations: IFCS, intracellular flow cytometry staining; FACS, fluorescence-activated cell sorting; CD4, cluster of differentiation 4; IFN-γ, interferon gamma; IL, interleukin; SI, stimulation index; SD, standard deviation.

Article Snippet: Then, the cells were permeabilized with Cytofix/Cytoperm™ kit (BD Pharmingen) and stained intracellularly for 30 mins at 4°C for cytokines IL-4-PE and IFN-γ-PE (BD Pharmingen, USA).

Techniques: Incubation, Staining, Flow Cytometry, Fluorescence, FACS, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Simultaneous Intramuscular And Intranasal Administration Of Chitosan Nanoparticles–Adjuvanted Chlamydia Vaccine Elicits Elevated Protective Responses In The Lung

doi: 10.2147/IJN.S218456

Figure Lengend Snippet: Cytokine secretion by splenocytes from vaccinated mice. Notes: ( A ) IFN-γ, ( B ) IL-2, ( C ) TNF-α, ( D ) IL-4, ( E ) IL-10 and ( F ) IL-17A levels were measured in the splenocyte supernatants of immunized mice by ELISA kits. Each bar indicates the mean ± SD of triplicates from 6 mice per group. *P < 0.05; **P < 0.01. Abbreviations: IFN-γ, interferon gamma; IL, interleukin; TNF-α, tumor necrosis factor alpha; ELISA, enzyme-linked immunosorbent assay; Th, helper T; SD, standard deviation.

Article Snippet: Then, the cells were permeabilized with Cytofix/Cytoperm™ kit (BD Pharmingen) and stained intracellularly for 30 mins at 4°C for cytokines IL-4-PE and IFN-γ-PE (BD Pharmingen, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Simultaneous Intramuscular And Intranasal Administration Of Chitosan Nanoparticles–Adjuvanted Chlamydia Vaccine Elicits Elevated Protective Responses In The Lung

doi: 10.2147/IJN.S218456

Figure Lengend Snippet: Cytokine levels in the lungs of immunized mice after the C. psittaci challenge. Notes: ( A ) IFN-γ, ( B ) TNF-α, ( C ) IL-4, ( D ) IL-6, ( E ) IL-10 and ( F ) IL-12 levels were detected in the lung homogenates of immunized mice by ELISA kits after the C. psittaci infection. Each bar represents the mean ± SD of the cytokine levels (pg/mL) in the lung homogenates from 6 mice per group in three independent experiments. *P < 0.05; **P < 0.01. Abbreviations: IFN-γ, interferon gamma; IL, interleukin; TNF-α, tumor necrosis factor alpha; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation.

Article Snippet: Then, the cells were permeabilized with Cytofix/Cytoperm™ kit (BD Pharmingen) and stained intracellularly for 30 mins at 4°C for cytokines IL-4-PE and IFN-γ-PE (BD Pharmingen, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Standard Deviation