Journal: Microbiome

Article Title: Amino acid availability influences Staphylococcus aureus interaction with the nasal microbiota in a strain- and community-specific manner

doi: 10.1186/s40168-026-02563-x

Figure Lengend Snippet: Species composition and structure of 11 nasal communities. A Schematic methodology used to determine nasal community structures and establish the strain collection LaCa. B, C Relative species abundance determined by 16S rRNA gene sequencing of nasal metagenomic DNA ( B ) or by isolation and enumeration of nasal species on plates ( C ), from the nasal microbiomes of eleven healthy volunteers ( A – K ). Each color designates one of the 20 most abundant species across all volunteers. Less abundant species were assigned to “Other species”. Species with a missing or unclear identification were assigned to “Not identified”. Below the bars, “ + ” indicates the volunteers in which S. aureus was detected by both techniques, and numbers correspond to the community state type (CST) assigned to the corresponding community. “a” refers to ambiguous S. aureus carriage (detected by only one method) or CST classification (balanced proportion of several species). Each bar displays the results of one replicate. See also Supplementary Fig. S1 and Tables S1, S2, and S3

Article Snippet: Briefly, for 16S rRNA gene amplicon sequencing, total microbial DNA was extracted from the sample (1 mL Amies medium in which nasal swabs were suspended, or 500 μL of bacterial suspension derived from SynCom cultures) using the HostZero Microbial DNA kit (Zymo Research), including the host DNA depletion step for nasal swab samples.

Techniques: Sequencing, Isolation

Journal: Microbiome

Article Title: Amino acid availability influences Staphylococcus aureus interaction with the nasal microbiota in a strain- and community-specific manner

doi: 10.1186/s40168-026-02563-x

Figure Lengend Snippet: S. aureus and SynComs dynamics in mono- and cocultivation. A Schematic methodology used to study synthetic communities in vitro and in vivo. B Relative species abundance determined by 16S rRNA gene sequencing from frozen SynComs (Input) or SynComs cultivated 72 h on SNM agar in isolation for two biological replicates (R1, R2). Each color designates a species. Less abundant species were assigned to “Other species”. Complete data is detailed in Supplementary Table S4. See also Supplementary Fig. 4. C Heatmap depicting the growth fold-change induced by synthetic culture with five different SynComs (A, B, C, J, K) on SNM agar, for four S. aureus strains. Sa A, Sa B, and Sa C originated from volunteers A, B, and C, respectively. Positive fold-changes (verdigris) and negative fold-changes (red) indicate increases or decreases in S. aureus cell numbers in synthetic culture relative to the monoculture, respectively. Each fold-change is based on four biological replicates. Significant changes are marked with stars (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, 2-way ANOVA). See also Supplementary Fig. S5. D. Barplots depicting the bacterial counts of S. aureus Sa A recovered from the noses of gnotobiotic mice, 7 days after nasal inoculation in isolation or in the presence of three different SynComs. Bars represent the median ± interquartile range from 5 to 6 animals per group. Dashed lines represent the nasal inoculum and the detection limit. Statistical significance is indicated by *** P adj < 0.001 (Kruskal–Wallis with Dunn’s correction)

Article Snippet: Briefly, for 16S rRNA gene amplicon sequencing, total microbial DNA was extracted from the sample (1 mL Amies medium in which nasal swabs were suspended, or 500 μL of bacterial suspension derived from SynCom cultures) using the HostZero Microbial DNA kit (Zymo Research), including the host DNA depletion step for nasal swab samples.

Techniques: In Vitro, In Vivo, Sequencing, Isolation, Co-Cultures

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Effects of systemic treatment with an anti-IL-16 or an anti-CD4 antibody on the hypernociception measured 2 days after sciatic ligation in male and female mice. a General schema of the experimental behavioural procedure. b , c The s.c. administration of an anti-IL-16 antibody 1 h before testing dose-dependently inhibited neuropathic thermal hyperalgesia ( b ; 1–10 µg/kg; n = 5–6) and mechanical allodynia ( c ; 0.01–1 µg/kg; n = 5–7) in male mice. d , e Inhibitory effect evoked in female mice after the s.c. administration of the anti-IL-16 antibody of neuropathic thermal hyperalgesia ( d ; 10 µ/kg; n = 5) and mechanical allodynia ( e ; 1 µg/kg; n = 6) 1 h before testing. f , g The s.c. administration of an anti-CD4 antibody 1 h before testing dose-dependently inhibited neuropathic thermal hyperalgesia ( f ; 1–10 µg/kg; n = 5–7) and mechanical allodynia ( g ; 0.01–0.1 µg/kg; n = 5–6) in male mice. h , i Inhibitory effect evoked in female mice after the s.c. administration of the anti-CD4 antibody of neuropathic thermal hyperalgesia ( h ; 10 µ/kg; n = 5) and mechanical allodynia ( i ; 0.1 µg/kg; n = 6–8) 1 h before testing

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Ligation

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Time course of anti-IL-16 and anti-CD4 antibodies on thermal hyperalgesia measured 2 days after sciatic ligation in mice. Effects of systemic administration of an anti-IL-16 or an anti-CD4 antibody on hypernociception measured 9 days after sciatic ligation. a Schema of time course of the study of the antihyperalgesic effects induced by the anti-IL-16 or anti-CD4 antibodies in mice submitted to nerve ligation 2 days before. b , c Withdrawal latencies measured in mice 2 days after nerve ligation by the unilateral hot plate test 0, 1, 2, 4 and 6 h after the s.c. administration of anti-IL16 antibody ( b ; 10 µg/kg; n = 6), an anti-CD4 antibody ( c ; 10 µg/kg; n = 6) or the corresponding IgG2bk isotype ( b , c ; IgG2bk 10 µg/kg; n = 6 in each case). Each point represents the mean and corresponding standard error. • p < 0.05, •• p < 0.01 compared with the measure done at time 0, before drug administration; ** p < 0.01 compared with right paws of mice treated with IgG2bк isotype at the corresponding time, two-way ANOVA followed by Tukey’s test

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Ligation, Hot Plate Test

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Effects on Il16 and Casp3 mRNA expression in DRGs and nerves two days after sciatic ligation. Measurement of caspase-3 activity and IL-16 immunofluorescence in DRGs of mice submitted to sham surgery or sciatic ligation. a – b Il16 mRNA expression measured by qRT-PCR in DRGs ( a ; n = 9) or sciatic nerves ( b ; n = 7–8) coming from mice submitted to the sham surgical procedure or to sciatic nerve ligation. c – d Casp3 mRNA expression measured by qRT-PCR in DRGs ( c ; n = 9) or sciatic nerves ( d ; n = 8–9) coming from mice submitted to the sham surgical procedure or to sciatic nerve ligation. e Caspase-3 activity measured in DRG homogenates coming from mice submitted to sham surgery or sciatic ligation 2 days before ( n = 4). Means and their corresponding standard errors are represented. ** p < 0.01 compared with the value obtained in mice submitted to sham procedure, Student’s t test. f Representative example of IL-16 labelling by immunofluorescence in a DRG coming from a mouse submitted to sham surgery (f1) or to nerve ligation 2 days before (f2). Some IL-16 + neurons are indicated with arrows. g Percentage of IL-16 + cells quantified in DRG sections coming from mice submitted to sham surgery or sciatic ligation ( n = 3). h Percentage of IL-16 + cells classified as large, medium or small in mice submitted to sham surgery ( n = 3) or sciatic ligation ( n = 3) 2 days before. In g and h , means and their corresponding standard errors are represented. * p < 0.05, ** p < 0.01 compared with the number of IL-16 + large DRG cells, one-way ANOVA followed by Tukey’s test. i Representative example of an image of DAPI (i1), IL-16 (i2) and peripherin (i3) labelling in DRG together with their merged image (i4) showing the colocalization of IL-16 and peripherin (yellow)

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Expressing, Ligation, Activity Assay, Immunofluorescence, Quantitative RT-PCR

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Measurement of caspase-3 activity and IL-16 immunofluorescence in sciatic nerves of mice submitted to sham surgery or sciatic ligation. a Caspase-3 activity measured in nerve homogenates coming from mice submitted to sham surgery or sciatic ligation 2 days before ( n = 3). b Representative example of IL-16 labelling by immunofluorescence in a nerve coming from a mouse submitted to sham surgery (b1) or nerve ligation (b2) 2 days before. The image in b2 corresponds to the proximal side of the ligature closest to DRG. The white arrow indicates the ligation site. ( c ) Examples of IL-16 labelling in the second (c1) and third (c2) more distal ligatures. The white arrow indicates the ligation site. d Percentage of IL-16 immunoreactive surface measured in nerve sections coming from mice submitted to sham surgery or sciatic ligation ( n = 5). e Bars of the upper graph represent the % of IL-16 immunoreactive surface measured in five 200-µm segments of sciatic nerves corresponding to the proximal area close to the first ligature ( n = 5). In control mice submitted to sham surgery, an equivalent area of intact nerve was analysed ( n = 5). Below, schematic draw showing the five sections in which the studied area was divided, being number 1 assigned to the segment closer to ligation. Green traits graphically represent IL-16 + axons. f IL-16 expression shown in a cross section performed at the first ligation of a sciatic nerve proximal to DRG. Bright field image without staining to visualize the histological structure with axon profiles and myelin sheaths (f1), DAPI staining (f2) and IL-16 labelling (f3) are shown. A merged image of f1, f2 and f3 showing a more precise localization of IL-16 immunofluorescence appears in f4. f5 shows details of f4, in which IL-16 labelling can observed in a myelinated axon (upper, yellow frame), a possible unmyelinated fibre (middle, blue frame) and a Schwann cell (lower, red frame). In ( a ), ( d ) and ( e ), means and their corresponding standard errors are represented. In ( a ) and ( d ) * p < 0.05; ** p < 0.01 compared with values obtained in mice submitted to sham surgery, Student’s t test. In ( e ) •• p < 0.01 compared with the corresponding segment in mice submitted to sham surgery; * p < 0.05 compared with segment number 1 of ligated nerves, two-way ANOVA followed by Tukey’s test

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Activity Assay, Immunofluorescence, Ligation, Control, Expressing, Staining

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Expression of IL-16 in axons and Schwann cells of injured sciatic nerves. Lack of effect of neutrophil depletion on thermal hyperalgesia and IL-16 labelling in nerves. ( a1 – a4 ) Representative examples of confocal images of cross sections of injured sciatic nerves labelled with DAPI (blue, a1 ), IL-16 (green, a2 ) and the neural marker βIII-tubulin (red, a3 ). The merged image shows the colocalization of IL-16 and βIII-tubulin (yellow, a4 ), and higher-magnification images of the yellow, blue and red boxes are shown in a5 ). The yellow box shows an axon double-positive for IL-16 and βIII-tubulin (yellow). The blue box contains two small-diameter axons, one of them double-positive for IL-16 and βIII-tubulin (yellow) and the other only positive for βIII-tubulin (red). The red box indicates a βIII-tubulin-positive axon surrounded by IL-16 immunoreactivity with a shape compatible with a Schwann cell (green). ( b1 – b4 ) Confocal images corresponding to cross sections in injured sciatic nerves labelled with DAPI (blue, b1 ), IL-16 (green, b2 ) and the Schwann cell marker S100β (red, b3 ). The merged image shows the colocalization of IL-16 and S100β (yellow, b4 ). Higher-magnification images of the cell contained in the red box appear in b5 , when labelled with IL-16 (green, left hand side), S100β (red, middle image) and the merged image with DAPI (yellow, right hand side). ( c1 ) Representative merged image of IL-16, Ly6G and DAPI immunofluorescence in a longitudinal section coming from an injured nerve showing the presence of some neutrophils in a square remarked by a white dashed line. ( c2 – c5 ) Higher-magnification images of cells contained in the box, when labelled with DAPI (blue, c2 ), IL-16 (green, c3 ), Ly6G (red, c4 ) and the merged image showing the colocalization of IL-16 + and Ly6G + cells (yellow, c5 ). ( d ) Schematic representation of the experimental procedure used to evaluate the functional effect of an anti-IL-16 antibody in neutrophil-depleted mice submitted to sciatic nerve ligation 2 days before. ( e ) Decreased withdrawal latencies are shown in neutrophil-depleted mice 2 days after nerve ligation. The s.c. administration of the anti-IL-16 antibody (10 µg/kg) reversed this thermal hyperalgesia measured by the unilateral hot plate test 1 h later. Means and their corresponding standard errors are represented, •• p < 0.01 compared with the contralateral paw; ** p < 0.01 compared with values obtained in injured mice treated with IgG2bκ, two-way ANOVA followed by Tukey’s test. ( f ) Representative immunofluorescent merged image of IL-16 + traits (green) and DAPI nuclear staining (blue) in a longitudinal section of the ligated sciatic nerve from a neutrophil-depleted mouse

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Expressing, Confocal, Marker, Immunofluorescence, Functional Assay, Ligation, Hot Plate Test, Staining

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Hypernociceptive effects evoked by the intra-sciatic (i.sc.) administration of IL-16 and inhibition of hypernociception by i.sc. injection of an anti-IL-16 antibody in mice with nerve ligation. ( a ) Photographs of the intra-sciatic injection procedure performed with 1% methylene blue. On the left hand side, the sciatic nerve of the anesthetized mouse is hold with a surgical forceps and the micropipette loaded with the dye can be observed just before its introduction into the nerve. At the upper right hand side, a detailed view of the tip of the micropipette introduced into the nerve and the dye can be observed along a segment of the nerve. In the lower part of the right hand side, the removed nerve appears placed on a graph millimetre paper. ( b–d ) Effects of the i.sc. administration of 0.2 ng of IL-16 1 h after injection. A timeline schema is represented in ( b ), the effects of IL-16 or saline administration on thermal hyperalgesia appears in ( c ) ( n = 5–6) and mechanical allodynia in ( d ) ( n = 5). ( e – g ) Effects of the i.sc. administration of 0.2 ng of IL-16 2 days after injection. A timeline schema is represented in ( e ), the effects of IL-16 or saline administration on thermal hyperalgesia appears in ( f ) ( n = 6) and mechanical allodynia in ( g ) ( n = 6). ( h – j ) Behavioural testing of the effects induced after the intra-sciatic administration of the anti-IL-16 antibody in mice submitted to nerve ligation 2 days before. Timeline schema of the procedure ( h ). Prevention of thermal hyperalgesia ( i ; n = 5) and mechanical allodynia ( j ; n = 5) following the i.sc. administration of an anti-IL-16 antibody (50 ng; 1 h before testing) in mice submitted to nerve ligation 2 days before. Means and their corresponding standard errors are represented. •• p < 0.01 compared with their contralateral, non-injured, paws of mice; * p < 0.05, ** p < 0.01 compared with right paws of mice treated with saline or IgG2bk, two-way ANOVA followed by Tukey’s test (c,f,i) or Kruskall-Wallis followed by Dunn’s test (d,g,j)

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Inhibition, Injection, Ligation, Saline, Behavioral Assay

Journal: Molecular Neurobiology

Article Title: Neural Expression of IL-16 Mediates Neuropathic Pain after Sciatic Injury in Mice

doi: 10.1007/s12035-026-06280-2

Figure Lengend Snippet: Inhibition of neuropathic hypernociception by i.sc. injection of an anti-CD4 antibody. Expression of CD4 receptors in sciatic nerves and up-regulation after nerve injury. Absence of CD3 labelling in the sciatic injured nerve and lack of effect of lymphocyte depletion on neuropathic thermal hyperalgesia. ( a – c ) Behavioural testing of the effects induced after the intra-sciatic administration of the anti-CD4 antibody in mice submitted to nerve ligation 2 days before. A timeline schema of the procedure is represented in ( a ). The i.sc. administration of an anti-CD4 antibody (50 ng; 1 h before testing) prevents thermal hyperalgesia ( b ; n = 5) and mechanical allodynia ( c ; n = 5) in mice submitted to nerve ligation 2 days before. In ( b and c ), means and their corresponding standard errors are represented, • p < 0.05, •• p < 0.01 compared with mice submitted to sham surgery; ** p < 0.01 compared with values obtained in injured mice treated with IgG2bκ, two-way ANOVA followed by Tukey’s test. ( d ) Representative example of CD4 labelling by immunofluorescence in a nerve coming from a mouse submitted to sham surgery ( d1 ) or nerve ligation ( d2 ) 2 days before. The image in d2 ) corresponds to the proximal side of the ligature closest to DRG. The white arrow indicates the ligation site. ( e ) Percentage of CD4 + immunoreactive surface measured in nerve sections coming from mice submitted to sham surgery or sciatic ligation ( n = 3). Means and their corresponding standard errors are represented, ** p < 0.01 compared with mice submitted to sham surgery, Student’s t test. ( f ) Immunofluorescence image showing a CD4 + cell located in the sciatic nerve compatible with a Schwann cell with its nucleus stained with DAPI, ( g1 – g4 ) CD4 expression in a cross section performed at the first ligation of a sciatic nerve proximal to DRG showing a myelinated axon. Bright field image showing the histological structure compatible with the axon profile and its myelin sheath ( g1 ), DAPI staining (blue, g2 ) and CD4 labelling (green, g3 ) are shown. The merged image ( g4 ) shows a more precise localization of CD4 immunofluorescence. ( h1 – h4 ) CD4 expression in a cross section performed at the first ligation of a sciatic nerve proximal to DRG showing CD4 labelling in a Schwann cell surrounding a myelinated axon. Bright field image without staining shows the histological structure compatible with a Schwann cell surrounding an axon ( h1 ), DAPI staining (blue, h2 ) and CD4 labelling (green, h3 ) are shown. The merged image ( h4 ) shows a more precise localization of CD4 immunofluorescence. ( i ) Representative examples of confocal images of cross sections of injured sciatic nerves labelled with DAPI (blue, i1 ), CD4 (green, i2 ) and the neural marker βIII-tubulin (red, i3 ). The merged image shows the colocalization of CD4 and βIII-tubulin in an axon ( i4 , yellow). ( j ) Representative examples of confocal images of injured sciatic nerves labelled with DAPI (blue, j1 ), CD4 (green, j2 ) and the neural marker βIII-tubulin (red, j3 ). The merged image ( j4 ) shows the labelling of an axon (red) surrounded by a CD4 + cell compatible with the shape of a Schwann cell (green). ( k ) Representative examples of confocal images of injured sciatic nerves labelled with DAPI (blue, k1 ), CD4 (green, k2 ), the Schwann cell marker S100β (red, k3 ). The merged image shows the colocalization of CD4 and S100β + Schwann cell ( k4 , yellow). ( l1 - l4 ) Immunofluorescence images of DAPI (blue) and CD3 (red) labelling in thymus cells ( l1 ) and longitudinal nerve sections coming from mice submitted to nerve ligation 2 days before ( l2 - l4 ). Contrasting with the clear-cut lymphocyte morphology observed in the thymus ( l1 ), no CD3 labelling is observed in nerve ( l2 - l4 ) and the sporadic red labelling signal that appeared in some nerve sections was not associated to DAPI-stained nuclei (Fig. l4 ). ( m - n ) Lack of effect of lymphocyte depletion on the hyperalgesic effect evoked by nerve ligation and on the antihyperalgesic effect evoked by the administration of an anti-IL-16 antibody. m ) Schematic representation of the experimental procedure used to evaluate the functional effect of an anti-IL-16 antibody in lymphocyte-depleted mice submitted to sciatic nerve ligation. n ) Decreased withdrawal latencies are shown in lymphocyte-depleted mice 2 days after nerve ligation. The s.c. administration of the anti-IL-16 antibody (10 µg/kg) reversed this thermal hyperalgesia measured by the unilateral hot plate test 1 h later. Means and their corresponding standard errors are represented, •• p <0.01 compared with the contralateral paw; ** p <0.01 compared with values obtained in injured mice treated with IgG2bκ, two-way ANOVA followed by Tukey’s test.

Article Snippet: Mouse recombinant IL-16 (Raybiotech, 230–30050-10) was dissolved in saline.

Techniques: Inhibition, Injection, Expressing, Ligation, Immunofluorescence, Staining, Confocal, Marker, Morphology, Functional Assay, Hot Plate Test, Behavioral Assay