How to improve communication with deaf children in the dental clinic

It may be difficult for hearing-impaired people to communicate with people who hear. In the health care area, there is often little awareness of the communication barriers faced by the deaf and, in dentistry, the attitude adopted towards the deaf is not always correct. A review is given of the basic rules and advice given for communicating with the hearing-impaired. The latter are classified in three groups ? lip-readers, sign language users and those with hearing aids. The advice given varies for the different groups although the different methods of communication are often combined (e.g. sign language plus lip-reading, hearing-aids plus lip-reading). Treatment of hearing-impaired children in the dental clinic must be personalised. Each child is different, depending on the education received, the communication skills possessed, family factors (degree of parental protection, etc.), the existence of associated problems (learning difficulties), degree of loss of hearing, age, etc

Realtime PCR probes and primers of NP-markers (from Applied Biosystems) and corresponding antibodies for protein analysis.

<p>NA: not applicable, no commercially available rat-specific anti-CD155; ND: PCR not determined; Rb: rabbit; Ms: mouse; IHC: immunohistochemical staining; FC: flow cytometry.</p

Immunostaining illustrated region-dependent and age-related changes in the expression of NP markers.

<p>(A) Basp1; (B) Ncdn; (C) Nrp-1; (D) CD221 in rat IVD tissues at different ages (1 m, 12 m and 21 m). Bar: 20 µm; m: month. (TIF)</p

Average fold-differences in relative mRNA levels of selected genes between both tissue regions (AF & NP of 1 m rat) and ages (1 m & 12 m of NP) analyzed by cDNA microarray (n = 4, p<0.01, fold-differences greater than or equal to 2 folds between groups by one-factor ANOVA).

<p>a. Fold-differences were nomalized to 1 m AF; b. Fold-differences were nomalized to 12 m NP; c. Brachyury T gene was not included in the rat genome 230 2.0 Array, therefore T2 was listed here. Bolded gene names/symbles indicate the targets selected for PCR and protein expression anylsis. –: fold not changed; m: month.</p

Immunostaining and flow cytometry detection for NP markers.

<p>Immunostaining (A, C, E) and flow cytometry (B, D, F) illustrated region-dependent and age-related changes in the expression of NP markers (A, B) CD24; (C, D) CD90; (E, F) Brachyury T in rat IVD tissues at different ages (1 m, 12 m and 21 m). Bar: 20 µm; m: month. Representative histograms of flow cytometry at left illustrate the relative fluorescence intensity of NP markers on X-axis for freshly isolated cells of 1 m rats (cell surface: CD24 and CD90; nucleus: T). The numbers appeared in each histogram indicate the percentage of positive fluorescence labeled cells and mean fluorescence intensity (MFI) for each cell type. (First left black line: isotype control, red line: AF cells, blue line: NP cells). (TIF)</p

Realtime RT-PCR for relative mRNA level (2^−ΔΔCt) of NP markers.

<p>(A) Basp1; (B) Ncdn; (C) Nrp-1; (D) Brachyury T; (E) CD155; (F) CD221 in rat IVD tissues at different ages (1 m, 12 m and 21 m). All values of fold-difference were normalized to AF tissue (1 m) for comparison between different tissue regions and ages. m: month. * p<0.05, ** p<0.01, two-factor ANOVA. (TIF)</p

Differentiation of iPSCs under hypoxic conditions <i>in vitro</i>.

<p>(A) General experimental overview. (B) Sulfated-glycosaminoglycan content normalized to DNA content (sGAG/DNA) in cell constructs (results shown as mean+SD, n=3 samples). Significant effects of treatment and time were detected using a two-way ANOVA and Tukey’s HSD (p<0.05). Treatment and time points denoted by different letters were found to be significantly different: uppercase letters for treatment; lowercase letters for time. (C) Total collagen content normalized to DNA (collagen/DNA) in cell constructs (results shown as mean+SD or –SD, n=3 samples). No significant effects of treatment on collagen production were detected via two-way ANOVA and Tukey’s HSD (p>0.05). (D) Cell constructs extracted from Transwelsl at the end of the culture (day 28) and representative images of histology and immunostaining revealed matrix accumulation (Saf O for glycosaminoglycan and type II collagen, Col II; Scale bar: 100 µm) and expression of NP-associated markers (vimentin, VIM; cytokeratin 5/8, CK and integrin α6, ITGα6) in cell constructs at days 28. Scale bar: 50 µm. NCCM: notochordal cell conditioned medium. (E) Representative images of histology (H and E) and immunostaining for above mentioned NP-associated markers and matrix proteins in the immature mouse IVD (1 month-old) tissue control (AF: anulus fibrosus; NP: nucleus pulposus).</p

Differentiation of enriched CD24⁺ iPSCs in a 3D culture system <i>in vitro</i>.

<p>(A) Cell constructs form both CD24^- and CD24⁺ fractions extracted from Transwells at the end of culture (day 28) and representative images of histology and immunostaining revealed cell morphology (H and E) and matrix accumulation (Saf O for glycosaminoglycan and type II collagen, Col II; Scale bar: 50 µm). (B) CD24⁺ cell construct and representative images for expression of NP-associated markers by immunostaining (integrin α6, ITGα6; laminin α5, LM α5; vimentin, VIM and cytokeratin 5/8, CK) in cell constructs at day 28. Scale bar: 20 µm).</p

Enrichment of CD24⁺ cell population from undifferentiated iPSCs via magnetic activated cell sorting (MACS).

<p>(A) General experimental overview. (B) Flow cytometry analysis for the expression of CD24 in presorted, “flow-through” and sorted cell fractions. Undifferentiated iPSCs incubated with CD24 antibody and magnetic beads were passed through a MACS column, yielding a CD24⁺ fraction of cells; in the “flow-through” fraction, ~ 5.1% of total cells on average were found to express CD24, which was less than that in the presorted fraction. The gated readouts represent the percentage of cells expressed CD24 in presorted cells and sorted fractions from one sorting experiment.</p

iPSC morphology and the expression notochordal-related transcriptional factors.

<p>(A) Undifferentiated iPSCs cultured on a feeder layer of mitotically inactivated mouse embryonic fibroblasts and formed colonies (top panels); iPSCs formed cell clusters of embryoid bodies (EBs, bottom panels) when promoted to undergo spontaneous differentiation in ultra-low attaching dishes (bars = 100 µm). (B) Relative mRNA levels for transcriptional factors (<i>Oct4, Noggin, Brachyury, Noto, Foxa2, Shh</i>) in EBs (3 days) normalized to undifferentiated iPSCs for each target gene. Data are averages of two replicates for cells collected from multiple EBs and undifferentiated culture.</p