Expression of trefoil factor 1 in the developing and adult rat ventral mesencephalon

Trefoil factor 1 (TFF1) belongs to a family of secreted peptides with a characteristic tree-looped trefoil structure. TFFs are mainly expressed in the gastrointestinal tract where they play a critical role in the function of the mucosal barrier. TFF1 has been suggested as a neuropeptide, but not much is known about its expression and function in the central nervous system. We investigated the expression of TFF1 in the developing and adult rat midbrain. In the adult ventral mesencephalon, TFF1-immunoreactive (-ir) cells were predominantly found in the substantia nigra pars compacta (SNc), the ventral tegmental area (VTA) and in periaqueductal areas. While around 90% of the TFF1-ir cells in the SNc co-expressed tyrosine hydroxylase (TH), only a subpopulation of the TH-ir neurons expressed TFF1. Some TFF1-ir cells in the SNc co-expressed the calcium-binding proteins calbindin or calretinin and nearly all were NeuN-ir confirming a neuronal phenotype, which was supported by lack of co-localization with the astroglial marker glial fibrillary acidic protein (GFAP). Interestingly, at postnatal (P) day 7 and P14, a significantly higher proportion of TH-ir neurons in the SNc co-expressed TFF1 as compared to P21. In contrast, the proportion of TFF1-ir cells expressing TH remained unchanged during postnatal development. Furthermore, significantly more TH-ir neurons expressed TFF1 in the SNc, compared to the VTA at all four time-points investigated. Injection of the tracer fluorogold into the striatum of adult rats resulted in retrograde labeling of several TFF1 expressing cells in the SNc showing that a significant fraction of the TFF1-ir cells were projection neurons. This was also reflected by unilateral loss of TFF1-ir cells in SNc of 6-hydroxylase-lesioned hemiparkinsonian rats. In conclusion, we show for the first time that distinct subpopulations of midbrain dopaminergic neurons express TFF1, and that this expression pattern is altered in a rat model of Parkinson's disease

Implementation and Evaluation of Resuscitation Training for Childcare Workers

BACKGROUND AND OBJECTIVE: Children spend a large amount of time in daycare centers or schools. Therefore, it makes sense to train caregivers well in first-aid measures in children. The aim of this study is to evaluate whether a multimodal resuscitation training for childcare workers can teach adherence to resuscitation guidelines in a sustainable way. MATERIALS AND METHODS: Caregivers at a daycare center who had previously completed a first-aid course received a newly developed multimodal resuscitation training in small groups of 7–8 participants by 3 AHA certified PALS instructors and providers. The 4-h focused retraining consisted of a theoretical component, expert modeling, resuscitation exercises on pediatric manikins (Laerdal Resusci Baby QCPR), and simulated emergency scenarios. Adherence to resuscitation guidelines was compared before retraining, immediately after training, and after 6 months. This included evaluation of chest compressions per round, chest compression rate, compression depth, full chest recoil, no-flow time, and success of rescue breaths. For better comparability and interpretation of the results, the parameters were evaluated both separately and summarized in a resuscitation score reflecting the overall adherence to the guidelines. RESULTS: A total of 101 simulated cardiopulmonary resuscitations were evaluated in 39 participants. In comparison to pre-retraining, chest compressions per round (15.0 [10.0–29.0] vs. 30.0 [30.0–30.0], p < 0.001), chest compression rate (100.0 [75.0–120.0] vs. 112.5 [105–120.0], p < 0.001), correct compression depth (6.7% [0.0–100.0] vs. 100.0% [100.0–100.0], p < 0.001), no-flow time (7.0 s. [5.0–9.0] vs. 4.0 s. [3.0–5.0], p < 0.001), success of rescue breaths (0.0% [0.0–0.0] vs. 100.0% [100.0–100.0], p < 0.001), and resuscitation score were significantly improved immediately after training (3.9 [3.2–4.9] vs. 6.3 [5.6–6.7], p < 0.001). At follow-up, there was no significant change in chest compression rate and success of rescue breaths. Chest compressions per round (30.0 [15.0–30.0], p < 0.001), no-flow time (5.0 s. [4.0–8.0], p < 0.001), compression depths (100.0% [96.7–100.0], p < 0.001), and resuscitation score worsened again after 6 months (5.7 [4.7–6.4], p = 0.03). However, the results were still significantly better compared to pre-retraining. CONCLUSION: Our multimodal cardiopulmonary resuscitation training program for caregivers is effective to increase the resuscitation performance immediately after training. Although the effect diminishes after 6 months, adherence to resuscitation guidelines was significantly better than before retraining

Co-expression of trefoil factor 1 (TFF1) and calcium-binding proteins in the ventral mesencephalon of adult rats.

<p>Double immunofluorescence stainings for TFF1 and the calcium-binding proteins calretinin (CR), calbindin (CB) or parvalbumin (PV). Note that many TFF1-immunoreactive (-ir) cells co-expressed CR (arrows), but not all CR-ir cells co-localized with TFF1 (arrowheads). Similarly, co-localization with CB was seen for a few TFF1-ir cells (arrows). As expected from the distribution pattern depicted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0076592#pone-0076592-g001" target="_blank">Figure 1</a> no co-localization was detected for TFF1-ir cells with PV (arrows). Scale bars middle panel: 50 µm, lower panel: 100 µm.</p

Detection of Trefoil factor 1 (TFF1) in nigrostriatal projection neurons by Fluorogold (FG) labelling.

<p>Representative photomicrographs of double immunofluorescence stainings for TFF1 and the retrograde tracer FG at the level of substantia nigra pars compacta (SNc) 10 days after intrastriatal FG injection. A subpopulation of TFF1-ir cells was found to co-express FG identifying these as projection neurons (arrows). Scale bars upper panel: 100 µm, lower panel: 50 µm.</p

Co-expression of trefoil factor 1 (TFF1) and tyrosine hydroxylase (TH) in the ventral mesencephalon of adult rats.

<p>Double immunofluorescence staining for TFF1 (red) and TH (green) in the ventral mesencephalon of adult rats. Note that nearly all TFF1-immunoreactive (-ir) cells co-localized with TH, while many TH-ir cells did not co-express TFF1. Scale bars: 1 mm (overview); 100 µm (magnification). Abbreviations: SNc, substantia nigra pars compacta; SNl, substantia nigra pars lateralis; VTA, ventral tegmental area.</p

Loss of trefoil factor 1 (TFF1) expressing cells in a rat model of Parkinson’s disease.

<p>Representative photomicrographs of tyrosine hydroxylase (TH; A) and TFF1 (B) in brain sections from adult rat ventral mesencephalon at 4 weeks after unilateral 6-hydroxydopamine (6-OHDA) lesion. The lesion resulted in a distinct loss of TH-ir neurons in right SN (A2) as compared to the contralateral, unlesioned control side (A1). Similarly, a reduction of TFF1-ir cells was detected on the lesioned side (B2) as compared to the intact control side (B1). This loss of TFF1-ir cells is better recognized on the enlarged images (B2). Scale bars: 1 mm (overview), 500 µm (magnification).</p

Quantification of trefoil factor 1 (TFF1) and tyrosine hydroxylase (TH) co-expressing cells in adult rats.

<p>Percentage of trefoil factor 1-immunoreactive (TFF1-ir) cells that co-localized with tyrosine hydroxylase (TH) (A) and percentage of TH-ir cells that co-localized with TFF1 (B) in the substantia nigra (SN) and ventral tegmental area (VTA) of adult rats. *: p<0.05 vs. corresponding VTA values, n = 4.</p

Quantification of trefoil factor 1 (TFF1) and tyrosine hydroxylase (TH) co-expressing cells during development.

<p>A) Quantification of trefoil factor 1-immunoreactive (TFF1-ir) cells co-expressing tyrosine hydroxylase (TH) and B) TH-ir cells co-expressing TFF1 in the substantia nigra (SN) of postnatal (P) day 7, 14, 21 and adult rats. The percentage of TH/TFF1 co-expressing cells was significantly higher for P7 and P14 rats compared to P21 rats, whereas there was no significant difference between the percentages of TFF1-ir cells co-expressing TH. Data are expressed as mean ± SEM (*: p<0.05, n = 4-7). C) Representative double immunofluorescence images of TFF1-ir and TH-ir cells in SN of P7, P14 and P21 rats (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0076592#pone-0076592-g002" target="_blank">Figure 2</a> for adult rats). Scale bar: 200 µm.</p

Trefoil factor 1 (TFF1) expression is restricted to neurons.

<p>Representative photomicrographs of double immunofluorescence stainings for TFF1 and astroglial or neuronal markers in the substantia nigra pars compacta (SNc) of adult rats. No co-localization was found of TFF1-positive cells (arrows) with the astroglial marker glial fibrillary acid protein (GFAP) (arrowheads). Notably, a substantial number of the TFF1-ir cells were demonstrated to co-express the pan neuronal marker NeuN (arrows), while some did not (arrowheads). Scale bar: 50 µm.</p