, 2013). In addition, mutations in a single causative gene may only be a portal to far greater molecular complexity. Thus, for example, FMR1, which is a neuronal polyribosome-associated RNA binding protein, has been shown to affect the translation of 842 mRNA transcripts ( Darnell et al., 2011) each with their own “downstream” biology; many of these individual targets are now being implicated for subtler contributions to complex, TSA HDAC order polygenic disease. The unexpected complexity of many monogenic brain disorders pales in comparison with the emerging complexity of common polygenic brain disorders, a challenge that is only now coming into view.

Because severe, highly penetrant mutations often produce marked decrements in reproductive fitness, they tend to be rare. In contrast, many common human illnesses result from the interaction of a large number of genes (polygenicity) in combination with nongenetic risk factors. Moreover, disease phenotypes tend to result from different combinations of genetic (and likely nongenetic) risk factors in different families and individuals. The use of the term VX-770 supplier “risk factor” rather than “cause” indicates that among polygenic disorders, any individual sequence variant (or

environmental factor) acts in a statistical rather than deterministic fashion. No single genetic variant is necessary or sufficient for the disorder phenotype and thus cannot be used to predict phenotype except in a probabilistic manner. Several important genetic results support polygenic models, for example, in schizophrenia and autism. The first is the finding that large numbers of common variants shape an individual’s disease risk. Statistical geneticists define a “polygene” from large constellations of common alleles that are observed (in one cohort) at slightly higher frequencies in schizophrenic patients than controls. When such alleles are subsequently evaluated in other cohorts, schizophrenic

patients are found to carry more such alleles (on average) than control subjects do. (Purcell et al., 2009). Within families, schizophrenic children of unaffected parents Rolziracetam also tend to have inherited more than the 50% of such alleles that they would be expected to have inherited by chance from parents heterozygous at the relevant loci (Ruderfer et al., 2011). These results suggest that one important component of genetic risk for a polygenic disorder such as schizophrenia arises from many small genetic nudges, rather than a single, hard shove. The polygenic model is also supported by rare alleles of larger effect. For example, a substantial minority of autistic patients (about 5%–10%), but only a small fraction of the general population (about 1%), have de novo deletions and duplications of large (>500 kb) genomic segments in their genomes.

Although one report demonstrated that human DCs can be transduced with ID-LVs [20], there was so far no information regarding their functionality in the stimulation of human T cell responses in vivo. Thus, here we further validated iDCs in order to address translationally relevant aspects regarding bio-safety and function. iDCs engineered with ID-LV expressing GM-CSF/IL-4 were characterized in vitro and in vivo. In addition, in order to evaluate a novel modality of ID-LV expressing a cytokine relevant for stimulation and/or expansion of NK cells and central memory T cells, we tested if human interferon alpha (IFN-α)

co-expressed with GM-CSF in monocytes would also result into iDCs. The combination of GM-CSF/IFN-α for the production of clinical DCs is currently being explored [21], 3-MA chemical structure but their co-expression in DCs via gene transfer has not been reported. This goal was achieved, and this new modality of iDC showed to be highly

viable and functional in vitro and in vivo. The construction of the vectors LV-GM-CSF-P2A-IL-4 (LV-G24), RRL-cPPT-CMV-pp65 (65 kDa phosphoprotein) and RRL-cPPT-CMV-fLUC (firefly luciferase) were previously described [10]. For the generation of the vector RRL-cPPT-CMV-GM-CSF-P2A-IFN-α (LV-G2α) overlapping-PCR was BMN 673 datasheet performed using cDNAs of human GM-CSF and human IFN-α (Origene technologies, Inc. Rockville, USA) as templates interspaced

with a 2A element of porcine teschovirus (P2A). The strategy of LV construction with P2A element was previously described [22]. Primers second used to generate the interspacing P2A element between GM-CSF, IFN-α were: P2A/IFN-α Forward 5′-GGATCCGGAGCCACGAACTTCTCTCTGTTAAAGCAAGCAGGAGACGTGGAAGAAAACCCCGGTCCTATGGCCTTGACCTTTGCTTTAC-3′ and P2A/GM-CSF Reverse: 5′-GTCTCCTGCTTGCTTTAACAGAGAGAAGTTCGTGGCTCCGGATCCCTCCTGGACTGGCTCCCAGCA-3′. The PCR products were digested with restriction enzymes XbaI and XmaI and inserted into the multiple cloning site of RRL-cPPT-CMV-MCS vector. The structural integrity of all constructs was confirmed by restriction digestion and sequencing analysis. Large scale lentivirus production was performed by transient co-transfection of human embryonic kidney 293T cells as formerly described [23]. 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin (100 U/ml) and streptomycin (100 mg/ml). The combination of the following packaging plasmids was used in the co-transfection: the plasmid containing the lentiviral vector expressing the cytokines, the plasmid expressing rev (pRSV-REV), the plasmid expressing gag/pol containing a D64V point mutation in the integrase gene (pcDNA3g/pD64V.4xCTE), and the plasmid encoding the VSV-G envelope (pMD.G).

, 2008). In this respect, we found a prominent

enhancement of the heat sensitivity of TRPM3 by the neurosteroid PS. In particular, our data indicate strong synergism between heat and PS at concentrations between 100 and 1000 nM, which is well within the range of plasma PS levels measured in adult humans (0.1–0.8 μM Havlíková et al., 2002). Plasma PS levels can rise to supramicromolar concentrations during parturition Selleck 3 Methyladenine and under various pathological conditions but also decreases with aging (Havlíková et al., 2002, Hill et al., 2001 and Schumacher et al., 2008), which may further influence heat sensitivity and pain through TRPM3. Clearly, further study is needed to elucidate the in vivo interplay between neurosteroid production and TRPM3 activity in normal and pathological conditions. In conclusion, we have identified TRPM3 as nociceptor Ibrutinib channel involved in acute heat sensing and inflammatory heat hyperalgesia,

and thus as a potential target for analgesic treatments. Human embryonic kidney cells, HEK293T, were grown in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% (v/v) human serum, 2 mM L-glutamine, 2 units/ml penicillin, and 2 mg/ml streptomycin at 37°C in a humidity-controlled incubator with 10% CO2. HEK293T cells were transiently transfected with murine TRPM3α2 (accession number AJ 544535) in the bicistronic pCAGGS/IRES-GFP vector, using Mirus TransIT-293 (Mirus corporation; Madison, WI, USA). Transfected cells were visualized by green fluorescence protein (GFP) expression, whereas GFP-negative cells from the same batch were used as controls. TG and DRG neurons from adult (postnatal weeks 8–12) male mice were isolated as described previously (Karashima et al., 2007). HEK293T cells stably transfected with TRPM3α2 were developed using the Flp-In System (Invitrogen). Trpm3−/− SB-3CT mice ( Figure S2), obtained from Lexicon Genetics (see http://www.informatics.jax.org/searches/accession_report.cgi?id=MGI:3528836), were generated using homologous recombination in 129SvEvBrd ES cells. ES cells were injected into blastocysts from C57BL/6J donor mice to generate chimeric animals, which were mated with C57BL/6J mice and genotyped

for the mutated allele. Heterozygotes were mated, resulting in Trpm3+/+, Trpm3+/−, and Trpm3−/− mice with the expected Mendelian distribution. Unless mentioned otherwise, paired Trpm3+/+and Trpm3−/− littermates were used in behavioral experiments. For comparison, we also used age-matched pure 129SvEvBrd (kindly provided by The Sanger Institute, Cambridge, UK) and C57BL/6J (Charles River) mice in behavioral experiments, as indicated in the text. Trpv1−/− mice in pure C57BL/6J background were obtained from The Jackson Laboratory (http://jaxmice.jax.org/strain/003770.html), and age- and weight-matched C57BL/6J mice were used as matched controls (Trpv1+/+). Trpv1−/− mice were mated with Trpa1−/− mice ( Kwan et al., 2006) to obtain Trpv1−/−/Trpa1−/− double-knockout mice.

These

results indicate that the axons of selleck screening library DGCs born at P15 are synaptically integrated by P23 to compete with mature axons. To examine the specificity of the effect of AraC, we used DG-S mice. In DG-S mice, tTA is expressed by mature DGCs (37.1% ± 1.4%; Figures 3B and 3C) and CA1 neurons (48.2% ± 3.3%; Figure S5). Using DG-S::TeTxLC-tau-lacZ mice, we addressed whether AraC specifically inhibits the elimination of inactive DG axons or also affects the elimination of inactive CA1 axons. If the effect of AraC is through the suppression of neurogenesis, the elimination of inactive CA1 axons should not be affected. In DG-S::TeTxLC-tau-lacZ mice, axons PD-332991 of TeTxLC-expressing DG and CA1 neurons were both eliminated between P15 and P25 (Figure 3F and Figure 8A). After P25, very few TeTxLC-expressing axons were observed in both regions. On the other hand, DG-S::tau-lacZ mice (no TeTxLC) maintained β-gal-expressing axons of DG and CA1 neurons at P25 (Figure 8C). Thus, in DG-S mice, DG and CA1 axons are refined in an activity-dependent

manner between P15 and P25. We administered AraC daily (i.p. injection) from P15 to P22 (8 days total) into DG-S::TeTxLC-tau-lacZ mice. AraC injections inhibited the elimination of TeTxLC-expressing DG axons, but not that of TeTxLC-expressing CA1 axons (Figure 8D). Therefore, the effect of AraC is specific to DG axons, which is consistent with the fact

that neurogenesis is specific to the DG in the hippocampus. Finally, using DG-S mice, we addressed whether the DG axon refinement is mainly achieved by competition between mature and young DG axons or also by competition between mature and mature axons. In DG-S mice, tTA is expressed only by 37.1% ± 1.4% of mature DGCs (Figures 3B and 3C). If competition between mature and mature neurons equally contributes to refinement, AraC injections should not inhibit inactive axon elimination in the DG-S::TeTxLC-tau-lacZ line, which contains both active and inactive mature DGCs. Interestingly, however, AraC injections (from P15 to P22) inhibited the elimination of TeTxLC-expressing DG axons in DG-S::TeTxLC-tau-lacZ mice (Figures second 8D and 8E) as effectively as in DG-A::TeTxLC-tau-lacZ mice (Figures 6D and 6E). This suggests that the activity-dependent competition of DG axons is largely between axons of mature and young neurons. To further confirm that young neurons drive inactive axon elimination in the DG, we utilized nestin-tk animals, in which herpes simplex virus thymidine kinase is expressed under the control of the nestin enhancer to drive expression in neural progenitors (Singer et al., 2009). In this animal, administration of ganciclovir efficiently and specifically kills neural progenitors.

This downward adjustment of synaptic currents occurs, at least partly, during sleep (see below). Synaptic plasticity can also offer energetic savings to synaptic transmission. Long-term depression of the cerebellar parallel fiber to Purkinje cell synapse, used to learn motor patterns, ultimately results in ∼85% of the synapses producing no postsynaptic current (Isope and Barbour, 2002). The existence of silent synapses is predicted theoretically for optimal

storage of information (Brunel et al., 2004) but also provides a massive decrease in the amount of energy used synaptically (Howarth et al., 2010). Increasingly, sleep is thought to play an energetically restorative role in the brain (Scharf et al., 2008). This theory coincides with most people’s experience of sleep but has found direct physiological support only recently. Dworak et al. selleck screening library (2010) reported that during sleep there is a transient increase in ATP level in cells of awake-active regions of the brain. This was suggested to fuel restorative biosynthetic processes in cells that, during the day, must use all of their energy on electrical and chemical signaling. This implies an energy consumption trade-off:

a high use of ATP on synapses during awake ISRIB research buy periods is balanced by more ATP being allocated to other tasks during sleep. Energy use in the awake state also increases due to synaptic potentiation. In the awake state (compared to sleep), GluR1 subunits of AMPA receptors are present at a higher level and in a more phosphorylated state (consistent with an increased synaptic strength), synaptic currents and spine numbers increase, and evoked neuronal responses are larger (Vyazovskiy et al., 2008; Maret et al., 2011). These changes

are reversed during sleep, presumably because of homeostatic Sodium butyrate plasticity as discussed above. Thus, sleep is essential for adjusting synaptic energy use. Finally, we turn to the pathological effects of disruptions to synaptic energetics. Since synapses account for the majority of energy use in the brain, any disorder of mitochondrial trafficking or function will inevitably affect synapses. Reciprocally, excessive glutamatergic synaptic transmission raises neuronal [Ca2+]i, which depolarizes mitochondria, reducing their ATP production and in extremis leading to cytochrome C release and the initiation of apoptosis. Increasingly, one or other of these mitochondrial dysfunctions is reported as contributing to brain disorders. Mitochondrial dysfunction may contribute to neuronal damage in Parkinson’s disease (Youle and Narendra, 2011). Dopaminergic neurons in the substantia nigra consume a significant amount of ATP to reverse the Ca2+ influx that mediates their pacemaking activity (Puopolo et al., 2007). Producing this ATP leads to oxidative stress (Guzman et al., 2010) that can uncouple or depolarize mitochondria.

We conducted

experiments to assess its impact on palmitoylation. NMDA treatment of granule cells reduces palmitoylation of PSD-95 by ∼70% (Figure 2H). Vorinostat in vitro This action reflects NMDA augmentation of NO formation, as it is largely reversed by treatment with L-VNIO. Moreover, we observe reciprocal changes in PSD-95 nitrosylation, suggesting that nitrosylation is responsible for the effects of NMDA-mediated NO formation (Figure 3I). To further explore the importance of endogenous NO in regulating PSD-95 palmitoylation, we made use of nNOS-deleted mice (Figure 3J). We confirm the marked reduction of PSD-95 palmitoylation in response to NMDA treatment. Levels of PSD-95 palmitoylation are significantly augmented in nNOS-deleted granule cells both in the absence and presence of NMDA treatment (Figure 3J). NMDA does signaling pathway elicit some decrease in PSD-95 palmitoylation in nNOS knockout mice, which may reflect the retention of alternatively spliced, catalytically active nNOS (Eliasson

et al., 1997) or compensatory mechanisms involving eNOS or iNOS (Huang and Fishman, 1996). Thus, diverse experimental approaches establish that in mammalian brain, endogenous NO under basal conditions and in response to NMDAR-mediated neurotransmission regulates the palmitoylation of PSD-95. One of the principal roles of PSD-95 palmitoylation is to regulate synaptic clustering of the protein. Thus, synaptic clustering of PSD-95 is abolished with C3 and C5 mutation, which prevents palmitoylation (Craven et al., 1999). Moreover, inhibition of palmitoylation with 2-bromopalmitate (2-BP) reduces PSD-95 synaptic clusters (El-Husseini et al., 2002). Bredt, Nicoll, and Fukata established that glutamate neurotransmission, acting via ionotropic receptors, diminishes PSD-95 clustering both by inhibiting palmitoylation (Noritake

et al., 2009) and by stimulating depalmitoylation (el-Husseini and Bredt, 2002). We wondered whether this process might involve influences of NO acting by displacing Cediranib (AZD2171) palmitate from PSD-95. Accordingly, we examined the effect of NMDA treatment upon PSD-95 clustering in cerebellar granule neurons and evaluated the influence of the nNOS inhibitor VNIO (Figure 4). PSD-95 is synaptic in localization as it is closely apposed to clusters of synapsin, a presynaptic marker, with statistically significant colocalization (Pearson correlation coefficient, 0.45) (Figure 4A). NMDA treatment reduces PSD-95 synaptic clusters by ∼40%, while VNIO significantly reverses this action. In contrast, synapsin clusters are unchanged. The partial reversal by VNIO may indicate that the NMDA effects involve other signaling systems in addition to NO, possibly associated with calcium entering cells through NMDA channels. Double labeling is specific, as demonstrated by omission of primary antibodies (Figure 5C). As previously reported, 2-BP reduces PSD-95 clustering (Figures 5A and 5B).

Fluorescence images were acquired by using a high-speed confocal laser scanning microscope (CSU22, Yokogawa, Japan), as described (Tanimura et al., 2009). Other details are described in Supplemental Experimental Procedures. Under anesthesia with chloral hydrate (350 mg/kg Cabozantinib price of body weight, i.p.), a glass pipette filled with 2–3 μl of 10% solution of DTR (3,000 molecular weight; Invitrogen, Carlsbad, CA) in PBS (pH 7.4) was inserted stereotaxically to the inferior olive. The tracer was injected by air pressure at 10 psi with 5 s intervals for 1 min. After 4 days of survival, mice were fixed by transcardial perfusion with 4% paraformaldehyde

in 0.1 M PB, and parasagittal cerebellar sections (50 μm in thickness) were cut. The DTR-labeled sections were incubated overnight with a mixture of goat calbindin antibody and guinea pig type II VGluT2 antibody (Miyazaki et al., 2003) followed by 2 hr incubation with a mixture of Alexa

488- (Invitrogen) and Cy5-labeled species-specific secondary antibodies. Images of triple labeling were taken with confocal laser-scanning microscope (FV1000, Olympus). Procedures for immunohistochemistry are described in Supplemental Experimental Procedures. To inhibit GAD activity or enhance GABAA receptor sensitivity to GABA in the cerebellum, we applied 3-MP or diazepam, respectively, by continuous infusion from Elvax implants prepared as described (Kakizawa et al., 2000, Kakizawa et al., 2003 and Kakizawa et al., 2005). For the implantation of an

Elvax piece, Epacadostat clinical trial mice at P10 or P17 were anesthetized with isoflurane, the skin over the cerebellum was cut, and the occipital bone and the dura over the cerebellar lobules 6–8 was removed. A small piece of Elvax containing drugs or vehicle was placed on the cerebellar surface and then the skin was sutured. Effects of chronic infusion of 3-MP or diazepam were examined electrophysiologically at P23–P40 within those the lobules 6–8. Throughout the text and figures, data are presented as means ± SEM. Statistical significance was assessed by Student’s t test or Mann-Whitney U test (for comparison of two independent samples), two-tailed paired-t test or Wilcoxon’s signed-rank test (for paired comparison of the same sample), depending on whether the data sets pass the normality test and equal variance test, unless otherwise mentioned in the text. For comparison of frequency distributions, Mann-Whitney U test was used. Statistical analysis was conducted with Sigma Stat 3.1 program. p value was described as p < 0.001 when the actual p value was smaller than 0.001. Differences between data sets were judged to be statistically significant if the p value was less than 0.05. We thank S. Kakizawa for helpful advice on preparation and implantation of Elvax, A. Koseki for mice genotyping and members of Kano’s lab for discussion.

Morphed probe trials. After the acquisition of the discrimination problem, performance was evaluated by using feature-ambiguous probe trials. These probe trials increased the difficulty of the discrimination task by increasing the similarity of the S+ and S−. Probe trials were created by morphing the S+ and S− into one another in 14 steps (Morpheus Photo Animator; ACD Systems, Saanichton, Canada). Thus, one stimulus was gradually morphed into the other, physically changing each stimulus from one step to

the next ( Figure 2). This morphing procedure is similar to procedures used in previous work with monkeys ( Bussey et al., 2003) and humans ( Lee et al., Selleckchem MDV3100 2005 and Shrager et al., 2006). Note that one stimulus was not blended into the other. Rather, the entire stimuli were gradually altered so that they became more alike. Probe level 1 consisted of the least amount of feature overlap (i.e., the two stimuli were quite distinct and most similar to the training stimuli). At level 14 the two stimuli contained substantial feature overlap and appeared quite similar ( Figure 2). During this phase of testing, DNA Damage inhibitor 80% of the trials were standard trials (training stimuli). The remaining 20% of the trials

were rewarded morphed probe trials. The order of the probe trials (levels 1–14) was pseudorandom with the constraint that each of the 14 difficulty levels had to be presented once before any one difficulty level could be repeated. This procedure ensured that data for probe trials accrued at the same rate for every difficulty level. This phase of testing continued until 150 probe trials were completed at each difficulty level. Thus, across this phase of testing each animal received 2,100

probe trials across the 14 different difficulty levels (150 × 14) and an additional 10,500 trials with the training stimuli. Surgery. Animals were assigned to a perirhinal lesion group or a normal control group based upon their trials-to-criterion score for the discrimination task (to create two equal groups). The intention was to and remove the entire perirhinal cortex bilaterally. For surgery, the rat was placed in a Kopf stereotaxic instrument and the incisor bar was adjusted until bregma was level with lambda. Bilateral excitotoxic perirhinal lesions were produced by local microinjections of ibotenate acid (IBO; Biosearch Technologies, San Rafael, CA). IBO was dissolved in 0.1 M phosphate-buffered saline to provide a solution with a concentration of 10 mg/ml, pH 7.4. IBO was injected at a rate of 0.1 μl/min with a 10 μl Hamilton syringe mounted on a stereotaxic frame and held with a Kopf microinjector (model 5000). The syringe needle was lowered to the target coordinate and left in place for 1 min before beginning the injection. After the injection, the syringe needle was left in place for a further 5 min to reduce the spread of IBO up the needle tract. A total of 0.

Both interface in V4 and both selectively shape networks in V4 (cf. Reynolds and Desimone, 2003 and Qiu et al., 2007). (Note that for the purposes of this review, although object “salience” may influence attention, we consider this part of the bottom-up process. Here, we use the term “attention” to refer to internally generated, top-down influences.) We frame our conception of V4 function in terms of “selection”. The visual attention literature commonly uses the term “select” to indicate either a region of space that is selected (spatial attention) or specific object features that are selected (feature http://www.selleckchem.com/products/Rapamycin.html attention). In the same vein,

objects in the visual scene “select” the neuronal networks in V4 that encode their features. We propose that these two “selection” processes share a common framework. More specifically, we propose that the functional architecture in V4 is the substrate through which both sets of influences are mediated and that, at the neural level, selective

modulation of networks in V4 may be fundamentally the same, albeit directed from different sources. Our perceptual system is continuously confronted with much more information than it can actively deal with. One way to reduce processing load is to select a fraction of the incoming visual information for scrutinized processing. Visual attention achieves this by focusing on a particular location in space (spatial attention) or on certain features of objects (feature attention). The ability to attend appropriately ATR inhibitor can be negatively affected by having other competing objects (distractors) in the visual field. In the biased competition model of visual attention (Bundesen, 1990, Desimone and Duncan, 1995 and Grossberg, 1980), attentional selection is achieved via a competition for neural resources; this competition can be biased in several ways. One source of this bias comes from involuntary, click here sensory-driven bottom-up mechanisms (e.g., salient attention-attracting stimuli). Another biasing mechanism is voluntary

attentional top-down feedback (e.g., internally generated goal-directed attention), which presumably originates in areas outside the visual cortex. The biased competition model states that only those stimuli that win the competition against surrounding distractor stimuli will have further access to higher order neural mechanisms linking percepts to mechanisms sustaining goal-directed actions including systems involved in memory, decision-making and generating motor plans (Desimone and Duncan, 1995, Luck et al., 1997 and Moran and Desimone, 1985). One goal of this review is to consider this integrative bottom-up and top-down view in the context of functional organization in V4. Spatial attention has often been characterized as a “spotlight” on a region in space where visual processes appear heightened (e.g., Posner, 1980).

It seems plausible that sedentary people have more benefit on their sleep after joining an exercise event than active people do.27 The participants of our study had a normal PA level at baseline. Therefore, it can be assumed that PA of longer duration, above the national recommendations, is needed for this activity level to improve in sleep quality, but also the higher general activity during the day reveals sleep-promoting effects. Furthermore,

the Baecke sport index from baseline did not correspond to improvements in sleep quality and therefore the program seems to be effective for both unfit and fit participants. In general, the regression analysis did not show any correspondence to the intensity of PA. Even though, the recommendations to the participants to be physically active on a moderate intensity level, there was a range from PLX-4720 order 7 to 17 in individual data of perceived exertion on the Borg scale. The previous research is ambiguous about whether the dose–response effect is due to increased doses of exercise intensity, duration, or both.17 At least from our analysis we can conclude that the intensity might be of less importance than the duration

of PA. Buman and King17 learn more suggested that a minimum of 16 weeks of intervention would be needed along with exercise doses that meet or exceed current PA recommendations to answer this question satisfactorily. In our study with an intervention time of 6 weeks we achieved an average 3.1 point reduction in the PSQI global score16 which is comparable to the findings of King et al.28 Thiamine-diphosphate kinase with an average reduction of 3.3 after a 16-week

moderate endurance exercise intervention. As Youngstedt8 mentioned, an important, but overlooked, consideration in assessing treatment efficacy may be ceiling and floor effects, which dictate that the greater the initial impairment in sleep, the greater the potential for improvement. In the regression analyses severity of sleep symptoms at baseline (PSQI and SF-B) are one of the predictors for the changes in sleep quality. Therefore, it can be assumed that the higher the sleep severity symptoms the more steps and exercise of longer duration has to be done to get improvements in sleep. With respect to PA-F, PA-D, and PA-I but also the length of the treatment, additional research is needed in this area to formally test dose–response effects for chronic exercise on sleep. The second aim of the study was to display the week-to-week variability of sleep quality and PA starting from a baseline week over the 6-week intervention period. Our data showed as expected an increase of PA due to the intervention program: PA-F increased from 2.6 times in the baseline week to an average of 4.2 times during the weeks of intervention, PA-D augmented from 176 min in the baseline week to 279 min during the weeks of intervention. In contrast, PA-I showed a slight but statistically not significant increase from 11.9 to 12.3 over time.