Evaluating the Safety of Intra-Articular Mitotherapy in the Equine Model: A Potential Novel Treatment for Osteoarthritis.

No current treatments available halt osteoarthritis progression in horses or humans. Intra-articular injection of mitochondria is a novel treatment that has the potential to improve cell metabolism and decrease inflammation, but safety of this treatment has yet to be established in the horse. Autologous blood-derived mitochondria isolated using a commercially available kit were injected into the left carpus joint of 3 horses which were monitored for 28 days. Horses received physical examinations, video recorded gait evaluations, joint diameter measurement, synovial fluid collection, and blood collection on day 0 (baseline prior to mitotherapy, day of mitochondria injection), 1, 3, 7, 14, and 28. Systemic inflammation was assessed via complete blood count, fibrinogen, and plasma serum amyloid A (SAA). Local inflammation was assessed via synovial fluid cytology and physical examination parameters. Physical exam parameters remained stable and no joint swelling was observed after mitotherapy. No change was noted in video recorded gait evaluations as determined by a blinded evaluator. Complete blood counts revealed no significant increase in white blood cells. SAA only increased mildly in 1 horse. Fibrinogen became slightly elevated above reference range in 2 horses at day 7, but later normalized. Mild increases in synovial fluid nucleated cell counts and total protein occurred on day 1 and 3, but resolved within 7 days without intervention. Autologous mitochondria injection into the equine intercarpal joint was well tolerated with no signs of inflammation. This safety information allows for future studies evaluating mitotherapy efficacy.

Focus meets motivation: When regulatory focus aligns with approach/avoidance motivation in creative processes.

According to Regulatory Focus Theory, two systems determine our strategies to pursue goals - the promotion and the prevention system. Individuals with a dominant promotion system focus on achieving gains, i.e., promoters, and individuals with a dominant prevention system focus on avoiding losses, i.e., preventers. Regulatory Fit Theory suggests that a fit between this focus and the situation causes superior performance and makes individuals feel right. We transfer the fit idea to the interaction of dominant regulatory focus (promotion vs. prevention) with motivational direction (approach vs. avoidance motivation). We investigated these interaction effects on individuals' performance and their experience within creativity workshops. In Study 1 (N 1 = 172), using multi-level analyses, we found that a promotion focus was associated with fluency and a prevention focus with elaborated ideas. This effect was stronger, when preventers also scored high on avoidance motivation. Further, preventers experienced more autonomy support and were more satisfied when they scored high on avoidance. Promoters high on approach motivation reported more autonomy support and more satisfaction than preventers high on approach motivation. For Study 2 (N 2 = 112), we used an experimental design: After measuring regulatory focus, we manipulated approach vs. avoidance motivation in creativity workshops. Using multi-level analyses, we did not find main or interaction effects on fluency or elaboration but we found interaction effects on participants' experience of the creativity workshop. Preventers were more satisfied when they received the avoidance condition. Promoters reported less autonomy support, lower satisfaction, and more perceived conflicts within their teams in the avoidance condition.

Clinical Development of Sphingosine as Anti-Bacterial Drug: Inhalation of Sphingosine in Mini Pigs has no Adverse Side Effects.

BACKGROUND/AIMS: Pulmonary infections with Pseudomonas aeruginosa (P. aeruginosa) or Staphylococcus aureus (S. aureus) are of utmost clinical relevance in patients with cystic fibrosis, chronic obstructive pulmonary disease, after trauma and burn, upon ventilation or in immuno-compromised patients. Many P. aeruginosa and S. aureus strains are resistant to many known antibiotics and it is very difficult or often impossible to eradicate the pathogens in patient´s lungs. We have recently shown that the sphingoid base sphingosine very efficiently kills many pathogens, including for instance P. aeruginosa, S. aureus or Acinetobacter baumannii, in vitro. In vivo experiments of our group on cystic fibrosis mice indicated that inhalation of sphingosine prevents or eliminates existing acute or chronic pneumonia with P. aeruginosa or S. aureus in these mice. We also demonstrated that sphingosine is safe to use for inhalation up to high doses, at least in mice. To facilitate development of sphingosine to an anti-bactericidal drug that can be used in humans for inhalation, safety data on non-rodents, larger animals are absolutely required. METHODS: Here, we inhaled mini pigs with increasing doses of sphingosine for 10 days and analyzed the uptake of sphingosine into epithelial cells of bronchi as well as into the trachea and lung and the systemic circulation. Moreover, we measured the generation of ceramide and sphingosine 1-phosphate that potentially mediate inflammation, the influx of leukocytes, epithelial cell death and disruption of the epithelial cell barrier. RESULTS: We demonstrate that inhalation of sphingosine results in increased levels of sphingosine in the luminal membrane of bronchi and the trachea, but not in systemic accumulation. Inhaled sphingosine had no side effects up to very high doses. CONCLUSION: In summary, we demonstrate that inhalation of sphingosine results in an increase of sphingosine concentrations in the luminal plasma membrane of tracheal and bronchial epithelial cells. The inhalation has no systemic or local side effects.

Honokiol, a constituent of Magnolia species, inhibits adrenergic contraction of human prostate strips and induces stromal cell death.

PURPOSE: Smooth muscle contraction and prostate growth are important targets for medical therapy of lower urinary tract symptoms (LUTS) in patients with benign prostatic hyperplasia. Honokiol and Magnolol are lignan constituents of Magnolia species, which are used in traditional Asian medicine. Here, we examined effects of honokiol and magnolol on contraction of human prostate tissue and on growth of stromal cells. METHODS: Prostate tissues were obtained from radical prostatectomy. Contraction of prostate strips was examined in organ bath studies. Effects in stromal cells were assessed in cultured immortalized human prostate stromal cells (WPMY-1). Ki-67 mRNA was assessed by reverse transcription-polymerase chain reaction, and proliferation by a fluorescence 5-ethynyl-2'-deoxyuridine assay. RESULTS: Honokiol (100µM) reduced noradrenaline-induced contractions, which was significant at 10 to 100µM noradrenaline. Honokiol reduced phenylephrine-induced contractions, which was significant at 3 to 100µM phenylephrine. Honokiol reduced electric field stimulation-induced contractions very slightly. In WPMY-1 cells, honokiol (24 hours) induced cell death. Magnolol (100µM) was without effects on contraction, and cellular viability. CONCLUSIONS: Honokiol inhibits smooth muscle contraction in the human prostate, and induces cell death in cultured stromal cells. Because prostate smooth muscle tone and prostate growth may cause LUTS, it appears possible that honokiol improves voiding symptoms.

The receptor antagonist picotamide inhibits adrenergic and thromboxane-induced contraction of hyperplastic human prostate smooth muscle.

Inhibition of prostate smooth muscle contraction is an important strategy for medical treatment of lower urinary tract symptoms (LUTS). Besides α1-adrenoceptors, prostate smooth muscle contraction is induced by activation of thromboxane (TXA2) receptors (TXA2-R). Here, we examined the effects of the TXA2-R antagonist picotamide on contraction of human prostate tissue. Prostate tissues were obtained from radical prostatectomy. The effects of picotamide (300 µM), L-665,240 (3 µM), and seratrodast (3 µM) on U46619-, electric field stimulation- (EFS-), phenylephrine-, and norepinephrine-induced contractions were studied in organ baths. Expression of TXA2-R and TXA2 synthase (TXS) was examined by fluorescence stainings. Picotamide, seratrodast, and L-655,240 inhibited concentration-dependent contractions induced by the TXA2 analog U46619. Picotamide, but not seratrodast or L-655,240, inhibited frequency-dependent contractions induced by EFS. Picotamide inhibited concentration-dependent contractions induced by norepinephrine or by the selective α1-adrenoceptor agonist phenylephrine. In prostate strips, where only submaximal contraction by a low dose of phenylephrine was induced, application of U46619 raised tone to maximum phenylephrine-induced tension. Immunoreactivity for TXA2-R and TXS was observed in the stroma and in epithelial cells of glands. Colocalization of both immunoreactivites was observed with the smooth muscle markers calponin and α-smooth muscle actin, with the epithelial marker pan-cytokeratin, and with prostate-specific antigen in the stroma and glands. The receptor antagonist picotamide inhibits α1-adrenergic, TXA2-mediated, and EFS-induced contractions in the human prostate. To the best of our knowledge, this is the first antagonist able to inhibit two different contraction systems in the prostate.

Role of structural order and excess energy on ultrafast free charge generation in hybrid polythiophene/Si photovoltaics probed in real time by near-infrared broadband transient absorption.

Despite the central role of light absorption and the subsequent generation of free charge carriers in organic and hybrid organic-inorganic photovoltaics, the precise process of this initial photoconversion is still debated. We employ a novel broadband (UV-Vis-NIR) transient absorption spectroscopy setup to probe charge generation and recombination in the thin films of the recently suggested hybrid material combination poly(3-hexylthiophene)/silicon (P3HT/Si) with 40 fs time resolution. Our approach allows for monitoring the time evolution of the relevant transient species under various excitation intensities and excitation wavelengths. Both in regioregular (RR) and regiorandom (RRa) P3HT, we observe an instant (<40 fs) creation of singlet excitons, which subsequently dissociate to form polarons in 140 fs. The quantum yield of polaron formation through dissociation of delocalized excitons is significantly enhanced by adding Si as an electron acceptor, revealing ultrafast electron transfer from P3HT to Si. P3HT/Si films with aggregated RR-P3HT are found to provide free charge carriers in planar as well as in bulk heterojunctions, and losses are due to nongeminate recombination. In contrast for RRa-P3HT/Si, geminate recombination of bound carriers is observed as the dominant loss mechanism. Site-selective excitation by variation of pump wavelength uncovers an energy transfer from P3HT coils to aggregates with a 1/e transfer time of 3 ps and reveals a factor of 2 more efficient polaron formation using aggregated RR-P3HT compared to disordered RRa-P3HT. Therefore, we find that polymer structural order rather than excess energy is the key criterion for free charge generation in hybrid P3HT/Si solar cells.

Investigation of two-beam-pumped noncollinear optical parametric chirped-pulse amplification for the generation of few-cycle light pulses.

We demonstrate a new and compact Phi-plane-pumped noncollinear optical parametric chirped-pulse amplification (NOPCPA) scheme for broadband pulse amplification, which is based on two-beam-pumping (TBP) at 532 nm. We employ type-I phase-matching in a 5 mm long BBO crystal with moderate pump intensities to preserve the temporal pulse contrast. Amplification and compression of the signal pulse from 675 nm - 970 nm is demonstrated, which results in the generation of 7.1-fs light pulses containing 0.35 mJ energy. In this context, we investigate the pump-to-signal energy conversion efficiency for TBP-NOPCPA and outline details for few-cycle pulse characterization. Furthermore, it is verified, that the interference at the intersection of the two pump beams does not degrade the signal beam spatial profile. It is theoretically shown that the accumulated OPA phase partially compensates for wave-vector mismatch and leads to extended broadband amplification. The experimental outcome is supported by numerical split-step simulations of the parametric signal gain, including pump depletion and parametric fluorescence.

Generation of sub-three-cycle, 16 TW light pulses by using noncollinear optical parametric chirped-pulse amplification.

We present a two-stage noncollinear optical parametric chirped-pulse amplification system that generates 7.9 fs pulses containing 130 mJ of energy at an 805 nm central wavelength and 10 Hz repetition rate. These 16 TW light pulses are compressed to within 5% of their Fourier limit and are carefully characterized by the use of home-built pulse diagnostics. The contrast ratio before the main pulse has been measured as 10(-4), 10(-8), and 10(-11) at t=-3.3 ps, t=-5 ps, and t=-30 ps, respectively. This source allows for experiments in a regime of relativistic light-matter interactions and attosecond science.