Objective:To describe how differing injector needles and delivery vehicles impact Autologous Muscle-Derived Cell (AMDC) viability when used for laryngeal injection.Methods:In this study, adult porcine muscle tissue was harvested and used to create AMDC populations. While controlling cell concentration (1 × 107 cells/ml), AMDCs including Muscle Progenitor Cells (MPCs) or Motor Endplate Expressing Cells (MEEs) were suspended in either phosphate-buffered saline or polymerizable (in-situ scaffold forming) type I oligomeric collagen solution. Cell suspensions were then injected through 23- and 27-gauge needles of different lengths at the same rate (2 ml/min) using a syringe pump. Cell viability was measured immediately after injection and 24- and 48-hours post-injection, and then compared to baseline cell viability prior to injection.Results:The viability of cells post-injection was not impacted by needle length or needle gauge but was significantly impacted by the delivery vehicle. Overall, injection of cells using collagen as a delivery vehicle maintained the highest cell viability.Conclusion:Needle gauge, needle length, and delivery vehicle are important factors that can affect the viability of injected cell populations. These factors should be considered and adapted to improve injectable MDC therapy outcomes when used for laryngeal applications.
Hypertension and breast cancer are two common diseases occurring in women. Clinical studies have shown increased breast cancer incidence in hypertensive women. Several lines of evidence demonstrate that G protein-coupled Receptor Kinase 4 (GRK4) could be a common risk factor for hypertension and breast cancer. This article reviews our current understanding of molecular mechanisms of GRK4 in hypertension and breast cancer.
Light exposure at night can disrupt the circadian timing of cellular processes and is associated with a broad range of health disorders. To spectrally engineer lighting which minimizes circadian disruption at night it is necessary to define the precise spectral sensitivity of the human circadian system. Prior attempts have used short monochromatic light exposures in dark-adapted human subjects, or in vitro dark-adapted isolated retina or melanopsin. However, humans spend virtually all their awake hours in a fully light-adapted state. Here we review the evidence for a narrow blue circadian sensitivity curve for light-adapted humans derived from experiments using spectral filtering of light sources, and comparisons of light sources with diverse spectral power distributions. This light-adapted Circadian Potency function permits the development of circadian-protective light for nocturnal use and circadian-entraining light for daytime use.
1. In the established classic UV-cAMP-MITF-dependent “tanning pathway”, UV radiation mediates an increase in the transcription factor p53 in human keratinocytes, thereby stimulating the induction of Pro-Opiomelanocortin (POMC) and secretion of the Alpha Melanocyte-Stimulating Hormone (αMSH). In turn MSH binds the Melanocortin 1 Receptor (MC1R) on melanocytes, which increases the expression of the Microphthalmia Associated Transcription Factor (MITF) directly regulating the transcription of Tyrosinase (TYR), Tyrosinase-Related Protein 1 and 2 (TRP1 and TRP2) and other pigmentation genes 1 (Figure 1A). Although pigmentation regulation by the MITF dependent pathway is well established, other pathways that are independent of MITF are not yet fully elucidated. Deciphering other melanogenesis mechanisms will facilitate understanding of pathogenesis associated with pigmentation disorders and the development of potential therapeutic options. Particularly, the question of how redox in mitochondria
Ultraviolet (UV) light and incompletely understood genetic and epigenetic variations determine skin color. Here we describe an UV- and microphthalmia-associated transcription factor (MITF)-independent mechanism of skin pigmentation. Targeting the mitochondrial redox-regulating enzyme nicotinamide nucleotide transhydrogenase (NNT) resulted in cellular redox changes that affect tyrosinase degradation. These changes regulate melanosome maturation and, consequently, eumelanin levels and pigmentation. Topical application of small-molecule inhibitors yielded skin darkening in human skin, and mice with decreased NNT function displayed increased pigmentation. Additionally, genetic modification of NNT in zebrafish alters melanocytic pigmentation. Analysis of four diverse human cohorts revealed significant associations of skin color, tanning, and sun protection use with various single-nucleotide polymorphisms within NNT. NNT levels were independent of UVB irradiation and redox modulation. Individuals with postinflammatory hyperpigmentation or lentigines displayed decreased skin NNT levels, suggesting an NNT-driven, redox-dependent pigmentation mechanism that can be targeted with NNT-modifying topical drugs for medical and cosmetic purposes.
Uterine leiomyosarcoma (LMS) is an aggressive tumor that presents a poor prognosis, high rates of recurrence, and metastasis. Because of its rarity, there is no information available concerning LMS molecular mechanisms of origin and development. Here, we assessed the expression profile of Hedgehog (HH) signaling pathway markers and the effects of their pharmacological inhibition on uterine smooth muscle (UTSM), leiomyoma, and LMS cells. Additionally, we also evaluated the effects of DNMTs inhibition on LMS cell behavior. Cell proliferation, migration and apoptosis rates were evaluated by MTT, Scratch, and Annexin V assays, respectively. RNA expression and protein levels were assessed by qRT-PCR and Western blot. We found that SMO and GLIs (1, 2, and 3) expression was upregulated in LMS cells, with increased nuclear levels of GLI proteins. Treatment with LDE225 (SMOi) and Gant61 (GLIi) resulted in a significant reduction in Glis protein levels in LMS (p < 0.05). Additionally, the expression of DNMT (1, 3a, and 3b), as well as GLI1 nuclear expression, was significantly decreased after treatment with HH inhibitor in LMS cells. Our results showed that blocking of SMO, GLI, and DNMTs is able to inhibit LMS proliferation, migration, and invasion. Importantly, the combination of those treatments exhibited a potentiated effect on LMS malignant features due to HH pathway deactivation.
When we aspire to cure cancer, we may need to search no further than a curable cancer, such as Germ Cell Tumor of the Testis (TGCT). After all, a germ cell is a primordial stem cell. Importantly, TGCT provides a classic stem cell model of cancer that teaches us some invaluable lessons about curing other intractable solid tumors. The intrinsic intratumoral heterogeneity of TGCT alludes to its stem-ness origin and nature. Which implicates the existence of putative lethal TGCT subtypes-the identification and detection of which may further enhance the cure rate and improve the therapeutic ratio of TGCT. In this Mini review, we discuss about the role of biologic insights, clinical lessons, and therapeutic strategies in drug and therapy development. We illustrate some clinical pearls and perils when it concerns drug versus therapy development in the cure and care of patients with TGCT. In many respects, we have cured more TGCT patients when we apply multimodal therapy rather than targeted therapy and integrated medicine rather than precision medicine. In principle and in practice, this is the implication of therapy versus drug development in improving the overall outcome and cure rate of patients with cancer.
O imperfecta (OI) is a congenital dysplasia of connective tissue characterized mainly by fragility and low bone density. The type I OI, a mild form of the disease, is associated with the quantitative decrease of type I collagen in the extracellular matrix, a characteristic also observed in the oim mice used in pre-clinical OI research. Previous studies have shown the efficiency of recombinant human growth hormone (GH) treatment for OI, both in animal and human models, reducing bone fragility, increasing bone density and stimulating α 1 and α 2 procollagen synthesis. This work aimed at using plasmid containing the murine GH (mGH) gene to treat oim heterozygous mice. Serological quantification of mGH by ELISA was carried out to evaluate the plasmid expression, bone density by DEXA and three-point flexion test were performed on the femurs to assess bone quality. In a short-term (3-day) trial, mGH levels of treated animals were 20.6±6.6 ng/mL, versus 3.3±2.2 ng/mL for those receiving saline and 2.3±1.3 ng/mL for the untreated wild-type group. A 90-day assay with plasmid applications at 0, 30 and 60 days was performed in parallel. The results of bone density showed greater effectiveness of the hormone during the first month of treatment, with a 34% increase in comparison with the saline group, whereas in the second month there was no significant statistical difference (p>0.05). The three-point bending test and the final analysis of bone density are currently being carried out.
