Oncopig bladder cancer cells recapitulate human bladder cancer treatment responses in vitro.

Introduction: Bladder cancer is a common neoplasia of the urinary tract that holds the highest cost of lifelong treatment per patient, highlighting the need for a continuous search for new therapies for the disease. Current bladder cancer models are either imperfect in their ability to translate results to clinical practice (mouse models), or rare and not inducible (canine models). Swine models are an attractive alternative to model the disease due to their similarities with humans on several levels. The Oncopig Cancer Model has been shown to develop tumors that closely resemble human tumors. However, urothelial carcinoma has not yet been studied in this platform. Methods: We aimed to develop novel Oncopig bladder cancer cell line (BCCL) and investigate whether these urothelial swine cells mimic human bladder cancer cell line (5637 and T24) treatment-responses to cisplatin, doxorubicin, and gemcitabine in vitro. Results: Results demonstrated consistent treatment responses between Oncopig and human cells in most concentrations tested (p>0.05). Overall, Oncopig cells were more predictive of T24 than 5637 cell therapeutic responses. Microarray analysis also demonstrated similar alterations in expression of apoptotic (GADD45B and TP53INP1) and cytoskeleton-related genes (ZMYM6 and RND1) following gemcitabine exposure between 5637 (human) and Oncopig BCCL cells, indicating apoptosis may be triggered through similar signaling pathways. Molecular docking results indicated that swine and humans had similar Dg values between the chemotherapeutics and their target proteins. Discussion: Taken together, these results suggest the Oncopig could be an attractive animal to model urothelial carcinoma due to similarities in in vitro therapeutic responses compared to human cells.

Biochar as a carbonaceous material to enhance soil quality in drylands ecosystems: A review.

Drylands are fragile environments that should be carefully managed to improve their quality and functions to achieve sustainable development. Their major problems involve low availability of nutrients and soil organic carbon content. Biochar effect on soil is a joint response of micro to nano sized biochar and soil characteristics. In this review, we attempt to carry out a critical analysis of biochar application to enhance dryland soil quality. Correlating the effects identified from its soil application, we explored the subjects that remains open in the literature. The relation of composition-structure-properties of biochar vary among pyrolysis parameters and biomass sources. Limitations in soil physical quality in drylands, such as low water-holding capacity, can be alleviated by applying biochar at a rate of 10 Mg ha-1 also resulting in beneficial effects on soil aggregation, improved soil porosity, and reduced bulk density. Biochar addition can contribute to the rehabilitation of saline soils, by releasing cations able to displaces sodium in the exchange complex. However, the recovery process of salt-affected soils might be accelerated by the association of biochar with another soil conditioners. This is a promising strategy especially considering the biochar alkalinity and variability in nutrients bioavailability to improve soil fertilization. Further, while higher biochar application rate (>20 Mg ha-1) might change soil C dynamics, a combination of biochar and nitrogen fertilizer can increase microbial biomass carbon in dryland systems. Other aspect of biochar soil application is the economic viability of scale-up production, which is mainly associate to pyrolysis process being biochar production the costliest stage. Nevertheless, the supplying of feedstock might also represent a great input on biochar final costs. Therefore, biochar-based technology is a big opportunity to improve fragile environments such as drylands, integrating sustainable technologies with regional development. Considering the specificity of application area, it might be a model of sustainable agricultural practices protecting the environment in a bioeconomic perspective.

Effectiveness and safety of rotary and reciprocating kinematics for retreatment of curved root canals: a systematic review of in vitro studies.

Objectives: This systematic review (register-osf.io/wg7ba) compared the efficacy and safety of rotary and reciprocating kinematics in the removal of filling material from curved root canals. Materials and Methods: Only in vitro studies evaluating both kinematics during retreatment were included. A systematic search (PubMed/MEDLINE, Scopus, and other databases, until January 2021), data extraction, and risk of bias analysis (Joanna Briggs Institute checklist) were performed. Efficacy in filling removal was the primary outcome. Results: The search resulted in 2,795 studies, of which 15 were included. Efficacy was measured in terms of the remaining filling material and the time required for this. Nine studies evaluated filling material removal, of which 7 found no significant differences between rotary and reciprocating kinematics. Regarding the time for filling removal, 5 studies showed no difference between both kinematics, 2 studies showed faster results with rotary systems, and other 2 showed the opposite. No significant differences were found in apical transportation, centering ability, instrument failure, dentin removed and extruded debris. A low risk of bias was observed. Conclusions: This review suggests that the choice of rotary or reciprocating kinematics does not influence the efficacy of filling removal from curved root canals. Further studies are needed to compare the kinematics safety in curved root canals.

Effects of oral wound on the neutrophil lineage in murine bone-marrow: Modulation mechanism hindered by chlorhexidine.

OBJECTIVE: The oral cavity undergoes frequent stress caused by repeated mechanical trauma, and the constant contact of the injured oral mucosa with bacteria leads to the production of various pro-inflammatory cytokines and chemokines. Neutrophils play essential roles in the acute inflammation against the invasive microbiota, and compromised neutrophil recruitment hinders the bacterial clearance and worsens periodontitis. In this study, we aimed to explore whether wounding at the oral cavity would have an impact on the neutrophil lineage, and, if so, whether microbial contamination of the wounded surface plays significant roles. METHODS: We developed a surgical model of an oral wound (palate wound), by a small incision in the hard palate of the mice. We also evaluated the effect of chlorhexidine on oral wound-induced neutrophilia of bone-marrow. RESULTS: We demonstrated an increased neutrophilia in the bone-marrow of the oral wound group, as well as decreasedex vivoneutropoietic potential, in both IL-3 and GM-CSF-driven bone-marrow cultures. Washing of the entire oral cavity with chlorhexidine before surgery abolished the bone-marrow neutrophilia in the oral wound group and increased neutropoiesis in culture, relative to the saline-treated oral wound control group. Co-located treatment (both chlorhexidine treatment and wound on the right side of the palate) resulted in significantly reduced bone-marrow neutrophilia, compared to the mismatched treatment (chlorhexidine treatment and wound on opposite sides of the palate). Neither neutrophilia nor decreased neutropoiesis were dependent on glucocorticoid signaling. CONCLUSIONS: The prophylactic use of chlorhexidine ameliorates the neutrophilic response on the bone marrow, restoring the neutrophil numbers.

Neuroprotective Effect of 3-[(4-Chlorophenyl)selanyl]-1-methyl-1H-indole on Hydrogen Peroxide-Induced Oxidative Stress in SH-SY5Y Cells.

Extensive data have reported the involvement of oxidative stress in the pathogenesis of neuropsychiatric disorders, prompting the pursuit of antioxidant molecules that could become adjuvant pharmacological agents for the management of oxidative stress-associated disorders. The 3-[(4-chlorophenyl)selanyl]-1-methyl-1H-indole (CMI) has been reported as an antioxidant and immunomodulatory compound that improves depression-like behavior and cognitive impairment in mice. However, the exact effect of CMI on specific brain cells is yet to be studied. In this context, the present study aimed to evaluate the antioxidant activity of CMI in H2O2-induced oxidative stress on human dopaminergic neuroblastoma cells (SH-SY5Y) and to shed some light into its possible mechanism of action. Our results demonstrated that the treatment of SH-SY5Y cells with 4 µM CMI protected them against H2O2 (343 µM)-induced oxidative stress. Specifically, CMI prevented the increased number of reactive oxygen species (ROS)-positive cells induced by H2O2 exposure. Furthermore, CMI treatment increased the levels of reduced glutathione in SH-SY5Y cells. Molecular docking studies demonstrated that CMI might interact with enzymes involved in glutathione metabolism (i.e., glutathione peroxidase and glutathione reductase) and H2O2 scavenging (i.e., catalase). In silico pharmacokinetics analysis predicted that CMI might be well absorbed, metabolized, and excreted, and able to cross the blood-brain barrier. Also, CMI was not considered toxic overall. Taken together, our results suggest that CMI protects dopaminergic neurons from H2O2-induced stress by lowering ROS levels and boosting the glutathione system. These results will facilitate the clinical application of CMI to treat nervous system diseases associated with oxidative stress.

Zinc(II), copper(II) and nickel(II) ions improve the selectivity of tetra-cationic platinum(II) porphyrins in photodynamic therapy and stimulate antioxidant defenses in the metastatic melanoma lineage (A375).

Tetra-cationic porphyrins with peripheral Pt (II) -bipyridyl complexes demonstrated a potential as photosensitizers to be used in photodynamic therapy (PDT). First-line transition metals, such as zinc (II), copper (II) and nickel (II), can be incorporated into the porphyrin nucleus, making this molecule more selective and more effective for this therapy in combating to tumor cells, such as metastatic melanoma. We characterized these derivatives to verify the improvement in selectivity of platinum (II) 4-PtTPyP porphyrins. Receptors such as LDL and endothelin (ERT-B) were investigated, as well as the binding affinity of two antioxidants: catalase model enzymes and superoxide dismutase. Human serum albumin (SAH) HSA binding properties have been verified. In addition, we evaluated the antitumor action of such metalloporphyrins in an in vitro cell viability. Our results demonstrated that porphyrins have significant antitumor potential when exposed to white light conditions. The affinity for the LDL receptor was better when compared to platinum porphyrin 4-PtTPyP without addition of metals and the affinity for the endothelin receptor was higher than the control used in this study. Still, the interaction with the HSA showed the possibility of this connection taking photosensitizers to places of interest, such as the delivery of medicines.

Toxicological evaluation of 3-(4-Chlorophenylselanyl)-1-methyl-1H-indole through the bovine oocyte in vitro maturation model.

The development of new bioactive molecules based on the molecular hybridization has been widely explored. In line with this, reliable tests should be employed to give information about the toxicology of these new molecules. In this sense, the use of in vitro tests is a valuable tool, especially the in vitro maturation of oocytes (IVM), which is an efficient resource to discover the potential toxicity of synthetic molecules. Thus, the aim of the present study was to evaluate the toxicological effects of the selenium-containing indolyl compound 3-(4-Chlorophenylselanyl)-1-methyl-1H-indole (CMI), on different quality parameters of bovine oocytes through the IVM. Different concentrations of the CMI compound (0, 25, 50, 100, 200 µM) were supplemented during the in vitro maturation process. After, the oocyte maturation rate, glutathione (GSH) levels, reactive oxygen species (ROS) levels, membrane, and mitochondrial integrity were evaluated. The results showed that the lowest concentration of CMI induced the highest GSH production (P < 0.05), an important marker of cytoplasmic quality and maturation. All treatments increased ROS production in relation to non-supplementation (P < 0.05). In addition, oocyte maturation was reduced only with the highest concentration of CMI (P < 0.05). Supplementation with CMI did not impact mitochondrial activity, integrity and cell membrane. To our knowledge, this is the first study that evaluates CMI on the oocyte in vitro maturation process. Importantly, our results did not find any toxic effect of CMI on bovine oocytes. CMI was efficient for cytoplasmic maturation by promoting an increase in the intracellular levels of glutathione.