Anodized Biomedical Stainless-Steel Mini-Implant for Rapid Recovery in a Rabbit Model

The study aimed to analyze the recovery period of the anodized 316L biomedical stainless steel (BSS) mini-implant through its implantation on femur of rabbit model. The 316L BSS mini-implant was modified by an electrochemical anodization approach with different voltages. The anodized samples were characterized via field-emission scanning electron microscopy, X-ray diffractometry, and X-ray photoelectron spectroscopy. The biocompatibility was assessed by cell culture assay. The anodized mini-implant was implanted on rabbit’s femur then evaluated histologically after 4 and 8 weeks. Analytical results indicated that the topography of the anodized mini-implant at 5 V for 5 min consisted of a dual (micro/nano) porous structure. Oxide film of Cr2O3 was formed on the surface of anodized mini-implant after anodizing with 5 V for 5 min. In vitro cell culture assay revealed that fibroblast cells (NIH-3T3) on the anodized samples were more firmly attached as compared with the control sample. Moreover, histological analysis demonstrated that the anodized mini-implant improved bone recovering at 4 weeks after implantation. Thus, this study suggests that the anodized 316L BSS mini-implant could be a potential choice as anchorage device for effective and efficient orthodontic treatment.

Quorum sensing in Aliivibrio wodanis 06/09/139 and its role in controlling various phenotypic traits

Background Quorum Sensing (QS) is a cell-to-cell communication system that bacteria utilize to adapt to the external environment by synthesizing and responding to signalling molecules called autoinducers. The psychrotrophic bacterium Aliivibrio wodanis 06/09/139, originally isolated from a winter ulcer of a reared Atlantic salmon, produces the autoinducer N-3-hydroxy-decanoyl-homoserine-lactone (3OHC10-HSL) and encodes the QS systems AinS/R and LuxS/PQ, and the master regulator LitR. However, the role of QS in this bacterium has not been investigated yet. Results In the present work we show that 3OHC10-HSL production is cell density and temperature-dependent in A. wodanis 06/09/139 with the highest production occurring at a low temperature (6 °C). Gene inactivation demonstrates that AinS is responsible for 3OHC10-HSL production and positively regulated by LitR. Inactivation of ainS and litR further show that QS is involved in the regulation of growth, motility, hemolysis, protease activity and siderophore production. Of these QS regulated activities, only the protease activity was found to be independent of LitR. Lastly, supernatants harvested from the wild type and the ΔainS and ΔlitR mutants at high cell densities show that inactivation of QS leads to a decreased cytopathogenic effect (CPE) in a cell culture assay, and strongest attenuation of the CPE was observed with supernatants harvested from the ΔlitR mutant. Conclusion A. wodanis 06/09/139 use QS to regulate a number of activities that may prove important for host colonization or interactions. The temperature of 6 °C that is in the temperature range at which winter ulcer occurs, plays a role in AHL production and development of CPE on a Chinook Salmon Embryo (CHSE) cell line.

Agronomic Performance in Low Phytic Acid Field Peas

Field pea is a pulse that delivers high protein content, slowly digestible starch and fiber, and many vitamins and minerals, including iron. Naturally occurring plant phytic acid molecules bind iron, lowering its availability for absorption during digestion. Two low phytic acid (lpa) pea lines, 1-2347-144 and 1-150-81, developed by our group had 15% lower yield and 6% lower seed weight relative to their progenitor cultivar. Subsequently, we crossed the two lpa lines and two cultivars, and derived 19 promising lpa pea breeding lines; here we document their agronomic performance based on 10 replicated field trials in Saskatchewan. Seventeen of these lpa lines yielded greater than 95% of the check mean (associated cultivars) and 16 were above 98% of the check mean for 1000 seed weight. The 19 lpa lines showed 27 to 55% lower phytic acid concentration than the check mean. Iron concentrations were similar in all the lpa lines and cultivars, yet the Caco-2 human cell culture assay revealed 14 of the 19 lpa lines had 11 to 55% greater iron bioavailability than check means. Thus, a single round of plant breeding has allowed for closing the gap in performance of low phytic acid pea.

Evidence from oyster suggests an ancient role for Pdx in regulating insulin gene expression in animals

Hox and ParaHox genes encode transcription factors with similar expression patterns in divergent animals. The Pdx (Xlox) homeobox gene, for example, is expressed in a sharp spatial domain in the endodermal cell layer of the gut in chordates, echinoderms, annelids and molluscs. The significance of comparable gene expression patterns is unclear because it is not known if downstream transcriptional targets are also conserved. Here, we report evidence indicating that a classic transcriptional target of Pdx1 in vertebrates, the insulin gene, is a likely direct target of Pdx in Pacific oyster adults. We show that one insulin-related gene, cgILP, is co-expressed with cgPdx in oyster digestive tissue. Transcriptomic comparison suggests that this tissue plays a similar role to the vertebrate pancreas. Using ATAC-seq and ChIP, we identify an upstream regulatory element of the cgILP gene which shows binding interaction with cgPdx protein in oyster hepatopancreas and demonstrate, using a cell culture assay, that the oyster Pdx can act as a transcriptional activator through this site, possibly in synergy with NeuroD. These data argue that a classic homeodomain-target gene interaction dates back to the origin of Bilateria.

Prospective Prediction of Clinical Drug Response in High Grade Gliomas Using an Ex Vivo 3D Cell Culture Assay

Background Clinical outcomes in high-grade glioma (HGG) have remained relatively unchanged over the last three decades with only modest increases in overall survival. Despite the validation of biomarkers to classify treatment response, most newly diagnosed (ND) patients receive the same treatment regimen. This study aimed to determine whether a prospective functional assay that provides a direct, live tumor cell-based drug response prediction specific for each patient could accurately predict clinical drug response prior to treatment. Methods A modified 3D cell culture assay was validated to establish baseline parameters including drug concentrations, timing, and reproducibility. Live tumor tissue from HGG patients were tested in the assay to establish response parameters. Clinical correlation was determined between prospective ex vivo response and clinical response in ND HGG patients enrolled in 3D-PREDICT (ClinicalTrials.gov Identifier: NCT03561207). Clinical case studies were examined for relapsed HGG patients enrolled on 3D-PREDICT, prospectively assayed for ex vivo drug response, and monitored for follow-up. Results Absent biomarker stratification, the test accurately predicted clinical response/non-response to temozolomide in 17/20 (85%, p = 0.007) ND patients within 7 days of their surgery, prior to treatment initiation. Test-predicted responders had a median overall survival post-surgery of 11.6 months compared to 5.9 months for test-predicted non-responders (p = 0.0376). Case studies provided examples of the clinical utility of the assay predictions and their impact upon treatment decisions resulting in positive clinical outcomes. Conclusion This study both validates the developed assay analytically and clinically and provides case studies of its implementation in clinical practice.

Roles of Stra8 and Tcerg1l in retinoic acid induced spermatogonial differentiation in mouse.

Retinoic acid (RA) induces spermatogonial differentiation, but the mechanism by which it operates remains largely unknown. We developed a germ cell culture assay system to study genes involved in spermatogonial differentiation triggered by RA. Stimulated by Retinoic Acid 8 (Stra8), a RA-inducible gene, is indispensable for meiosis initiation, and its deletion results in a complete block of spermatogenesis at the pre-leptotene/zygotene stage. To interrogate the role of Stra8 in RA mediated differentiation of spermatogonia, we derived germ cell cultures from the neonatal testis of both wild type and Stra8 knock-out mice. We provide the first evidence that Stra8 plays a crucial role in modulating the responsiveness of undifferentiated spermatogonia to RA and facilitates transition to a differentiated state. Stra8-mediated differentiation is achieved through downregulation of a large portfolio of genes and pathways, most notably including genes involved in the spermatogonial stem cell self-renewal process. We also report here for the first time the role of Transcription Elongation Regulator-1 Like (Tcerg1l) as a downstream effector of RA-induced spermatogonial differentiation.

Differences of palbociclib and ribociclib in terms of cell death pathways; a cell culture assay

Differences of palbociclib and ribociclib in terms of cell death pathways; a cell culture assay The contribution of adding cyclin dependent kinase 4/6 (CDK4/6) inhibitors to hormonotherapy in relation with the survival was indicated for the treatment of metastatic breast cancer which had positive hormone receptor and negative HER2. In this study, it was planned to indicate two different CDK4/6 inhibitors (palbociclib and ribociclib) providing CDK4/6 inhibition on the cell lines (MCF-7 and BT474) having specifications of luminal-A and luminal-B, the molecular sub-types of positive hormone receptor of breast cancer, and to reveal the molecular differences and to compare cytotoxicity data and necrosis mechanisms in the cell culture experiments. It was planned to determine the differences of resistance mechanism of new combinations. MCF-7 and BT474 cell lines were handled with Palbociclib and Ribociclib individually. The possible cytotoxicity, apoptosis and authopagy effects and the levels of apoptotic proteins were examined. It was indicated that Palbociclib and Ribociclib had cytotoxic effects in both breast cancer cells. In regard of comparing from the point of intracellular pathways, it was determined that the effect of Palbociclib decreased in the increased dosage but that the effect of Ribociclib on cell indicated by means of apoptosis. It was found that palbociclib induced autophagy in the autophagy experiment conducted on the decrease of apoptotic activity with increasing doses. Even though they worked on the same pathway, Palbociclib and Ribociclib utilized different mechanisms to kill the cells and new evidences were obtained that the resistance mechanism may be different from each other according to the treatment.

Surface Characteristics and Cell Adhesion Behaviors of the Anodized Biomedical Stainless Steel

In this study, an electrochemical anodizing method was applied as surface modification of the 316L biomedical stainless steel (BSS). The surface properties, microstructural characteristics, and biocompatibility responses of the anodized 316L BSS specimens were elucidated through scanning electron microscopy, X-ray photoelectron spectroscopy, X-ray diffractometry, transmission electron microscopy, and in vitro cell culture assay. Analytical results revealed that the oxide layer of dichromium trioxide (Cr2O3) was formed on the modified 316L BSS specimens after the different anodization modifications. Moreover, a dual porous (micro/nanoporous) topography can also be discovered on the surface of the modified 316L BSS specimens. The microstructure of the anodized oxide layer was composed of amorphous austenite phase and nano-Cr2O3. Furthermore, in vitro cell culture assay also demonstrated that the osteoblast-like cells (MG-63) on the anodized 316L BSS specimens were completely adhered and covered as compared with the unmodified 316L BSS specimen. As a result, the anodized 316L BSS with a dual porous (micro/nanoporous) oxide layer has great potential to induce cell adhesion and promote bone formation.

In Vivo Wound Healing and In Vitro Anti-Inflammatory Activity Evaluation of Phlomis russeliana Extract Gel Formulations

The air-dried aerial parts of Phlomis russeliana (Sims) Lag. Ex Benth. was extracted by methanol and fractionated by n-hexane, dichloromethane, and ethyl acetate, respectively. The wound healing properties of P. russeliana extract gel was evaluated using the in vivo excisional wound model using Balb-c mice. Initially, the P. russeliana methanol extract showed LOX inhibitory activity at IC50 = 23.2 µg/mL, whereas the DPPH• assay showed IC50 = 0.89 mg/mL, and the ABTS• assay showed IC50 = 0.99 mg/mL, respectively. In addition, a remarkable anti-inflammatory activity was observed in the cell culture assay. Thereafter, activity-guided fractionation was performed by LOX enzyme inhibition assays, and the structures of the two most active fractions were revealed by both GC–FID and GC/MS analyses, simultaneously. Phytol and 1-heptadecanoic acid were characterized as the active constituents. Moreover, the P. russeliana extract gel formulation was applied for in vivo tests, where the new gel formulation supported the in vitro anti-inflammatory activity findings. As a conclusion, this experimental results support the wound healing evidence based on the ethnobotanical application of Phlomis species with further potential.