Putative protective effect of Cadmium chloride highdiluted solution on LLC-PK1 cell intoxicated by high concentration of this same metal: an isopathic in vitro assay.

Cadmium is an important toxic environmental heavy metal. Several studies have demonstrated that a major site of cadmium toxicity in humans and in other animals is the proximal tubule of the kidney. A well established model for nefrotoxicity is the use of in vitro technique with proximal tubule epithelial cell lines, as LLC-PK1. Herein, we have the intention to study the possible protective effect of highdiluted CdCl2 solutions. In a blinding way, LLC-PK1 cells were pre-treated with highdiluted cadmium chloride in the potencies 10 cH, 15 cH and 20cH. After 4 days, these cells have received CdCl2 in a pre-determined toxic concentration. The cell viability was assessed by MTT assay. We have identified a protective effect of two CdCl2 highdiluted solutions, 10 cH and 20 cH, when cells were intoxicated by sublethal CdCl2 concentration. The results indicate that probably the highdilutions have an expressive action on cells in sublethal intoxication.

FORMULATION AND EVALUATION OF FLOATING IN SITU GEL OF OMEPRAZOLE MAGNESIUM FOR ORAL DRUG DELIVERY SYSTEM

Objective: Omeprazole magnesium is indicated for the treatment of erosive esophagitis associated with gastroesophageal reflux disease. It is one of the highly prescribed proton pump inhibitor in the management of peptic ulcer diseases. The therapeutic concentration of a drug in blood can be maintained for a prolonged period of time by administering it in the form of in situ floating gel dosage form. Omeprazole magnesium undergoes degradation at a low pH of the esophagus and stomach; it is therefore given as in situ gel, so, there is minimum contact with acidic pH. Methods: Omeprazole magnesium suspension prepared using various polymers and floating agents in varying concentrations. Several evaluation tests including dissolution test to ensure the release of the drug from formulation by in vitro technique, color and homogeneity, in vitro floating duration, in vitro gelling capacity, drug content determination, pH of the formulation, and floating lag time were studied. Results: All formulations demonstrated good Fourier-transform infrared compliance and no interaction between drug, polymer, and other excipients. The study’s findings show that the formulation F6 showed the best results. Conclusion: The developed formulation was a viable alternative conventional solution by virtue of its ability to enhance bioavailability through its longer gastric residence time and ability to sustain drug release as well as the advantage of floating and pH which minimize the degradation of omeprazole magnesium which is easily degraded by acidic environment.

Rapid in vitro quantification of TDP-43 and FUS mislocalisation for screening of gene variants implicated in frontotemporal dementia and amyotrophic lateral sclerosis

Identified genetic mutations cause 20% of frontotemporal dementia (FTD) and 5-10% of amyotrophic lateral sclerosis (ALS) cases: however, for the remainder of patients the origin of disease is uncertain. The overlap in genetic, clinical and pathological presentation of FTD and ALS suggests these two diseases are related. Post-mortem, ~ 95% of ALS and ~ 50% of FTD patients show redistribution of the nuclear protein TDP-43 to the cytoplasm within affected neurons, while ~ 5% ALS and ~ 10% FTD show mislocalisation of FUS protein. We exploited these neuropathological features to develop an unbiased method for the in vitro quantification of cytoplasmic TDP-43 and FUS. Utilising fluorescently-tagged cDNA constructs and immunocytochemistry, the fluorescence intensity of TDP-43 or FUS was measured in the nucleus and cytoplasm of cells, using the freely available software CellProfiler. Significant increases in the amount of cytoplasmic TDP-43 and FUS were detectable in cells expressing known FTD/ALS-causative TARDBP and FUS gene mutations. Pharmacological intervention with the apoptosis inducer staurosporine and mutation in a secondary gene (CYLD) also induced measurable cytoplasmic mislocalisation of endogenous FUS and TDP-43, respectively. These findings validate this methodology as a novel in vitro technique for the quantification of TDP-43 or FUS mislocalisation that can be used for initial prioritisation of predicted FTD/ALS-causative mutations.

In Vitro Genotoxic Effects of Sarcocystis gigantea Cystizoites Acetone Powder Extract on Sheep Lymphocytes

The toxin of the protozoan intracellular parasite of sheep Sarcocystis gigantea is associated with many clinical and pathological signs. The aim of the study was to investigate In Vitro various chromosomal aberrations due to sarcocystosis infection. Macrocysts of Sarcocystis gigantea were isolated from local karadi sheep, homogenized with glass Dounce homogenizer; acetone powder was prepared from it and used in various concentration to investigate the chromosomal aberration in vitro against sheep lymphocytes. The direct effects of parasite cystizoites acetone powder revealed various genotoxicity effects. These effects included chromosomal aberration (Isogap, Breaks and Dicentrics) and chromatids aberration (Gap and Deletion). It had also an effect on the mitotic index of the lymphocyte cells division. These genotoxicities were studied for the first time with in vitro technique using sheep lymphocytes. These results reflected that Sarcocystis gigantean parasite could cause structural and internal aberration in the chromosomes of their hosts.

The Influence of the Implant Macrogeometry on Insertion Torque, Removal Torque, and Periotest Implant Primary Stability: A Mechanical Simulation on High-Density Artificial Bone

Background: The primary stability is a determinant clinical condition for the success of different dental implants macro-design in different bone density using a validated and repeatable in vitro technique employing solid rigid polyurethane blocks. Materials and Methods: Five implants 3.8 × 13 mm2 for each macro-design (i.e., IK—tapered; IC—cylindric; and IA—active blade shape) were positioned into 20- and 30- pounds per cubic foot (PCF) polyurethane blocks. Bucco-lingual (BL) and mesial-distal (MD) implant stability quotient score (ISQ) was assessed by resonance frequency analysis while, insertion/removal torques were evaluated by dynamometric ratchet. Results: IC implants shown better primary stability in terms of ISQ compared to IA and IK in lower density block (20 PCF), while IK was superior to IA in higher density (30 PCF). IC shown higher removal torque in 30-PCF compared to IA and IC. Conclusions: The study effectiveness on polyurethane artificial bone with isotropic symmetry structure showed that the implants macro-design might represent a key factor on primary stability, in particular on low-density alveolar bone. Clinicians should consider patients features and implant geometry during low-density jaws rehabilitation. Further investigations are needed to generalize these findings.

Somatic Embryogenesis in Olive

The olive is a fruit tree species economically very important in countries of the Mediterranean basin. Somatic embryogenesis is a powerful in vitro technique with multiple applications in different fields, including breeding programs performed by both classical and innovative procedures. This editorial paper presents a special issue focused on “Somatic embryogenesis in olive”. In this manuscript, the conceptual framework of the special issue is established and the contributions are summarized and put into context. Finally, the main bottlenecks limiting the practical applicability of somatic embryogenesis are identified and the future research prospects are discussed.