In vitro adhesion of Bacillus sp. and Enterobacter sp. probiotics on intestinal epithelial cells of red tilapia (Oreochromis sp.) and the application effects on the fish growth and survival rate

This research aimed to determine the adhesion of Bacillus sp. (PCP1) and Enterobacter sp. (JC10) on intestinal epithelial cells of red tilapia (Oreochromis sp.) and the effect of the probiotics application in feed on fish growth, survival rate, and feed conversion ratio. In vitro adhesion test was performed by using 108 cells/ml of bacteria, and 105 cells/ml of epithelial cells for 1 hour of incubation. The probiotics were added to the fish pellet with the dose of 5 x 104 CFU/g of feed with four treatments, including probiotic application every three days, seven days, without probiotic, and commercial probiotic application every three days. Each treatment consists of three replications. Red tilapia is maintained for 30 days in fiberglass ponds. The feed is given two times per day with a dose of 5 % of the biomass. The adhesion experiment results showed that Bacillus sp. (PCP1) and Enterobacter sp. (JC10) have adherence abilities higher than the commercial probiotics. The application of probiotics in tilapia for one month did not affect the fish growth, survival rate, and feed conversion ratio (P > 0.05). Probiotic application in a longer period is needed to be addressed.

Differences in a Single Extracellular Residue Underlie Adhesive Functions of Two Zebrafish Aqp0s

Aquaporin 0 (AQP0) is the most abundant lens membrane protein, and loss of function in human and animal models leads to cataract formation. AQP0 has several functions in the lens including water transport and adhesion. Since lens optics rely on strict tissue architecture achieved by compact cell-to-cell adhesion between lens fiber cells, understanding how AQP0 contributes to adhesion would shed light on normal lens physiology and pathophysiology. We show in an in vitro adhesion assay that one of two closely related zebrafish Aqp0s, Aqp0b, has strong auto-adhesive properties while Aqp0a does not. The difference appears to be largely due to a single amino acid difference at residue 110 in the extracellular C-loop, which is T in Aqp0a and N in Aqp0b. Similarly, P110 is the key residue required for adhesion in mammalian AQP0, highlighting the importance of residue 110 in AQP0 cell-to-cell adhesion in vertebrate lenses as well as the divergence of adhesive and water permeability functions in zebrafish duplicates.

Polishing the Therapy of Onychomycosis Induced by Candida spp.: Amphotericin B–Loaded Nail Lacquer

Onychomycosis induced by Candida spp. has several limitations regarding its treatment. Nail lacquers display the potential to overcome these drawbacks by providing therapeutic compliance and increasing local drug bioavailability. Thus, this work aimed to produce a nail lacquer loaded with Amphotericin B (AmB) and evaluate its performance. The AmB-loaded nail lacquer was produced and preliminarily characterized. An AmB quantification method was developed. Stability, drug release, permeability and anti-Candida activity assays were conducted. The analytical method validation met the acceptance criteria. The drug loading efficiency was 100% (0.02 mg/g of total product), whereas the AmB stability was limited to ≅ 7 days (≅ 90% remaining). The nail lacquer displayed a drying time of 187 s, non-volatile content of around 20%w/w, water-resistance of approximately 2%w/w of weight loss and satisfactory in vitro adhesion. Moreover, the in vitro antifungal activity against different Candida spp. strains was confirmed. The AmB release and the ex vivo permeability studies revealed that AmB leaves the lacquer and permeates the nail matrix in 47.76 ± 0.07% over 24 h. In conclusion, AmB-loaded nail lacquer shows itself as a promising extemporaneous dosage form with remarkable anti-Candida activity related to onychomycosis.

The adhesion of clots in wounds contributes to hemostasis and can be enhanced by coagulation factor XIII

The adhesion of blood clots to wounds is necessary to seal injured vasculature and achieve hemostasis. However, it has not been specifically tested if adhesive failure of clots is a major contributor to rebleeding and what mechanisms prevent clot delamination. Here, we quantified the contribution of adhesive and cohesive failure to rebleeding in a rat model of femoral artery injury, and identified mechanisms that contribute to the adhesive strength of bulk clots in a lap-shear test in vitro. In the rat bleeding model, the frequency of clot failures correlated positively with blood loss (R = 0.81, p = 0.014) and negatively with survival time (R = − 0.89, p = 0.0030), with adhesive failures accounting for 51 ± 14% of rebleeds. In vitro, adhesion depended on fibrinogen and coagulation factor XIII (FXIII), and supraphysiological FXIII improved adhesive strength. Furthermore, when exogenous FXIII was topically applied into the wound pocket of rats, eleven adhesive failures occurred between eight rats, compared to seventeen adhesive failures between eight untreated rats, whereas the number of cohesive failures remained the same at sixteen in both groups. In conclusion, rebleeding from both adhesive and cohesive failure of clots decreases survival from hemorrhage in vivo. Both endogenous and exogenous FXIII improves the adhesive strength of clots.

Bisphenol A impaired cell adhesion by altering the expression of adhesion and cytoskeleton proteins on human podocytes

Bisphenol A (BPA), a chemical -xenoestrogen- used in food containers is present in the urine of almost the entire population. Recently, several extensive population studies have proven a significant association between urinary excretion of BPA and albuminuria. The alteration of glomerular podocytes or "podocytopathy" is a common event in chronic albuminuric conditions. Since many podocytes recovered from patients' urine are viable, we hypothesized that BPA could impair podocyte adhesion capabilities. Using an in vitro adhesion assay, we observed that BPA impaired podocyte adhesion, an effect that was abrogated by Tamoxifen (an estrogen receptor blocker). Genomic and proteomic analyses revealed that BPA affected the expression of several podocyte cytoskeleton and adhesion proteins. Western blot and immunocytochemistry confirmed the alteration in the protein expression of tubulin, vimentin, podocin, cofilin-1, vinculin, E-cadherin, nephrin, VCAM-1, tenascin-C, and β-catenin. Moreover, we also found that BPA, while decreased podocyte nitric oxide production, it lead to overproduction of ion superoxide. In conclusion, our data show that BPA induced a novel type of podocytopathy characterizes by an impairment of podocyte adhesion, by altering the expression of adhesion and cytoskeleton proteins. Moreover, BPA diminished production of podocyte nitric oxide and induced the overproduction of oxygen-free metabolites. These data provide a mechanism by which BPA could participate in the pathogenesis and progression of renal diseases.

Formulation Development of Rotigotine Transdermal System Using Dot-Matrix Technology

The purpose of this research was to prepare and evaluate drug-in-adhesive type patches of rotigotine using dot-matrix technology, which is the new generation of drug-in-adhesive transdermal delivery system (TDS) that deliver drug therapy through less patch surface area and without compromising adhesion. Preformulation studies, like solubility in permeation enhancers, compatibility study, transmission study, uptake study, and crystallization study of rotigotine in various pressure-sensitive adhesive (PSA) polymers were performed. Transdermal system was prepared by solvent casting method. Central composite design (CCD) was chosen for optimization of the formulation. Design of experiment (DoE) was used to study the impact of critical formulation parameters, like silicone adhesive concentration, povidone K29/32 concentration, and propylene glycol concentration. Crystallization study of rotigotine in different PSAs suggested that crystal inhibitor is required to load drugs above 5%. Selection of optimum batch was made using a constraint-based graphical optimization technique. The optimum batch exhibited desired in vitro adhesion parameters, like peel, tack, shear, and permeation rate, which is suitable for 3 days’ wear properties and desired permeation rate. The optimum batch was evaluated for appearance, weight of matrix, thickness, % assay of drug content, in vitro adhesion testing, cold flow study, and ex vivo skin permeation studies. Backing film Scotchpak 9730 and release liner Scotchpak-1022 was selected based on transmission and uptake study of rotigotine. Stability study indicates that developed formulation remains stable. The present research confirms the feasibility of developing rotigotine transdermal system using novel technology.

Design, Development and Delivery of Rasagiline Mesylate from Monolithic Drug in Adhesive Matrix Patches

The purpose of this research was to prepare and evaluate monolithic drug-in-adhesive type patches of Rasagiline Mesylate (RM) containing penetration enhancer and having seven day wear property. Preformulation studies like solubility in permeation enhancers, compatibility study, transmission study, uptake study and crystallization study of Rasagiline Mesylate in various pressure sensitive adhesive polymers were performed. Transdermal system was prepared by solvent casting method. The effects of various permeation enhancers (Propylene Glycol, Oleic Acid, Isopropyl Palmitate, and lauryl lactate) on the ex-vivo transcutaneous absorption of Rasagiline Mesylate through human cadaver skin were evaluated by modified Franz diffusion cell system. Ex-vivo transcutaneous absorption of prepared transdermal patch was performed using different concentration of Lauryl lactate (3%, 5%, and 7%). In-vitro Adhesion testing (Peel, tack shear etc.) was performed on different dry GSM (Grams per Square Meter) of patch like 80GSM, 100 GSM and 150 GSM. The final transdermal patches were tested for appearance, weight of matrix, thickness, % assay of drug content, in-vitro adhesion testing, cold flow study and ex-vivo skin permeation studies. Based on crystallization study and adhesion testing, Durotak-4098 (14% drug concentration) was selected as pressure sensitive adhesive. Patch containing Lauryl lactate showed highest cumulative permeation compared to other permeation enhancers. The patch containing 5% laurel lactate showed greater transdermal flux (2.36 µg/cm2 /hr). Patch with 150 dry GSM showing promising adhesion properties. Backing film Scotchpak 9723 and release liner Saint Gobain 8310 was selected based on transmission and uptake study of Rasagiline Mesylate. Stability study indicates that developed formulation remains stable. In conclusion, the present research confirms the practicability of developing Rasagiline Mesylate transdermal system.

Bioactivity of Juniperus communis essential oil and post-distillation waste: Assessment of selective toxicity against food contaminants

Previously chemically characterized Juniperus communis essential oil (EO) and post-distillation waste (PDW) were tested for cytotoxicity and antimicrobial activity against food contaminants. Microdilution assay showed that PDW induced moderate antifungal (minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values, ranging between 0.118-0.900 mg mL-1), and an antibacterial effect against Listeria monocytogenes (MIC and minimum bactericidal concentration (MBC) were 0.39 and 0.74 mg mL-1, respectively). Combinations of EO/PDW with selected antibiotics induced synergistic antilisterial activity in the checkerboard assay. The MTT assay determined that cytotoxicity against colon cancer cells was high for the EO but negligible for PDW (IC50 values were 0.087-0.106 and 1.450-6.840 mg mL-1, respectively). The selectivity indices indicated high selectivity of PDW against tested fungi and L. monocytogenes. In the adhesion-inhibition assay, PDW reduced in vitro adhesion of L. monocytogenes to colon cells (29-62% of inhibition). In conclusion, PDW exhibited an antimicrobial effect against important food spoilage and poisoning fungi and L. monocytogenes, and also reduced in vitro adhesion of L. monocytogenes to colon cells. The results indicate that J. communis PDW could be considered as natural preservative against food spoilage and poisonous fungi, and as an adjuvant to conventional therapy of listeriosis.