Administration of a polyphenol-enriched feed to farmed sea bass (Dicentrarchus labrax L.): Kidney melanomacrophages response.

The reinforcement of the defense mechanism of fish, through the administration of immunostimulants, is considered as a promising alternative to vaccines. Natural immunostimulants such as polyphenols, flavanoids, pigments and essential oils can modulate the innate immune response. In lower vertebrates, melano-macrophage centres, i.e. clusters of pigment-containing cells forming the extracutaneous pigment system, are wide-spread in the stroma of the haemopoietic tissue, mainly in kidney and spleen. In fishes, melano-macrophage centres play an important role in the immune response against antigenic stimulants and pathogens. In the present study, we evaluated the effect of a polyphenol-enriched diet on the health status of European sea bass (Dicentrarchus labrax L.). Farmed sea bass were administered a feed containing a phytocomplex, rich in catechins and epigallocatechins, which was obtained from the seeds of Canosina Nero di Troia Vitis vinifera and mixed with conventional feed at two different concentrations. The effects of such a diet were investigated in juvenile and commercial size samples, i.e. undergoing a short- and long-term period of diet, respectively, focusing on their extracutaneous pigmentary system and, in more detail, on the enzymatic activities leading to melanin biosynthesis. Our results show that prolonged dietary treatments with higher concentration of polyphenols might modulate tyrosinase activity and gene expression in commercial size fishes. An increase of melano-macrophage activity is correlated to a stimulation of cytoprotective functions against antigenic stimulants and pathogens, as an expression of a robust and protective adaptive immune response.

Physical-chemical Stability of the Extemporaneous Paracetamol Oral Suspension in Puccini Base.

This study describes a new method for the preparation of extemporaneous paracetamol-based suspensions for pediatric and adult patients. This method allows the preparation of extemporaneous suspensions up to concentrations of 50 mg/mL by using a liquid base, named "Puccini". A high-pressure liquid chromatographic method was developed and validated for the determination of chemical stability of paracetamol when the formulations were stored at 4°C and 25°C. The chemical stability of the active pharmaceutical ingredient in the base was demonstrated for more than 90 days. Visual analyses of the formulations showed a phenomenon of precipitation at both storage temperatures, but the simple agitation of the formulations before its use re-established the formation of homogeneous suspensions.

Extemporaneous Topical Minoxidil Solutions for the Treatment of Androgenetic Alopecia- Stability Studies and Incorporation Tests of Active Pharmaceutical Ingredients in Aloplus Total Base.

Minoxidil is one of the most employed active pharmaceutical ingredients for the  treatment of androgenetic alopecia. The authors propose a new method for production of minoxidil lotions using Aloplus Total. The latter is a propylene glycol-free liquid base in which the presence of hydroxypropyl-ß-cyclodextrin and ethanol allows the solubilization of high drug amounts. Minoxidil intrinsic solubility in the base was determined, and a comprehensive chemical and physical stability study was conducted on 8% w/w minoxidil lotions. Incorporation tests of different active pharmaceutical ingredients that can be combined to 5% w/w minoxidil were also carried out. The analyses showed that minoxidil intrinsic solubility in the new base was 85.93 mg/mL ± 4.17 mg/mL (8.64% w/w ± 0.42% w/w) at 25°C, and the topical lotions were found to be physically and chemically stable for more than 180 days when stored at 25°C or 40°C. Incorporation tests of several active pharmaceutical ingredients also were successful, indicating that Aloplus Total is a liquid vehicle also useful for the preparation of minoxidil-based topical lotions for a synergistic treatment of androgenetic alopecia.

Mucoadhesive Budesonide Solution for the Treatment of Pediatric Eosinophilic Esophagitis.

Eosinophilic Esophagitis is an antigen-mediated inflammatory disease characterized by thickening of the esophageal wall, leading to dysphagia, vomiting, reflux, and abdominal pain. This disease can be treated with a therapeutic approach ranging from diet to pharmacological therapy. Jorveza® (budesonide) and Dupixent® (dupilumab) are treatments for Eosinophilic Esophagitis approved by the European Medicines Agency in adults but not in children. Budesonide-based extemporaneous oral liquid suspensions could be prepared for pediatric use. The main limit of this formulation is that budesonide needs a longer residence time on the esophageal mucosa to solubilize and diffuse in it to exert its local anti-inflammatory effect. Herein, we propose the development of an extemporaneous mucoadhesive oral budesonide solution for the pediatric population. A liquid vehicle containing hydroxypropyl-beta-cyclodextrin as a complexing agent and carboxymethylcellulose sodium as a mucoadhesive excipient was used to prepare budesonide-based formulations. A stable solution at a concentration of 0.7 mg/mL was successfully prepared and characterized. The formulation showed rheological and mucoadhesive properties suitable for an Eosinophilic Esophagitis local prolonged treatment. In this way, pharmacists can prepare stable budesonide-based mucoadhesive solutions, providing both patients and physicians with a new therapeutic option for Eosinophilic Esophagitis pediatric treatment.

Hydrophilic Ready-to-use Vehicles and Compounded Topical Medications.

Pharmaceutical compounding has a crucial role for the health of patients, allowing the preparation of formulation out of market or for a personalized therapy. This study is aimed to conduct a screening of possible ready-to-use hydrophilic vehicles for the preparation of topical dosage forms. Incorporation tests of several active pharmaceutical ingredients were performed, and the physical stability of the extemporaneous formulations was assessed by performing an accelerated centrifuge test. The results showed that it was possible to realize several physically stable topical medications without using special equipment or instruments, guaranteeing a fast and repeatable preparation process. The goal of this work is to provide compounding pharmacists a table that summarizes some of the possible vehicles that can be used for the formulation of topical treatments.

Lipophilic Ready-to-use Vehicles and Compounded Topical Medications.

The preparation of formulations that are not currently on the market or prepared for customized therapy is possible by pharmaceutical compounding. In this study, incorporation tests of some active pharmaceutical ingredients in five ready-touse lipophilic semisolid vehicles were performed, and the physical stability of the prepared extemporaneous formulations was assessed by performing an accelerated centrifuge test. The results demonstrated that it was possible to formulate physically stable topical medications without using special equipment or instruments, ensuring a fast, efficient, and repeatable preparation process. The objective of this work was to provide to compounding pharmacists a table that summarizes some of the semisolid lipophilic vehicles, such as creams water/oil, and ointments, that can be used for the formulation of topical treatments.

Physicochemical Stability of the Extemporaneous Ibuprofen Oral Suspension in "Wagner" Base.

This article discusses a new method for the preparation of extemporaneous ibuprofen-based suspensions for use in paediatric patients. This method allows the preparation of extemporaneous suspensions up to concentrations of 200 mg/5 mL by using a liquid base named "Wagner." A comprehensive physicochemical stability study was conducted on the formulation at a drug concentration of 200 mg/5 mL by performing high-pressure liquid chromatography and Turbiscan analyses. Chromatographic analyses of the samples demonstrated the chemical stability of the active pharmaceutical ingredient in the base for more than 90 days when the formulations were stored at 4°C and 25°C. Visual and optical analyses evidenced a reversible, slightly creaming phenomenon when the formulations were stored at 4°C or 25°C restoring the initial suspension by simply shaking.

Stability of Omeprazole Extemporaneous Oral Solution in Chopin Base.

Omeprazole is the progenitor of proton pump inhibitors. It is used for the treatment of ulcer and gastroesophageal reflux in dosages ranging from 10 mg/day to 40 mg/day, calibrated according to the patient's age and body weight. In this study, the authors provide a report on the preparation of an extemporaneous liquid formulation of omeprazole using fast oral solution Chopin a hydroxypropyl-?-cyclodextrin liquid base (pH 8 to 9) that is able to solubilize the drug. A solubility study of the drug in the liquid vehicle and a physical-chemical stability study of the 1-mg/mL formulation at 4°C and 25°C were performed. Analyses were carried out by using a high-pressure liquid chromatographic analytical method. Results showed that the intrinsic solubility of the drug in Chopin base was 5.33 mg/mL ± 0.23 mg/mL at 25°C and that omeprazole was chemically stable when the formulation was stored at 4°C over a period of 3 months, while its shelf life at 25°C was only 9 days. This study has demonstrated that the resulting liquid formulation is suitable for all patients, in particular children or adults who are unable to take other pharmaceutical dosage forms, which overcomes the limitations of the medicines currently available on the market.

Extemporaneous Topical Minoxidil Solutions for the Treatment of Alopecia: Stability Studies and Incorporation Tests of Active Ingredients in ALOPLUS FAST Base.

Minoxidil is a vasodilator drug generally employed for the treatment of various forms of alopecia. In this article, the authors propose an alternative to the formulation reported in the British Pharmacopoeia for the realization of topical minoxidil -based solutions using ALOPLUS FAST. This liquid vehicle is an ethanol- and propylene glycol-free base which allows the complete solubilization of minoxidil, thanks to the presence of hydroxypropyl-ß-cyclodextrin. Solubility and chemical stability studies of the active ingredient in the formulation at a concentration of 5% w/w and physical stability studies of this extemporaneous preparation are reported. Incorporation tests of various active pharmaceutical ingredients that can be combined with minoxidil for alopecia synergic treatment have been carried out. Analyses were performed by using a high-pressure liquid chromatography analytical method. The results showed that the intrinsic solubility of the drug in the liquid base was 62.37 mg/mL ± 0.85 mg/mL (5.24 w/w ± 0.07% w/w) at 25°C; minoxidil was chemically stable in ALOPLUS FAST; and the formulation was physically stable for more than six months, under different storage conditions. Incorporation tests of several active pharmaceutical ingredients in 2% to 4% w/w minoxidil formulations were successful as well.

Stability of Diazepam Enema Extemporaneous Formulation in Manzoni Base.

The authors provide a report on the preparation of an extemporaneous liquid formulation of diazepam enema using an alcohol-free base solution for use by both pediatric and adult patients. A solubility study using this base and the chemical stability of the drug in this formulation was performed by using a high-pressure liquid chromatography analytical method. Results of this study showed that the intrinsic solubility of the drug in this liquid base was 6.03 mg/mL ± 0.06 mg/mL and that diazepam was chemically stable when the formulation was stored at 25°C over a period of 3 months.

From oil to microparticulate by prilling technique: Production of polynucleate alginate beads loading Serenoa Repens oil as intestinal delivery systems.

Natural oils that are rich in biologically active polyunsaturated fatty acids have many health benefits but have insufficient bioavailability and may oxidize in the gastrointestinal tract. For these reasons and to improve the handling as well, the possibility of incorporating a natural oil, extracted from Serenoa Repens fruits (SR-oil), in alginate-based beads was investigated. SR-oil has been used from centuries in both traditional and modern medicine for various nutraceutical or therapeutic purposes such as, in both sexes, as a general tonic, for genitourinary problems, to increase sexual vigor, as a diuretic or to treat in male lower urinary tract symptoms and benign prostatic hyperplasia. In this study, alginate-based beads prepared by vibration technology, also known as prilling technique, were explored as SR-oil delivery systems. Twenty-seven different formulations (F1-F27) were produced starting from stable emulsions for the period of the production. The formulations having spheroid shape (sfericity factor <0.07), high formulation yield (>90%) and high encapsulation efficiency (EE% > 80) were selected for further characterizations. Gas chromatographic analysis revealed a high loading of lauric acid as principal component of SR-oil allowing to calculate the content of total fatty acids (>50%) into the beads. Swelling behavior and release features were also studied at different pH values. The swelling of the beads and their SR-oil release were negligible for the first 2 h in simulated gastric fluid (pH 1.2), and appreciable in simulated intestinal fluid (pH 6.8). The release data were fitted by various equations to define the release kinetic mechanism. In addition, the selected formulation (F16) was stable to the oxidation not only during the formulation process, but also after 3 months of storage at room temperature. In summary, these polynucleate alginate beads, produced by prilling technique, are promising systems for improving the intestinal specific delivery and bioavailability of health-promoting bioactive SR-oil.

In Vitro Selection of Probiotics, Prebiotics, and Antioxidants to Develop an Innovative Synbiotic (NatuREN G) and Testing Its Effect in Reducing Uremic Toxins in Fecal Batches from CKD Patients.

We aimed to develop an innovative synbiotic formulation for use in reducing dysbiosis, uremic toxins (e.g., p-cresol and indoxyl sulfate), and, consequently, the pathognomonic features of patients with chronic kidney disease (CKD). Twenty-five probiotic strains, belonging to lactobacilli and Bifidobacterium, were tested for their ability to grow in co-culture with different vegetable (pomegranate, tomato, and grapes) sources of antioxidants and prebiotics (inulin, fructo-oligosaccharides, and ß-glucans). Probiotics were selected based on the acidification rates and viable cell counts. Inulin and fructo-oligosaccharides reported the best prebiotic activity, while a pomegranate seed extract was initially chosen as antioxidant source. The investigation was also conducted in fecal batches from healthy and CKD subjects, on which metabolomic analyses (profiling volatile organic compounds and total free amino acids) were conducted. Two out of twenty-five probiotics were finally selected. After the stability tests, the selective innovative synbiotic formulation (named NatuREN G) comprised Bifidobacterium animalis BLC1, Lacticaseibacillus casei LC4P1, fructo-oligosaccharides, inulin, quercetin, resveratrol, and proanthocyanidins. Finally, NatuREN G was evaluated on fecal batches collected from CKD in which modified the viable cell densities of some cultivable bacterial patterns, increased the concentration of acetic acid and decane, while reduced the concentration of nonanoic acid, dimethyl trisulfide, and indoxyl sulfate.

An Innovative Synbiotic Formulation Decreases Free Serum Indoxyl Sulfate, Small Intestine Permeability and Ameliorates Gastrointestinal Symptoms in a Randomized Pilot Trial in Stage IIIb-IV CKD Patients.

Proteolytic dysbiosis of the gut microbiota has been recognized as both a typical feature of chronic kidney disease (CKD) and a risk factor for its progression. Blood accumulation of gut-derived uremic toxins (UTs) like indoxyl sulfate (IS) and p-cresyl sulfate (PCS), intestinal permeability and constipation are typical features accompanying CKD progression and triggering chronic inflammation. In order to verify the efficacy of the innovative synbiotic formulation NATUREN G® in modulating the levels of circulating UTs, intestinal permeability and gastrointestinal symptoms, we set up a randomized, single-blind, placebo-controlled, pilot trial in stage IIIb-IV CKD patients and in healthy controls. Two-month administration of the synbiotic resulted in a decrease of free IS, as compared with the placebo-treated arm, only in the CKD group. The other UTs did not significantly change, although different trends in time (increase in the placebo arm and decrease in the synbiotic arm) were observed. Moreover, after supplementation, reduction of small intestinal permeability and amelioration of abdominal pain and constipation syndromes were observed only in the CKD group. The obtained results suggest the specificity of action of NATUREN G® in CKD and justify further validation in a wider study population.

Stability data of extemporaneous suspensions of hydroxychloroquine sulphate in oral liquid bases after tablet manipulation.

Hydroxychloroquine is a well-known anti-malarial and anti-rheumatic drug that has garnered recently unprecedented attention as potential therapeutic agent against virus infections. Hydroxychloroquine sulphate is authorized in the EU as film-coated tablets (i.e. Plaquenil®), which cannot be administered to non-cooperative patients, such as those in intensive care units or, more in general, unable to swallow solid dosage forms. Therefore, the hospital pharmacist must manipulate the solid dosage form for the preparation of suspension, even if it can strongly affect the product quality. In this scenario, it is crucial to offer useful information and advice to assist hospital pharmacists in their activity. The data presented in this article suggest that extemporaneous suspensions of hydroxychloroquine sulphate in oral liquid bases after tablet manipulation are stable for at least 30 days.

The hydroxypropyl-ß-cyclodextrin-minoxidil inclusion complex improves the cardiovascular and proliferative adverse effects of minoxidil in male rats: Implications in the treatment of alopecia.

The efficacy of minoxidil (MXD) ethanolic solutions (1%-5% w/v) in the treatment of androgenetic alopecia is limited by adverse reactions. The toxicological effects of repeated topical applications of escalating dose (0.035%-3.5% w/v) and of single and twice daily doses (3.5% w/v) of a novel hydroxypropyl-ß-cyclodextrin MXD GEL formulation (MXD/HP-ß-CD) and a MXD solution were investigated in male rats. The cardiovascular effects were evaluated by telemetric monitoring of ECG and arterial pressure in free-moving rats. Ultrasonographic evaluation of cardiac morphology and function, and histopathological and biochemical analysis of the tissues, were performed. A pharmacovigilance investigation was undertaken using the EudraVigilance database for the evaluation of the potential cancer-related effects of topical MXD. Following the application of repeated escalating doses of MXD solution, cardiac hypertrophy, hypotension, enhanced serum natriuretic peptides and K+ -ion levels, serum liver biomarkers, and histological lesions including renal cancer were observed. In addition, the administration of a twice daily dose of MXD solution, at SF rat vs human = 311, caused reductions in the systolic, diastolic, and mean blood pressure of the rats (-30.76 ± 3%, -28.84 ± 4%, and -30.66 ± 5%, respectively, vs the baseline; t test P < .05). These effects were not reversible following washout of the MXD solution. Retrospective investigation showed 32 cases of cancer associated with the use of topical MXD in humans. The rats treated with MXD HP-ß-CD were less severely affected. MXD causes proliferative adverse effects. The MXD HP-ß-CD inclusion complex reduces these adverse effects.

Characterization of minoxidil/hydroxypropyl-ß-cyclodextrin inclusion complex in aqueous alginate gel useful for alopecia management: Efficacy evaluation in male rat.

Solid inclusion complex between hydroxypropyl-ß-cyclodextrin (HP-ß-CD) and minoxidil (MXD) was prepared by freeze-drying and characterized by yield, drug loading and dissolution rate. Moreover, the complex was formulated as alginate gel (GEL HP-ß-CD)/MXD 3.5% w/w). The efficacy of the novel GEL HP-ß-CD)/MXD 3.5% w/w and of MXD 3.5% w/w ethanolic/propylene-glycol solution (MXD solution) were evaluated by monitoring the hair growth of dorsal skin 1-4 weeks after depilation followed by histological analysis and gene expression in skin biopsies in male rat. Patch-clamp experiments and cell-dehydrogenase activity (CDA) were performed to evaluate the capability of the formulations to activate "in vitro" the ATP-sensitive K+-channels (KATP) and their effects on cell viability in skin fibroblasts. After 3 weeks, the MXD solution and MXD/HP-ß-CD GEL enhanced the hair growth, respectively, of 80.1 ±â€¯2% and 84.3 ±â€¯4% vs controls. After 4 weeks, the MXD/HP-ß-CD GEL significantly enhanced the hair length and bulb diameter vs others groups. The MXD/HP-ß-CD GEL significantly enhanced the mRNA levels of the SUR2 and Kir6.1 subunits of the KATP channels and AKT2 vs other groups. The AR gene was down-regulated vs controls following the treatment with either MXD formulations. Either MXD (10-4 M) formulations were effective in potentiating the KATP currents. The MXD solution and its vehicle after 9 h of incubation time, but not MXD/HP-ß-CD, reduced CDA in fibroblasts. In sum, the MXD/HP-ß-CD GEL shows a favorable profile following topical long-term use.

Alginate-Based Hydrogel Containing Minoxidil/Hydroxypropyl-ß-Cyclodextrin Inclusion Complex for Topical Alopecia Treatment.

Cutaneous minoxidil (MXD) formulations were developed with the intent to reduce the side effects of the cosolvents propylene glycol and ethanol, frequently used in commercial MXD solutions. Completely aqueous alginate-based hydrogels were investigated and MXD aqueous solubility was improved using inclusion complexes with hydroxypropyl-ß-cyclodextrin (HP-ß-CD) at 2 different molar substitution degree (MS), namely 0.65 and 0.85. HP-ß-CD MS 0.65 was selected for its improved solubilizing ability toward MXD. At concentration of 39% w/v, this cyclodextrin increased the intrinsic aqueous solubility of MXD of about 22-fold. The calculated complexation constant was 2309 ± 20 M-1, and the inclusion process was spontaneous and enthalpically driven. Nuclear magnetic resonance studies (Job plot, 1H, 2D correlations spectroscopy, nuclear overhauser effect spectroscopy, and rotating-frame overhauser enhancement spectroscopy) confirmed the stoichiometry 1:1 between MXD and HP-ß-CD providing information about the exact geometry of the inclusion complex. Rheological and in vitro release studies performed on the formulation loaded with MXD 3.5% w/w proved that the inclusion complex increased the viscosity of the hydrogel modulating the release of the free drug. Furthermore, the hydrogel formulation facilitate MXD to permeate into the skin and did not damage epidermis, suggesting that these completely aqueous MXD delivery systems can be proposed as alternative formulations to commercial solutions.

Corrigendum to "Immune Profile of Obese People and In Vitro Effects of Red Grape Polyphenols on Peripheral Blood Mononuclear Cells".

[This corrects the article DOI: 10.1155/2017/9210862.].

Immune Profile of Obese People and In Vitro Effects of Red Grape Polyphenols on Peripheral Blood Mononuclear Cells.

The in vitro ability of polyphenols, extracted from red grape, to modulate peripheral blood mononuclear cell responses has been evaluated in 20 obese (Ob) people. With regard to cytokine release in response to phorbol myristate acetate (PMA), levels of interleukin-2 (IL-2), interferon-γ (IFN-γ), IL-4, IL-10, and IL-17 were higher in the Ob than in healthy (H) subjects. Vice versa, IL-21 concentrations were detected only in H people but they were undetectable in the Ob counterpart. In general terms, levels of IL-1ß, IL-6, IL-8, and tumor necrosis factor-α were higher in Ob people when compared to H controls. On the other hand, polyphenols did not modify IFN-γ, IL-4, and IL-17 levels. However, an increase in IL-2 was observed in H individuals, whereas its levels were decreased in the Ob counterpart. Polyphenols significantly increased IL-10 release from H donors, whereas a trend to increase was observed in Ob people. In addition, polyphenols were able to significantly increase levels of H IL-21, while this was not the case in Ob people. Since IL-21 is an inducer of Th17 cells, it is likely that polyphenols may suppress the sources of this cytokine via production of IL-10. Accordingly, polyphenols decreased IL-1ß and IL-6 release in comparison to H controls.

Administration of a Polyphenol-Enriched Feed to Farmed Sea Bass (Dicentrarchus labrax L.) Modulates Intestinal and Spleen Immune Responses.

Farmed fish are exposed to a continuous antigenic pressure by microbial and environmental agents, which may lead to a condition of chronic inflammation. In view of the notion that polyphenols, largely contained in fruits and vegetables, are endowed with antioxidant and anti-inflammatory activities, farmed sea bass (Dicentrarchus labrax L.) have been administered with red grape polyphenol-enriched feed. Polyphenols were extracted from the seeds of Canosina Nero di Troia Vitis vinifera and mixed with conventional feed at two different concentrations (100 and 200 mg/kg, resp.). Fish samples collected at days 223 and 273, respectively, were evaluated for intestinal and spleen cytokine release as well as for spleen macrophage (MØ) and melanomacrophage center (MMC) areas and distribution. Data will show that in treated fish decrease of intestinal interleukin- (IL-) 1ß and IL-6 and increase of splenic interferon- (IFN-) γ occur. On the other hand, in the spleen reduction of MØ number seems to parallel increase in MMCs. Collectively, these data suggest that polyphenol-administered sea bass generate lower levels of intestinal proinflammatory cytokines, while producing larger amounts of spleen IFN-γ, as an expression of a robust and protective adaptive immune response. Increase of MMCs corroborates the evidence for a protective spleen response induced by feed enriched with polyphenols.