Green Processing of Black Raspberry Pomace: Application of Sonotrode-Based Extraction Technique and Particles from Gas-Saturated Solutions (PGSS) Technology.

The aim of this study was to develop, for the first time, anthocyanin-enriched fractions from black raspberry pomace (BRP) using ultrasound-assisted extraction (UAE) via sonotrode and the Particles from Gas-Saturated Solutions (PGSS) process. UAEs with different amplitudes and sonication times were evaluated and showed relevant effects on the yields of target analytes. The raspberry pomace extracts were formulated in a powder form by PGSS using glyceryl monostearate as a carrier at different extract-to-carrier ratios of 1:11, 1:5, and 1:3. The effects of all variables were evaluated in terms of extraction yield, total phenolic content, and encapsulation yield. UAE was strongly affected by amplitude, and the highest amplitude (100%) provided the best results for extraction yield and total phenolics. HPLC of UAE extracts and powders was utilized for quantification of polyphenol compounds, showing cyanidin-3-rutinoside as a main compound, followed by cyanidin-3-glucoside, rutin, ellagic acid, and gallic acid. These results show that these time-efficient and high-performance techniques enable the production of natural fractions from industrial BRP with acceptable characteristics to be used for the development of nutraceuticals and different food formulations.

Significance tests for R2 of out-of-sample prediction using polygenic scores.

The coefficient of determination (R2) is a well-established measure to indicate the predictive ability of polygenic scores (PGSs). However, the sampling variance of R2 is rarely considered so that 95% confidence intervals (CI) are not usually reported. Moreover, when comparisons are made between PGSs based on different discovery samples, the sampling covariance of R2 is required to test the difference between them. Here, we show how to estimate the variance and covariance of R2 values to assess the 95% CI and p value of the R2 difference. We apply this approach to real data calculating PGSs in 28,880 European participants derived from UK Biobank (UKBB) and Biobank Japan (BBJ) GWAS summary statistics for cholesterol and BMI. We quantify the significantly higher predictive ability of UKBB PGSs compared to BBJ PGSs (p value 7.6e-31 for cholesterol and 1.4e-50 for BMI). A joint model of UKBB and BBJ PGSs significantly improves the predictive ability, compared to a model of UKBB PGS only (p value 3.5e-05 for cholesterol and 1.3e-28 for BMI). We also show that the predictive ability of regulatory SNPs is significantly enriched over non-regulatory SNPs for cholesterol (p value 8.9e-26 for UKBB and 3.8e-17 for BBJ). We suggest that the proposed approach (available in R package r2redux) should be used to test the statistical significance of difference between pairs of PGSs, which may help to draw a correct conclusion about the comparative predictive ability of PGSs.

Frequency of Use of Fixed-Combination Eye Drops by Patients with Glaucoma at Multiple Private Practices in Japan.

Purpose: To investigate the current use of fixed-combination eye drops by patients with glaucoma in Japan. Patients and Methods: A total of 5303 outpatients (5303 eyes) with glaucoma or ocular hypertension were enrolled in the survey at 78 private practices from March 8 to 14, 2020. The medications they used were investigated. The use of fixed-combination medications was analyzed, stratified by the number of medications used. The results were compared to those of the survey performed in 2016 via χ 2 tests. Results: Fixed-combination medications were used by 55.6%, 79.8%, 84.9%, 91.3%, and 94.1% of patients in the two-, three-, four-, five, and six-medications groups. The use of prostaglandin analog/ß-blocker (PG/ß), carbonic anhydrase inhibitor/ß-blocker (CAI/ß), and α-2-adrenergic agonist/ß-blocker (α2/ß) fixed-combination medications were 42.8%, 12.0%, and 0.8% in the two-medications group; 36.2%, 41.6%, and 2.0% in the three-medications group; and 23.5%, 60.1%, and 1.3% in the four-medications group, respectively. The most commonly used fixed-combination medications were PG/ß in the two-medications group and CAI/ß in the groups using three or more medications. Compared to those in the survey performed in 2016, there were increases (both P < 0.0001) in the use of PG/ß fixed-combination medication in the two- (28.7% to 42.8%) and three-medications (21.7% to 36.2%) groups, and none in the use of CAI/ß fixed-combination medication. There was little use of α2/ß fixed-combination medication, probably because it was launched only three months before the survey. Conclusion: The proportion of fixed-combination medications increased as the number of medications per patient increased.

Pharmaceutical amorphous solid dispersion: A review of manufacturing strategies.

Amorphous solid dispersions (ASDs) are popular for enhancing the solubility and bioavailability of poorly water-soluble drugs. Various approaches have been employed to produce ASDs and novel techniques are emerging. This review provides an updated overview of manufacturing techniques for preparing ASDs. As physical stability is a critical quality attribute for ASD, the impact of formulation, equipment, and process variables, together with the downstream processing on physical stability of ASDs have been discussed. Selection strategies are proposed to identify suitable manufacturing methods, which may aid in the development of ASDs with satisfactory physical stability.

A supercritical fluid technology for liposome production and comparison with the film hydration method.

Liposomes were produced by an innovative method using supercritical carbon dioxide as a dispersing agent. A quality by design strategy was used to find optimal production conditions with specific parameters (lipid concentration, dispersion volume, agitation rate, temperature and pressure) allowing the production of liposomes with predicted physicochemical characteristics (particles size and PdI). Two conditions were determined with specific production parameters. It was shown that these two conditions allowed the production of liposomes of different compositions and that most of the liposome formulations had size and dispersity in accordance with the prediction values. The condition involving the higher lipid concentration showed a higher variability in terms of size and dispersity. However, this variability remained acceptable. This innovative supercritical method allowed the production of liposomes with physicochemical characteristics similar to those obtained by the conventional thin film hydration method. This new supercritical carbon dioxide method easily scalable in GMP conditions is a one-step production method contrarily to conventional methods which generally need an additional step as extrusion to homogenize the size of liposomes.

Performance of Chinese medical postgraduate students in literature searching: a two-center survey and analysis.

BACKGROUND: To explore the performance status of Chinese postgraduate medical students in literature searching. METHODS: A self-designed online questionnaire was used to assess the literature search performance of postgraduate students (PGSs) from the classes of 2016, 2017, 2018, and 2019 from two medical colleges. The items of the questionnaire mainly included: the demographic characteristics of the PGSs, methods of literature review, literature reading habits, and use of literature. We also designed a self-assessed score that ranged from the lowest 1 point to the highest 5 points. RESULTS: A total of 902 PGSs (482 male, average age: 29.4±5.8 years old, working time range: 0-10 years, average 3.7±2.4 years) completed the questionnaire. Most PGSs investigated literature only at the work tasks (632, 70.1%) and writing papers (571, 63.3%) stages. Of the PGSs, 542 (60.1%) PGSs searched literature frequency (≥1 paper/week), and 114 (12.6%) did not perform advanced searches, and some had no knowledge of advanced search techniques at all. Most PGSs had not read more than 100 Chinese articles or English articles before. Most PGSs were used to read articles from the most authoritative journals (665, 73.7%) or high impact factor (IF) (540, 59.9%). PGSs (845, 93.7%) only read the full text of articles they deemed important. Of the PGSs, 441 (48.9%) did not use literature management tools. For self-assessed score of literature searching and reading skills, the mean was 2.1 (standard deviation, 0.8). Reading literature efficiently (710, 78.7%) and tracking recent literatures (615, 68.2%) were the two needed literature skills reported. CONCLUSIONS: Chinese medical PGSs still have room for improvement in relation to literature investigation. Intensive training in literature searching should be given to improve their performance.

Modeling of the Production of Lipid Microparticles Using PGSS® Technique.

Solid lipid microparticles (SLMPs) are attractive carriers as delivery systems as they are stable, easy to manufacture and can provide controlled release of bioactive agents and increase their efficacy and/or safety. Particles from Gas-Saturated Solutions (PGSS®) technique is a solvent-free technology to produce SLMPs, which involves the use of supercritical CO2 (scCO2) at mild pressures and temperatures for the melting of lipids and atomization into particles. The determination of the key processing variables is crucial in PGSS® technique to obtain reliable and reproducible microparticles, therefore the modelling of SLMPs production process and variables control are of great interest to obtain quality therapeutic systems. In this work, the melting point depression of a commercial lipid (glyceryl monostearate, GMS) under compressed CO2 was studied using view cell experiments. Based on an unconstrained D-optimal design for three variables (nozzle diameter, temperature and pressure), SLMPs were produced using the PGSS® technique. The yield of production was registered and the particles characterized in terms of particle size distribution. Variable modeling was carried out using artificial neural networks and fuzzy logic integrated into neurofuzzy software. Modeling results highlight the main effect of temperature to tune the mean diameter SLMPs, whereas the pressure-nozzle diameter interaction is the main responsible in the SLMPs size distribution and in the PGSS® production yield.

Lidocaine-Loaded Solid Lipid Microparticles (SLMPs) Produced from Gas-Saturated Solutions for Wound Applications.

The delivery of bioactive agents using active wound dressings for the management of pain and infections offers improved performances in the treatment of wound complications. In this work, solid lipid microparticles (SLMPs) loaded with lidocaine hydrochloride (LID) were processed and the formulation was evaluated regarding its ability to deliver the drug at the wound site and through the skin barrier. The SLMPs of glyceryl monostearate (GMS) were prepared with different LID contents (0, 1, 2, 4, and 10 wt.%) using the solvent-free and one-step PGSS (Particles from Gas-Saturated Solutions) technique. PGSS exploits the use of supercritical CO2 (scCO2) as a plasticizer for lipids and as pressurizing agent for the atomization of particles. The SLMPs were characterized in terms of shape, size, and morphology (SEM), physicochemical properties (ATR-IR, XRD), and drug content and release behavior. An in vitro test for the evaluation of the influence of the wound environment on the LID release rate from SLMPs was studied using different bioengineered human skin substitutes obtained by 3D-bioprinting. Finally, the antimicrobial activity of the SLMPs was evaluated against three relevant bacteria in wound infections (Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa). SLMPs processed with 10 wt.% of LID showed a remarkable performance to provide effective doses for pain relief and preventive infection effects.

Microencapsulation of rice bran oil using pea protein and maltodextrin mixtures as wall material.

In this work, the encapsulation of rice bran oil extracted using supercritical CO2 has been studied. In the first stage, the emulsification process by high pressure homogenization was studied and optimized. The effect of the working pressure (60-150 MPa), the composition of the carrier (mixtures of pea protein isolate (PPI) and maltodextrin (MD), from 50 to 90% of PPI) and the carrier to oil ratio (2-4) on the emulsion droplet size (EDS) was studied. To minimize the EDS, moderate pressures (114 MPa), a carrier composed mainly by PPI (64%) and carrier to oil ratios around 3.2 were required. The emulsion obtained in the optimal conditions (EDS = 189 ± 3nm) was dried using different technologies (spray-drying, PGSS-drying and freeze drying). The supercritical CO2 based drying process (PGSS) provided spherical particles that resulted in the smallest average size (but broader distribution) and lower encapsulation efficiency (53 ± 2%).

Development of innovative medical devices by dispersing fatty acid eutectic blend on gauzes using supercritical particle generation processes.

In order to limit bacterial infections during wound treatment, it is interesting to consider the concept of loading medical devices with antibacterial agents. With this in mind, an innovative system with thermosensitive properties was produced: loading a commercially available gauze with a fatty acid eutectic blend based on lauric acid (LA) and myristic acid (MA). This eutectic blend presents a melting point near physiological temperature, which together with its antibacterial properties make an appealing alternative in biomedical applications. At room temperature, the properties and the efficacy of the eutectic blend loaded onto gauzes are preserved, whereas at physiological temperature the eutectic blend undergoes a phase change that facilitates its diffusion from the gauze. The loading of the eutectic blend onto gauzes was performed using two different supercritical fluid technologies, namely, particle from gas saturated solutions (PGSS) and a derived version of rapid expansion of supercritical solution (D-RESS). The PGSS led to a heterogeneous dispersion of the eutectic blend in the gauze, whereas the D-RESS process led to the formation of a homogeneous dispersion along the surface of the gauze. Additionally, with D-RESS no phase separation of the eutectic blend occurred and the cytotoxicity was greatly improved compared with PGSS without compromising the antibacterial properties of the fatty acid eutectic blend. Hence, the present study highlights the potential use of the flexible D-RESS process to load the fatty acid eutectic blend with antibacterial properties onto medical devices in a controllable way. Overall, the effects produced by the loaded gauzes suggest the enormous potential of the developed technology in health-related areas.

Gas-saturated solution process to obtain microcomposite particles of alpha lipoic acid/hydrogenated colza oil in supercritical carbon dioxide.

Alpha lipoic acid (ALA), an active substance in anti-aging products and dietary supplements, need to be masked with an edible polymer to obscure its unpleasant taste. However, the high viscosity of the ALA molecules prevents them from forming microcomposites with masking materials even in supercritical carbon dioxide (scCO2). Therefore, the purpose of this study was to investigate and develop a novel production method for microcomposite particles for ALA in hydrogenated colza oil (HCO). Microcomposite particles of ALA/HCO were prepared by using a novel gas-saturated solution (PGSS) process in which the solid-dispersion method is used along with stepwise temperature control (PGSS-STC). Its high viscosity prevents the formation of microcomposites in the conventional PGSS process even under strong agitation. Here, we disperse the solid particles of ALA and HCO in scCO2 at low temperatures and change the temperature stepwise in order to mix the melted ALA and HCO in scCO2. As a result, a homogeneous dispersion of the droplets of ALA in melted HCO saturated with CO2 is obtained at high temperatures. After the rapid expansion of the saturated solution through a nozzle, microcomposite particles of ALA/HCO several micrometers in diameter are obtained.

Supercritical fluid precipitation of ketoprofen in novel structured lipid carriers for enhanced mucosal delivery--a comparison with solid lipid particles.

Structured lipid carriers based on mixture of solid lipids with liquid lipids are the second generation of solid lipid particles, offering the advantage of improved drug loading capacity and higher storage stability. In this study, structured lipid carriers were successfully prepared for the first time by precipitation from gas saturated solutions. Glyceryl monooleate (GMO), a liquid glycerolipid, was selected in this work to be incorporated into three solid glycerolipids with hydrophilic-lipophilic balance (HLB) ranging from 1 to 13, namely Gelucire 43/01™, Geleol™ and Gelucire 50/13™. In general, microparticles with a irregular porous morphology and a wide particle size distribution were obtained. The HLB of the individual glycerolipids might be a relevant parameter to take into account during the processing of solid:liquid lipid blends. As expected, the addition of a liquid lipid into a solid lipid matrix led to increased stability of the lipid carriers, with no significant modifications in their melting enthalpy after 6 months of storage. Additionally, Gelucire 43/01™:GMO particles were produced with different mass ratios and loaded with ketoprofen. The drug loading capacity of the structured lipid carriers increased as the GMO content in the particles increased, achieving a maximum encapsulation efficiency of 97% for the 3:1 mass ratio. Moreover, structured lipid carriers presented an immediate release of ketoprofen from its matrix with higher permeation through a mucous-membrane model, while solid lipid particles present a controlled release of the drug with less permeation capacity.

Particle Formation and Product Formulation Using Supercritical Fluids.

Traditional methods for solids processing involve either high temperatures, necessary for melting or viscosity reduction, or hazardous organic solvents. Owing to the negative impact of the solvents on the environment, especially on living organisms, intensive research has focused on new, sustainable methods for the processing of these substances. Applying supercritical fluids for particle formation may produce powders and composites with special characteristics. Several processes for formation and design of solid particles using dense gases have been studied intensively. The unique thermodynamic and fluid-dynamic properties of supercritical fluids can be used also for impregnation of solid particles or for the formation of solid powderous emulsions and particle coating, e.g., for formation of solids with unique properties for use in different applications. We give an overview of the application of sub- and supercritical fluids as green processing media for particle formation processes and present recent advances and trends in development.

Reprogenetics: Preimplantational genetics diagnosis.

Preimplantational Genetics Diagnosis (PGD) is requested by geneticists and reproductive specialists. Usually geneticists ask for PGD because one or both members of the couple have an increased genetic risk for having an affected offspring. On the other hand, reproductive specialists ask for embryo aneuploidy screening (PGS) to assures an euploid embryo transfer, with the purpose to achieve an ongoing pregnancy, although the couple have normal karyotypes. As embryonic aneuploidies are responsible for pre and post implantation abortions, it is logical to considerer that the screening of the embryonic aneuploidies prior to embryo transfer could improve the efficiency of the in vitro fertilization procedures. Nevertheless, it is still premature to affirm this until well-designed clinical trials were done, especially in women of advanced age where the rate of embryos with aneuploidies is much greater. Although the indications of PGD are similar to conventional prenatal diagnosis (PND), PGD has less ethical objections than the PND. As with the PGD/PGS results only unaffected embryos are transferred, both methods can avoid the decision to interrupt the pregnancy due to a genetic problem; this makes an important difference when compared to conventional prenatal diagnosis.

Evaluation of P(L)LA-PEG-P(L)LA as processing aid for biodegradable particles from gas saturated solutions (PGSS) process.

A series of biodegradable P(L)LA-PEG1.5 kDa-P(L)LA copolymers have been synthesized and compared as processing aid versus Poloxamer 407 (PEO-PPO-PEO), in the formulation of protein encapsulated microparticles, using supercritical carbon dioxide (scCO2). Bovine serum albumin (BSA) loaded microcarriers were prepared applying the particles from the gas saturated solutions (PGSS) technique using scCO2 and thus, avoiding the standard practice of organic solvent encapsulation. Four triblock copolymers were synthesized and characterized, particularly in terms of thermal properties and behaviour when exposed to scCO2. The effects of the inclusion of these copolymers in the formulation of poly(α-hydroxy acids) based microparticles - e.g. poly(D,L-lactic-co-glycolic acid) (PLGA) and poly(D,L-lactide) (PLA) - were analysed in terms of yield, particle size, morphology and drug release. The use of P(L)LA-PEG1.5 kDa-P(L)LA triblock copolymers were found to increase the yield of the PGSS-based process and to decrease the size of the microparticles produced, in comparison with the formulation containing the Poloxamer 407. Moreover the microparticles formulated with the triblock copolymers possessing the higher hydrophobic character were able to maintain a controlled drug release profile.

Practice of rational drug uses in a rural area of 24 pgs(s) in West Bengal.

Rational drug use is a function of prescription practices having medical, social, and economic implications. A household level cross-sectional study was undertaken to determine the prescribing practices and extent of rational therapy in the rural area of 24 pgs (s) South district. Following multistage stratified random sampling, 250 households were selected for the study from 24 pgs South, West Bengal, India. Medical care received by morbid persons was noted as per prescription details, wherever available. The results indicate that of the total of 485 morbid episodes, 302 were treated at some healthcare facility and prescriptions were available for 137 (45.36%) only. Injection use was seen in 9.49% of the prescriptions. In all, 29.20% prescriptions contained at least one antibiotic. This observation was more or less the same among the private and public practitioners (34%). The average number of drugs found was 3.153 per prescription. Overall 63.51% prescriptions were found to be irrational. The prescribing practices and use of drugs have been found to be truly unsatisfactory.