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Haemorrhagic shock is a leading cause of death worldwide. Blood transfusions can be used to treat patients suffering severe blood loss but donated red blood cells (RBCs) have several limitations that limit their availability and use. To solve the problems associated with donated RBCs, several acellular haemoglobin-based oxygen carriers (HBOCs) have been developed to restore the most important function of blood: oxygen transport. One promising HBOC is the naturally extracellular haemoglobin (i.e. erythrocruorin) of Lumbricus terrestris (LtEc). The goal of this study was to maximise the portability of LtEc by lyophilising it and then testing its stability at elevated temperatures. To prevent oxidation, several cryoprotectants were screened to determine the optimum formulation for lyophilisation that could minimise oxidation of the haem iron and maximise recovery. Furthermore, samples were also deoxygenated prior to storage to decrease auto-oxidation, while resuspension in a solution containing ascorbic acid was shown to partially reduce LtEc that had oxidised during storage (e.g. from 42% Fe3+ to 11% Fe3+). Analysis of the oxygen equilibria and size of the resuspended LtEc showed that the lyophilisation, storage, and resuspension processes did not affect the oxygen transport properties or the structure of the LtEc, even after 6 months of storage at 40 °C. Altogether, these efforts have yielded a shelf-stable LtEc powder that can be stored for long periods at high temperatures, but future animal studies will be necessary to prove that the resuspended product is a safe and effective oxygen transporter in vivo. [ABSTRACT FROM AUTHOR]

This study aimed to prepare a combined self-nanoemulsifying and self-assembled cubic nanoparticles (SNE/SAC) lyophilized tablet eliciting biphasic release pattern escorted with enhanced bioavailability for drugs hampered with slow dissolution and poor absorption. The antimuscarinic Darifenacin hydrobromide (DRF) was selected as a model drug used to treat overactive bladder-associated nocturia. The DRF-SNE/SAC lyophilized tablet was prepared so that upon reconstitution a mixture of DRF-loaded cubic nanoparticles and nanoemulsion dispersion is obtained. The nanoemulsion portion is responsible for the fast release followed by controlled release of the remaining dose loaded in cubic nanoparticles. A comparative pharmacokinetic study adopting randomized crossover design in male albino rabbits versus marketed product Frequefenacine® tablet was performed. Half of the dose (52.05% ± 4.21%) was rapidly released in the first 4 h followed by sustained release of the remaining drug where (90.16% ± 8.85%) was released in 24 h. The tested system showed 2.45 folds higher % relative bioavailability and 1.57 folds higher Cmax with 1.62 longer residence time relative to reference product. The results endow the ability of the developed DRF-SNE/SAC lyophilized tablet to be considered as a propitious approach for the treatment of overactive bladder-associated nocturia without midnight dose administration. [ABSTRACT FROM AUTHOR]

INTRODUCTION. A key priority in maintaining a collection of microorganisms is to ensure the stability of characteristics and the viability of microbial cultures during their storage and transport. In addition, some applications of collection strains as control samples require accurate data on the number of viable microbial cells in each sample. Therefore, it is necessary to develop and implement an analytical procedure for predicting the guaranteed shelf life of test strains.AIM. This study aimed to predict the guaranteed shelf life for test strains in a variety of primary packaging by assessing changes in microbial viability under accelerated storage conditions, with the Salmonella enterica subsp. enterica serovar Abony NCTC 6017 strain as a model organism.MATERIALS AND METHODS. The study used lyophilised samples of the S. enterica subsp. enterica serovar Abony NCTC 6017 strain deposited in the National Collection of Pathogenic Microorganisms (NCPM) at the Scientific Centre for Expert Evaluation of Medicinal Products of the Ministry of Health of the Russian Federation. The studied primary packaging types included vacuum-filled borosilicate glass ampoules and 2R lyophilisation vials. The samples were tested for a number of quality attributes (loss on drying, viable cell count, cell viability, colony morphology, and biochemical identification) and subjected to accelerated shelf-life testing at ele­vated temperatures (35–65 °С).RESULTS. The study did not show any significant differences in the quality of lyophilised samples depending on the type of primary packaging. The authors experimentally determined rate constants for the loss of viability in microbial cultures during storage at elevated temperatures and calculated the rate constants for the storage and transport temperatures and for different types of primary packaging. The predicted time to viable cell count reduction to 10% of the initial level was 19 years for vials and 25 years for ampoules, and the predicted time to 50% viability was 5.8 years for vials and 7.6 years for ampoules.CONCLUSIONS. The results of this study confirm the applicability of different primary packaging (ampoules and vials) for the lyophilisation and storage of microbial test strains. The data obtained can guide further research and contribute to the development of recommendations for the storage of lyophilised strains in various types of packaging.

Globally there is a shortage of available donor corneas with only 1 cornea available for every 70 needed. A large limitation to corneal transplant surgery is access to quality donor tissue due to inadequate eye donation services and infrastructure in many countries, compounded by the fact that there are few available long-term storage solutions for effectively preserving spare donor corneas collected in countries with a surplus. In this study, we describe a novel technology termed low-temperature vacuum evaporation (LTVE) that can effectively dry-preserve surplus donor corneal tissue, allowing it to be stored for approximately 5 years, shipped at room temperature, and stored on hospital shelves before rehydration prior to ophthalmic surgery. The dry-preserved corneas demonstrate equivalent biological characteristics to non-dried donor tissue, with the exception that epithelial and endothelial cells are removed and keratocytes are rendered non-viable and encapsulated within the preserved extracellular matrix. Structure and composition of the dried and rehydrated corneas remained identical to that of non-dried control corneas. Matrix-bound cytokines and growth factors were not affected by the drying and rehydration of the corneas. The ability to preserve human donor corneas using LTVE will have considerable impact on global corneal supply; utilisation of preserved corneas in lamellar keratoplasties, corneal perforations, ulcers, and tectonic support, will allow non-preserved donor tissue to be reserved for where it is truly required. [ABSTRACT FROM AUTHOR]

Replicating the intricate architecture of native tissues remains a significant challenge in tissue engineering. Ice-templated biomimetic scaffolds possess controlled porosity that conveniently resembles the native parenchyma of many tissues. In this study, we investigate the relationship between the porous architecture of lyophilised collagen scaffolds and key processing parameters during production. We establish a predictive model that correlates specific lyophilisation conditions with the resulting pore sizes. Systematic variations in the freeze-drying conditions resulted in scaffolds with average pore sizes ranging from 46 μm to 251 μm, effectively matching the length scale of extracellular matrix features found in native tissues. We introduce the concept of heat flux density (HFD) at equilibrium as a metric for quantifying latent heat extraction efficiency during the freezing process. Our findings reveal a power law relationship between HFD at equilibrium and pore size, with an exponent of −0.44. This approach provides a non-destructive and non-intrusive method for precisely controlling pore architecture, advancing the potential for creating scaffolds that closely emulate the complex structures of native tissues.

At present, society has embraced the fact apropos population aging and climate changes, that demand, amongst others, innovative pharmaceutical technologies, emphasising the development of patient-specific delivery systems and thus the provision of efficient and sustainable drugs. Protein drugs for subcutaneous administration, by allowing less frequent application, represent one of the most important parts of the pharmaceutical field, but their development is inevitably faced with obstacles in providing protein stability and suitable formulation viscosity. To gain further knowledge and fill the gaps in the already constructed data platform for the development of monoclonal antibody formulations, we designed a study that examines small model proteins, i.e., bovine serum albumin. The main aim of the presented work is to evaluate the effect of protein concentrations on critical quality attributes of both, pre-lyophilised liquid formulations, and lyophilised products. Through the study, the hypothesis that increasing protein concentration leads to higher viscosity and higher reconstitution time without affecting the stability of the protein was confirmed. The most important finding is that sucrose plays a key role in the lyophilisation of investigated protein, nevertheless, it can be predicted that, to ensure the beneficial effect of mannitol, its amount has to prevail over the amount of sucrose.

Summary: This research aimed to produce water and ethanol extract of the medicinal mushroom Coriolus versicolor, to lyophilise the extracts and to determine the influence of the lyophilised extracts, as a complete substitute for the MSG additive, on the sensory properties of industrially produced dehydrated vegetable soup. Based on SEM analysis, it can be observed that there are differences in the microstructure between the water and ethanol extracts. In the water extract, a great heterogeneity is observed in terms of structural characteristics. Water extract showed identical antimicrobial activity (15.7 mm) against Enterococcus faecalis compared to tetracycline, while the ethanol extract had identical antimicrobial activity (14.0 mm) compared to chloramphenicol. Both extracts can be competitive (P < 0.05) with BHT in terms of capturing DPPH radicals. According to the results of the sensory analysis, it can be noted that all analysed soups were well received (quality <70%) by the evaluators. [ABSTRACT FROM AUTHOR]

Schistosomosis is a prevalent zoonotic parasitic disease affecting both humans and animals on a global scale, with an estimated 165 million cattle and 200 million people impacted worldwide. Eventhough, serological methodologies designed for the identification of specific antibodies targeting parasitic antigens are esteemed for their high sensitivity, there are criticisms due to their incapacity to reliably indicate active infection, inability to correlate with the intensity of infection and lack of specificity. Enhancing the specificity of serological assays presents a significant challenge, primarily attributable to the identification and synthesis of specific antigens. In addressing these limitations, recombinant technology with specific immunogenic proteins as candidate antigens emerges as a viable alternative. In this study, induction of 22.6 kDa recombinant tegument protein of Schistosoma spindale was achieved using 0.6 mM concentration of IPTG at 37°C for four hours. Nickel chelating affinity chromatography was employed for protein purification, yielding maximum protein concentration at 75mM elution. Subsequently, the dialysis technique was employed to remove contaminants, while lyophilisation method was employed for protein concentration. The protein concentration postdialysis was measured at 0.220 mg/mL, while lyophilisation resulted in a concentration of 2mg/mL. [ABSTRACT FROM AUTHOR]

Traumatic musculoskeletal injuries require advanced therapeutic intervention to heal effectively. Regenerative medicine research has aimed to address this by using biomaterials to deliver gene therapeutic nanoparticles (NPs) to the injury site to direct healing. However, clinical translation has proven challenging due to the short shelf-life of NPs and requirements for cold storage conditions. Thus, this study aimed to investigate lyophilisation as a process to formulate 'off-the-shelf' NPs that can be incorporated into biomaterial scaffolds at the point of use and can be stored and transported at ambient temperatures. To this end, NPs consisting of a non-viral delivery vector, glycosaminoglycan-enhanced transduction (GET) peptide, complexed with plasmid DNA (pDNA), were formulated at three charge ratios (CRs - 6, 9, 12) and lyophilised. Firstly, the effects of lyophilisation on NP physicochemical properties were investigated; it did not affect NP size, polydispersity or charge. Next, the ability of the lyophilised NPs to express the pDNA cargo in mesenchymal stem cell (MSC) 2D monolayer culture was assessed. Transfection with lyophilised NPs at each CR promoted stable transgene expression and furthermore, once lyophilised, transgene expression could be maintained following long-term storage at room temperature. Transfection with lyophilised GET-pSOX9 NPs also significantly increased MSC-mediated articular cartilage matrix deposition in methacrylated hyaluronic acid (MeHA)-collagen type II (Col II) injectable hydrogel scaffolds, highlighting the therapeutic potential of this NP formulation. In conclusion, this study outlines an effective method for formulating 'off-the-shelf' NPs for regenerative medicine applications that could be applied to the musculoskeletal system as well as other tissues.

A high-resolution infrared (IR) camera was used for temperature measurements in a pharmaceutical formulation (mannitol/sucrose solution, 4:1%, m/m) during a freeze-drying process. The temperature was measured simultaneously at the surface as well as vertically (e.g., in-depth) along the side of custom-made cuvettes equipped with a germanium (Ge) window. Direct imaging during 45 h from −40 °C to 40 °C took place every 60 s on the surface and on the side with 0.28 × 0.28 mm per IR-pixel providing 2700 thermograms. The spatial resolution per cuvette was approximately 4225 pixels for the surface view (without a probe) and 6825 IR-pixels for the side view. Temperature effects and gradients due to the presence of a Pt100 and a LyoRx-probe in the pharmaceutical formulation were clearly visible and were quantified during the freezing step as well as the primary and secondary drying stages. The temperature was about 3.5 K higher during primary drying as compared to the temperature measured in the same material in adjacent cuvettes without probes. During secondary drying, evaporative cooling of upper layers was clearly visible. [ABSTRACT FROM AUTHOR]

Bioactive peptides (BPs) are molecules of paramount importance with great potential for the development of functional foods, nutraceuticals or therapeutics for the prevention or treatment of various diseases. A functional BP-rich dairy product was produced by lyophilisation of bovine milk fermented by the autochthonous strains Lactococcus lactis subsp. lactis ZGBP5-51, Enterococcus faecium ZGBP5-52 and Enterococcus faecalis ZGBP5-53 isolated from the same artisanal fresh cheese. The efficiency of the proteolytic system of the implemented strains in the production of BPs was confirmed by a combined high-throughput mass spectrometry (MS)-based peptidome profiling and an in silico approach. First, peptides released by microbial fermentation were identified via a non-targeted peptide analysis (NTA) comprising reversed-phase nano-liquid chromatography (RP nano-LC) coupled with matrix-assisted laser desorption/ionisation-time-of-flight/time-of-flight (MALDI-TOF/TOF) MS, and then quantified by targeted peptide analysis (TA) involving RP ultrahigh-performance LC (RP-UHPLC) coupled with triple-quadrupole MS (QQQ-MS). A combined database and literature search revealed that 10 of the 25 peptides identified in this work have bioactive properties described in the literature. Finally, by combining the output of MS-based peptidome profiling with in silico bioactivity prediction tools, three peptides (75QFLPYPYYAKPA86, 40VAPFPEVFGK49, 117ARHPHPHLSF126), whose bioactive properties have not been previously reported in the literature, were identified as potential BP candidates. [ABSTRACT FROM AUTHOR]

Summary: The aim of this research was to produce a lyophilised water and ethanol extract from Fuscoporia torulosa mushroom, to determine the antimicrobial and antioxidant potential of the extracts and in industrial conditions to produce a new, functional product – Bio Soups, enriched with lyophilised extracts derived from fruiting body of F. torulosa mushroom, at the same time without use of synthetic additives. The application of the lyophilised extracts in the industrial production of dehydrated soups showed positive results. Such soups are characterised with higher (P < 0.05) antimicrobial and antioxidant activity, compared to conventional soups, while it has been proven constant biological potential and stability up to the 90th day of production. This is the first dehydrated soup enriched with lyophilised extract of medicinal mushroom, since its production does not require special conditions, and it possesses exceptional functional properties, this product could have a many times greater market potential compared to conventional soups. [ABSTRACT FROM AUTHOR]

A high-resolution infrared (IR) camera was used for temperature measurements in a pharmaceutical formulation (mannitol/sucrose solution, 4:1%, m/m) during a freeze-drying process. The temperature was measured simultaneously at the surface as well as vertically (e.g., in-depth) along the side of custom-made cuvettes equipped with a germanium (Ge) window. Direct imaging during 45 h from −40 °C to 40 °C took place every 60 s on the surface and on the side with 0.28 × 0.28 mm per IR-pixel providing 2700 thermograms. The spatial resolution per cuvette was approximately 4225 pixels for the surface view (without a probe) and 6825 IR-pixels for the side view. Temperature effects and gradients due to the presence of a Pt100 and a LyoRx-probe in the pharmaceutical formulation were clearly visible and were quantified during the freezing step as well as the primary and secondary drying stages. The temperature was about 3.5 K higher during primary drying as compared to the temperature measured in the same material in adjacent cuvettes without probes. During secondary drying, evaporative cooling of upper layers was clearly visible.

Abstract A holistic approach was taken to characterise lyophilisers at both laboratory and commercial scale to design a focused validation strategy for commercialising parenteral drug products. Vial heat transfer coefficients (Kv) and equipment mass transfer boundaries were generated for a Lyostar II and three commercial scale IMA Lyomax lyophilisers. K v studies were performed using gravimetric methodologies. K v calculated for the Lyostar II was equivalent to the commercial equipment at 133 µBar however trended higher below 133 µBar and lower above 133 µBar potentially impacting primary drying product temperature during scale-up depending on the chamber pressure recipe set point. Kv profiles were consistent within and across the commercial equipment. Edge effect was most prominent at commercial scale with minimal shielding of the edge vials in contrast to the presence of a metal ring around the vial pack in the Lyostar II. Equipment capability studies for mass transfer showed commercial scale equipment could achieve lower chamber pressure and greater sublimation rates when compared to the Lyostar II. Furthermore, differences were also measured between large-scale lyophilisers based on condenser orientation (horizontal vs vertical). The results demonstrate greater equipment capability of the two-storey vertical configuration with respect to choked flow regime. Worst-case locations within a commercial lyophiliser were identified providing rationale for reduced sampling for product shelf-mapping locations. This work provides guidance on execution of commercial scale characterisation studies and application of the data to enhance scale-up, technical transfer and focused process validation strategies.

Elderflower extracts are known to be a source of valuable substances that show a wide spectrum of biological activity, including antibacterial and antiviral properties, which demonstrate a degree of effectiveness against SARS CoV-2. In this work, the influence of fresh inflorescence stabilisation methods (freezing, air drying, and lyophilisation) and extraction parameters on the composition and antioxidant properties of the extracts were studied. Wild elderflower plants growing in the Małopolska Region of Poland were studied. Antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging ability and ferric-reducing antioxidant power assays. The total phenolic content was determined using the Folin-Ciocalteu method and the phytochemical profile of the extracts was analysed using HPLC. The obtained results showed that the best method for the stabilisation of elderflower was lyophilisation, and the determined optimal maceration parameters were 60% methanol as a solvent and a process time of 1–2 days. [ABSTRACT FROM AUTHOR]

The fundamental purpose of this study was to develop a stable lyophilised finasteride nanosystem (FNS-NS) for topical delivery. The FNS-NS was fabricated using an ultrasonication technique. The impact of two different cryoprotectants on the physicochemical characteristics of FNS-NS before and after lyophilisation was thoroughly investigated. The lyophilised FNS-NS had spherical shape with particle size lied between 188.6 nm ± 4.4 and 298.7 nm ± 4.7, low PDI values (0.26 ± 0.02 to 0.32 ± 0.02) and zeta potential ranging from −38.3 to +53.3 mV. The confocal laser microscopy depicted a comparatively higher cellular internalisation achieved for undecorated FNS-NS with respect to its chitosan-decorated counterpart. The lyophilised FNS-NS was stable for 90 days at proper storage conditions. The FNS-NS with 15% trehalose had appropriate physicochemical attributes that could be a promising carrier for topical delivery to treat androgenic alopecia. [ABSTRACT FROM AUTHOR]

The efficient extraction of phenols from grapes is an important step for their reliable quantification. The aim was to optimise the lyophilisation process and the extraction of phenols from grape skins and seeds. The phenol extraction yield from lyophilised tissues was investigated with different accelerated solvent extraction (ASE) operating conditions. Skins and seeds were separated from frozen berries and lyophilised without being ground. The weight loss during lyophilisation was followed daily. Phenols were extracted from lyophilised, cryo-ground seeds and skins with ASE at room temperature and 10.3 MPa using 80% aqueous acetone and 60% aqueous methanol. The effects of ASE operational parameters (the number of extraction cycles (ECs) and static time (ST) duration) were investigated. The yield of extracted phenols was evaluated spectrophotometrically by determining total phenolic index at 280 nm (TPI). The weight of skins and seeds significantly dropped after 24 h of lyophilisation and continued to decrease, although not significantly, up until the 9th day. The optimal lyophilisation time was estimated to be 3 days and 5 days for skins and seeds, respectively. The phenol extraction yield was significantly affected after changes of ASE conditions. Based on TPI, the optimal ASE conditions were as follows: (i) lyophilised seeds—eight ECs with 10 min ST using aqueous acetone and then four ECs with 20 min ST using aqueous methanol; (ii) lyophilised skins—eight ECs with 1 min ST using aqueous acetone and then one EC with 20 min ST using aqueous methanol. [ABSTRACT FROM AUTHOR]

Liquid erythrocyte diagnostic preparations have a practical disadvantage; i.e., long-distance transportation involving possible non-compliance with cold-chain requirements may result in a complete loss of biological activity. A lyophilisation technology is necessary to ensure that the preparations retain their original properties for a long time. The aim of the work was to develop a protective medium and conditions for lyophilisation to stabilise the erythrocyte diagnostic preparation of tularaemia immunoglobulin. Materials and methods: Gelatin, thiourea, trehalose, sucrose, dextran, and Tween 80 were used as excipients for protective media. The authors used nine strains of homologous and heterologous microorganisms of different genera and species to control the lyophilised diagnostic preparation sensitivity and specificity. Evaluation of the main stability-related quality attributes (appearance of the dried preparation, loss on drying, solubility, appearance after reconstitution, appearance after settling, sensitivity, specificity) considered the temperatures specific to the climatic zones where the in vitro diagnostics is intended to be marketed and used. Results: The authors developed protective stabilising media with different compositions, used them in freeze-drying of the preparation and carried out control testing. The most promising was the lyophilisation medium containing a smaller amount of ingredients —6% of dextran, 0.06% of Tween 80 and up to 0.01% of sodium azide—as it was the simplest one to prepare and ensured complete preservation of the quality attributes. The authors carried out practical evaluation of lyophilisation procedures, and the 12–14-hour procedure proved to be the most cost-effective. Conclusions: The results of long-term, or real time, and accelerated stability testing of the lyophilised diagnostic preparation demonstrated the possibility of two-year storage at a labelled temperature of 2–8 °C, as well as at elevated and low temperatures of 30±2 °С and –18 °С, respectively. The tests showed no negative effects of the temperatures on the controlled quality attributes.

Aims Filoviruses encompass highly pathogenic viruses placing significant public health burden on countries affected. Efforts for improved diagnostics and surveillance are needed. The requirement for high-containment can be circumvented by using pseudotype viruses (PV), which can be handled safely, in tropism, drug screening, vaccine evaluation, and serosurveillance studies. We assessed the stability and functionality after long-term storage of lyophilised filovirus pseudotypes for use in neutralisation assays. Methods and results We generated a panel of filovirus lentiviral pseudotypes followed by lyophilisation and storage in different conditions. Next, we reconstituted and tested PVs in infection experiments and pseudotype neutralisation assays where possible. Lyophilised Ebola and Marburg PVs retained production titres for at least two years when stored at +4˚C or less. Lyophilised Ebola PVs performed similarly to non-lyophilised PVs in neutralisation assays after reconstitution. When stored at high temperatures (+37˚C), lyophilised PVs did not retain titres after 1-month storage, however, when lyophilised using pilot-scale facilities EBOV PVs retained titres and performed as standard in neutralisation assays after on 1-month storage at 37˚C. Conclusions Filovirus PVs are amenable to lyophilisation and can be stored for at least 2 years in a household fridge to be used in antibody assays. Lyophilisation performed in the right conditions would allow transportation at room temperature, even in warmer climates. [ABSTRACT FROM AUTHOR]

At the moment, there are no scientific publications devoted to the technological aspects of production of immunoglobulin solid dosage forms. The aim of the study was to review Russian and foreign literature on production of immunoglobulin solid dosage forms, and present the results of the authors’ own research. The authors analysed data of the National Register of Medicines of the Russian Federation as of mid-2021 on the authorised medicines with a generic name ‘globulin in a solid dosage form’, and summarised their characteristics. They reviewed data on the qualitative and quantitative composition of excipients used in lyophilisation, preparation of tablets and capsules. A number of examples were used to illustrate the effect of technological parameters of immunoglobulin solid form production on the quality of the finished products. It was demonstrated that the production of solid forms of immunoglobulin products prevents aggregation and fragmentation of proteins during storage, which affect the product’s specific activity, and also help to preserve the product’s target characteristics for a longer period of time as compared to liquid dosage forms of immunoglobulins. The results of the study may be used as a basis for development of a manufacturing technology for solid forms of immunoglobulin products.

Determinations concerning the glycoalkaloids content were made over four potato varieties (Hansa, Sieglinde, Nicola and Linda) obtained from a private German source, country in which potatoes are cultivated and consumed frequently. Potatoes have been cultivated in classic conditions, as well as in biodynamic conditions.The glycoalkaloids complex and their aglycones had been obtained through a sequence of operations, like extractions with several specific solvents at reflux and in backward flow, hydrolysis, lyophilisation etc.The determinations concerning the glycoalcaloids content of the four studied potato varieties were accomplished comparatively for both culture modes, in classic and biodynamic conditions.

Selection of lactic acid bacteria (LAB) for probiotic effect and preservation of LAB by lyophilized have been studied. The two LAB strains belonging to Lactobacillus plantarum and Lactobacillus pentosus producing bacteriocins in inhibitory concentration for colibacilli were selected from a total of 17 tested strains. The selected strains were further tested for their ability to survive the preservation process by lyophilisation. Two methods of preserving LAB by lyophilisation were tested and the viability of cells in lyophilized products was monitored in order to obtain products used as starter cultures. The presence of inhibition zones in mixt cultures indicates the ability of LAB to inhibit the growth of E. coli bacteria due to the synthesis of bacteriocins. During three years period, the viability of lyophilized LAB was monitored. Obtained data indicate that starter cultures of LAB can be stored in the lyophilized state for at least three years in dark and cool place, and the inoculation rate can be increased after this period by 50% if the starter culture was preserved and stored by applying the described techniques.

Probiotics are extensively used in animal nutrition to improve feed conversion parameters, to boost the immune response and ultimately to improve animal health and welfare. The aim of our study was to obtain a lyophilised product for use as supplement in poultry feeding and to determine the optimal concentration required for inclusion in feed. In our experiment we have used the following human isolated probiotic bacteria: Lactococcus lactis CMGB 31, Lactobacillus paracasei CMGB 18, Lactococcus lactis CMGB 33, Lactobacillus rhamnosus CMGB 34 and Lactococcus lactis CMGB 32. The strains were preserved by freezing at - 80ºC. After lyophilisation, using skimmed milk and MRS mixture as preservative and MRS broth as growth media the recorded concentration was of 1·109 CFU/g lyophilized product. This product can now be used as probiotic supplement in poultry feed at a concentration of 1 g/kg feed.

Faecal microbiota transplant (FMT) is an established and effective treatment for recurrent Clostridioides difficile infection (CDI) and has many other potential clinical applications. However, preparation and quality of FMT is poorly standardised and clinical studies are hampered by a lack of well-defined FMT formulations that meet regulatory standards for medicines. As an alternative to FMT suspensions for administration by nasojejunal tube or colonoscopy, which is invasive and disliked by many patients, this study aimed to develop a well-controlled, standardised method for manufacture of lyophilised FMT capsules and to provide stability data allowing storage for extended time periods. Faecal donations were collected from healthy, pre-screened individuals, homogenised, filtered and centrifuged to remove dietary matter. The suspension was centrifuged to pellet bacteria, which were resuspended with trehalose and lyophilised to produce a powder which was filled into 5 enteric-coated capsules (size 0). Live-dead bacterial cell quantitative PCR assay showed <10 fold viable bacterial load reduction through the manufacturing process. No significant loss of viable bacterial load was observed after storage at -80 °C for 36 weeks (p = 0.24, n = 5). Initial clinical experience demonstrated that the capsules produced clinical cure in patients with CDI with no adverse events reported (n = 7). We provide the first report of a detailed manufacturing protocol and specification for an encapsulated lyophilised formulation of FMT. As clinical trials into intestinal microbiota interventions proceed, it is important to use a well-controlled investigational medicinal product in the studies so that any beneficial results can be replicated in clinical practice. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] During the filling process of a biopharmaceutical drug product (DP), a liquid DP film might creep up the inner vial wall which is barely discernible, appears as milky-white haze after lyophilisation and is known as fogging. Creeping and fogging are mainly dependent on the primary packaging material surface and its hydration, vial preparation process as well as DP composition. The occurrence of both can impede visual inspection and might lead to DP rejection. Hence, our studies focused on the early detection of liquid solution and glass vial surface interaction directly after filling. For a fast and highly sensitive evaluation a novel video-based analysis was used. To our knowledge, this is the first time a MATLAB®-algorithm-based video analysis was applied to quantitatively determine creeping time-resolved. Furthermore, creeping in dependence of vial processing sites, surfactant type and concentration, filling temperature, and vial format were investigated. The results were verified using orthogonal conventional methods such as surface tension, wetting behaviour, and contact angle measurements, as well as ToF-SIMS, ICP-MS, and SEM. Additionally, the methods applied were assessed regarding their cross-validation capability. The observations indicate that the vial preparation process can have a pronounced impact on alteration of the glass vial surface and related creeping behaviour of the filled solution. [ABSTRACT FROM AUTHOR]

A holistic approach was taken to characterise lyophilisers at both laboratory and commercial scale to design a focused validation strategy for commercialising parenteral drug products. Vial heat transfer coefficients (Kv) and equipment mass transfer boundaries were generated for a Lyostar II and three commercial scale IMA Lyomax lyophilisers. Kv studies were performed using gravimetric methodologies. Kv calculated for the Lyostar II was equivalent to the commercial equipment at 133 µBar however trended higher below 133 µBar and lower above 133 µBar potentially impacting primary drying product temperature during scale-up depending on the chamber pressure recipe set point. Kv profiles were consistent within and across the commercial equipment. Edge effect was most prominent at commercial scale with minimal shielding of the edge vials in contrast to the presence of a metal ring around the vial pack in the Lyostar II. Equipment capability studies for mass transfer showed commercial scale equipment could achieve lower chamber pressure and greater sublimation rates when compared to the Lyostar II. Furthermore, differences were also measured between large-scale lyophilisers based on condenser orientation (horizontal vs vertical). The results demonstrate greater equipment capability of the two-storey vertical configuration with respect to choked flow regime. Worst-case locations within a commercial lyophiliser were identified providing rationale for reduced sampling for product shelf-mapping locations. This work provides guidance on execution of commercial scale characterisation studies and application of the data to enhance scale-up, technical transfer and focused process validation strategies. [ABSTRACT FROM AUTHOR]

Colostrum, the first food given to mammals after birth, plays a critical role in newborn nutrition. It is characterized by its high nutrient content, presence of immunoglobulins, and functional compounds. This study collected and freeze-dried colostrum from four different animals (Karayaka sheep, Saanen goats, Danish red cows and buffalo). Chemical and functional properties (total phenolic content and antioxidant capacity), mineral, as well as mineral content, amino acid composition, and fatty acid profile of the dried samples, were determined. Nutritional properties (protein efficiency ratio (PER), biological value (BV) and net protein utilization (NPU)) of powdered colostrum were also determined. It was found that buffalo colostrum had higher fat and protein content, while goat colostrum had high antioxidant activity. Buffalo and goat colostrums had a biological value (BV) of 94.96–97.78. Major minerals were more abundant in cow colostrum and minor minerals were more abundant in sheep colostrum. In terms of essential fatty acids, sheep and goat colostrums stood out. In conclusion, this comprehensive analysis of the composition of colostrum from different species sheds light on the different nutrient profiles and potential health benefits and provides a theoretical basis for the development of commercial products from goat, sheep, cow and buffalo colostrum. [ABSTRACT FROM AUTHOR]

Supercooling during the freezing of pharmaceutical solutions often leads to suboptimal freeze-drying results, such as long primary drying times or a collapse in the cake structure. Thermal treatment of the frozen solution, known as annealing, can improve those issues by influencing properties such as the pore size and collapse temperature of the lyophilisate. In this study we aimed to show that annealing causes a rearrangement of water molecules between ice crystals, as well as between the freeze-concentrated amorphous matrix and the crystalline ice phase in a frozen binary aqueous solution. Ice crystal sizes, as well as volume fractions of the crystalline and amorphous phases of 10% (w/w) sucrose and trehalose solutions, were quantified after annealing using freeze-drying microscopy and image labelling. Depending on the annealing time and temperature, the amorphous phase was shown to decrease its volume due to the crystallisation of vitreous water (i.e., glassy state relaxation) while the crystalline phase was undergoing coarsening (i.e., Ostwald ripening). These results allow, for the first time, a quantitative comparison of the two phenomena. It was demonstrated that glassy state relaxation and Ostwald ripening, although occurring simultaneously, are distinct processes that follow different kinetics. [ABSTRACT FROM AUTHOR]

The coupling of an infrared (IR) camera to a freeze dryer for monitoring of the temperature of a pharmaceutical formulation (sucrose/mannitol solution, 4:1%, m/m) during freeze-drying has been exploited further. The new development allows monitoring of temperatures simultaneously at the surface as well as vertically, (e.g., in depth) along the side using custom-made cuvettes. The IR camera was placed on the chamber roof of a process-scale freeze dryer. Monitoring of cuvettes containing the formulation took place from above where one side of each cuvette was equipped with a germanium window. The Ge-window was placed next to an IR mirror having a 45° angle. The long-wave infrared radiation (LWIR) coming from the inside of the cuvette was reflected upwards toward the IR camera. Accurate recording of the temperature along the cuvettes' depth profile was therefore possible. Direct imaging from −40 °C to 30 °C took place every 60 s on the surface and on the side with a 2 × 2 mm resolution per IR pixel for 45 h resulting in 2700 thermograms. Results are presented for freeze-drying of a pharmaceutical formulation as a function of time and spatially for the entire side (depth) of the cuvette. As the sublimation process was progressing, the spatial resolution (84 IR pixels for the side-view and 64 pixels for the surface-view) was more than sufficient to reveal lower temperatures deeper down in the material. The results show that the pharmaceutical formulation (a true solution at the onset) dries irregularly and that the sublimation front does not progress evenly through the material. During secondary drying, potential evaporative cooling of upper layers could be detected thanks to the high thermal and spatial resolution. [ABSTRACT FROM AUTHOR]

Some studies have found that the nutritional values of stingless bee honey (SBH) may be similar if not more than normal honey, prompting the Malaysian government to promote it as a superfood. However, SBH does not fulfil the Codex Standard for Honey (CODEX STAN 12-19811) in terms of moisture content and the lack of protein to be analysed with Internal Standard Carbon Isotope Ratio Analysis (ISCIRA). Hence, a lyophilization process was introduced prior to stable carbon isotope analysis of SBH to address both of these issues. It was found that once moisture content was decreased to a level below 20 % for 19 SBH samples, the percentage increment of protein extracted from the samples varied between 6 and 385 % relative to protein extracted from SBH before lyophilization with nine samples found to be adulterated. Caution is necessary when lyophilizing the SBH as significant isotope shifts were seen for SCIRA and ISCIRA values. Nevertheless, the carbon isotope shifts did not change the final outcome of the 'pass' or 'fail' of the adulteration result. Overall, the removal of water from SBH is required but caution is necessary as carbon isotope shifts were observed as SBHs underwent the lyophilization process. [ABSTRACT FROM AUTHOR]

The pulmonary administration of drugs offers advantages over administration by intravenous injection. The present research work was to prepare polymeric micelles nanomicelles containing Doxofylline as dry powder for inhalation. In order to bypass the drawbacks of predictable preparations, nanotechnology-based drug delivery systems for pulmonary administration and pulmonary targeting have been oppressed. The work was aimed towards formulating the Doxofylline loaded Soluplus nanomicelles with polymer Soluplus by film hydration technique in different ratios. The nanomicelles so prepared were characterized for its particle size, ζ-potential, XRD, SEM, drug content and drug release rate. Soluplus Doxofylline Polymeric Micelles showed mean size of 70.73nm and Zeta Potential is - 8.71. The Drug Entrapment Efficiency is 87.7% and The Drug Loading Capacity is 10%. In-Vitro study reveals the drug release is 44%. Conclusion: The Doxofylline-Soluplus Polymeric Nanomicelles could have the significant value in the treatment of Asthama and COPD. [ABSTRACT FROM AUTHOR]

The Ni/Y2O3 catalyst showed high catalytic activity. Based on this, the aim of this study was to create Ni/Y2O3 nanocomposites powder with two innovative technologies, Ultrasonic Spray Pyrolysis (USP) and lyophilisation. In the USP process, thermal decomposition of the generated aerosols in an N2/H2 reduction atmosphere caused a complete decomposition of the nickel (II) nitrate to elemental Ni, which became trapped on the formed Y2O3 nanoparticles. The Ni/Y2O3 nanocomposite particles were captured via gas washing in an aqueous solution of polyvinylpyrrolidone (PVP) in collection bottles. PVP was chosen for its ability to stabilise nano-suspensions and as an effective cryoprotectant. Consequently, there was no loss or agglomeration of Ni/Y2O3 nanocomposite material during the lyophilisation process. The Ni/Y2O3 nanocomposite powder was analysed using ICP-MS, SEM-EDX, and XPS, which showed the impact of different precursor concentrations on the final Ni/Y2O3 nanocomposite particle composition. In a final step, highly concentrated Ni/Y2O3 nanocomposite ink (Ni/Y2O3 > 0.140 g/mL) and test coatings from this ink were prepared by applying them on a white matte photo paper sheet. The reflection curve of the prepared Ni/Y2O3 nanocomposite coating showed a local maximum at 440 nm with a value of 39% reflection. Given that Ni is located on the surface of the Ni/Y2O3 nanocomposite in the elemental state and according to the identified properties, tests of the catalytic properties of this coating will be performed in the future. [ABSTRACT FROM AUTHOR]

The efficient extraction of phenols from grapes is an important step for their reliable quantification. The aim was to optimise the lyophilisation process and the extraction of phenols from grape skins and seeds. The phenol extraction yield from lyophilised tissues was investigated with different accelerated solvent extraction (ASE) operating conditions. Skins and seeds were separated from frozen berries and lyophilised without being ground. The weight loss during lyophilisation was followed daily. Phenols were extracted from lyophilised, cryo-ground seeds and skins with ASE at room temperature and 10.3 MPa using 80% aqueous acetone and 60% aqueous methanol. The effects of ASE operational parameters (the number of extraction cycles (ECs) and static time (ST) duration) were investigated. The yield of extracted phenols was evaluated spectrophotometrically by determining total phenolic index at 280 nm (TPI). The weight of skins and seeds significantly dropped after 24 h of lyophilisation and continued to decrease, although not significantly, up until the 9th day. The optimal lyophilisation time was estimated to be 3 days and 5 days for skins and seeds, respectively. The phenol extraction yield was significantly affected after changes of ASE conditions. Based on TPI, the optimal ASE conditions were as follows: (i) lyophilised seeds—eight ECs with 10 min ST using aqueous acetone and then four ECs with 20 min ST using aqueous methanol; (ii) lyophilised skins—eight ECs with 1 min ST using aqueous acetone and then one EC with 20 min ST using aqueous methanol.

The use of dried cheese in food production and food preparation sectors is becoming increasingly widespread. Dried cheese is preferred because of their longer shelf life at room temperatures, smaller packaging sizes, practical use, and their ease of homogenization in food products. Various technologies are currently being used for the drying of cheese and efforts are still being made to develop new methods. Freeze drying is a highly preferred technique for the production of dried cheese due to high quality of the final product. Instant removal of water and low process temperatures ensure an effective prevention of chemical and microbial deterioration. In this study, White, Kashar and Tulum cheeses were dried by freeze-drying method. Physicochemical analyses were carried out on the first day in order to determine the initial quality of cheese samples. The moisture and salt content were highest in White cheese, titratable acidity and fat content were highest in Tulum Cheese. Freze dried cheese samples were stored in room conditions for 6 months and changes in microbial populations such as mesophilic aerobic bacteria, yeast and moulds, lactic acid bacteria, Staphylococci spp. and coliform group bacteria were determined. The results showed that Tulum Cheese contained highest microbial counts in general. The number of bacteria was 7.71±0.10 log cfu/g in Tulum Cheese, 5.70±0.07 log cfu/g in White cheese. Total counts of yeasts and molds were 7.37±0.06 log cfu/g for Tulum cheese and 4.92±0.07 log cfu/g for Kashar Cheese. Statistical analysis was performedby using Minitab Statistical Package Program to evaluate the results of microbiological analysis on dried cheese samples.

The development of new multicomponent nanoparticles is gaining increasing importance due to their specific functional properties, i.e., synthesised new complex concentrated nanoparticles (CCNPs) in the form of powder using ultrasonic spray pyrolysis (USP) and lyophilisation from the initial cast Ag20Pd20Pt20Cu20Ni20 alloy, which was in the function of the material after its catalytic abilities had been exhausted. Hydrometallurgical treatment was used to dissolve the cast alloy, from which the USP precursor was prepared. As a consequence of the incomplete dissolution of the cast alloy and the formation of Pt and Ni complexes, it was found that the complete recycling of the alloy is not possible. A microstructural examination of the synthesised CCNPs showed that round and mostly spherical (not 100%) nanoparticles were formed, with an average diameter of 200 nm. Research has shown that CCNPs belong to the group with medium entropy characteristics. A mechanism for the formation of CCNPs is proposed, based on the thermochemical analysis of element reduction with the help of H2 and based on the mixing enthalpy of binary systems.

PurposeMetronidazol and vancomycin were long the two best options against Clostridioides (formerly Clostridium) difficile infections (CDI). Now, the cost of new drugs such as fidaxomicin directs us towards alternative treatment options, such as faecal microbiota transplant (FMT). Its effectiveness is similar to fidaxomicin. There are questions regarding its safety, but the biggest challenges are prejudice and inconvenience. Most protocols refer to FMT applied in the form of a solution. We investigated different modalities of FMT.MethodsInstead of using nasoenteric tubes or colonoscopy, we place frozen or lyophilised stool in non-coated, size “00”, hard gelatine capsules or enterosolvent, size “0” capsules.ResultsWe found that non-coated, size “00”, hard gelatine capsules are appropriate for conducting FMT. Capsules containing lyophilised supernatant with a low number of bacteria have been proven to be non-inferior to other FMT modalities. The primary cure rate in the supernatant group was 93.75%, and 66.67% in the sediment group. The overall cure rate was 82.14%. Depending on the protocol, 4–7 capsules are sufficient per patient. Capsules can be stored for up to one year at -20°C.ConclusionsFMT is a feasible alternative to antibiotic treatments in CDI. Our method makes the process flexible and less inconvenient to patients. Long storage time allows a consistent supply of capsules, while small volume and formulation make the procedure tolerable.

Selection of lactic acid bacteria (LAB) for probiotic effect and preservation of LAB by lyophilized have been studied. The two LAB strains belonging to Lactobacillus plantarum and Lactobacillus pentosus producing bacteriocins in inhibitory concentration for colibacilli were selected from a total of 17 tested strains. The selected strains were further tested for their ability to survive the preservation process by lyophilisation. Two methods of preserving LAB by lyophilisation were tested and the viability of cells in lyophilized products was monitored in order to obtain products used as starter cultures. The presence of inhibition zones in mixt cultures indicates the ability of LAB to inhibit the growth of E. coli bacteria due to the synthesis of bacteriocins. During three years period, the viability of lyophilized LAB was monitored. Obtained data indicate that starter cultures of LAB can be stored in the lyophilized state for at least three years in dark and cool place, and the inoculation rate can be increased after this period by 50% if the starter culture was preserved and stored by applying the described techniques. [ABSTRACT FROM AUTHOR]

Purpose: Metronidazol and vancomycin were long the two best options against Clostridioides (formerly Clostridium) difficile infections (CDI). Now, the cost of new drugs such as fidaxomicin directs us towards alternative treatment options, such as faecal microbiota transplant (FMT). Its effectiveness is similar to fidaxomicin. There are questions regarding its safety, but the biggest challenges are prejudice and inconvenience. Most protocols refer to FMT applied in the form of a solution. We investigated different modalities of FMT. Methods: Instead of using nasoenteric tubes or colonoscopy, we place frozen or lyophilised stool in non-coated, size "00", hard gelatine capsules or enterosolvent, size "0" capsules. Results: We found that non-coated, size "00", hard gelatine capsules are appropriate for conducting FMT. Capsules containing lyophilised supernatant with a low number of bacteria have been proven to be non-inferior to other FMT modalities. The primary cure rate in the supernatant group was 93.75%, and 66.67% in the sediment group. The overall cure rate was 82.14%. Depending on the protocol, 4–7 capsules are sufficient per patient. Capsules can be stored for up to one year at -20°C. Conclusions: FMT is a feasible alternative to antibiotic treatments in CDI. Our method makes the process flexible and less inconvenient to patients. Long storage time allows a consistent supply of capsules, while small volume and formulation make the procedure tolerable. [ABSTRACT FROM AUTHOR]

Objectives The main objective of the present study was to develop a nanoparticulate drug delivery system that can protect insulin against harsh conditions in the gastrointestinal (GI) tract. The effects of the following employed techniques, including lyophilisation, cross-linking and nanoencapsulation, on the physicochemical properties of the formulation were investigated. Methods We herein developed a nanocarrier via ionotropic gelation by using positively charged chitosan and negatively charged Dz13Scr. The lyophilised nanoparticles with optimal concentrations of tripolyphosphate (cross-linking agent) and ß-cyclodextrin (stabilising agent) were characterised by using physical and cellular assays. Key findings The addition of cryoprotectants (1% sucrose) in lyophilisation improved the stability of nanoparticles, enhanced the encapsulation efficiency, and ameliorated the pre-mature release of insulin at acidic pH. The developed lyophilised nanoparticles did not display any cytotoxic effects in C2C12 and HT-29 cells. Glucose consumption assays showed that the bioactivity of entrapped insulin was maintained post-incubation in the enzymatic medium. Conclusions Freeze-drying with appropriate cryoprotectant could conserve the physiochemical properties of the nanoparticles. The bioactivity of the entrapped insulin was maintained. The prepared nanoparticles could facilitate the permeation of insulin across the GI cell line. [ABSTRACT FROM AUTHOR]

One of the main distinguishing features of bacteria belonging to the Cellulomonas genus is their ability to secrete multiple polysaccharide degrading enzymes. However, their application in biomass deconstruction still constitutes a challenge. We addressed the optimisation of the xylanolytic activities in extracellular enzymatic extracts of Cellulomonas sp. B6 and Cellulomonas fimi B-402 for their subsequent application in lignocellulosic biomass hydrolysis by culture in several substrates. As demonstrated by secretomic profiling, wheat bran and waste paper resulted to be suitable inducers for the secretion of xylanases of Cellulomonas sp. B6 and C. fimi B-402, respectively. Both strains showed high xylanolytic activity in culture supernatant although Cellulomonas sp. B6 was the most efficient xylanolytic strain. Upscaling from flasks to fermentation in a bench scale bioreactor resulted in equivalent production of extracellular xylanolytic enzymatic extracts and freeze drying was a successful method for concentration and conservation of the extracellular enzymes, retaining 80% activity. Moreover, enzymatic cocktails composed of combined extra and intracellular extracts effectively hydrolysed the hemicellulose fraction of extruded barley straw into xylose and xylooligosaccharides. Key points: • Secreted xylanase activity of Cellulomonas sp. B6 and C. fimi was maximised. • Biomass-induced extracellular enzymes were identified by proteomic profiling. • Combinations of extra and intracellular extracts were used for barley straw hydrolysis. [ABSTRACT FROM AUTHOR]

Background: Faecal microbiota transplantation (FMT) is an effective treatment of recurrent Clostridioides difficile infection (rCDI) and is being applied experimentally in other diseases. Encapsulated administration may be equivalent in efficacy to delivery through other routes. Methods: A systematic review was undertaken of studies using encapsulated FMT up to 26 October 2020. Data on indication, clinical outcomes, safety, treatment protocol and capsule preparation were collected and reported. Pooled rates of clinical efficacy in rCDI were calculated using random-effects meta-analysis. The impact of single variables on clinical efficacy was evaluated using univariate meta-regression. Results: A total of 35 studies reporting the treatment of 960 patients with encapsulated FMT for eight different indications met the inclusion criteria. Most studies (n = 18, 51%) and patients (n = 755, 79%) were from studies on rCDI. Cure rates after single and multiple courses of treatments with encapsulated FMT in rCDI were 85% (95% CI: 82%-88%) and 93% (95% CI: 88%-96%) respectively. The treatment outcome was not significantly affected by dose, number of delivered capsules, anaerobic/aerobic processing, single/multi-donor treatment, lyophilisation, or any other single factor in the production or delivery of encapsulated FMT. Promising but non-comparable results from the treatment of ulcerative colitis and multidrug-resistant organisms were reported. Conclusions: Encapsulated FMT is an effective and safe treatment of rCDI, with cure rates comparable to FMT delivered through other routes. The treatment is effective despite variations in donor screening, preparation and treatment protocol. For other indications, the role of FMT capsules is still not sufficiently examined, although some studies show promising results. Plain Language Summary: Transfer of faecal material through capsules in the treatment of various diseases. Evidence for clinical efficacy The bacteria and other microorganisms of the gut is different in patient with various diseases in comparison with healthy subjects. Therefore, ways to change the microorganisms of the gut in a beneficial direction has been the subject of various research projects within recent years. Faecal microbiota transplantation often referred as FMT is a method of transferring microorganisms from healthy donors to patients with various diseases and is seen as one way to change the microbial community of the gut in a beneficial direction. Faecal microbiota transplantation can be performed in different ways such as through endoscopy, enemas or capsules. The transfer through capsules is preferred by the patients and has advantages since it can be administered long-term and can be delivered to the patients in their home. In this paper, we evaluated all accessible research reporting treatment with encapsulated faecal microbiota transplantation in the treatment of various diseases. We report the following major findings: -Treatment with capsules is safe when guidelines for screening donors and testing faecal material is followed. -The treatment is highly effective in the treatment of recurrent C. difficile infection, a disease with high mortality often caused by repeated antibiotic treatments. The treatment was effective in 596 of 723 patients following one course of capsule treatment. -Faecal microbiota transplantation delivered through capsules is as effective as treatment delivered through other routes in the treatment of C. difficile infection. -The treatment is effective in the treatment of C. difficile infection across studies and countries, despite great differences in the ways the capsules were prepared and delivered. -Increasing the amount of faecal material used in the production did not affect the efficacy of the treatment. -There are promising results in the treatment of other diseases such as liver disease, inflammatory bowel disease and the treatment of multi-drug resistant bacteria. [ABSTRACT FROM AUTHOR]

The cleaning effectiveness of agar gels for copper stain removal from marble is mainly related to the gel morphology and to the possibility of copper coordination. • Different agar gel formulations were tested for copper stain removal from marble. • Hydrogel lyophilisation allowed to systematically compare gel formulations. • The relationship between gel morphology and effectiveness was assessed. • AgarArt 1% with TAC was the most effective gel for copper removal from marble. • EDTA in gels leads to a broader pore size distribution and to a lower gel strength. The effectiveness of Agar gels for copper stain removal from marble surfaces was systematically studied. Gels with different agar concentrations (1, 3, 5%) and different chelating agents used as additives (ethylenediaminetetraacetic acid, EDTA, and ammonium citrate tribasic, TAC) were tested on laboratory marble specimens for different contact times (30 and 60 min). For better characterization, hydrogels were lyophilised and cryogels were obtained. Systematic comparison of different formulations was feasible on cryogels and performed in terms of: (i) the morphological properties, by field-emission scanning electron microscopy (FE-SEM); (ii) the type of Cu(II)-complexes formed and their quantitative comparison by electron paramagnetic resonance (EPR) spectroscopy; (iii) the total amount of copper removed from marble surfaces, by Inductively Coupled Plasma–Optical Emission Spectrometry (ICP-OES). AgarArt 1% with TAC exhibited the highest effectiveness for copper stain removal after 60 min contact (431 μg/cm2). Such a good cleaning effectiveness can be ascribed to the co-presence of the following properties: efficient metal coordination, which is related to the additive presence, and favourable gel morphology, related both to the gel concentration and to the additive type. In fact, it was observed that both the low gel concentration and the presence of TAC are related to a narrow pore size distribution in gels, besides the possibility of copper coordination. The presence of EDTA results in a broader pore size distribution and in a lower gel strength, with respect to gels with TAC. Thus, a new procedure for studying gels was proposed, which allows to optimize the conditions for metal stain removal from built heritage. [ABSTRACT FROM AUTHOR]

Amino acids are not only used as buffering agents in lyophilisation, but also exhibit cryo- and lyoprotecting characteristics. L-Arginine based lyophilisates were tested regarding their ability to stabilise a monoclonal antibody (mAb) at different residual moisture (RM) levels. Arginine base was formulated with citric, hydrochloric, lactobionic, phosphoric, and succinic acid for pH adjustment. Lyophilisates with less than 0.5% and approx. 2.5% RM were stored for up to 6 months at 40 °C. The mAb aggregation in arginine in combination with hydrochloric acid and succinic acid was similar or even less compared to a sucrose reference formulation. Arginine in combination with citric acid, lactobionic acid, and phosphoric acid resulted in lower protein stability. Overall, arginine formulations with high RM levels resulted in better protein stabilisation despite decreased glass transition temperatures (T g). Whereas we detected mAb glycation in the sucrose based formulations, this chemical reaction did not occur in arginine based formulations. Arginine hydrochloride and succinate, especially at high RM levels, could be promising alternatives to sucrose for stabilisation of mAb in lyophilisates. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) enables highly specific silencing of potential target genes for treatment of pulmonary pathologies. The intracellular RNAi pathway can be activated by cytosolic delivery of small interfering RNA (siRNA), inducing sequence-specific gene knockdown on the post-transcriptional level. Although siRNA drugs hold many advantages over currently applied therapies, their clinical translation is hampered by inefficient delivery across cellular membranes. We previously developed hybrid nanoparticles consisting of an siRNA-loaded nanosized hydrogel core (nanogel) coated with Curosurf®, a clinically used pulmonary surfactant (PS). The latter enhances both particle stability as well as intracellular siRNA delivery, which was shown to be governed by the PS-associated surfactant protein B (SP-B). Despite having a proven in vitro and in vivo siRNA delivery potential when prepared ex novo , clinical translation of this liquid nanoparticle suspension requires the identification of a long-term preservation strategy that maintains nanoparticle stability and potency. In addition, to achieve optimal pulmonary deposition of the nanocomposite, its compatibility with state-of-the-art pulmonary administration techniques should be evaluated. Here, we demonstrate that PS-coated nanogels can be lyophilized, reconstituted and subsequently nebulized via a vibrating mesh nebulizer. The particles retain their physicochemical integrity and their ability to deliver siRNA in a human lung epithelial cell line. The latter result suggests that the functional integrity of SP-B in the PS coat towards siRNA delivery might be preserved as well. Of note, successful lyophilization was achieved without the need for stabilizing lyo- or cryoprotectants. Our results demonstrate that PS-coated siRNA-loaded nanogels can be lyophilized, which offers the prospect of long-term storage. In addition, the formulation was demonstrated to be suitable for local administration with a state-of-the-art nebulizer for human use upon reconstitution. Hence, the data presented in this study represent an important step towards clinical application of such nanocomposites for treatment of pulmonary disease. [ABSTRACT FROM AUTHOR]

The treatment of wastewaters is crucial to maintain the ecological status of receiving waters, and thereby guarantee the protection of aquatic life and human health. Wastewater quality evaluation is conventionally based on physicochemical parameters, but increasing attention has been paid to integrate physicochemical and biological data. Nevertheless, the regulatory use of fish in biological testing methods has been subject to various ethical and cost concerns, and in vitro cell-based assays have thus become an important topic of interest. Hence, the present study intends: (a) to evaluate the efficiency of two different sample pre-concentration techniques (lyophilisation and solid phase extraction) to assess the toxicity of municipal effluents on rat cardiomyoblast H9c2(2-1) cells, and (b) maximizing the use of the effluent sample collected, to estimate the environmental condition of the receiving environment. The gathered results demonstrate that the H9c2(2-1) sulforhodamine B-based assay is an appropriate in vitro method to assess biological effluent toxicity, and the best results were attained by lyophilising the sample as pre-treatment. Due to its response, the H9c2(2-1) cell line might be a possible alternative in vitro model for fish lethal testing to assess the toxicity of municipal effluents. The physicochemical status of the sample suggests a high potential for eutrophication, and iron exceeded the permissible level for wastewater discharge, possibly due to the addition of ferric chloride for wastewater treatment. In general, the levels of carbamazepine and sulfamethoxazole are higher than those reported for other countries, and both surpassed the aquatic protective values for long-term exposure. Image, graphical abstract [ABSTRACT FROM AUTHOR]