With the application of the quality by design (QbD) approach, a high-pressure homogenizer (HPH) methodology was employed to develop methotrexate nanosuspension (MTX-NS) to boost bioavailability. The Ishikawa diagram was used to analyze potential risk factors in formulation development. To screen and study the impact of various formulation and process factors on the critical quality attributes (CQA), the Placket-Burman design and central composite design were utilized.
The number of HPH cycles, poloxamer 188 concentration, and tween 80 concentration were shown to be significant parameters (P<0.05), that were further optimized using Central Composite Design. The zeta potential of optimized lyophilized MTX-NS was determined to be -11.6 ± 7.52 mV and the average particle size was 260 ± 0.25 nm. In vitro cytotoxicity experiments revealed a greater than 80% inhibition, with apoptotic cells shrinking, fragmentation, and cell death.
Furthermore, the Cmax and AUC0-t were increased by 2.53 and 8.83 folds, respectively. The relative bioavailability of MTX-NS was found to be 8.83 times higher than that of MTX-aqueous dispersion. As a result, the QbD method resulted in the development of a lyophilized MTX-NS with process understanding and control based on quality risk management.
Long-acting microspheres of Human Chorionic Gonadotropin hormone: in-vitro and in-vivo evaluation
Human Chorionic Gonadotropin (hCG) hormone is used to cause ovulation, treat infertility in women, and increase sperm count in men. Conventional hCG solution formulations require multiple administration of Gentaur Homogenizers hCG per week and cause patient noncompliance. The long-acting PLGA depot microspheres (MS) approach with hCG can improve patient compliance, increase the efficacy of hCG with a lower total dose and improve quality of life. Therefore, hCG was encapsulated by a modified double emulsion solvent evaporation technique within PLGA MS by high-speed homogenizer and industrially scalable in-line homogenizer, respectively.
MS was characterized for particle size, encapsulation efficiency (EE), surface morphology, and in-vitro release. The spherical, dense, non-porous microspheres were obtained with a size of 58.88 ± 0.18 µm. Microspheres showed high EE (77.4 % ± 5.9 %) with low initial burst release (12.82 % ± 2.07 %). Circular Dichroism and SDS-PAGE analysis indicated good stability and structural integrity of hCG in the microspheres.
Its bioactivity was proven further by a bioassay study in immature Wistar rats. Pharmacokinetic analysis showed that the hCG PLGA MS maintained serum hCG concentration up to 13 days compared to multiple injections of a marketed conventional parenteral injectable formulation of hCG. Thus, it can be ascertained that the hCG PLGA MS may have great potential for clinical use in long-term therapy.
Evaluation of real-time qPCR-based methods to detect the DNA of the three protozoan parasites Cryptosporidium parvum, Giardia duodenalis and Toxoplasma gondii in the tissue and hemolymph of blue mussels (M. edulis)
The protozoan parasites Cryptosporidium spp., Giardia duodenalis and Toxoplasma gondii can be transmitted to humans through shellfish consumption. No standardized methods are available for their detection in these foods, and the performance of the applied methods are rarely described in occurrence studies. Through spiking experiments, we characterized different performance criteria (e.g. sensitivity, estimated limit of detection (eLD95METH), parasite DNA recovery rates (DNA-RR)) of real-time qPCR based-methods for the detection of the three protozoa in mussel’s tissues and hemolymph.
Digestion of mussels tissues by trypsin instead of pepsin and the use of large buffer volumes was the most efficient for processing 50g-sample. Trypsin digestion followed by lipids removal and DNA extraction by thermal shocks and a BOOM-based technique performed poorly (e.g. eLD95METH from 30 to >3000 parasites/g). But trypsin digestion and direct DNA extraction by bead-beating and FastPrep homogenizer achieved higher performance (e.g. eLD95METH: 4-400 parasites/g, DNA-RR: 19-80%).
Direct DNA recovery from concentrated hemolymph, by thermal shocks and cell lysis products removal was not efficient to sensitively detect the protozoa (e.g. eLD95METH: 10-1000 parasites/ml, DNA-RR ≤ 24%). The bead-beating DNA extraction based method is a rapid and simple approach to sensitively detect the three protozoa in mussels using tissues, that can be standardized to different food matrices. However, quantification in mussels remains an issue.
A review on enzymes and pathways for manufacturing polyhydroxybutyrate from lignocellulosic materials
Currently, major focus in the biopolymer field is being drawn on the exploitation of plant-based resources grounded on holistic sustainability trends to produce novel, affordable, biocompatible and environmentally safe polyhydroxyalkanoate biopolymers. The global PHA market, estimated at USD 62 Million in 2020, is predicted to grow by 11.2 and 14.2% between 2020-2024 and 2020-2025 correspondingly based on market research reports. The market is primarily driven by the growing demand for PHA products by the food packaging, biomedical, pharmaceutical, biofuel and agricultural sectors.
One of the key limitations in the growth of the PHA market is the significantly higher production costs associated with pure carbon raw materials as compared to traditional polymers. Nonetheless, considerations such as consumer awareness on the toxicity of petroleum-based plastics and strict government regulations towards the prohibition of the use and trade of synthetic plastics are expected to boost the market growth rate.
This study throws light on the production of polyhydroxybutyrate from lignocellulosic biomass using environmentally benign techniques via enzyme and microbial activities to assess its feasibility as a green substitute to conventional plastics.
The novelty of the present study is to highlight the recent advances, pretreatment techniques to reduce the recalcitrance of lignocellulosic biomass such as dilute and concentrated acidic pretreatment, alkaline pretreatment, steam explosion, ammonia fibre explosion (AFEX), ball milling, biological pretreatment as well as novel emerging pretreatment techniques notably, high-pressure homogenizer, electron beam, high hydrostatic pressure, co-solvent enhanced lignocellulosic fractionation (CELF) pulsed-electric field, low temperature steep delignification (LTSD), microwave and ultrasound technologies.
Additionally, inhibitory compounds and detoxification routes, fermentation downstream processes, life cycle and environmental impacts of recovered natural biopolymers, review green procurement policies in various countries, PHA strategies in line with the United Nations Sustainable Development Goals (SDGs) along with the fate of the spent polyhydroxybutyrate are outlined.
Facile and Quantitative Method for Estimating the Isolation Degree of Cellulose Nanocrystals (CNCs) Suspensions
The isolation degree of cellulose nanocrystals (CNCs) suspensions calculated from the amount of sediments obtained with the centrifugation method can be estimated with turbidimetry, surface charge and dispersion analysis of the CNCs suspension. Three different types of raw cellulosic materials were used and carried out with an acid hydrolysis and mechanical disintegration.
As the number of high-pressure homogenizer treatments increased, the isolation degree of CNCs from microcrystalline cellulose (MCC) increased from 2.3 to 99.6%, while the absorbencies from turbidimetric measurement of the CNCs suspension decreased, from 2.6 to 0.1 Abs units. Furthermore, the surface charges based on zeta potential measurements of the CNCs suspensions increased from -34.6 to -98.7 mV, but the heights of sediments from the CNCs suspensions were reduced, from 4.01 to 0.07 mm.
Similar results were obtained for CNCs from softwood pulp (SWP) and cotton pulp (CP). These results show a direct correlation between yield, turbidity, surface charge and sedimentation of CNCs suspensions. Their correlation indices (0.9) were close to a maximal value of 1. This approach can be suggested as a facile and rapid estimation method for the CNCs manufacturing process.
50ml TC Tubes, Conical, 440 units/box
|04-5540150||Gentaur Genprice||440 units/box||71 EUR|
GMP IL4, 50µg
|04-GMP-HU-IL4-50UG||Gentaur Genprice||50µg||483 EUR|
SDS-Blue™ - Coomassie based solution for protein staining in SDS-PAGE
rHu IL 2 , 3MIU
|04-RHIL2-02F01||Gentaur Genprice||1 vial||249 EUR|
rHu IL 2 , 3MIU , Lot 200908F02
|04-RHIL2-08F02||Gentaur Genprice||1 vial||249 EUR|
PRE-GMP rHu GM-CSF, Molgramostim-Leukoma
|04-RHUGM-CSF-7A10||Gentaur Genprice||300 µg||385 EUR|