Ales Franc

The Effect of Mycotoxin Deoxynivalenol on Haematological and Biochemical Indicators and Histopathological Changes in Rainbow Trout (Oncorhynchus mykiss)

Deoxynivalenol (DON), produced by the Fusarium genus, is a major contaminant of cereal grains used in the production of fish feed. The effect of mycotoxin deoxynivalenol on rainbow trout (Oncorhynchus mykiss) was studied using a commercial feed with the addition of DON in a dose of 2 mg/kg feed. The fish (n = 40) were exposed to the mycotoxin for 23 days. The trout were divided into two groups, control and experimental groups. Control groups were fed a commercial feed naturally contaminated with a low concentration of DON (225 μg/kg feed); experimental groups were fed a commercial feed with the addition of DON (1964 μg/kg feed). Plasma biochemical and haematological indices, biometric parameters, and histopathological changes were assessed at the end of the experiment. The experimental groups showed significantly lower values in MCH (P < 0.05). In biochemical indices, after 23-day exposure, a significant decrease in glucose, cholesterol (P < 0.05), and ammonia (P < 0.01) was recorded in the experimental group compared to the control group. Our assessment showed no significant changes in biometric parameters. The histopathological examination revealed disorders in the caudal kidney of the exposed fish. The obtained data show the sensitivity of rainbow trout (O. mykiss) to deoxynivalenol.

Influence of Process Parameters on Content Uniformity of a Low Dose Active Pharmaceutical Ingredient in a Tablet Formulation According to GMP

Abstract The article describes the development and production of tablets using direct compression of powder mixtures. The aim was to describe the impact of filler particle size and the time of lubricant addition during mixing on content uniformity according to the Good Manufacturing Practice (GMP) process validation requirements. Processes are regulated by complex directives, forcing the producers to validate, using sophisticated methods, the content uniformity of intermediates as well as final products. Cutting down of production time and material, shortening of analyses, and fast and reliable statistic evaluation of results can reduce the final price without affecting product quality. The manufacturing process of directly compressed tablets containing the low dose active pharmaceutical ingredient (API) warfarin, with content uniformity passing validation criteria, is used as a model example. Statistic methods have proved that the manufacturing process is reproducible. Methods suitable for elucidation of various properties of the final blend, e.g., measurement of electrostatic charge by Faraday pail and evaluation of mutual influences of researched variables by partial least square (PLS) regression, were used. Using these methods, it was proved that the filler with higher particle size increased the content uniformity of both blends and the ensuing tablets. Addition of the lubricant, magnesium stearate, during the blending process improved the content uniformity of blends containing the filler with larger particles. This seems to be caused by reduced sampling error due to the suppression of electrostatic charge.

Preparation of pellets with controlled release of glucose as prevention of hypoglycaemia in paediatric patients.

Hypoglycaemic episodes represent serious and frequent complications of type 1 and 2 diabetes. Theoretically, the risk of hypoglycaemic states can be affected by a dosage form based on a food supplement containing a delayed release formulation of glucose. The release of glucose should compensate for balance the peak effect of an antidiabetic treatment. In clinical practice, a diet with fibre and grains is recommended and patients are broadly educated in the topic of low and high glycaemic indexes to achieve the same effect. However, a precisely-timed release of carbohydrates can favourably target expected hypoglycaemia and concurrently decrease carbohydrate content. To study the possibility of preparing the dosage form with controlled-release carbohydrates, a dosage form of pellets containing four osmotically active substances coated by a membrane created of ethylcellulose was prepared. These pellets can be administered in a mixture with liquid or semisolid food. The resulting dissolution profiles for selected compositions showed that delayed release can be achieved for 120, 240 and 360min in vitro, representing an ideal delay for clinical purposes.

Quality by design approach: antioxidant activity of the tablets containing cornelian cherry fruits in relation to their composition and physical properties

Abstract The aim of this study was to prepare tablets containing ground fruits of cornelian cherry (Cornus mas L.) with high antioxidant capacity. The experiment was planned and evaluated on Design of Experiment (DoE) principle using Multivariate Data Analysis (MVA) as modern tools used in Quality by Design (QbD) approach. Various tableting mixtures with three different particle sizes of the plant material (up to 800 μm, more than 800 μm and their mixture) and percentage of silicon dioxide (1, 3 and 5%) were prepared. Tablets with a diameter of 10 mm and mass of 400 mg were subsequently produced from these mixtures using two compression forces (C1=7 kN and C2=14 kN). Principal Component Analysis (PCA) and Multiple Linear Regression (MLR) with response surface methodology were used to find the influential process-formulation parameters and describe their optimal settings. Finally, it is possible to say that the increasing level of silicon dioxide and the decreasing particle size of ground cornelian cherry lead to prolongation of disintegration time and increase of radial hardness and abrasion loss. Maximal antioxidant activity was obtained using 5% amount of silicon dioxide, the largest particle size and the low compression force.

Effect of T-2 toxin-contaminated diet on common carp (Cyprinus carpio L.).

ABSTRACT The T‐2 toxin, a fungal metabolite produced by Fusarium molds, occurs in a range of agriculture products. Reduced availability of fish meal has led to increasing use of cereals as a source of protein in commercial aquaculture feeds, which has increased the potential for mycotoxin contamination. The purpose of this study was to investigate toxicity of T‐2 toxin intake in common carp (Cyprinus carpio L.) using haematological, biochemical and immunological parameters and oxidative stress indices. In a four‐week feeding trial, fish were fed a commercial diet with 5.3 mg/kg T‐2 toxin added. Ingestion of contaminated diet did not lead to mortality of fish, probably due to lower feed intake. On the other hand, it significantly affected haematological variables such as haematocrit, haemoglobin, red blood cell counts leading to anemia and white blood cell counts leading to leukopenia due to lymphopenia. Plasma glucose concentration and alanine amino transferase activity showed a significant increase while triglycerides concentration decreased. Activity of ceruloplasmin was significantly decreased in plasma. Further, liver glutathione S‐transferase activity was significantly increased and catalase activity decreased, in parallel with a significant increase in caudal kidney catalase activity and a decrease in glutathione peroxidase activity. Finally, lipid peroxidation (detected as malondialdehyde) was significantly increased in the liver and caudal kidney. Changes in non‐specific immune response and cytokine levels in head kidney indicated immune system sensitivity to T‐2 toxin. Overall, the results demonstrate that this feed‐borne mycotoxin is able to induce anaemia and oxidative stress and cause changes in the immune response of common carp. HIGHLIGHTSThe aim of the study was to investigate toxicity of T‐2 toxin intake in common carp.Feeding trial led to changes in haematological and biochemical parameter.Lipid peroxidation was significantly increased in the liver and caudal kidney.Exposition to T2 led to changes in non‐specific immune response.Exposition to T2 led to changes in mRNA expression for selected cytokines.

Coated pellets with delayed-release glucose for prevention of hypoglycemic episodes.

Abstract Patients tend to prevent hypoglycemia by excessive saccharide intake leading to poorer glycemic control with potentially fatal consequences. This problem could be resolved by means of pellets with glucose release delayed by 120–360 min as a compensation of the antidiabetic drug peak effect. No glucose is released before; hence there is no risk of hyperglycemia and secondary complications. The pellets contain glucose in combination with an osmotically active ingredient and are coated with an ethylcellulose dispersion, which forms an insoluble semipermeable membrane and ensures delayed release. The release of glucose was assessed using dissolution and high-performance liquid chromatography. Dissolution profiles indicated the possibility of achieving the requested lag time using a combination of adequate compositions and coating concentrations. Lag times of 60, 240 and 360 min were achieved. The sample containing carboxymethyl starch was found to be most suitable for the intent of this work.

Interdiction of hypoglycemia in diabetic children by multiparticulate dosage form with controlled glucose release.

Abstract Patients tend to evade the occurrence of hypoglycemic episodes by excessive carbohydrate intake. Glucose pellets with delayed release in the time of the maximum effect of insulin can not only prevent hypoglycemia but also eliminate the preventive carbohydrate intake. The pellets can be administered in a mixture with semisolid food. The cores containing glucose in combination with osmotically active agents (croscarmellose sodium, carmellose sodium, polyethylene glycol, or carboxymethyl starch) were prepared by extrusion–spheronization and coated with 15% water ethylcellulose dispersion (Surelease® B NF) in Wurster column (Medipo, Havlíčkův Brod, Czech Republic) into four coating levels (12.5, 25, 35, and 50%). Mean particle size is 0.63–0.73 for cores and 0.82–0.98 for coated pellets. Cores and coated pellets have excellent or good flow properties according to Hausner ratio and Carr index. Aspect ratio ranges from 1.78 to 2.17 for cores and from 1.73 to 2.31 for coated pellets. Dissolution was performed using pH-independent method and method with continual change of pH. The suitable pH-independent release was achieved in the samples containing carboxymethyl starch or polyethylene glycol. Glucose release is enabled by a membrane rupture caused by core swelling. It can be, therefore, assumed that the glucose release profile will not be affected by food or transit time.

Influence of concentration and type of microcrystalline cellulose on the physical properties of tablets containing Cornelian cherry fruits

Abstract The aim of this study was to find the optimal tablet composition with maximum content of dried fruits (Cornus mas L.). The effect of three different concentrations (12.5, 25 and 50 %) of two types of microcrystalline cellulose (Avicel® PH 101 and Avicel® PH 200) and three different compression pressures (20, 60 and 100 MPa) on the physical properties of tablet blends and tablets was studied. Tablets containing 50 % Avicel® PH 101 compressed under 100 MPa were found to have the best physical properties. This combination of composition and compression pressure resulted in stable tablets even after storage under accelerated stability conditions (6 months, 40 °C and 75 % RH).

Biphasic dissolution method for quality control and assurance of drugs containing active substances in the form of weak acid salts

Abstract Substances in the form of weak acid salts have been found to be problematic for dissolution testing. Their absorption can start only after they are turned into the form of an acid following the gastric passage although they were administered in the form of a salt. Due to poor solubility, they cannot be tested in acidic gastric environment for a biased dissolution profile. The biphasic dissolution method is promising for overcoming this obstacle. Tablets with warfarin clathrate sodium salt in two concentrations and two different particle size distributions were tested as a suitable model for finding the medium and process conditions of dissolution. The dissolution method based on the use of the upper organic layer (1-octanol) and the lower aqueous layer 0.1 mol L−1 HCl) was found suitable and discriminatory for tablets containing active substances in the form of salts of weak acids. The method also reflects physical differences in the quality of used substances.

Pellet patented technology for fast and distinct visual detection of cholinesterase inhibitors in liquids

Graphical abstract Figure. No Caption available. HighlightsDouble‐coated pellets for detection of cholinesterase inhibitors were prepared.Lower amount of enzyme for a formulation of the detection system was needed.A more distinctive color transition during the detection in liquids was achieved. ABSTRACT The main objective of the presented research was to prepare an innovative carrier as a filler for detection tubes in the form of double‐coated pellets with a very significant color transition during the detection of cholinesterase inhibitors such as nerve agents, organophosphorus or carbamate insecticides in liquids that is observable visually and also spectrophotometrically at 412 nm. The pellet cores were prepared by the extrusion/spheronization method. Consecutively, two different coats were applied on the pellet cores in the coating device using the Wurster column method. To increase the color change intensity, the second semipermeable coat based on Eudragit® RL was applied on top of the first coat, which was formed by butyrylcholinesterase immobilized in hydroxypropyl methylcellulose. Prepared samples differing in thickness of the second coat were evaluated for their quality parameters, enzymatic activity and inhibition. The detection mechanism was based on the standard Ellman’s colorimetric reaction. It was observed that the semipermeable coat prevented leaching of the enzyme into the solution and led to an increased intensity of color transition from white – yellow to white – deep yellow/orange, thus enabling a more accurate visual detection. This system allows easy, rapid and safe identification of cholinesterase inhibitors in liquids, especially chemical warfare agents.