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The aim of the study was to develop direct-compress-tablets of microencapsulated probiotic Lactobacillus casei 01 in soy protein-alginate microparticles and excipients able to provide probiotic delivery near the colon. Considering their physicochemical properties, all series of tablets prepared met the requirements of the Ph Eur 9.0. The compaction of the probiotic loaded microparticles caused viability decrease up to 1.5 log cycles. The tablets containing Methocel K100M showed higher potential for preserving probiotic viability in simulated gastrointestinal fluids within 4 h and retarding its release in the intestinal stage, maintaining the required minimum of viable probiotic cells above 107 CFU per gram of tablet. In addition, acceptable storage stability (viability of probiotic above 106 CFU/g) at 25?C/60% relative humidity for 42 days was observed. In conclusion, novel tablet dosage forms of microencapsulated L. casei 01 were prepared with high potential for preserving probiotic viability in simulated gastrointestinal fluids and retarding its release in the lower intestine. Further research is needed to optimize the formulation and process parameters in order to obtain tablets with probiotic viability over long storage periods.

This study aimed to develop a probiotic/synbiotic ayran with a sufficient number of live bacteria Lactobacillus casei 01 (L. casei 01) during the product shelf-life. Under conditions of refrigerated storage for 15 days, probiotic survival and organic acid (lactic, acetic, propionic and butyric acid) production were evaluated. Simultaneously, pH, titratable acidity, total solids, and carbohydrate, protein, fat and mineral contents of the samples were examined. Overall results of the chemical analyses indicated the quality of the prepared samples to be comparable with the conventional ayran used as a matrix for probiotic/synbiotic supplementation. L. casei 01 survived better in ayran containing synbiotic chitosan–Ca–alginate microparticles compared to samples enriched with non-encapsulated probiotic alone or with synbiotic (L. casei 01 + oligofructose-enriched inulin), followed by increased production of lactic, acetic and propionic acid. Probiotic/synbiotic ayran may contribute to a healthy diet for consumers, while the sample enriched with microencapsulated formulation is advantageous in terms of probiotic activity.

The aim of this study was to develop chitosan (CS) microparticulated mucoadhesive drug delivery system (DDS) with improved therapeutic performance and biological responce. Ionotropic gelation/spray drying process was used for preparation of doxycycline hyclate (DOXY) loaded low and medium molecular weight (LMw and MMw) CS/sodium tripolyphosphate microparticles (CS/TPP MPs), further coated with ethyl cellulose (EC) using coacervation/solvent displacement technique. The relevant physico-chemical and biopharmaceutical properties were optimized using experimental design approach. Both coated and uncoated CS/TPP MPs showed high mucoadhesive potential and did not affect the viability of the tested epithelial cell line. The MPs induced slow and gradual apoptotic response in murine macrophage cell line RAW 264.7 and the observed effect depended upon formulation type and MP concentration. Biological effect of the CS-based MPs observed in our experiments point to synergism of the biological response of the carrier with the anti-inflammatory effect of DOXY.

Herein synchronous occurrence of Hodgkin lymphoma and secondary myelodysplastic syndrome in a 60 year old male patient with small cell lung cancer treated with combined chemotherapy (carboplatin and paclitaxel) and radiotherapy is presented. The objective of this report is to stress the importance of documenting and monitoring adverse drug reactions that arise from chemotherapy. After four years of treatment with the combined chemotherapy, the patient presented inguinal lymphadenopathy and enlarged lymph nodes and histopathology rapport was suggestive for plasmacytoid variant of Castleman disease. Three years later, biopsy of lymph node was performed and diagnosis of Hodgkin lymphoma – mixed cellularity has been established. Molecular analyses revealed presence of dominant monoclonal population of the immunoglobulin genes in the oligo/monoclonal background. Bone marrow biopsy findings suggested secondary myelodysplasia and revealed signs of hematopoietic cells dismaturation with signs of megaloblastic maturation of the erytropoetic lineage, appearance of ALIP (abnormal localization of immature precursors) in the myeloid lineage and dysplastic megakaryocytes. In addition, an increased level of polyclonal plasmacytes (lambda vs kappa was 60%:40%) was found. Hodgkin lymphoma and MDS occurring after 4 years of carboplatin/paclitaxel therapy might be contributed to the accumulation of alkylator-related DNA damage. This emphasize the need of outlining a monitoring plan regarding development of secondary leukemia and other malignant hematological proliferations should be outlined in the protocols.

New therapeutic strategies against inflammatory bowel disease (IBD) consider the usage of probiotics, prebiotics and synbiotics as beneficial for the intestinal microbial balance. Limitations of such an approach are addressed into difference in survival, persistence, colonization and variable effects among different probiotic strains, lack in understanding of probiotic mechanisms of action, as well the complex etiology of IBD. The anti-inflammatory activity of Lactobacillus casei 01 (L. casei 01) was assessed in trinitrobenzenesulphonic (TNBS) acid model of rat colitis when the probiotic was used alone and/or in combination with oligofructose-enriched inulin (Synergy 1), and as synbiotic (L. casei 01+Synergy 1) loaded chitosan-Ca-alginate microparticles; all suspended in ayran. The results from the probiotic/synbiotic treatments (8.5-8.9log CFU g-1L. casei 01 and 1.5% Synergy 1) have shown reduction in the colonic damage and increased lactobacilli counts in feces. Lactobacilli translocation to sterile extra-intestinal organs demonstrated acceptable safety of the probiotic strain used. The best effect at reducing inflammation and lesions associated with a significant decline in myeloperoxidase (MPO) activity was observed in rats that received synbiotic microparticles. This finding suggests colon targeted delivery of the probiotics/synbiotics, as an advantageous approach in prevention and treatment of IBD.

Ciprofloxacin is a widely used fluoroquinolone antibiotic. In this work, a comprehensive evaluation of MP2 and DFT with different functionals and basis sets was carried out to select the most suitable level of theory for the study of the NMR properties of ciprofloxacin. Their relative predictive capabilities were evaluated comparing the theoretically predicted and experimental spectral data. Our computational results indicated that in contrast to the solid state, the molecule of ciprofloxacin does not exist as a zwitterion in gaseous state. The results of the calculations of the chemical shifts most close to the experimental were obtained with B3LYP/aug-cc-pVDZ. The F-C coupling constants were calculated systematically with different DFT methods and several basis sets. In general, the calculations of the coupling constants with the BHandH computational method including the applied in this work 6-311++G**, EPRII, and EPRIII basis sets showed a good reproducibility of the experimental values of the coupling constants.

The study highlights the current progress in the development of coumarin scaffolds for drug discovery as novel anticancer agents in metastatic breast cancer. Eight compounds, combining the coumarin core and five membered heterocycles (isoxazoles and thiazoles) in hydrazinyldiene- -chroman-2,4-diones, were characterized in terms of a potential antiproliferative effect on bone (SCP1833) and lung (SCP4175) metastatic breast cancer cell lines using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Cell viability was evaluated after 48 and 72 h of treatment and the 50 % inhibitory concentrations were determined. The results demonstrated dose- and time-dependent activity, with the most potent molecules having a thiazole moiety, without or with additional methyl group(s) attached to the carbon(s) at position(s) 5 and/or 4 in the thiazole ring. These molecules possessed significantly higher potency against both test cell lines compared to 4-hydroxycoumarin

Lactobacillus casei 01 was co-encapsulated with the prebiotic oligofructose-enriched inulin at different concentrations to investigate the efficiency of the prebiotic for improving the probiotic viability. Prebiotic effect on the probiotic survival under microencapsulation conditions by spray- and freeze-drying and storage stability of encapsulated living cells at 4 °C during period of 8 weeks was evaluated. Adhesiveness of L. casei 01 loaded microparticles to pig mucin was investigated in vitro to estimate the role of microencapsulation for improving the cell adhesion ability. The microparticles produced with 3% w/w oligofructose-enriched inulin showed higher initial count, while oligofructose-enriched inulin applied at 1.5% w/w resulted in better protection of L. casei 01 under storage conditions. Further, it has been observed significantly increased pig mucin binding to microparticles compared to free probiotic cells in buffer solutions simulating GI conditions, during 24 h incubation. Hence, cell microencapsulation beside enhanced viability may allow prolonged residence time of the probiotic cells in the lower intestine through excellent muco-adhesive properties of the encapsulating materials. The results suggest synbiotic chitosan-Ca-alginate microparticles as convenient delivery system capable to ensure effective cell concentration in the lower intestine where probiotic colonization is dominant.

The specific aims of this study were to select the favorable prebiotic for L casei-01 as well as the suitable inoculum of the probiotic for fermented soy with ACE-inhibitory potential. For that purpose the metabolic activity of L casei-01 in soymilk supplemented with (1.5% w/v) prebiotics Fructooligosaccharide (FOS) or oligofructose enriched inulin (Synergy 1) was assessed. The evaluated parameters were: pH, viable cell counts, proteolysis, organic acid production and inhibition of angiotensine converting enzyme activity (IACE). The cell growth of L casei-01 reached the recommended therapeutic level of 9.58 ± 0.035 log cfu mL-1 for low inoculum samples (0.005 and 0.01%w/v) and 11.543 ± 0.13 log cfu mL-1 for high inoculum samples (0.075 and 0.1%w/v) regardless of the prebiotic used. The lower pH during fermentation, faster cell growth and superior proteolysis in Synergy 1 samples indicated better utilization of that prebiotic vs. FOS. The hydrolysis depended on the prebiotic used, showing higher values in Synergy 1 samples. The faster proteolysis was confirmed by SDSPAG electrophoresis. The Mw of polypeptides in the synbiotic end-products were lower than 30kD. The observed values for inhibition of ACE activity were app. 71, 74, 77 and 78% for inoculum rates of 0.005, 0.01, 0.075 and 0.1% w/v, respectively. Based on the results obtained in our study, the prebiotic Synergy 1 (1.5% w/v) and L. casei-01 at inoculum of 0.01% w/v for low dose and 0.075% w/v for high dose were considered more favorable for the production of synbiotic soy drink with antihypertensive potential.

Lactobacillus casei loaded chitosan-Ca-alginate microparticles enriched with the prebiotic fructooligosaccharide were prepared using spray-drying method associated with the polymers complexation and cross-linking with calcium. The concentrations of the formulation factors of alginate, chitosan and CaCl2 were optimized using 23 full factorial design. Experiments showed that microparticles with favorable physicochemical properties and high probiotic viability during preparation and storage could be obtained when 40 mg/g sodium alginate, 5 mg/g chitosan and 50 mg/g CaCl2 is used. Stability of L. casei during microencapsulation was identified by FTIR spectroscopy. The viability of the probiotic in the optimal formulation of synbiotic microparticles remained above the therapeutic minimum during incubation of 24 hours in simulated gastrointestinal conditions (7.67?0.4 log cfu/g) as well as after 3 months of cold storage (8.1?0.6 log cfu/g). High viability of L. casei was maintained during 6 weeks of cold storage when carrot juice was enriched with encapsulated cells. The effective preservation of L. casei into synbiotic microparticles provided production of new non-dairy functional food as an alternative of the population who is at risk of lactose intolerance.

Possible synergistic effect of tamoxifen (2 μM) and hydrazinyldiene-chroman-2,4-diones (10-100 μM) was examined with an aim to create more effective treatment for ER+ breast cancer. Anti-breast cancer effect has been evaluated on the proliferation of MCF-7 breast adenocarcinoma cells using MTT and alamarBlue assays. Cell viability was evaluated after 48h-treatment and the ICs50 of the coumarin derivatives were determined. The apoptotic effect was evaluated by detection of PARP cleavage and reduced activity of the survival kinase Akt. The results demonstrated dose-dependent activity, with a percent of growth inhibition after combination treatment being significantly higher (53% to 79%, 10 μM and 100 μM, respectively) than the one in the cell lines treated with tamoxifen (29% to 37%) and the synthesized coumarin derivatives alone (11% to 68%, 10 μM and 100 μM, respectively). The ICs50 of the synthesized compounds significantly decreased in synergy with tamoxifen (33% to 51%). Coumarin derivative having thiazole moiety with additional methyl groups attached to the carbons at positions 5 and 4 in the thiazole ring showed to be the most potent, with IC50 20 µM when administered alone and 10 µM in synergy with tamoxifen. The levels of phospho-Thr308 Akt were down-regulated by the combination treatment, pointing to tyrosine kinase phosphorylation inhibition. In conclusion, the novel coumarin derivatives enhance the activity of tamoxifen and this combination may be suitable for prevention of ER+ breast cancer or development of related compounds. Further studies are needed to elucidate precisely the type of receptor involved in the activity and the mechanism of action.

In our previous work Box-Behnken experimental design was applied for formulation optimization of the thermoreversible mucoadhesive in situ vaginal hydrogels with propolis and optimized batches were identified. Optimized batches of bioartificial polymer hybrid composites (chitosan, Lutrol® F-127 and Lutrol® F-68 mixture)) (CP1, CP2, CP3) were prepared using so-called cold method. Formulation P3 (chitosan free) was prepared in order to evaluate the effect of chitosan on the physico-chemical and biopharmaceutical properties of the polymer hybrid composites (gels). The pH values of the gels were 4-4.5. The gelation temperature for all formulations was in a range of 29-33 o C. Total flavonoids content was above 95%. Increase in concentration of Lutrol® F-127 and Lutrol® F-68/Lutrol® F-127 ratio lead to a higher viscosity values and slower gel erosion/dissolution. The presence of chitosan increased gel viscosity and hence slow-down erossion/dissoluiton. Propolis release rate was the highest in P3 which released propolis within 5 h, corresponding to time of complete erosion. The same correlation between erosion process and drug release rate was observed in CP1-CP3, where prolonged propolis release for more than 10 h was achieved. Microbiological quality was in accordance with the requirements of Ph. Eur. 7. All formulations demonstrated adequate stability at 5 ± 3 °C during 6 months. Based on overall results it can be anticipated that bioartificial blended bioinspired polymer hybrid composites for propolis vaginal delivery could represent intelligent delivery systems with physicochemical and biopharmaceutical properties in favor or efficacious and safe therapy of vaginal infections.

A fast and simple liquid chromatography–electrospray ionization tandem mass spectrometry method for determination of indapamide in human whole blood was developed and validated. The sample extraction of indapamide from human whole blood was achieved using automated solid-phase extraction. Chromatographic separation was performed on Kinetex C18 column (100 × 2.1 mm, 1.7 µm particle size) using acetonitrile and 2 mm ammonium formate in ratio 90:10 (v/v) as a mobile phase. The mass spectrometer was operated in the multiple reaction monitoring mode using positive electrospray ionization for indapamide and the internal standard (zolpidem tartarate). The total run time was 2.5 min. The present method was found to be linear in the concentration range of 1–50 ng/mL with the coefficient of determination 0.9987. The absolute recoveries of indapamide were 90.51–93.90%. The method was validated according the recommendations for validation of bioanalytical methods of European Medicines Agency guideline and was successfully used to analyze human whole blood samples for application in a pharmacokinetic study. Copyright © 2013 John Wiley & Sons, Ltd.

Polymer hybrid composites for propolis vaginal delivery were developed and optimized in term of gelling temperature (Tg) using experimental design studies. Formulation of intelligent delivery system, with in situ gelling and controlled release properties, was carried out by bioartificial blending of chitosan and poloxamers, Lutrol F127 and Lutrol F68. Response-surface, Box-Behnken, experimental design was applied for determination of key factors influencing Tg. Influence of Lutrol F127 concentration, Lutrol F68/Lutrol F127 mass ratio and volume of simulated vaginal fluid was evaluated. Derived correlations enabled formulation optimization in term of Tg (32 °C). The model was cross-validated. Confirmation report and relative error percent of predicted and experimental values were obtained.

Simple, automated SPE procedure combined with fast LC-MS/MS chromatographic separation resulted in obtaining high-throughput LC-MS/MS method for determination of indapamide in serum. The SPE procedure was performed on polymeric mixed-mode sorbent and the analytes were quantitated using electrospray ionization in positive mode. The recovery of indapamide and internal standard were 89.25- 90.36% and 79.10%, respectively. Experimentally it was confirmed that the matrix effect had a negligible effect of ionization efficiency. The validation data showed that the proposed method provides accurate and reproducible results in range of 0.50-50 ng/mL. In addition, a comparison was made between the method for determination of indapamide in serum and in blood (develop in our previously work) regarding the extraction procedure and matrix effect.

This article presents a novel formulation for preparation of Lactobacillus casei 01 encapsulated in soy protein isolate and alginate microparticles using spray drying method. A response surface methodology was used to optimise the formulation and the central composite face-centered design was applied to study the effects of critical material attributes and process parameters on viability of the probiotic after microencapsulation and in simulated gastrointestinal conditions. Spherical microparticles were produced in high yield (64%), narrow size distribution (d50=9.7 µm, span=0.47) and favourable mucoadhesive properties, with viability of the probiotic of 11.67, 10.05, 9.47 and 9.20 log CFU/g after microencapsulation, 3 h in simulated gastric and intestinal conditions and four-month cold storage, respectively. Fourier-transform infrared spectroscopy confirmed the probiotic stability after microencapsulation, while differential scanning calorimetry and thermogravimetry pointed to high thermal stability of the soy protein isolate-alginate microparticles with encapsulated probiotic. These favourable properties of the probiotic microparticles make them suitable for incorporation into functional food or pharmaceutical products.

This study evaluates the effects of previously synthesized hydrazinyldiene-chroman-2,4-diones on cell proliferation and apoptosis, cell cycle distribution and migration capacity of MCF-7 breast cancer cells in synergy with doxorubicin. Physicochemical properties of the synthesized compounds were correlated with their structure and activity. Significant cell viability decrease in comparison with the effect of doxorubicin alone and the reference 4-hydroxycoumarin was observed when combination treatment comprising doxorubicin and the title compounds was applied. Synergistic effect with doxorubicin was also observed in down-regulation of phospho-Thr308Akt levels, confirming reduced proliferation and increased apoptosis. Combined treatment increased the percentage of cells arrested at the G2/M stage. Additive inhibition of cell migration was also observed, pointing to the possibility of reducing the risk of metastases. With their solubility profile and log D7.4, all the synthesized compounds follow Lipinski’s rule of five for good permeability (absorption) potential.

This study aims to evaluate the influence of different concentrations of fructooligosaccharide (FOS) as prebiotic on functional and physicochemical properties of Lactobacillus casei loaded chitosan-Ca-alginate microparticles prepared by spray-drying method, after preparation and 6-month storage at 4 °C. Prebiotic compound positively influenced the viability of L. casei during preparation of the microparticles and in storage conditions, while no significant difference of the probiotic viability was observed when different concentrations of FOS in a range of 1.5 - 5% w/w were applied. Having in regard that non-significant improvement in the functional and physicochemical properties of the microparticles with increased content of the FOS was observed, the concentration of 1.5% w/w was recommended as an optimal for preparation of synbiotic loaded microparticles.

C-C bond formation reactions between (benzamidomethyl)triethylammonium chloride (1) and some carbon nucleophiles (2a-f) were performed smoothly in aqueous media, under mild reaction conditions and ambient temperature. Mono-C-alkyl (3a, 3b, 3c) and di-C-alkyl (3d, 3e, 3f) derivatives were obtained without using a catalyst. Crystals of the products were easily isolated by simple filtration, in moderate yields and with no apparent contamination due to the formation of O-alkyl products.

Background: Elevated lipoprotein (a) [Lp(a)] is an independent risk factor for coronary artery disease (CAD). However, there is little evidence about the therapeutic efficacy of different lipid-lowering agents in reducing Lp(a). Objective: The primary objective of our study was to test the effect of different therapeutic treatment strategies on elevated Lp(a) levels, specifically to compare rosuvastatin versus atorvastatin alone or in combination with niacin or fibrates. The secondary objective was to analyze the occurrence of potential adverse effects. Methods: It was a prospective, single-center, interventional study. Patients with CAD, or high CAD risk, with increased Lp(a), >50 mg/dL, were included in the study. Lp(a), total cholesterol (C), triglycerides (TGs), high-density lipoprotein cholesterol and low-density lipoprotein cholesterol (LDL-C), apolipoprotein (Apo) A1, Apo B, and enzymes of myocyte and hepatic injury were comparatively analyzed between 4 lipid-lowering treatment strategies: rosuvastatin (R group) 40 mg, atorvastatin (A group) 80 mg, atorvastatin 40 mg add-on micronized fenofibrate (A+F group), and atorvastatin 40 mg add-on 1 g extended-release niacin (A+ERN group). Comparison was made for their lipid lowering therapeutic efficacy, primarily on Lp(a), and their safety profile. Results: A total of 87 patients, 61 ± 12 years old, were analyzed. The main risk factors were obesity (64.7%) and hypertension (64.6%). Men were more often smokers (odds ratio [OR] = 5.1) and had CAD (OR = 2.8), but lower total C (206.9 ± 32.9 vs 238.6 ± 47.9 mg/dL, P = 0.002) and LDL-C (136.5 ± 18.2 vs 160.9 ± 30.9 mg/dL, P = 0.000). Mean Lp(a) was 94.6 ± 39.6 mg/dL, without significant gender difference. There were 25 patients in the R group, 22 in the A group, and 20 each in the A+F and A+ERN groups. Significant reduction in all lipid fractions in all treatment groups was reported after 6 months. The average reduction of Lp(a) was 15.9 ± 21.0 mg/dL, with 18.2 ± 24.8 (P = 0.001) in the R group and similar values in the A+F and A+ERN groups (17.3 ± 10.4, P = 0.001, and 19.5 ± 10.9, P = 0.001, respectively), and the lowest in the A group (11.24 ± 22.91, P = 0.032). No adverse effects were observed in any of the treatment groups. Conclusions: When compared with atorvastatin, it seems that rosuvastatin can achieve a statistically more significant decrease of Lp(a). The efficacy of atorvastatin on the Lp(a) optimization can be increased by adding either fibrate or ERN. Given in recommended doses, all agents were well tolerated.

Herein we report the synthesis of different derivatives of (fluoro)quinolones norfloxacin, ciprofloxacin and pipemidic acid, by incorporating (benzoylamino)methyl on the free nitrogen of the pyperazinyl moiety. The compounds were structurally characterized by 1D and 2D NMR, FTIR and high-resolution mass spectroscopy. In addition, their physicochemical properties were a matter of interest to be correlated with their structure and antimicrobial activity in vitro. Their antimicrobial activities were screened against Gram-positive, Gram-negative bacteria and C. albicans. Higher distribution coefficients and consequently lower water solubility were determined for all synthesized compounds than the ones of the corresponding leading compounds. Inconsequential correlations between the lipophilicity of the compounds and MIC were observed, suggesting that passive diffusion is not the only mechanism for their penetration into bacterial cells. Further studies are needed to determine how substitutions in the (fluoro)quinolone moiety affect the primary target(s), substrate behavior in respect to bacterial transporters and overall bioavailability.

Chitosan-Ca-alginate microparticles for colon-specific delivery and controlled release of 5-aminosalicylic acid after peroral administration were prepared using spray drying method followed by ionotropic gelation/polyelectrolyte complexation. Physicochemical characterization pointed to the negatively charged particles with spherical morphology having a mean diameter less than 9 microm. Chitosan was localized dominantly in the particle wall, while for alginate, a homogeneous distribution throughout the particles was observed. (1)H NMR, FTIR, X-ray and DSC studies indicated molecularly dispersed drug within the particles with preserved stability during microencapsulation and in simulated in vivo drug release conditions. In vitro drug release studies carried out in simulated in vivo conditions in respect to pH, enzymatic and salt content confirmed the potential of the particles to release the drug in a controlled manner. The diffusional exponents according to the general exponential release equation indicated anomalous (non-Fickian) transport in 5-ASA release controlled by a polymer relaxation, erosion and degradation. Biodistribution studies of [(131)I]-5-ASA loaded chitosan-Ca-alginate microparticles, carried out within 2 days after peroral administration to Wistar male rats in which TNBS colitis was induced, confirmed the dominant localization of 5-ASA in the colon with low systemic bioavailability.

Viability of L. casei during fermentation in soymilk and freeze-dried soymilk; effect of cryoprotectant, rehydration and storage temperature The aim of the work was to investigate the behaviour of L. casei and the effect of sorbitol on its viability during fermentation in soymilk drink. Values for pH, ranging from 6.82 to 3.42 in the soymilk drink without sorbitol and from 6.74 to 3.41 in the drink with sorbitol were noted during 72 h of fermentation at 25oC. The corresponding values for titratable acidity ranged from 0.071% to 0.758% and from 0.073% to 0.761%, respectively. Soymilk was found to support the growth of L. casei with improvement in viability for 0.24 log at the end of fermentation when sorbitol was added. Survival of L. casei and the effectiveness of sorbitol in improving viability during freeze-drying, subsequent rehydration and during a 5-week period of storage under different temperatures were also investigated. After freeze-drying, L. casei exhibited a survival percent of approximately 46%. Sorbitol improved the viability of L. casei by 0.51 log immediately after freeze-drying and by 1.30 log and 0.47 log during five weeks of storage at 25oC and 4oC, respectively. Further study revealed that the freeze-dried fermented soymilk rehydrated at 45oC was optimum for the recovery of L. casei with improvement in recovery for 0.68 log when sorbitol was added. A higher percent of survival was noted when the dried soymilk was stored at 4oC than at 25oC with improved viability at the end of 5 weeks storage for approximately 6 log for drinks with and without sorbitol. Fermented dried soymilk with sorbitol afforded significant tolerance of L. casei to acid stress. Generally, a stable probiotic diary product was prepared in which the concentration of L. casei remained above therapeutic level of 107 cfu/ml.

Certain variations in the process parameters (emulsification time, surfactant concentration) were performed in order to prepare BSA-loaded gelatin microspheres with particle size ranging from 1 to 10 µm and high loading efficiency using a procedure originally employed by Tabata and Ikada. In vitro degradation and drug release studies in the presence of trypsin and collagenase, respectively, were performed in order to evaluate the potential of gelatin microspheres as regulated and sustained release systems for oral vaccination. Degradation data showed that the preparation procedure had provided prolonged degradation in the presence of both enzymes, suggesting complete in vivo degradation. Exponential dependence of the amount of drug released on time was evidenced. The diffusion coefficients were superior to 0.5 indicating the Case II anomalous Fickian diffusion, except for the particles smaller than 5 µm where in the presence of collagenase the transition to Super Case II transport was observed due to the higher rate of polymer degradation and BSA diffusion through the matrix. The mathematical modeling of drug release showed a biphasic release pattern in the presence of both enzymes, where the rate constants for the initial time release confirmed the influence of the particle size and/or enzymatic degradation rate on the drug release rate.

Liposome dispersions and liposome gel formulations for topical administration were evaluated as modified release delivery systems for 5-fluorouracil. Drug substance has been entrapped in the internal aqueous compartment of liposomes during the preparation. The concentration of 5-fluorouracil in the hydration medium was varied and the effect on the liposome characteristics was considered. Liposome gel formulations were prepared by incorporation of liophylized liposomes into a structured vehicle of chitosan. The decrease of the amount of aqueous phase bearing total drug quantity (drug/aqueous phase ratio from 1:100, 1:60, 1:40) led to an increase of the percentage of liposome-entrapped drug, and decreased percentage of drug release, while particle size analysis showed no changes in vesicle size. Liposome gel formulations showed initially a higher drug release rate in comparision with liposome dispersions, which could be related to the release of “free” 5-fluorouracil, leaked from liposomes due to the process of liophylization. This was followed by slower release (after 1.5 hour) as a result of the influence of the viscosity of the gel matrix.

We report the synthesis of [(3-chlorobenzamido)methyl]triethylammonium chloride in a reaction of N-(chloromethyl)-3-chlorobenzamide and triethylamine in dry acetone. The structure of the newly synthesized compound was characterized with1H-NMR,13C-NMR, FTIR and Mass spectroscopy. Macedonian Academy of Sciences and Arts/07-520/15 Bulgarian Academy of Sciences/BAS-MANU/2014–2016 DFG/EXC62/REBIRTH

In this study, hybrid silica xerogel particles were developed as carriers of budesonide (BDS) for efficient local treatment of inflammatory bowel diseases (IBD). Organically modified silica particles (ORMOSILs) were prepared by co-condensation of 3-aminopropyltriethoxysilane (APTES) and tetraethyl orthosilicate (TEOS) by an ambient temperature acid catalysed sol-gel process followed by spray-drying. Formulation for preparation of BDS-loaded particles was optimized and their physicochemical parameters and drug release profiles were evaluated in vitro. Optimal formulation had a small particle size (mean diameter of 1.45±0.02μm) with unimodal narrow size distribution and high encapsulation efficiency (98.0 ± 1.85%). Due to the positive surface charge originated from amino group of APTES, ORMOSILs showed excessive mucoadhesiveness in comparison to native TEOS particles. The drug release decreased with increasing pH from 2.0 to 7.4. In order to avoid undesirable erroneous performance in the upper GI tract, particles were additionally coated with Eudragit(®) FS 30D, as a barrier to the drug release at pH range from 2.0 to 7.0. After Eudragit(®) FS 30D coating, the release of BDS in acidic media was sustained, while no significant differences in drug release were observed at pH 7.4. In conclusion, pH-responsive ORMOSILs showed great potential for efficient BDS delivery to the colon region.

The effect of the buffer system on the stability of tetracaine hydrochloride in eyedrop formulation was evaluated. Eyedrop formulations containing tetracaine hydrochloride 0.5%(w/v) were prepared using different buffer systems (acetate, phosphate and borate buffer) under a constant pH of 5.4, and a buffer concentration of 0.06M. Long-term tests at 26 o C and accelerated stability tests at elevated temperature (45, 50, 60 o C) over a period of 168 days were carried out by following the macroscopic view, pH, sterility, content of tetracaine hydrochloride and detection of the degradation products. Also, values of the constant of degradation rate at different temperatures and t90% were calculated. The phosphate and acetate buffers provided satisfactory stability of tetracaine hydrochloride eyedrops, while borate buffer was not sufficient to maintain the pH value of the solution.

Certain variations in the process parameters (emulsification time, surfactant concentration) were performed in order to prepare BSA-loaded gelatin microspheres with high loading efficacy and particle size ranging from 1 to 10 microm using a procedure originally employed by Tabata and Ikada. The mathematical modelling of drug release in the presence of collagenase showed a biphasic release pattern, where the rate constant for the initial time release confirmed the influence of the particle size and/or enzymatic degradation rate on drug release rate.

Biodistribution studies of radiolabelled [131I]BSA loaded gelatin microspheres were carried out on BALB/c mice after peroral administration. To two groups, the radiolabelled [131I]BSA gelatin microspheres with different mean particle size, 1.196+/-1.961 and 7.028+/-1.231 microm were administered orally. To the control group, a solution of [131I]BSA was also orally administered. Biodistribution was followed periodically within 15 days as a percent of total radioactivity present in stomach, small intestine with Peyer's patches and mesentery, colon with Peyer's patches, appendix and mesentery, liver, spleen, blood, kidney, lungs and heart. The biodistribution data confirmed that uptake in mice into Peyer's patches and passage to the liver and spleen via the mesentery lymph supply and nodes, increased with decreasing particle size.

(Benzoylamino)methyl 4-acetyloxybenzoate (3) was obtained in a reaction of benzamidomethylation of 4-acetyloxybenzoic acid (2) with (benzamidomethyl)­triethyl­ammonium chloride (1).

(Benzoylamino)methyl 4-hydroxybenzoate (“Benzamidomethylparaben”) (3) was obtained from a reaction of 4-hydroxybenzoic acid (2) with a dioxane suspension of (benzamidomethyl)triethylammonium chloride (1). The phenolic group in 2 cannot be benzamidomethylated with 1 in aqueous media.

Liposomal hydrogel formulations of lidocaine HCl, suitable for topical application, have been prepared and drug release properties in vitro have been evaluated. Liposomes composed of Soya lechitin and cholesterol, with lidocaine HCl, entrapped in the inner water compartment, were prepared by simple hydration method. Topical liposomal gel formulations were prepared by incorporation of liposomes into a structured vehicle (hydrogels of Carbopol 940 in concentration of 1.5, 1.75 and 2%). High percentage of encapsulated drug in liposomes has been obtained (over 70%). Liposomal gel formulations provided prolonged drug release rate. The concentration of gelling agent in a range 1.5–2.0% affected the release rate slightly. In vitro release data showed that release kinetic can be described as diffusion-controlled, while liposomes act as reservoir systems for continuous delivery of drug. Proposed formulations provided stable percentage of entrapped drug and drug release within an examination period of 3 weeks.

In this paper, the synthesis and structure of a novel pyrano[3,2-c]coumarin, which was obtained unexpectedly in a reaction of phosphoryl chloride and acenocoumarol, are presented. The chemical structure of the novel compound was elucidated by a detailed spectroscopic analysis based mainly on 1D and 2D NMR techniques. The structure was finally confirmed with a single-crystal X-ray analysis. The B3LYP/6-311+G** method correctly reproduces the bond lengths, bond angles, torsion angles, and other experimental spectroscopic data, which can be useful for investigating the characteristics of some structurally related molecules. Additionally, the title compound was obtained in high yields by performing a dehydratation reaction of acenocuomarol with acetic anhydride.

In this study, the probiotic Lactobacillus casei was microencapsulated using the method of spray-drying combined with polyelectrolyte complexation of alginate, fructooligosaccharide and chitosan, and cross-linking with calcium chloride, followed by freeze-drying. Survival rate and physicochemical properties of the prepared microparticles were evaluated. In addition, viability of Lactobacillus casei in simulated gastric and intestinal juices was investigated. Positively charged microparticles with average size of 11.08±1.1 μm and high cell viability of 10.98±0.11 log cfu/g were prepared. The synbiotic microparticles were stable during exposure to simulated gastric and intestinal juices, while release of viable cells above the therapeutic value (8.31±0.14 log cfu/g) in the simulated colonic pH was observed.The presented method for microencapsulation of synbiotics shows potential for effective protection of viable probiotic cells during exposure to harsh environmental conditions.

In this note, two procedures for the synthesis of (benzoylamino)methyl 4-[(benzoylamino)methoxy]benzoate ( 3 ) are presented. The first procedure is carried out in dioxane/water using benzoylaminomethyl-4-hydroxybenzoate, while the second one employs a suspension of 4-hydroxybenzoic acid in dioxane. In both procedures, benzamidomethyl triethylammonium chloride is used for the benzamidomethylation reaction. Keywords: benzamidomethyl ester; synthesis Methyl 4-methoxybenzoate (also known as methyl anisate) is a white crystalline powder, soluble in alcohol and ether, but insoluble in water. In the nature, it occurs as a volatile compound in mushroom species and plants [1–5]. These types of esters are used as pharmaceutical intermediates and take part in many organic syntheses [6,7]. For example, a new imaging compound, [(125)I]iodoDPA-713, was synthesized in several steps from methyl 4-methoxybenzoate as a tool for quantification of inflammation in preclinical models [6]. Nowadays, methyl 4-methoxybenzoate has application in the flavor and perfume industry as synthetic flavoring substance due to its sweet herbal anis aroma, impressing lilac or magnolia [8,9]. In this note, the synthesis of a new compound, (benzoylamino)methyl 4-[(benzoylamino)methoxy]-benzoate (

Methylparabene (2) was simply benzamidomethylated with (benzamidomethyl)- triethylammonium chloride (1) in aqueous medium to afford methyl 4-(benzamido- methoxy)benzoate (3) in high yield. The title compound was characterized by elemental analysis, FT-IR, 1 H-NMR and 13 C-NMR spectroscopy.