Short Review on the Biological Activity of Cyclodextrin-Drug Inclusion Complexes Applicable in Veterinary Therapy.

Cyclodextrins (CDs) are a family of carrier molecules used to improve the pharmacokinetic parameters of therapeutic molecules. These cyclic oligosaccharides have medical and pharmaceutical applications by being able to form inclusion complexes with molecules that are poorly soluble in water. The benefits of these complexes are directed towards improving the chemical and biological properties-i.e., solubility, bioavailability, stability, non-toxicity and shelf life of drug molecules. Since the 1960s, the first inclusion complexes used in therapeutics were those with α-, ß- and γ-CD, which proved their usefulness, but had certain degrees of particularly renal toxicity. Currently, to correct these deficiencies, ß-CD derivatives are most frequently used, such as sulfobutylether-ß-CD, hydroxypropyl-ß-CD, etc. Therefore, it is of interest to bring to the attention of those interested the diversity of current and potential future clinical applications of inclusion complexes in veterinary medicine and to present the contribution of these inclusion complexes in improving drug efficacy. The most important biological activities of ß-CD complexed molecules in the veterinary field are summarized in this short review.

Toxicity Assessment of Long-Term Exposure to Non-Thermal Plasma Activated Water in Mice.

Non-thermal plasma activated water (PAW) has recently emerged as a powerful antimicrobial agent. Despite numerous potential bio-medical applications, studies concerning toxicity in live animals, especially after long-term exposure, are scarce. Our study aimed to assess the effects of long-term watering with PAW on the health of CD1 mice. PAW was prepared from distilled water with a GlidArc reactor according to a previously published protocol. The pH was 2.78. The mice received PAW (experimental group) or tap water (control group) daily for 90 days as the sole water source. After 90 days, the following investigations were performed on the euthanatized animals: gross necropsy, teeth mineral composition, histopathology, immunohistochemistry, hematology, blood biochemistry, methemoglobin level and cytokine profile. Mice tolerated PAW very well and no adverse effects were observed during the entire period of the experiment. Histopathological examination of the organs and tissues did not reveal any structural changes. Moreover, the expression of proliferation markers PCNA and Ki67 has not been identified in the epithelium of the upper digestive tract, indicating the absence of any pre- or neoplastic transformations. The results of our study demonstrated that long-term exposure to PAW caused no toxic effects and could be used as oral antiseptic solution in dental medicine.

New insights into human hair: SAXS, SEM, TEM and EDX for Alopecia Areata investigations.

BACKGROUND: Alopecia areata (AA) is a T-cell-mediated autoimmune disease and affects up to 2% of the population. There is a need for a more profound and rigorous understanding of the structure and composition of human hair affected by AA in order to manage this disease. The aim of this article is to understand the effects of AA on the structure and composition of human hair. METHODS: Several physico-chemical investigation methods, such as Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Energy-Dispersive X-ray Spectroscopy (EDX), and microbeam Small Angle X-ray Scattering (SAXS), were used to analyze human hair samples obtained from healthy donors and patients with AA. RESULTS: SEM revealed more severe hair surface defects for the white regrown hair (W-AA) samples. TEM showed the presence of air-like vesicles located in the endocuticle of regrown hair. Analysis of ultrathin sections of W-AA showed the existence of empty vesicles and smaller melanin granules compared to control samples. SAXS demonstrated that unaffected hair of patients with AA (B-AA) and W-AA melanin aggregates are different in their sizes and shapes compared to the control samples. EDX data showed that W-AA elemental composition was significantly different from the other sample groups. Our study showcases promising non-invasive techniques for a better and more accurate understanding of changes in the internal structure and composition of hair affected by AA.

In vitro activity of echinocandins against 562 clinical yeast isolates from a Romanian multicentre study.

The study presents the echinocandin susceptibility profile of a multi-centre collection of pathogenic yeast isolates from Romanian tertiary hospitals. The 562 isolates were identified using ID32C strips, MALDI-TOF MS and DNA sequencing. Minimal inhibitory concentrations (MICs) of caspofungin (CAS), micafungin (MCA), and anidulafungin (ANI) were assessed and interpreted according to EUCAST guidelines. Minimal fungicidal concentrations (MFC) were determined by plating content from the clear MIC wells. The activity was considered fungicidal at MFC/MIC ≤ 4. The three echinocandins had strongly correlated MICs and high percentages of MIC essential agreement. Most often, MCA had the lowest MICs, followed by CAS and ANI. Against C. parapsilosis and C. kefyr, CAS had the lowest MIC values. The MIC50 values were between 0.03 and 0.25 mg/l, except C. parapsilosis. The MIC90 values were usually one dilution higher. MFCs and MICs were weakly correlated. ANI and MCA had the lowest MFC values. The MFC50 values were between 0.06 and 0.5 mg/l, except C. parapsilosis, C. guilliermondii, and C. dubliniensis. The MFC90 values were usually two dilutions higher. Based on EUCAST breakpoints, 47 isolates (8.4%) were resistant to at least one echinocandin, most often ANI. Most resistant isolates were of C. albicans, C. glabrata, and C. krusei. There were 17 isolates (3%) resistant to echinocandins and fluconazole and most belonged to the same three species. MCA and ANI had the highest rates of fungicidal activity. The high rates of echinocandin resistance and significant multidrug resistance make prophylaxis and empiric therapy difficult.

Drug-induced gingival hyperplasia - experimental model.

Several causes of gingival hyperplasia are known, the most widely accepted being the drug-induced gingival augmentation, a side effect associated mainly with three classes of drugs: anticonvulsants (Phenytoin), immunosuppressants (Cyclosporine A), and various calcium channel blockers (Nifedipine, Verapamil, Diltiazem). We studied the effect of Cyclosporine A (CsA) and Nifedipine on gingival fibroblasts extracted from the rat gum. Gingival fibroblasts were isolated from 6-week-old male rats weighing 150-170 g, from gingival explants, and grown in a specific culture medium consisting of Dulbecco's Modified Eagle's Medium (DMEM) supplemented with antibiotic and 10% fetal bovine serum (FBS). The medium was also supplemented with CsA (1 µg÷mL) and Nifedipine (3 mM). We also used a control group that received no treatment. The cells were photographed at 7, 14 and 30 days, with a Nikon Eclipse TE300 phase contrast microscope. For cell viability evidence, we used immunofluorescence (flow cytometry) with a FACS (fluorescence-activated cell sorting) Calibur device and its software. We noticed that the proliferation of these cells increased with the period of drug administration, and the subsequent morphological changes that occurred were related to the presence of drug accumulations in the cell cytoplasm. Modern molecular techniques are needed to shed some light upon the pathogenesis of drug induced gingival overgrowth and, thereby, provide novel information for preventative and effective future therapeutic strategies.

An original method for producing acetaldehyde and diacetyl by yeast fermentation

Abstract In this study a natural culture medium that mimics the synthetic yeast peptone glucose medium used for yeast fermentations was designed to screen and select yeasts capable of producing high levels of diacetyl and acetaldehyde. The presence of whey powder and sodium citrate in the medium along with manganese and magnesium sulfate enhanced both biomass and aroma development. A total of 52 yeasts strains were cultivated in two different culture media, namely, yeast peptone glucose medium and yeast acetaldehyde-diacetyl medium. The initial screening of the strains was based on the qualitative reaction of the acetaldehyde with Schiff's reagent (violet color) and diacetyl with Brady's reagent (yellow precipitate). The fermented culture media of 10 yeast strains were subsequently analyzed by gas chromatography to quantify the concentration of acetaldehyde and diacetyl synthesized. Total titratable acidity values indicated that a total titratable acidity of 5.5 °SH, implying culture medium at basic pH, was more favorable for the acetaldehyde biosynthesis using strain D15 (Candida lipolytica; 96.05 mg L-1 acetaldehyde) while a total titratable acidity value of 7 °SH facilitated diacetyl flavor synthesis by strain D38 (Candida globosa; 3.58 mg L-1 diacetyl). Importantly, the results presented here suggest that this can be potentially used in the baking industry.

An original method for producing acetaldehyde and diacetyl by yeast fermentation.

In this study a natural culture medium that mimics the synthetic yeast peptone glucose medium used for yeast fermentations was designed to screen and select yeasts capable of producing high levels of diacetyl and acetaldehyde. The presence of whey powder and sodium citrate in the medium along with manganese and magnesium sulfate enhanced both biomass and aroma development. A total of 52 yeasts strains were cultivated in two different culture media, namely, yeast peptone glucose medium and yeast acetaldehyde-diacetyl medium. The initial screening of the strains was based on the qualitative reaction of the acetaldehyde with Schiff's reagent (violet color) and diacetyl with Brady's reagent (yellow precipitate). The fermented culture media of 10 yeast strains were subsequently analyzed by gas chromatography to quantify the concentration of acetaldehyde and diacetyl synthesized. Total titratable acidity values indicated that a total titratable acidity of 5.5°SH, implying culture medium at basic pH, was more favorable for the acetaldehyde biosynthesis using strain D15 (Candida lipolytica; 96.05mgL-1 acetaldehyde) while a total titratable acidity value of 7°SH facilitated diacetyl flavor synthesis by strain D38 (Candida globosa; 3.58mgL-1 diacetyl). Importantly, the results presented here suggest that this can be potentially used in the baking industry.

Etiologic Agents and Antifungal Susceptibility of Oral Candidosis from Romanian patients with HIV-infection or type 1 diabetes mellitus.

This is the first Romanian investigation of oral candidosis in patients suffering of HIV-infection or type 1 diabetes mellitus (T1DM). Candida albicans was the dominant species in both types of isolates: n = 14 (46.7%) in T1DM, n = 60 (69.8%) in HIV. The most frequent non-albicans Candida spp. were Candida kefyr (n = 6; 20%) in T1DM and Candida dubliniensis (n = 8; 9.3%) in HIV. Resistance to fluconazole was detected only in the HIV non-albicans Candida group (n = 8; 9.3%). All isolates were susceptible to VOR. The experimental drug MXP had MIC values equal or close to the ones of VOR. Echinocandin resistance was more frequent than azole resistance.

Heparin-anthranoid conjugates associated with nanomagnetite particles and their cytotoxic effect on cancer cells.

The paper describes a methodology for preparing monodisperse, water-soluble magnetite nanoparticles, coated with heparin and loaded with 4,5-dihydroxy-9,10-dioxoanthracene-2-carboxylic acid (Rhein), able to be used as a drug delivery system for cancer chemotherapy. Upon preparation, nanoparticles structure and morphology were investigated. The surface charge and the equivalent dimensions of the nanoparticles dispersed in water were measured, as a function of the suspension pH. The concentration of the drug into the nanoparticles shell, and the drug release profile was determined. The functionality of Rhein-loaded heparin-coated magnetic nanoparticles was assessed by monitoring their cytotoxic effect on cultured human tumor hepatocyte cell line, HepG2, using MTT assay. We found that upon exposure of HepG2 cells to Rhein-loaded heparin-coated nanoparticles, the cell viability was drastically reduced (to approximately 10%) as compared to that of the cells exposed to the free drug, indicating the potential of these magnetite nanoparticles to be used in cancer therapy.