Analysis of the Influence of Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus Strains on Changes in the Hexachlorobenzene Content in Fermented Mare Milk during Refrigerated Storage.

(1) Background: Hexachlorobenzene (HCB) is a persistent organic pollutant that is possibly carcinogenic to humans. It is still found in the environment, humans and animals, and in foods, including milk and dairy products; (2) Methods: The influence of the probiotic cultures Lacticaseibacillus rhamnosus LCR and Lactiplantibacillus plantarum subsp. plantarum LP on the possibility of effecting the biodegradation of HCB in dairy products fermented from mare milk was investigated, taking into account the product storage time (maximum 21 days). HCB content was determined using the GC/MS method; (3) Results: A strong negative Pearson correlation (p < 0.05) was found between HCB concentration and the refrigeration storage time of the fermented beverages. The highest HCB reduction was observed in milk fermented with both Lacticaseibacillus rhamnosus LCR and Lactiplantibacillus plantarum subsp. plantarum LP (78.77%), while the lowest was noted when only Lactiplantibacillus plantarum subsp. plantarum LP was used (73.79%); (4) Conclusions: This pilot study confirmed that probiotics commonly used to give products health-promoting properties can also contribute to reducing the content of undesirable substances, and the bacterial cultures used might provide an alternative method for reducing HCB residues in fermented drinks.

PEG-liposomal doxorubicin as a potential agent for canine metastatic osteosarcoma - in vitro and ex ovo studies.

Introduction: Appendicular osteosarcoma (OSA) is a highly aggressive and metastatic primary bone tumour in dogs. Standard therapy is amputation and adjuvant chemotherapy (e.g. with doxorubicin). Liposomal drug delivery may augment therapeutic efficacy and reduce negative side effects. Polyethylene glycol (PEG)-liposomal doxorubicin treats human metastatic cancers effectively. The study aimed was to evaluate PEG-liposomal doxorubicin's inhibitory effect on canine metastatic proliferation and migration in vitro. It also aimed to appraise the drug's extravasation inhibition in vivo using the human medicine-proven chick embryo chorioallantoic membrane ex ovo model. Material and Methods: The canine D-17 OSA cell line was cultured and inoculated with decreasing concentrations of PEG-liposomal doxorubicin and conventional doxorubicin in a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) test of cell viability, proliferation and cytotoxicity. Flow cytometry with Annexin V and Draq 7 staining confirmed the MTT test results, indicating dead, early and late apoptotic, and live cells. The inhibitory effect of the two preparations on cancer cell migration was investigated with a wound-healing assay. Culture plates seeded with cells were prepared. The cell monolayer was scratched and images of cells migrating to the scratch were captured at 0 h, 12 h and 24 h. Also, embryos were removed from three-day-incubated fertilised chicken eggs. On the 12th day, labelled D-17 cells were injected into each embryo. Embryos in one group received 100 µL of phosphate-buffered saline as controls, those in another group 30 µg/mL of PEG-liposomal doxorubicin, and those in the last group 6 µg/mL of conventional doxorubicin. The effectiveness of the intravascular administration of the D-17 cells was confirmed under a microscope. Results: PEG-liposomal doxorubicin inhibited the migration of canine OSA cells more effectively than conventional doxorubicin (P ≤ 0.05). The ex ovo model showed that both drugs had similar impacts on canine metastatic OSA. Conclusion: The liposomal form of the drug may be considered a potentially effective compound in canine metastatic OSA; nevertheless, further in vivo studies are essential to confirm this hypothesis.

Gold Nanoparticles Inhibit Extravasation of Canine Osteosarcoma Cells in the Ex Ovo Chicken Embryo Chorioallantoic Membrane Model.

Canine osteosarcoma (OS) is an aggressive bone tumor with high metastatic potential and poor prognosis, mainly due to metastatic disease. Nanomedicine-based agents can be used to improve both primary and metastatic tumor treatment. Recently, gold nanoparticles were shown to inhibit different stages of the metastatic cascade in various human cancers. Here, we assessed the potential inhibitory effect of the glutathione-stabilized gold nanoparticles (Au-GSH NPs) on canine OS cells extravasation, utilizing the ex ovo chick embryo chorioallantoic membrane (CAM) model. The calculation of cells extravasation rates was performed using wide-field fluorescent microscopy. Transmission electron microscopy and Microwave Plasma Atomic Emission Spectroscopy revealed Au-GSH NPs absorption by OS cells. We demonstrated that Au-GSH NPs are non-toxic and significantly inhibit canine OS cells extravasation rates, regardless of their aggressiveness phenotype. The results indicate that Au-GSH NPs can act as a possible anti metastatic agent for OS treatment. Furthermore, the implemented CAM model may be used as a valuable preclinical platform in veterinary medicine, such as testing anti-metastatic agents.

The Characteristics of Quark Cheese Made from Buttermilk during Refrigerated Storage.

The dairy industry releases huge amounts of by-products. One of them is buttermilk, obtained during butter production. This by-product is characterized by high nutritional and technological value and is finding more and more applications in food production. This study aimed to produce and analyze the characteristics of quark cheese obtained entirely from buttermilk during 3-week refrigerated (4 ± 1 °C) storage. Four kinds of sour buttermilk were used: two from industrial butter production, and another two from butter production at laboratory scale. Laboratory buttermilk differs in the kind of starter culture used in the production. The evaluation of cheese quality properties included physicochemical analyses, texture measurement, and sensory assessment. The results showed that the kind of buttermilk used in production influences the acidity, total solids, textural characteristics, and fat content of the obtained quark cheeses. All obtained cheeses had very high sensory quality throughout the storage period. The study indicates that buttermilk may be successfully used as a substitution for milk in quark cheese production.

Enhanced Cytotoxic Effect of Doxorubicin Conjugated to Glutathione-Stabilized Gold Nanoparticles in Canine Osteosarcoma-In Vitro Studies.

Osteosarcoma (OSA) is the most common malignant bone neoplasia in humans and dogs. In dogs, treatment consists of surgery in combination with chemotherapy (mostly carboplatin and/or doxorubicin (Dox)). Chemotherapy is often rendered ineffective by multidrug resistance. Previous studies have revealed that Dox conjugated with 4 nm glutathione-stabilized gold nanoparticles (Au-GSH-Dox) enhanced the anti-tumor activity and cytotoxicity of Dox in Dox-resistant feline fibrosarcoma cell lines exhibiting high P-glycoprotein (P-gp) activity. The present study investigated the influence of Au-GSH-Dox on the canine OSA cell line D17 and its relationship with P-gp activity. A human Dox-sensitive OSA cell line, U2OS, served as the negative control. Au-GSH-Dox, compared to free Dox, presented a greater cytotoxic effect on D17 (IC50 values for Au-GSH-Dox and Dox were 7.9 µg/mL and 15.2 µg/mL, respectively) but not on the U2OS cell line. All concentrations of Au-GSH (ranging from 10 to 1000 µg/mL) were non-toxic in both cell lines. Inhibition of the D17 cell line with 100 µM verapamil resulted in an increase in free Dox but not in intracellular Au-GSH-Dox. The results indicate that Au-GSH-Dox may act as an effective drug in canine OSA by bypassing P-gp.

Buttermilk ice cream-New method for buttermilk utilization.

Buttermilk, the by-product of butter production, due to good technological features and excellent nutritional and health-promoting properties finds more and more applications in food industry. Considerable amount of polar lipids causes that buttermilk exhibits emulsifying and stabilizing effect and may be used to improve the product quality. The study aimed to design new kind of ice cream, in which all milk is substituted by buttermilk. Within the study, we compared physicochemical parameters, color, texture, and sensory properties of control milk ice cream (C), ice cream from sweet buttermilk (SB), and ice cream from cultured buttermilk (CB). Ice cream was tested on the production day, and some characteristics were tested also after 14 and 28 days of storage at -18 ± 1°C. The study showed that samples of ice cream from cultured buttermilk had the highest acidity and were the most resistant to melting. The samples did not differ in over-run value. The use of buttermilk influenced the texture of ice cream and product from sweet buttermilk had the highest stickiness during the storage. The color analysis showed that the highest lightness parameter had ice cream from cultured buttermilk, while samples from sweet buttermilk had the most greenish-yellow characteristics. All the obtained products had good sensory characteristics, only cultured buttermilk ice cream slightly deteriorated after 28 storage days. Good quality properties cause that buttermilk may be successfully used as substitution of milk in ice-cream formula and may improve its quality by exhibiting of some emulsifying stabilizing effect.

Probiotic fermented beverages based on acid whey.

Acid whey is a byproduct of cheesemaking that is difficult to use because of its low pH and less-favorable processing properties compared with rennet whey. The aim of this study was to evaluate the qualities of fermented beverages made using acid whey. In manufacturing the beverages, we used probiotic cultures Lactobacillus acidophilus LA-5 or Bifidobacterium animalis ssp. lactis BB-12. The production process included combining pasteurized acid whey with UHT milk, unsweetened condensed milk, or skim milk powder. We introduced milk to enrich casein content and obtain a product with characteristics similar to that of fermented milk drinks. The products were stored under refrigerated conditions (5 ± 1°C) for 21 d. During storage, we assessed the beverages' physicochemical properties and organoleptic characteristics. The properties of the beverages depended on their composition, microbial culture, and storage time. Beverages containing L. acidophilus had higher acidity, which increased during storage; the acidity of samples containing B. animalis was more stable. Beverages made with skim milk powder (La1 and Bb1) had higher acetaldehyde content, but this parameter decreased in all samples during storage. The hardness of the samples did not change during storage and was highest in beverage La3, made from whey, condensed milk, and L. acidophilus. Beverage La2, made from whey, milk, condensed milk, and L. acidophilus, had the best sensory properties. The whey beverages we developed provided a good medium for the probiotic bacteria; bacteria count throughout the storage period exceeded 8 log cfu/mL, distinctly higher than the minimum therapeutic dose.

Characteristics of lactose-free frozen yogurt with κ-carrageenan and corn starch as stabilizers.

Frozen yogurt is a type of dairy product that is considered to be a more healthful alternative to conventional ice cream due to its lower fat content and the presence of viable lactic acid bacteria. Lactose-free products are a growing trend in the dairy industry, and lactose-free yogurts and ice creams can both be found on the market. However, lactose-free frozen yogurt has not yet reached the market. In this study, we aimed to investigate the effect of adding κ-carrageenan (0.05, 0.1, and 0.15%) and corn starch (1, 2, and 3%) on acidity, texture, viscosity, overrun, melting properties, color attributes, and sensory characteristics of lactose-free frozen yogurts. Lactose was reduced by enzymatic hydrolysis during the fermentation process. The effectiveness of the hydrolysis was measured by HPLC, and lactose was reduced to 0.05% after 80 min of incubation with the enzyme. The addition of stabilizers did not change overrun and melting properties of frozen yogurt, but it did affect pH, titratable acidity, and color parameters. The product with 0.15% κ-carrageenan had the highest hardness and stickiness values. Moreover, κ-carrageenan had a positive effect on sensory attractiveness of lactose-free frozen yogurt, and it reduced the coarse texture in comparison with the control without stabilizers. A lactose-free frozen yogurt with good quality and nutritional characteristics was produced, particularly with the use of κ-carrageenan as stabilizer.

Thermal Dependence of Optical Parameters of Thin Polythiophene Films Blended with PCBM.

The main purpose of this work is to show the thermal dependence of the refractive and extinction indices of conjugated polymer films used in optoelectronics devices. Herein, we present the results of optical investigations performed for the following polymers: poly(3-hexylthiophene) (P3HT), poly(3-octylthiophene) (P3OT), and their blends with [6,6]-phenyl C61 butyric acid methyl ester (PCBM). For our analysis, we chose well-known polythiophenes such P3HT and P3OT, often used in photovoltaic cells. Our addition of PCMB to the polythiophenes allows their conversion efficiency to be increased. This paper presents the results of our investigation determining the spectral dispersion of optical constants in a wavelength range of 190⁻1700 nm by using spectroscopic ellipsometry (SE). Furthermore, we show the temperature dependence of the refractive indices of polythiophene films for a heating and a cooling process in the temperature range 25⁻130 °C. Additionally, thermo-optic coefficients and an optical gap were established and are presented in the paper, followed by a discussion on the conditions of the thermal stability of polythiophene blends and reversibility issues in thermal processes. Our paper presents a new and fresh analysis of depolarization beams after their reflection from the studied films. The paper presents the results of thermo-optical studies of polymer blends which have not been included in previously published works.

3D chick embryo chorioallantoic membrane model as an in vivo model to study morphological and histopathological features of feline fibrosarcomas.

BACKGROUND: The chick embryo chorioallantoic membrane (CAM) model is well described in human medicine as a cost-effective, easy to perform preclinical oncological model for observing pro- and antiangiogenic response, tumor biology and metastasis. The main objective of this article was to present the modification of the CAM assay in order to evaluate tumor growth from two feline fibrosarcoma cell lines (FFS1, FFS3) and describe their morphological and histopathological features. RESULTS: The authors described morphological and histopathological features of two feline fibrosarcoma cell lines (FFS1 and FFS3) grown on the CAM. Tumors from the FFS1 cell line showed high malignancy (grade III), while tumors from the FFS3 cell line were grade II. Proliferation markers (Ki-67 and PCNA) were determined and the positive correlation between PCNA and tumor grade (r = 0.8247; p < 0.001) was demonstrated, as opposed to Ki-67. CONCLUSIONS: The results obtained indicate that PCNA may be helpful to evaluate the tumor grade, better than Ki-67, for feline fibrosarcomas. However, further investigations of proliferation marker, in bigger number of feline spontaneous fibrosarcomas and feline fibrosarcomas grown on the CAM from different cell lines, are needed to confirm these observations.

Experimental tumor growth of canine osteosarcoma cell line on chick embryo chorioallantoic membrane (in vivo studies).

The chick embryo chorioallantoic membrane (CAM) model is extensively used in human medicine in preclinical oncological studies. The CAM model has several advantages: low cost, simple experimental approach, time saving and following "3R principles". Research has shown that the human osteosarcoma cell lines U2OS, MMNG-HOS, and SAOS can form tumors on the CAM. In veterinary medicine, this has been described only for feline fibrosarcomas, feline mammary carcinomas and canine osteosarcomas. However, in case of canine osteosarcomas, it has been shown that only non-adherent osteosarcoma stem cells isolated from KTOSA5 and CSKOS cell lines have the ability to form microtumors on the CAM after an incubation period of 5 days, in contrast to adherent KTOSA5 and CSKOS cells. In the presented study, we have proven that the commercial adherent canine osteosarcoma cell line (D-17) can form vascularized tumors on the CAM after the incubation period of 10 days.

Assessment of health risk from organochlorine xenobiotics in goat milk for consumers in Poland.

The occurrence of organochlorine xenobiotics in goat milk is a one of bioindicators of environmental pollution, and, consequently, food contamination. This study estimates contamination level of goat milk produced at two organic farms in years 2009-2013. Analysis covered determination of 18 organochlorine pesticides, including HCH isomers (α, ß, γ, δ), DDT and its metabolites, endosulfan and its derivatives, and methoxychlor. Pesticide content was determined using GCMS method. The detected levels of organochlorine residues in goat milk were low, in most cases below 8 ng g(-1) lipids. Among HCH isomers, γ- and ß-HCH occurred in the highest concentrations, up to 4.85 ng g(-1) lipids. While among DDT metabolites p,p'-DDD dominated, up to 7.86 ng g(-1) lipids. The detected residues were below the current maximum residue limits (MRLs) for the pesticides. Considering the average milk consumption in Poland, the goat milk from both farms was safe for consumers' health. The lifetime average daily dose (LADD) for the sum of the compounds under study ranged within 1.73 × 10(-5)-1.06 × 10(-4) mg kg bw(-1) d(-1) and were well below the acceptable daily intake (ADI) for particular compounds. This was also confirmed by the values of hazard quotient (HQ), which were very low and ranged within 3.42 × 10(-3)-5.55 × 10(-2).

Mercury bioaccumulation in cartilaginous fishes from Southern New England coastal waters: contamination from a trophic ecology and human health perspective.

This study examined total mercury (Hg) concentrations in cartilaginous fishes from Southern New England coastal waters, including smooth dogfish (Mustelus canis), spiny dogfish (Squalus acanthias), little skate (Leucoraja erinacea), and winter skate (Leucoraja ocellata). Total Hg in dogfish and skates were positively related to their respective body size and age, indicating Hg bioaccumulation in muscle tissue. There were also significant inter-species differences in Hg levels (mean ± 1 SD, mg Hg/kg dry weight, ppm): smooth dogfish (3.3 ± 2.1 ppm; n = 54) > spiny dogfish (1.1 ± 0.7 ppm; n = 124) > little skate (0.4 ± 0.3 ppm; n = 173) âˆ¼ winter skate (0.3 ± 0.2 ppm; n = 148). The increased Hg content of smooth dogfish was attributed to its upper trophic level status, determined by stable nitrogen (δ(15)N) isotope analysis (mean δ(15)N = 13.2 ± 0.7‰), and the consumption of high Hg prey, most notably cancer crabs (0.10 ppm). Spiny dogfish had depleted δ(15)N signatures (11.6 ± 0.8‰), yet demonstrated a moderate level of contamination by foraging on pelagic prey with a range of Hg concentrations, e.g., in order of dietary importance, butterfish (Hg = 0.06 ppm), longfin squid (0.17 ppm), and scup (0.11 ppm). Skates were low trophic level consumers (δ(15)N = 11.9-12.0‰) and fed mainly on amphipods, small decapods, and polychaetes with low Hg concentrations (0.05-0.09 ppm). Intra-specific Hg concentrations were directly related to δ(15)N and carbon (δ(13)C) isotope signatures, suggesting that Hg biomagnifies across successive trophic levels and foraging in the benthic trophic pathway increases Hg exposure. From a human health perspective, 87% of smooth dogfish, 32% of spiny dogfish, and <2% of skates had Hg concentrations exceeding the US Environmental Protection Agency threshold level (0.3 ppm wet weight). These results indicate that frequent consumption of smooth dogfish and spiny dogfish may adversely affect human health, whereas skates present minimal risk.

Assessment of daily intake of organochlorine pesticides from milk in different regions of Poland.

The common occurrence of organochlorine compounds in the environment, food and human tissues may constitute a serious threat to human health. The method of gas chromatography-mass spectroscopy was used to determine the content of pesticides in 15 samples of raw cow's milk from different regions of Poland. The results revealed high levels of p,p'-DDE, p,p'-DDT, heptachlor and aldrin. The studied milk contained lindane in average concentrations within the maximum limits. Although in 20% of all samples tested, the concentration of lindane exceeded permissible limits, while in 15% of samples the content of Σ DDT was too high. But the average daily consumption of milk containing organochlorine pesticides poses no direct threat to human health, because daily intake (DI) for all compounds were below the acceptable daily intake (ADI). Attention should be paid to the exposure of consumers to pesticide residues from other dairy foods.