Clinoptilolite has antiviral, antibacterial, anti-inflammatory, antidiabetic, and anticancer properties due to its biological activities. In various cancer cell culture studies, it has been reported effective against tumour cells and gave positive results in treatment of various tumours in dogs. No study was found on the effects of the nanoparticulate form, nanoclinoptilolite, on cancer cells. The aim of this study was to determine its cytotoxic and apoptotic effects in canine osteosarcoma (OSA) cell culture. Doses at 50% inhibitory concentration were determined by measuring the dose- and duration-dependent cytotoxicity of nanoclinoptilolite on canine D-17 osteosarcoma cells by methylthiazol tetrazolium (MTT) test for 24 h, 48 h, and 72 h. Murine caspase-3 and -7 activity and expression levels of the BAX and BCL2 genes were measured using RT-PCR to investigate the apoptotic effect. Nanoclinoptilolite decreased cell viability and induced caspase-3- and -7-mediated apoptosis in treated canine OSA cells. Furthermore, its application to canine OSA cells downregulated the expression of BCL2 and upregulated the expression of proapoptotic BAX. Clinoptilolite, which was previously demonstrated to have anticancer properties, decreased cell viability effectively and rapidly and increased the apoptotic cell ratio in a novel use in nanoparticle form, exhibiting this effect by increasing the BAX/BCL2 ratio.

Although hyaluronic acid (HA) has been developed as a nanoparticle (NP; 320-400 nm) for a drug delivery system, the tissue targeting efficacy and the pharmacokinetics of HA-NPs are not yet fully understood. After a dose of 5 mg/kg of cyanine 5.5-labeled HA-NPs or HA-polymers was intravenously administrated into mice, the fluorescence was measured from 0.5 h to 28 days. The HA-NPs fluorescence was generally stronger than that of HA-polymers, which was maintained at a high level over 7 days in vivo, after which it gradually decreased. Upon ex vivo imaging, liver, spleen, kidney, lung, testis and sublingual gland fluorescences were much higher than that of other organs. The fluorescence of HA-NPs in the liver, spleen and kidney was highest at 30 min, where it was generally maintained until 4 h, while it drastically decreased at 1 day. However, the fluorescence in the liver and spleen increased sharply at 7 days relative to 3 days, then decreased drastically at 14 days. Conversely, the fluorescence of HA-polymers in the lymph node was higher than that of HA-NPs. The results presented herein may have important clinical implications regarding the safety of as self-assembled HA-NPs, which can be widely used in biomedical applications.

Temporal and subcellular distributions of hyaluronic acid (HA) as a degradable nanoparticle (NP) in animals were investigated to determine if HA-NP could be utilized as an appropriate drug delivery system. After mice were intravenously injected with 5 mg/kg of Cy5.5-labeled HA-NP sized 350-400 nm or larger HA-polymers, the fluorescence intensity was measured in all homogenized organs from 0.5 h to 28 days. HA-NP was greatly detected in spleen, liver and kidney until day 28, while it was maintained at low levels in other organs. HA-polymer was observed at low levels in all organs. HA-NP quantities in spleen and liver were reduced until day 3, but increased sharply between days 3 and 7, then decreased again, while their HA-polymers were maintained at low levels until day 28. In kidneys, both HA-NP and HA-polymer showed high levels after 0.5 h of administration, but steadily decreased until day 28. According to ultrastructural analyses, HA-NP was engulfed in Kupffer cells of liver and macrophages of spleen and kidney at day 1 and was accumulated in the cytoplasm of kidney tubular cells at day 7. Overall, these findings suggest that HA-NP could be considered a desirable drug carrier in the liver, kidney, or spleen.

OBJECTIVE To compare ultracentrifugation, precipitation, and membrane affinity chromatography methods for isolation of extracellular vesicles (EVs) from canine plasma samples and to identify suitable reference genes for incorporation into a quantitative reverse transcription PCR assay of microRNA expression in plasma EVs of healthy dogs. ANIMALS 6 healthy Beagles. PROCEDURES Plasma samples were obtained from each dog, and EVs were isolated from 0.3 mL of these samples via ultracentrifugation, precipitation, and membrane-affinity chromatographic methods. Nanoparticle tracking analysis was performed to determine the concentration and size distribution of EVs isolated by the ultracentrifugation method. Expression levels (cycle threshold values) of 4 microRNAs (let-7a, miR-16, miR-26a, and miR-103) were then compared by means of quantitative reverse transcription PCR assay. Three statistical programs were used to identify the microRNAs most suitable for use as reference genes. RESULTS Results indicated that ultracentrifugation was the most stable of all 3 methods for isolating microRNAs from 0.3 mL of plasma. Nanoparticle tracking revealed that EV samples obtained by the ultracentrifugation method contained a mean ± SD of approximately 1.59 × 10(10) vesicles/mL ± 4.2 × 10(8) vesicles/mL. Of the 4 microRNAs in plasma EVs isolated by ultracentrifugation, miR-103 was the most stable. CONCLUSIONS AND CLINICAL RELEVANCE The ultracentrifugation method has potential as a stable method for isolating EVs from canine plasma samples with a high recovery rate, and miR-103 may provide the most stable reference gene for normalizing microRNA expression data pertaining to plasma EVs isolated by ultracentrifugation.

Many scolicidal agents have been used to destroy fertile protoscolices, but these scolicidal agents have side effects, highlighting the need for research on effective and non-toxic replacement scolicidal agents. Gold nanoparticles (AuNPs) are biocompatible and non-toxic. The current study examined the effects of AuNPs in killing the protoscolices of Echinococcus granulosus in vitro using eosin staining. The protoscolices were treated with 0.2, 0.4, 0.8, or 1.0 mg/mL of AuNPs for 15, 30, 45, or 60 minutes. A concentration of 1.0 mg/mL was the most efficient in killing the protoscolices after 60 minutes exposure, reaching 96%, followed by 0.8 mg/mL (84.5%), whereas 0.4 and 0.2 mg/mL of AuNPs achieved a death rate of 76.8% and 68.5%, respectively. The loss of the protoscolices was lower at shorter exposure times with the same concentration of AuNPs and increased as the AuNP concentration was increased at the same exposure time. Significant differences were found between the different groups compared to the control group.

Cryptosporidium is a primary cause of waterborne epidemics, despite being previously considered only an opportunistic pathogen. The disease is associated with significant economic losses in humans and animals that are brought on by diarrhea, which frequently causes dehydration. Contact with diseased people or animals, as well as polluted water, is the major cause of infection. Different drugs are used to control the parasites. Nitazoxanide (NTZ), which is an anti-protozoan and anti-viral drug, can be used to control helminths, viruses, and protozoan parasites as a broad-spectrum antibiotic and has been approved by the food and drug authority (FDA). However, the problem is the development of resistance over a period of time in these parasites. Nanoparticles have received significant attention as possible anti-parasitic agents in recent years. By directing medications to specific cellular locations, targeted drug delivery minimizes the side effects of medications. Nanoparticles have demonstrated effectiveness against different Cryptosporidium species. Nanoparticles loaded with NTZ are found to be an effective remedy for C. parvum in young ones and decrease the oocyst count shed in the stools. Additionally, silver nanoparticles have proven to be effective against C. parvum by releasing silver ions that breach the cell wall of the oocyst, causing the escape of intracellular contents and the destruction of sporozoites within the oocyst. Implementing tiny particles for the purification of consuming water from Cryptosporidium is an economical and environmentally sustainable process. However, the use of nanoparticles in medicine requires more research. [J Adv Vet Anim Res 2023; 10(4.000): 704-719]

Objective: This study assessed the effect of cellulose sheets fortified with Natamycin-loaded algi¬nate nanoparticles on the growth of toxigenic Aspergillus flavus and aflatoxin production on the superficial layer of Egyptian Romy cheese after 12 weeks of maturation. Materials and Methods: Toxigenic A. flavus (GenBank accession No. MT645073) was inoculated into the outer surface of Egyptian Romy cheese (at 5 log CFU/gm) and wrapped with a cellulose sheet fortified with Natamycin-loaded alginate nanoparticles. Unwrapped control contaminated Romy wheels were made as well as non-contaminated wrapped cheese wheels for sensory eval¬uation. Romy cheese wheels were stored at a temperature similar to commercial methods for 12 weeks. Fungal counts were enumerated during this time, and enzyme-linked immune sorbent assay detected aflatoxin after the 4th week of maturation storage. Results: In cheese samples covered with cellulose sheets containing Natamycin-loaded alginate nanoparticles, the fungal count was reduced by 2 log approximately in contrast to control samples after the 2nd week of storage. However, within the 8th week of storage, the greatest significant reduction (p < 0.05) was seen where fungal growth was hindered entirely to the end of the rip¬ening period. The mean values for taste, color, flavor, and overall acceptability were 4, 4.7, 4.09, and 4.3, respectively. Furthermore, in the treated samples, the total aflatoxin concentration was decreased by 78.6% relative to the untreated control one. Conclusion: Using cellulose sheets fortified with Natamycin-loaded alginate nanoparticles in Egyptian Romy cheese wrapping could be an effective way of controlling A. flavus and subsequent aflatoxin production without influencing the typical taste, color, flavor, and overall appearance of traditional Romy cheese. [J Adv Vet Anim Res 2021; 8(1.000): 58-63]