Unveiling of the antileishmanial activities of Linalool loaded zinc oxide nanocomposite through its potent antioxidant and immunomodulatory effects.

This study aimed to produce linalool loaded zinc oxide nanocomposite (LZNPs) and assess its in vitro and in vivo antileishmanial effects against Leishmania major. LZNPs was produced through the synthesis of an ethanolic solution containing polyvinyl alcohol. The average size of LZNPs was determined to be 105 nm. The findings indicated that LZNPs displayed significant (p < 0.01) antileishmanial effects on promastigotes and amastigotes. Following exposure of promastigotes to LZNPs, there was a notable rise in the percentage of early and late apoptotic cells from 9.0 to 57.2 %. The gene expression levels of iNOS, IFN-γ, and TNF-α in macrophages were upregulated in a dose-dependent approach following exposure to LZNPs. LZNPs alone and in conjunction with glucantime (Glu) resulted in a reduction in the diameter and parasite load of CL lesions in infected mice. Treatment of the CL-infected mice with LZNPs at 25 and 50 mg/kg mainly in combination with Glu-reduced the tissue level of malondialdehyde (MDA), increased both gene and protein expression of the antioxidant enzymes as well as raised the expression level of IFN-γ and IL-12 cytokines, whereas caused a significant reduction in the expression level of IL-4. The present study shows that LZNPs has potent antileishmanial effects and controls CL in a mice model through its antioxidant and immunomodulatory properties. Further investigation, especially in clinical trials, could explore the potential use of this nanocomposite in managing and treating CL.

Potent In Vitro and In Vivo Effects of Stachys lavandulifolia Methanolic Extract against Toxoplasma gondii Infection.

The present study aimed to evaluate the in vitro, in vivo, and safety of Stachys lavandulifolia Vahl. methanolic extract (SLME) against acute toxoplasmosis caused by Toxoplasma gondii RH strain in mice. METHODS: MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to evaluate the in vitro effect of the SLME on T. gondii tachyzoites. Totally, 72 male BALB/c mice (40 mice for in vivo evaluation of SLME and 32 mice for its toxicity effects on liver and kidney serum enzymes) were used for the present investigation. At first, 40 mice were orally pre-treated with the SLME at doses of 25, 50, and 75 mg/kg/day for two weeks. Mice were checked daily, and the rate of survival and the mean number of tachyzoites were recorded. Liver lipid peroxidation (LPO) and nitric oxide (NO) levels, the effects on kidney and liver function, as well as the expression level of the proinflammatory cytokines such as interleukin-1ß (IL-1ß) and interferon-γ (IFN-γ), were studied by the quantitative real-time PCR. Flow cytometry analysis was performed on the effects of SLME on the detection of apoptotic and necrotic cells in T. gondii tachyzoites. RESULTS: The SLME at the concentrations 75 and 150 µg/mL completely killed the tachyzoites after 2 hr of incubation. SLME at 25, 50, and 75 mg/kg/day increased the survival rate of infected mice by the sixth, seventh, and eighth days, respectively. SLME also significantly (p < 0.05) decreased the LPO and NO levels and upregulated the IL-1ß and IFN-γ mRNA gene expression levels, whereas no considerable change was observed in the serum level of kidney and liver enzymes. Flow cytometry analysis revealed the prompted early and late apoptosis after exposure to T. gondii tachyzoites with various concentrations of SLME. CONCLUSION: We found the relevant in vitro anti-Toxoplasma effects of SLME against T. gondii. Moreover, the results confirmed the promising in vivo prophylactic effects of SLME. SLME provokes the innate immune system, induces apoptosis, modulates the proinflammatory cytokines, and inhibits hepatic injury in infected mice. With all these descriptions, further surveys are required to support these findings and elucidate this plant's possible mechanisms of action.

Therapeutic Potential and Safety of the Cinnamomum zeylanicum Methanolic Extract Against Chronic Toxoplasma gondii Infection in Mice.

Background: This experimental study determined the in vitro, in vivo, and toxicity effects of Cinnamomum zeylanicum methanolic extract (CZME) against Toxoplasma gondii infection. Methods: The in vitro activity of CZME T. gondii tachyzoites was studied by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Infected mice were treated with CZME for two weeks at doses of 20, 40, and 60 mg/kg/day. Then, the therapeutic effects of CZME were evaluated by assessing the mean number and mean size of T. gondii tissue cysts, oxidant-antioxidant enzymes, pro-inflammatory cytokines, and mRNA expression levels of bradyzoite surface antigen 1 (BAG1) by real-time PCR. Results: CZME significantly (p <0.001) increased the mortality rate of parasites in a dose- and time-dependent response. The mean number of intracellular tachyzoites was significantly reduced after CZME therapy. The treatment of infected mice with CZME resulted in a significant (p <0.001) downregulation of BAG1 and the level of lipid peroxidation (LPO) and nitric oxide (NO) as oxidative stress markers. However, a considerable rise (p <0.05) was found in the levels of antioxidant markers such as glutathione peroxidase (GPx), catalase enzyme (CAT), and superoxide dismutase enzyme activity (SOD). In a dose-dependent response, after treatment of infected mice with CZME, the level of pro-inflammatory cytokines of IFN-γ, IL-1ß, and IL-12 was considerably elevated. CZME had no significant cytotoxicity on Vero cells, with a 50% cytotoxic concentration of 169.5 ± 5.66 µg/ml. Conclusion: The findings confirmed the promising therapeutic effects of CZME on chronic toxoplasmosis in mice. Nevertheless, further investigations must confirm these results, elucidate its precise mechanisms, and examine its effectiveness in human volunteers.

Role of Aromatherapy as a Natural Complementary and Alternative Therapy in Cardiovascular Disease: A Comprehensive Systematic Review.

Background: The purpose of this study was a comprehensive review of studies on the effect of aromatherapy with plant essential oils on the improvement of some conditions, for example, anxiety, stress, sleep quality, fatigue, and pain in people with cardiovascular disease. Materials and Methods: We carried out this systematic review based on the instructions of Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Ethical agreement was not necessary as main data have not been collected. During March 2022, we searched the main English databases, for example, Google Scholar, Web of Sciences, EMBASE, EBSCO, ScienceDirect, Scopus, and PubMed/MEDLINE, with limitation to human clinical trials. For this study, no time limit was applied for the publication of articles. Results: Out of 1380 papers, 52 papers up to March 2022 were eligible for review in this systematic review. Based on the obtained results, the most widely used medicinal plants for aromatherapy in patients with cardiovascular diseases were Lavandula angustifolia (lavender, 55.7%), Rosa damascena (Damask rose, 11.5%), and Mentha piperita (peppermint, 5.8%), respectively. Most studies have been performed on the effect of aromatherapy on coronary angiography (21 papers, 40.4%), followed by artery bypass graft surgery (14 studies, 26.9%), and cardiac patients (5 studies, 9.6%). Most studies on the effect of aromatherapy in cardiovascular diseases were performed on anxiety (31 papers, 59.6%), sleep quality (8 studies, 15.4%), and hemodynamic parameters (6 studies, 11.5%), respectively. Conclusion: This study systematically reviewed the effects of aromatherapy in patients with cardiovascular diseases. The review of studies showed that lavender, Damask rose, and peppermint are the most frequents plants used for aromatherapy, whereas they significantly improved some illnesses and conditions, especially anxiety and sleep quality. Therefore, it can be concluded that cardiologist can used aromatherapy as a natural complementary and alternative therapy particularly with lavender, Damask rose, and peppermint to improve quality of life and some conditions such as anxiety and sleep quality.

Antiparasitic effects of Elettaria cardamomum L. essential oil and its main compounds, 1-8 Cineole alone and in combination with albendazole against Echinococcus granulosus protoscoleces.

Background: The present investigation aims to determine the chemical structure and protoscolicidal effects of Elettaria cardamomum L. essential oil (ECEO) and its main compounds 1-8 cineole alone and along with albendazole (ALZ) against Echinococcus granulosus protoscoleces in vitro and ex vivo. We also decided to evaluate some cellular mechanisms such as the apoptotic activity and the permeability of plasma membrane of protoscoleces treated with ECEO and 1-8 cineole. Methods: Hydatid cyst protoscoleces were divided into seven groups including protoscoleces treated with ECEO 50 µl/mL (T1), protoscoleces treated with ECEO 100 µl/mL (T2), protoscoleces treated with ECEO 200 µl/mL (T3), protoscoleces treated with 1-8 cineole 100 µg/mL (T4), protoscoleces treated with 1-8 cineole 200 µg/mL (T5), protoscoleces treated with 1-8 cineole 100 µg/mL + albendazole 50 µg/mL (T6), and protoscoleces treated with 1-8 cineole 200 µg/mL + albendazole ALZ-50 µg/mL (T7). The viability of protoscoleces were recorded by eosin staining examination. Moreover, the induction of apoptosis and the plasma membrane permeability of the protoscoleces treated with ECEO and 1-8 cineole were evaluated. Results: The highest protoscolicidal effect of ECEO was observed at the dose of 200 µl/ml (T3). 1,8-Cineole alone and combined with ALZ, particularly at the dose of 200 µg/ml (T5 and T7), destroyed the 100% protoscolices after 10 min incubation. The ECEO (T1-T3) and 1-8 cineole alone (T4 and T5) and in combination with ALZ (T6 and T7) took longer to display their protoscolicidal effect ex vivo. The obtained results of relative fuorescent items exhibited that the protoscoleces incubated with ECEO and 1,8-Cineole, alter the permeability of plasma membrane by Sytox Green with increasing the concentration. The findings revealed exhibited that ECEO and 1,8-Cineole increasingly and dose-dependently induced activation of caspase-3 enzyme ranging from 6.8 to 23.3%. Conclusion: Our obtained results revealed that ECEO and its main compound, 1,8-Cineole exhibited the potent protoscolicidal in vitro and ex vivo; and if more research is done on their efficacy and toxicity in animal models and even clinical setting, it can be suggested as a protoscolicidal agent to use during hydatid cyst surgery.

Molecular Epidemiology of Theileria annulata in Cattle from Two Districts in Punjab (Pakistan).

The present study was designed to report the molecular prevalence of T. annulata in cattle blood samples collected from Punjab in Pakistan. A total of 428 cattle blood samples were collected from Districts Lodhran (n = 218) and Dera Ghazi Khan (n = 210). The prevalence of T. annulata was determined by the amplification of a fragment from its cytochrome b gene and parasite prevalence was significantly higher (p = 0.03) in the blood samples of cattle collected from Dera Ghazi Khan (70/210; 33%) as compared to Lodhran (52/218; 24%). Presence of T. annulata was also confirmed by the amplification of a fragment from their 30 kDa gene. The amplified PCR products of both genes were confirmed by DNA sequencing and these partial DNA sequences were submitted to GenBank. Phylogenetic analysis revealed that amplified partial gene sequences resembled previously reported T. annulata sequences in cattle from India, China, Iran, Tunisia, Turkey and Egypt. The incidence of T. annulata infection was higher in Sahiwal cattle (p = 0.04) than the other enrolled cattle breed from Dera Ghazi Khan. Female cattle from Lodhran (p = 0.02), while males (p = 0.02), animals housed in close compounds (p = 0.04), animals with a tick burden (p = 0.005) and farms with only cattle (p = 0.01) in Dear Ghazi Khan were found to be more susceptible to T. annulata infection. We recommend that large-scale tick and tick-borne disease control strategies be implemented in both districts under investigation, especially in Dera Ghazi Khan.

A recent update about seroprevalence of ovine neosporosis in Northern Egypt and its associated risk factors.

Neospora caninum (Family: Sarcocystidae) is an obligate intracellular protozoan. It is one of the most critical abortifacients in ruminants. The seroprevalence of antibodies against N. caninum and its risk factors was investigated among 430 sheep from four North Egyptian governorates, Alexandria, Gharbia, Menofia, and Qalyubia, during the period from 2017 to 2018. Generally, the overall prevalence rate of N. caninum among sheep was 8.6%. The logistic regression analysis for the obtained data revealed that N. caninum increased significantly with age (OR = 2.4, 95% CI: 8.4-18.7) of the ewe (OR = 3.3, 95% CI: 7.6-14.9), particularly among sheep in contact with dogs (OR = 4.9, 95% CI: 7.5-14.3). Besides, locality, season, and pregnancy status of examined sheep had no significant effect on the appearance of N. caninum infection. the present findings confirm the presence of N. caninum among sheep in Egypt which probably play a role in reproductive failure in sheep. Therefore, sanitary measures and monitoring of the infection should be implemented to reduce the spreading of the infection.

Chitosan-Based Nanomaterials as Valuable Sources of Anti-Leishmanial Agents: A Systematic Review.

BACKGROUND: The current chemotherapy agents against various forms of leishmaniasis have some problems and side effects, including high toxicity, high cost, and the emergence of resistant strains. Here, we aimed to review the preclinical studies (in vitro and in vivo) on the anti-leishmanial activity of chitosan and chitosan-based particles against Leishmania spp. METHODS: This study was conducted based on the 06-PRISMA guidelines and registered in the CAMARADES-NC3Rs Preclinical Systematic Review and Meta-Analysis Facility (SyRF) database. Various English databases such as PubMed, Google Scholar, Web of Science, EBSCO, ScienceDirect, and Scopus were used to find the publications related to the anti-leishmanial effects of chitosan and its derivatives and other pharmaceutical formulations, without a date limitation, to find all the published articles. The keywords included "chitosan", "chitosan nanoparticles", "anti-leishmanial", "Leishmania", "leishmaniasis", "cutaneous leishmaniasis", "visceral leishmaniasis", "in vitro", and "in vivo". The language for data collection were limited to English. RESULTS: Of 2669 papers, 25 papers, including 7 in vitro (28.0%), 7 in vivo (28.0%), and 11 in vitro/in vivo (44.0%) studies conducted up to 2020 met the inclusion criteria for discussion in this systematic review. The most common species of Leishmania used in these studies were L. major (12, 48.0%), L. donovani (7, 28.0%), and L. amazonensis (4, 16.80%). In vivo, the most used animals were BALB/c mice (11, 61.1%) followed by hamsters (6, 33.3%) and Wistar rats (1, 5.5%), respectively. In vitro, the most used Leishmania form was amastigote (8, 44.4%), followed by promastigote (4, 22.2%), and both forms promastigote/amastigote (6, 33.3%). CONCLUSION: According to the literature, different types of drugs based on chitosan and their derivatives demonstrated considerable in vitro and in vivo anti-leishmanial activity against various Leishmania spp. Based on the findings of this review study, chitosan and its derivatives could be considered as an alternative and complementary source of valuable components against leishmaniasis with a high safety index. Nevertheless, more investigations are required to elaborate on this result, mainly in clinical settings.

Determination of Seroprevalence of Contagious Caprine Pleuropneumonia and Associated Risk Factors in Goats and Sheep Using Classification and Regression Tree.

Classification and Regression Tree (CART) analysis is a potentially powerful tool for identifying risk factors associated with contagious caprine pleuropneumonia (CCPP) and the important interactions between them. Our objective was therefore to determine the seroprevalence and identify the risk factors associated with CCPP using CART data mining modeling in the most densely sheep- and goat-populated governorates. A cross-sectional study was conducted on 620 animals (390 sheep, 230 goats) distributed over four governorates in the Nile Delta of Egypt in 2019. The randomly selected sheep and goats from different geographical study areas were serologically tested for CCPP, and the animals' information was obtained from flock men and farm owners. Six variables (geographic location, species, flock size, age, gender, and communal feeding and watering) were used for risk analysis. Multiple stepwise logistic regression and CART modeling were used for data analysis. A total of 124 (20%) serum samples were serologically positive for CCPP. The highest prevalence of CCPP was between aged animals (>4 y; 48.7%) raised in a flock size ≥200 (100%) having communal feeding and watering (28.2%). Based on logistic regression modeling (area under the curve, AUC = 0.89; 95% CI 0.86 to 0.91), communal feeding and watering showed the highest prevalence odds ratios (POR) of CCPP (POR = 3.7, 95% CI 1.9 to 7.3), followed by age (POR = 2.1, 95% CI 1.6 to 2.8) and flock size (POR = 1.1, 95% CI 1.0 to 1.2). However, higher-accuracy CART modeling (AUC = 0.92, 95% CI 0.90 to 0.95) showed that a flock size >100 animals is the most important risk factor (importance score = 8.9), followed by age >4 y (5.3) followed by communal feeding and watering (3.1). Our results strongly suggest that the CCPP is most likely to be found in animals raised in a flock size >100 animals and with age >4 y having communal feeding and watering. Additionally, sheep seem to have an important role in the CCPP epidemiology. The CART data mining modeling showed better accuracy than the traditional logistic regression.