Salwa Sami Younis

Toxoplasmosis is an immunologically complex disease, particularly in immunocompromised patients. Although there are several therapeutic regimens for such disease, the majority of them have many drawbacks. Therefore, it is of utmost importance to improve the current regimen in an effort to achieve a well-tolerated therapy while also enhancing the host immune response. Famous for their immunomodulatory effect, Lactobacillus delbrueckii and Lactobacillus fermentum probiotics were chosen to be evaluated in this study as an adjuvant therapy against the virulent RH Toxoplasma gondii (T. gondii) strain. Experimental mice were divided into control and treated groups. The control group was further subdivided into two groups: group I: 10 uninfected mice and group II: 20 infected untreated mice. The treated experimental group was subdivided into three groups (20 mice each); group III: sulfamethoxazole-trimethoprim (SMZ-TMP) treated, group IV: probiotics treated, and group V: SMZ-TMP combined w...

To assess the molluscicidal effect of the eco-friendly green synthesized neem silver nanoparticles (neem-Ag NPs) against Biomphalaria alexandrina, the snail intermediate host for Schistosoma mansoni, and their cercaricidal potential. Methods: Methanol extracts from neem fruits were used for green synthesis of neem-Ag NPs. The neem-Ag NPs were characterized using UV-visible absorption spectra, dynamic laser light scattering technique, and transmission electron microscopy. The potential molluscicidal effect against adult and juvenile Biomphalaria alexandrina and the effect of the sub-lethal concentration on hatching of snail eggs and Schistosoma mansoni cercariae were evaluated. Results: The surface plasmon resonance of neem-Ag NPs showed a sharp absorption peak at λ max = 518 nm together with multiple peaks. The hydrodynamic diameter was (77.15±34.53) nm, the polydispersity index (0.338±0.000) and the zeta-potential-14.07 mV. Moreover, transmission electron microscopy showed that the average size of the nanoparticles was (27±2) nm. Agglomeration was evident and a light-colored capping layer could be seen coating the nanoparticles. Juvenile snails (LC 50 : 0.83 ppm) were more susceptible to neem-Ag NPs than adults (LC 50 : 1.07 ppm). In addition, neem-Ag NPs and neem at LC 50 concentrations inhibited the egg-hatching of snails and showed cercaricidal activity in a timedependent manner. Conclusions: Neem-Ag NPs have lethal activities against Biomphalaria alexandrina snails and their eggs, as well as Schistosoma mansoni cercariae. Hence, neem-Ag NPs could be a potential agent to control schistosomiasis.

The impact of the laboratory induced Schistosoma mansoni with decreased PZQ sensitivity on the biological performance of its different developmental stages and the concomitant structural changes of adult worms' total proteins were investigated. PZQ exposed snails showed stoppage of cercarial shedding for eight weeks followed by progressive significant reduction of cercarial production along four successive weeks. In the vertebrate host, in comparison to Schistosoma mansoni susceptible isolate, inoculated cercariae with decreased PZQ sensitivity led to an evident decrease in male to female ratio associated with significant reduction in tissue egg counts and significant increase in dead egg percentage. Significant reduction in the fecundity was also determined. Interestingly, eggs from adult worms with decreased PZQ sensitivity showed two unique features as they found to be smaller and more spherical in addition to the observation of hourglass shaped miracidium in about 10% of the detected mature eggs. Proteomic analysis of adult worms with decreased sensitivity to PZQ using mass spectrometry revealed up-regulation of Ca 2+ ATPase 2 and Hsp70. This study can point to the increase incidence of the neuroschistosomiasis due to the small size eggs of Schistosoma mansoni with reduced PZQ sensitivity. These worms can also impact the epidemiology in the field. The study can also provide help to elucidate underlying potential molecular mechanisms of resistance that could lead to possible strategies to reverse drug resistance.

Background: Previous studies have reported involvement of Toxoplasma gondii (T. gondii) infections in the pathogenesis of some autoimmune diseases, such as polymyositis, rheumatoid arthritis, autoimmune thyroiditis, and Crohn’s disease. However, data on the association between T. gondii infections and Type 1 diabetes mellitus (T1DM) are still controversial. Therefore, in the present study, we aimed to investigate the pancreatic pathological changes in mouse models with acute and chronic toxoplasmosis and their association with T1DM. Materials and Methods: Three groups (10 mice each) of male Swiss Albino mice were used. One group of mice was left uninfected, whereas the second and third groups were infected with the acute virulent T. gondii RH strain and the chronic less virulent Me49 T. gondii strain, respectively. T. gondii-induced pancreatic pathological changes were evaluated by histopathological examination of pancreatic tissues. Moreover, the expression of insulin, levels of ca...

Background: Toxoplasmosis is a deleterious parasitic disease with harmful impact on both humans and animals. The present study was carried out to evaluate the antiparasitic effect of chloroquine (CQ), spiramycin (SP), and combination of both against the highly virulent RH HXGPRT (−) strain of Toxoplasma gondii (T. gondii) and to explore the mechanisms underlying such effect. Methods: We counted the tachyzoites in the peritoneal fluid and liver smears of mice and performed scanning and transmission electron microscopy and immunofluorescence staining of tachyzoites. Moreover, relative caspase 3 gene expression was measured by real time polymerase chain reaction of liver tissues and immunoassay of antiapoptotic markers [B cell lymphoma-2 (Bcl-2) and X-chromosome linked inhibitor of apoptosis (XIAP)] and interferon gamma (IFN-γ) was done in liver tissues by ELISA. In addition, we estimated serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) and performed histopathological examination of liver sections for scoring of inflammation. Results: We found that both CQ and CQ/SP combination significantly reduced parasitic load in the peritoneal fluid and liver smears, induced apical disruption of tachyzoites, triggered host cell apoptosis through elevation of relative caspase 3 gene expression and suppression of both Bcl-2 and XIAP. Also, they upregulated IFN-γ level, reduced serum AST and ALT, and ameliorated liver inflammation. Conclusions: Either of CQ and CQ/SP combination was more effective than SP alone against T. gondii with the CQ/ SP combination being more efficient. Therefore, adding CQ to other anti-Toxoplasma therapeutic regimens may be considered in future research.

Introduction: Praziquantel (PZQ) is the only commercially available drug for schistosomiasis. The current shortage of alternative effective drugs and the lack of successful preventive measures enhance its value. The increase in the prevalence of PZQ resistance under sustained drug pressure is, therefore, an upcoming issue. Objective: To overcome the tolerance to PZQ using nanotechnology after laboratory induction of a Schistosoma mansoni isolate with reduced sensitivity to the drug during the intramolluscan phase. Materials and methods: Shedding snails were treated with PZQ doses of 200 mg/kg twice/ week followed by an interval of one week and then repeated twice in the same manner. The success of inducing reduced sensitivity was confirmed in vitro via the reduction of cercarial response to PZQ regarding their swimming activity and death percentage at different examination times. Results: Oral treatment with a single PZQ dose of 500 mg/kg in mice infected with cercariae with reduced sensitivity to PZQ revealed a non-significant reduction (35.1%) of total worm burden compared to non-treated control mice. Orally inoculated PZQencapsulated niosomes against S. mansoni with reduced sensitivity to PZQ successfully regained the pathogen's sensitivity to PZQ as evidenced by measuring different parameters in comparison to the non-treated infected animals with parasites with reduced sensitivity to PZQ. The mean total worm load was 1.33 ± 0.52 with a statistically significant reduction of 94.09% and complete eradication of male worms. We obtained a remarkable increase in the percentage reduction of tissue egg counts in the liver and intestine (97.68% and 98.56%, respectively) associated with a massive increase in dead eggs and the complete absence of immature stages. Conclusion: PZQ-encapsulated niosomes restored the drug sensitivity against laboratoryinduced S. mansoni adult worms with reduced sensitivity to PZQ.

The impact of the laboratory induced Schistosoma mansoni with decreased PZQ sensitivity on the biological performance of its different developmental stages and the concomitant structural changes of adult worms' total proteins were investigated. PZQ exposed snails showed stoppage of cercarial shedding for eight weeks followed by progressive significant reduction of cercarial production along four successive weeks. In the vertebrate host, in comparison to Schistosoma mansoni susceptible isolate, inoculated cercariae with decreased PZQ sensitivity led to an evident decrease in male to female ratio associated with significant reduction in tissue egg counts and significant increase in dead egg percentage. Significant reduction in the fecundity was also determined. Interestingly, eggs from adult worms with decreased PZQ sensitivity showed two unique features as they found to be smaller and more spherical in addition to the observation of hourglass shaped miracidium in about 10% of the detected mature eggs. Proteomic analysis of adult worms with decreased sensitivity to PZQ using mass spectrometry revealed up-regulation of Ca 2+ ATPase 2 and Hsp70. This study can point to the increase incidence of the neuroschistosomiasis due to the small size eggs of Schistosoma mansoni with reduced PZQ sensitivity. These worms can also impact the epidemiology in the field. The study can also provide help to elucidate underlying potential molecular mechanisms of resistance that could lead to possible strategies to reverse drug resistance.

Toxoplasma gondii is a zoonotic intracellular protozoan parasite and its therapeutic limitations are one of its major problems. L-citrulline is an organic compound that has beneficial effects on many diseases. The purpose of this study was to assess the impact of L-citrulline, alone or in combination with sulfamethoxazole-trimethoprim (SMZ-TMP) on acute toxoplasmosis caused by Toxoplasma gondii RH virulent strain. In our study, 60 Swiss albino mice were divided into two main groups; the control group and the infected treated group, which was subdivided into group IIa: infected treated with L-citrulline, group IIb: infected treated with SMZ-TMP, and group IIc: infected treated with L-citrulline combined with SMZ-TMP. The effects of treatment were assessed by parasitological study, electron microscopic study of tachyzoites, and histopathological study of the liver. Moreover, ELISA measurement of the serum level of Interferon-gamma, Interleukin 10, Nitric oxide, and apoptotic markers was used. It was noticed that L-citrulline combined with SMZ-TMP significantly increased the survival time of infected mice with a significant decrease in the number of tachyzoites compared to the other groups. Moreover, it increased the levels of measured cytokines and serum anti-apoptotic proteins Bcl-2 and improved the extent of liver cell damage associated with a decrease in inflammatory infiltration. In conclusion, L-citrulline supplementation was found to be effective against acute toxoplasmosis, especially when combined with SMZ-TMP as it has multifactorial mechanisms; nitric oxide production, anti-inflammatory, anti-apoptotic, and immune stimulator.

The preventative effect of triclosan (TS) and TS liposomal nanoparticles was studied on the early establishment of chronic infection with Toxoplasma gondii (T. gondii). Swiss albino mice were orally infected with 10 cysts of avirulent ME49 strain of T. gondii, and 2 weeks later they were orally treated with dual daily doses of 200 mg/kg and 120 mg/kg TS and TS liposomes for 30 days; respectively. Effect of TS and TS liposomes was parasitologically and ultrastructurally evaluated, versus infected non-treated control. Their safety was biochemically assessed. Parasitologically, both TS and TS liposomes induced significant reduction in mice mortality, brain parasite burden and infectivity of cysts obtained from the brains of treated mice. Ultrastructurally, scanning electron microscopy of cysts obtained from infected mice treated with either TS or TS liposomes showed surface irregularities, protrusions and depressions. Transmission electron microscopy revealed disintegration of the cyst wall and vacuolation of the bradyzoites with disintegration of plasma membranes of both cysts and bradyzoites whether treated with TS or TS liposomes. Biochemical study reflected the safety of the TS and TS liposomes. Therefore, TS proved an effective, promising and safe preventive drug against early establishment of chronic toxoplasmosis. Loading TS on liposomes marginally enhanced its efficacy against T. gondii cysts yet allowed its use in a lower dose.

Schistosoma spp. Host-parasite interaction: One mystery concerning the intra-mammalian life of Schistosoma adults is its adaptation to the anaerobic glucose metabolism and lactate production. This was explained lately by the presence of Schistosoma aquaporins that are membrane-bound proteins for water transport. Aquaporins capability to transport lactate across the cell membrane was also elucidated thus solving the mystery in Schistosoma adaptation and survival [2]. Identification of novel diagnostic biomarkers: Several micro RNA (miRNA) types were isolated from sera of patients with schistosomiasis, e.g. miR-124-3p, Bantam miRNA, miR-2C-3p, miR-3488 and miR2a-5p. Since miRNAs are secreted from Schistosoma extracellular vesicles, they are considered diagnostic biomarkers for diagnosis and therapeutic monitoring [3,4]. In 2022, advances in bioinformatics held a revolution in the diagnosis of schistosomiasis. Genomic datasets of S. haematobium and S. japonicum allowed the investigators to compare between both genomes utilizing gene ontology. Notably, the latter is a bioinformatics approach unifying genetic representation across a species followed by further in silico identification of identical proteins. Accordingly, distinct S. japonicum genes were demonstrated and enlisted as potential diagnostic markers for infection by Asian schistosomiasis [5]. Additionally, a study carried out in Nigeria demonstrated the role of bioinformatics in the design of S. haematobium diagnostic kit depending on in silico determination of high immunogenic Schistosoma proteins [6]. Identification of drug targets and vaccine candidates: Based on advances in schistosome genomics, a family of promising schistosome vaccine antigens were suggested. The best vaccine candidates identified against schistosomiasis are protein molecules expressed on the tegument surface or secreted by schistosomula such as Sm29, and Sm-p80 that attracted much attention. Notably, Sm29 is a glycosyl-phosphatidyl inositol (GPI)anchored protein on Schistosoma tegument and the recombinant Sm29 vaccine exhibited Th1 immunity induction, and ~50% pathology reduction [7]. Besides, Sm-p80 is a subunit of the tegument cysteine protease, calpain that plays a crucial role in immune evasion. The recombinant Sm-p80 proved to be a promising vaccine candidate that achieved 80% improvement in liver pathology and a prominent Th1 response [8]. In addition to its role as a druggable molecule, investigating acetylcholinesterase (AChE) as a vaccine candidate against schistosomiasis was elucidated by omics and bioinformatics. Accordingly, two isoforms of AChE were identified in S. mansoni. The first was incorporated in the muscle layer to interact with heparin, i.e., a drug target. The other was GPIanchored to the surface membrane, i.e., a potential vaccine candidate [9] .

To assess the molluscicidal effect of the eco-friendly green synthesized neem silver nanoparticles (neem-Ag NPs) against Biomphalaria alexandrina, the snail intermediate host for Schistosoma mansoni, and their cercaricidal potential. Methods: Methanol extracts from neem fruits were used for green synthesis of neem-Ag NPs. The neem-Ag NPs were characterized using UV-visible absorption spectra, dynamic laser light scattering technique, and transmission electron microscopy. The potential molluscicidal effect against adult and juvenile Biomphalaria alexandrina and the effect of the sub-lethal concentration on hatching of snail eggs and Schistosoma mansoni cercariae were evaluated. Results: The surface plasmon resonance of neem-Ag NPs showed a sharp absorption peak at λ max = 518 nm together with multiple peaks. The hydrodynamic diameter was (77.15±34.53) nm, the polydispersity index (0.338±0.000) and the zeta-potential-14.07 mV. Moreover, transmission electron microscopy showed that the average size of the nanoparticles was (27±2) nm. Agglomeration was evident and a light-colored capping layer could be seen coating the nanoparticles. Juvenile snails (LC 50 : 0.83 ppm) were more susceptible to neem-Ag NPs than adults (LC 50 : 1.07 ppm). In addition, neem-Ag NPs and neem at LC 50 concentrations inhibited the egg-hatching of snails and showed cercaricidal activity in a timedependent manner. Conclusions: Neem-Ag NPs have lethal activities against Biomphalaria alexandrina snails and their eggs, as well as Schistosoma mansoni cercariae. Hence, neem-Ag NPs could be a potential agent to control schistosomiasis.

Introduction: Praziquantel (PZQ) is the only commercially available drug for schistosomiasis. The current shortage of alternative effective drugs and the lack of successful preventive measures enhance its value. The increase in the prevalence of PZQ resistance under sustained drug pressure is, therefore, an upcoming issue.Objective: To overcome the tolerance to PZQ using nanotechnology after laboratory induction of a Schistosoma mansoni isolate with reduced sensitivity to the drug during the intramolluscan phase.Materials and methods: Shedding snails were treated with PZQ doses of 200 mg/kg twice/ week followed by an interval of one week and then repeated twice in the same manner. The success of inducing reduced sensitivity was confirmed in vitro via the reduction of cercarial response to PZQ regarding their swimming activity and death percentage at different examination times.Results: Oral treatment with a single PZQ dose of 500 mg/kg in mice infected with cercariae with reduced se...

Background:Toxoplasma gondii (T. gondii) is an opportunistic parasite that causes serious diseases in humans, particularly immunocompromised individuals and pregnant women. To date, there are limited numbers of therapeutics for chronic toxoplasmosis which necessitate the discovery of effective and safe therapeutics. In the present study, we aimed to evaluate the antitoxoplasmosis potential of ginger extract in mice with experimentally induced chronic toxoplasmosis. Results: Treatment with ginger extract significantly reduced cysts count in the brains of T. gondii-infected mice with a marked alleviation of edema and inflammation, and a reversal of neuronal injury. Moreover, ginger extract treatment reduced inflammation in liver and lungs and protected hepatocytes from infection-induced degeneration. Consistently, apoptosis was significantly mitigated in the brains of ginger extract-treated mice compared to infected untreated animals or spiramycin-treated animals. Methods: Four groups...

Efficacy of triclosan (TS) and TS-loaded liposomes against the virulent strain of Toxoplasma gondii (T. gondii) was evaluated. Swiss albino mice were intraperitoneally infected with 10(4) tachyzoites of RH HXGPRT(-) strain of T. gondii, then were orally treated with 150 mg/kg TS or 100 mg/kg TS liposomes twice daily for 4 days. Mice mortality, peritoneal and liver parasite burdens, viability, infectivity and ultrastructural changes of peritoneal tachyzoites of infected treated mice were studied, in comparison with those of infected non-treated controls. Drug safety was biochemically assessed by measuring liver enzymes and thyroxin. Both TS and TS liposomes induced significant reduction in mice mortality, parasite burden, viability and infectivity of tachyzoites harvested from infected treated mice. Scanning electron microscopy of treated tachyzoites showed distorted shapes, reduced sizes, irregularities, surface protrusions, erosions and peeling besides apical region distortion. Transmission electron microscopy showed that treated tachyzoites were intracellularly distorted, had cytoplasmic vacuolation, discontinuous plasma membranes, nuclear abnormalities and disrupted internal structures. Besides, in TS liposomes-treated subgroup, most tachyzoites were seen intracellularly with complete disintegration of the parasite plasma and nuclear membranes, with complete destruction of the internal structures. Biochemical safety of TS and TS liposomes was proven. Accordingly, TS can be considered as a promising alternative to the standard therapy for treating acute murine toxoplasmosis. Liposomal formulation of TS enhanced its efficacy and allowed its use in a lower dose.