Nano chloroquine delivery against Plasmodium berghei NK65 induced programmed cell death in spleen

Objective:To compare the protective effects of chitosan-trypolyphosphate(CS-TPP) nanoparticle conjugated chloroquine(CQ) with effect of CQ alone on the reversal of splenic damages and induction of apoptosis.Methods:Different researches have been carried out to explore the potential role of chitosan based drug delivery system against parasitic diseases.After successive Plasmodium berghei NK65 parasiste infection by intraperitoneal injection in Swiss mice and subsequent parasite development,the ROS generation,anti-apoptotic and pro apoptotic protein levels in spleen were measured.To analyze caspases,flow cytometry study was performed with annexin 桋-FITC and with PI staining.Results:The results revealed that ROS mediated caspase 3 and 9 activation and the induction of apoptosis occurred during the parasitic infection.However,CS-TPP conjugated CQ was relatively better in reversing the splenic damage compared with similar effects of CQ alone.Conclusions:This study indicates that Plasmodium berghei NK65 induces apoptosis in the spleen.The study further shows that CS-TPP nanoparticles conjugation with CQ have positive influence on the recovery of damaged host's system towards maintenance of normal homeostasis,and this is shown to be selective to CS-TPP conjugated CQ treated animals only.

In vivo combination therapy of chloroquine and stigmasterol against Plasmodium berghei induced pathologies in mice

Objective:To investigate the effect of combination therapy of chloroquine and stigmasterol on Plasmodium berghei(thereafter referred to as P.berghei)malaria-induced organ pathologies.Methods:Thirty five mice weighing 20-30 g were placed into seven groups of five mice each and distributed as uninfected administered 100 mg/kg BW stigmasterol,uninfected administered only feed and water ad libitum,infected with P.berghei and-administered 50 mg/kg BW stigmasterol,100 mg/kg BW stigmasterol,100 mg/kg BW stigmasterol plus 5 mg/kg BW chloroquine,and 5 mg/kg BW chloroquine.The last group of mice served as P.berghei infected and not treated control.The levels of parasitemia,packed cell volume,and other biochemical parameters were measured.Results:Combination therapy of P.berghei infection with stigmasterol and chloroquine did not significantly(P>0.05)reduce parasitemia level while stigmasterol treatment alone significantly(P<0.05)reduced the parasitemia level.However,the P.berghei induced anemia was decreased significantly(P<0.05)upon treatment with a combination of chloroquine and stigmasterol as well as with stigmasterol alone.Furthermore,the combination of chloroquine and stigmasterol significantly(P<0.05)decreased the activities of serum alanine aminotransferase,aspartate aminotransferase,urea and spleen total proteins levels in P.berghei mice in comparison with the untreated group.Treatment of P.berghei infected mice with stigmasterol alone and in combination with chloroquine significantly(P<0.05)increased the level of serum creatinine while serum and spleen malondialdehyde levels were significantly(P<0.05)decreased.Levels of glutathione in spleen and kidney were insignificantly(P>0.05)altered upon treatment with both doses of stigmasterol as well as the combination therapy.Conclusions:This study concluded that the combination of stigmasterol and chloroquine could combat anemia and some organ pathologies associated with P.berghei infection.

In vivo combination therapy of chloroquine and stigmasterol against Plasmodium berghei induced pathologies in mice

Objective:To investigate the effect of combination therapy of chloroquine and stigmasterol on Plasmodium(P.)berghei malaria-induced organ pathologies.Methods:Totally 35 mice weighing 20-30g were placed into 7 groups of 5 mice each and distributed as uninfected administered 100 mg/kg BW stigmasterol,uninfected administered only feed and water ad libitum,infected with P.berghei and administered 50 mg/Kg BW stigmasterol,100 mg/kg BW stigmasterol,100 mg/kg BW stigmasterol plus 5 mg/kg BW chloroquine,and 5 mg/kg BW chloroquine.The last group of mice served as P.berghei infected and not treated control.The levels of parasitemia,packed cell volume,and other biochemical parameters were measured.Results:Combination therapy of P.berghei infection with stigmasterol and chloroquine did not significantly(P>0.05)reduce parasitemia level while stigmasterol treatment alone significantly(P<0.05)reduced the parasitemia level.However,the P.berghei induced anemia was decreased significantly(P<0.05)upon treatment with a combination of chloroquine and stigmasterol as well as with stigmasterol alone.Furthermore,the combination of chloroquine and stigmasterol significantly(P<0.05)decreased the activities of serum alanine aminotransferase,aspartate aminotransferase,urea and level of spleen total proteins in P.berghei infected mice in comparison with the untreated group.Treatment of P.berghei infected mice with stigmasterol alone and in combination with chloroquine significantly(P<0.05)increased the level of serum creatinine while serum and spleen malondialdehyde levels were significantly(P<0.05)decreased.Levels of glutathione in spleen and kidney were insignificantly(P>0.05)altered upon treatment with both doses of stigmasterol as well as the combination therapy.Conclusions:This study concluded that the combination of stigmasterol and chloroquine could combat anemia and some organ pathologies associated with P.berghei infection.

Investigation of the Efficacy of Home-Based Oral Chloroquine Treatment among Under-Five Children with Plasmodium falciparum Malaria in Some Parts of Jos, Plateau State, Nigeria

Background: Nigeria is currently a malaria endemic country with an estimated 76% of her population at risk of contracting malaria [1]. According to a study in Nigeria, the first line of action mothers took when their children under 5 years have malaria showed that over 50% of them used non-prescription drugs they have at home or bought from pharmacy stores. And 60% of the most commonly used drugs for malaria treatment were chloroquine [2]. Many recent studies have demonstrated re-emergence of chloroquine-sensitive P. falciparum, suggesting a possible role in future malaria control [3]. Objective: The aim of this study was to investigate the effect of home-based oral chloroquine treatment among children under 5 years with Plasmodium falciparum malaria attending Jos University Teaching Hospital and OLA Hospital in Jos Metropolis. Method: This is a cross-sectional study of 93 malaria and non-malaria children. Malaria diagnosis was carried out using microscopical examination of Leishman’s stained thick and thin blood films, P. falciparum parasitemia was assessed by standard microscopy techniques and complete blood count was done using Beckman Coulter Analyzer. Results: The body temperature on admission was significantly lower (p &#730;C ± 0.07&#730;C) than in the three malaria groups. The mean body temperature of chloroquine treated children with malaria was significantly lower (p Conclusion: The results obtained in this study demonstrate that there was significant positive impact of chloroquine treatment on Plasmodium falciparum parasitemia and degree of anemia in children under 5 years with Plasmodium falciparum in Jos Metropolis.

Noscapine shows antimalarial activity against Plasmodium falciparum 3D7,its clinical isolate Pf140/SS,and Plasmodium berghei ANKA

Objective:To evaluate the antimalarial activity of noscapine against Plasmodium falciparum 3D7 strain(Pf3D7),its clinical isolate(Pf140/SS),and Plasmodium berghei ANKA(PbA).Methods:Using ring-stage survival assay,phenotypic assessments,and SYBR-green-based fluorescence assay,the antimalarial activities of noscapine were assessed compared with dihydroartemisinin(DHA)in in vivo and in vitro studies.In addition,hemolysis and cytotoxicity tests were carried out to evaluate its safety.RT-PCR assay was also conducted to determine the effect of noscapine on papain-like cysteine protease Plasmodium falciparum falcipain-2(PfFP-2).Results:The antimalarial efficacy of noscapine against Pf3D7 and Pf140/SS was comparable to DHA,with IC50 values of(7.68±0.88)and(5.57±0.74)nM/mL,respectively,and>95%inhibition of PbA infected rats.Noscapine also showed a safe profile,as evidenced by low hemolysis and cytotoxicity even at high concentrations.Moreover,PfFP-2 expression was significantly inhibited in both noscapine-treated Pf3D7 and Pf140/SS(P<0.01).Conclusions:Noscapine has antimalarial properties comparable to standard antimalarial DHA with better safety profiles,which may be further explored as a therapeutic candidate for the treatment of malaria.

Comparative study of chloroquine and quinine on malaria rodents and their effects on the mouse testis

Objective:To evaluate the effects of quinine and chloroquine against mole mice infected with Plasmodium berghei and their adverse effects on the mice testes.Methods:In this study,48 adult male mice,(20-23 g).aged 8 to 12 weeks were divided into four groups.This study was carried out from December 2009 until May 2010 in the School of Public Health,Tehran University of Medical Sciences.Results:The results showed that 58.33%of mice treated with chloroquine were completely recovered.Parasitemia was 4%on day 8 when compared to that on day 0,whereas it was 9%on day 9.There was no orchitis found in this group.The mortality of mice after exposing to quinine on day 5 was 8.3%,whereas from day 10 to day 14 it was 91.7%.We found 73%orchitis occurred in quinine treated group.There was a significant difference between quinine and chloroquine effects on the parasite and also mice testes(P<0.05).Conclusions:In this study,It can be concluded that male mice have full resistance to the quinine.Quinine does not only make male mice recover completely,but also cause inflammation on mice testicles tissue.

Interleukin-33 exerts pleiotropic immunoregulatory effects in response to Plasmodium berghei ANKA(PbA)infection in mice

Objective:To determine the involvement and the modulatory effects of IL-33 during Plasmodium berghei ANKA(PbA)infection.Methods:PbA infection in male ICR mice was utilized as a model of malaria.Systemically circulating IL-33 levels were determined in blood plasma by enzyme-linked immunosorbent assay(ELISA).After 24 hours post-inoculation of PbA,recombinant IL-33 and ST2,and antibodies against IL-33 and IgG treatments were administered daily for 3 days.Tissue expression and localization of IL-33 were assessed in organs generally affected by malaria via immunohistochemistry.Moreover,histopathological examination was performed to assess the effects of the treatments.Results:The levels of systemic IL-33 were elevated at the critical phase of PbA infection.Likewise,immunohistochemical analysis revealed a significant upregulation of IL-33 expression at the critical phase in the brain,lungs,and spleen of PbA-infected mice as compared to healthy controls.Treatment with IL-33 protected against experimental cerebral malaria development and reduced pathological features in the brain and lungs of the PbA-infected mice.Conclusions:A potential critical role and involvement of IL-33 in PbA infection may hint at the resolution of immunopathological sequelae associated with malaria.

Antimalarial potential of kolaviron,a biflavonoid from Garcinia kola seeds,against Plasmodium berghei infection in Swiss albino mice

Objective:To investigate the antimalarial potential of kolaviron(KV),a biflavonoid fraction from Garcinia kola seeds,against Plasmodium berghei(P.berghei)infection in Swiss albino mice.Methods:The study consists of seven groups of ten mice each.Groups I,II and III were normal mice that received com oil.KV1 and chloroquine(CQ),respectively.Groups IV,V,ⅥandⅦwere infected mice that received corn oil.CQ,KYI and KV2.respectively.CQ.KY1 and KV2were given at 10-,100-and 200-mg/kg daily,respectively for three consecutive days.Results:Administration of KV1 and KV2 significantly(P<0.05)suppressed P.berghei-infection in the mice by 85%and 90%.respectively,while CQ produced 87%suppression relative to untreated infected group after the fifth day of treatment.Also,KV2 significantly(P<0.05)increased the mean survival time of the infected mice by 175%.The biflavonoid prevented a drastic reduction in HCV from day4 of treatment,indicating its efficacy in ameliorating anaemia.Significant(P<0.05)oxidative stress assessed by the elevation of serum and hepatic malondialdehydewere observed in unlrealed P.berghei-infected mice.Specifically,senum and hepatic malondialdehyde levels increased by93%and 78%,resjiectively in the unlrealed infecled mice.Furlhennore,antioxidant indices,viz;superoxide dismutase.catalase,glutathione-s-transferasc.glualhione peroxidase and reduced gluathione decreased significantly(P<0.05)in the tissues of untreated P.berghei-infected mice.KV significantly(P<0.05)ameliorated the P.berghei-induced decrease in antioxidant status of the infected mice.Conclusions:This study shows that kolaviron,especially at 200 mg/kg,has high antimalarial activities in P.berghei-infected mice,in addition to its known antioxidant properties.

Nauclea latifolia aqueous leaf extract eliminates hepatic and cerebral Plasmodium berghei parasite in experimental mice

Objective: To assess the effects of hot water leaf extract of Nauclea latifolia(N. latifolia) on antioxidant status, lipid peroxidation values and parasite levels in hepatic and brain tissue of experimental mice(BALB/c) infected with Plasmodium berghei(P. berghei) malaria.Methods: Forty nine mice were divided into seven groups(n = 7) and used for the study. Group A(control) were given 0.2 m L/kg phosphate buffer saline; Group B mice were infected with P. berghei and treated with phosphate buffer saline. Groups C and D mice were also infected but treated with 200 and 300 mg/kg body weight of leaf extract respectively. Groups E and F mice were not infected, but received 200 and 300 mg/kg of leaf extract respectively. Group G mice were infected and treated with chloroquine(5 mg/kg). Liver and brain tissues of mice were prepared for both biochemical assay and microscopic examination. Results: Results showed that P. berghei malaria infection induced oxidative stress in both liver and brain tissues as evidenced by the significant(P < 0.05) decrease in antioxidants: superoxide dismutase, reduced glutathione and catalase. These reductions perhaps caused compromise in membrane integrity as indicated by the significant increase in lipid peroxidation product malondialdhyde. Malaria parasites were also identified in these tissues. However, N. latifolia treatment eliminated the parasites in tissues and protected them from oxidative damage even better than chloroquine treatment did, whose anti-malarial potency also cleared tissue parasites. The measurement of protection by N. latifolia against damage was strengthened by the insignificant micro structural alterations.Conclusions: The bioactive phytochemical(s) in N. latifolia should be structured and the mechanism(s) of its antimalarial tendency should be further investigated.

Immunogenicity and immunizing protection effect of GAMA gene DNA vaccine on Plasmodium berghei

Objective: To explore the effect of immunogenicity and immunizing protection of GAMA gene DNA vaccine, which was related with merozoite, ookinete and sporozoite invasion. Methods:Gene fragments were obtained using PCR technique and eukaryotic expression vector(containing immunostimulatory sequence) was built. BALB/c mice were divided into PBS control group, empty vector control group and study group and were immunized at week 0, 3 and 6 respectively. Blood was collected 2 weeks after each immunization and serum was separated to detect the Ig G, Ig G1 and Ig G2 a levels. Spleen of mice was obtained for preparation of splenic mononuclear cell and the cytokine IL-4 and IFN-αlevels were detected. Indirect immunofluorescence and western blot were employed to verify the specificity of antiserum. Sporozoite and merozoite invasion were used respectively to detect the immune protective effect 2 weeks after the third immunization. Ookinete conversion rate in vitro and oocyst numbers of mosquito stomach were observed to evaluate the transmission-blocking levels. Results: In GAMA DNA vaccine group: antiserum could be combined with recombinant protein specifically and green fluorescence signals of merozoite, ookinete and sporozoite were observable, while specific fragments and fluorescence signals were not observable in empty vector group. Compared with control group, specific Ig G in DNA vaccine immunity group significantly increased(P<0.01), and Ig G1 and Ig G2 a all increased(P<0.01). IL-4, IFN-αcontent in study group significantly increased, compared with control group(P<0.01). GAMA DNA vaccine immunity could not obviously block the erythrocyte-stage infection(caused by sporozoite invasion); compared with control group, liver worm load was slightly reduced(P<0.05), and antiserum ookinete numbers(cultured in vitro) had no significant difference with oocyst numbers of mosquito stomach in DNA vaccine group. Conclusions:GAMA has good antigenicity, which could stimulate the body to produce specific immune responses; while DNA vaccine immunity could not play a good protective effect, the effect of which is only limited to the slight reduction of liver worm load, and has no obvious erythrocytestage protective effect and transmission-blocking effect. Therefore, trying other immunization strategies for further research on the value of GAMA(as multi-stage antigen vaccine and multistage combined vaccine components of the life-cycle of plasmodium) is necessary.

Bioinformatics analysis and prediction for structure and function of nitric oxide synthase and similar proteins from Plasmodium berghei

ObjectiveTo search and analyze nitric oxide synthase (NOS) and similar proteins from Plasmodium berghei(Pb).MethodsThe structure and function of nitric oxide synthase and similar proteins from Plasmodium berghei were analyzed and predicted by bioinformatics.ResultsPbNOS were not available, but nicotinamide adenine dinucleotide 2′–phosphate reduced tetrasodium (NADPH)–cytochrome p450 reductase(CPR) were gained. PbCPR was in the nucleus of Plasmodium berghei, while 134aa–229aa domain was localize in nucleolar organizer. The amino acids sequence of PbCPR had the closest genetic relationship with Plasmodium vivax showing a 73% homology. The tertiary structure of PbCPR displayed the forcep–shape with wings, but no wings existed in the tertiary structure of its' host, Mus musculus(Mm). 137aa–200aa, 201aa–218aa, 220aa–230aa, 232aa–248, 269aa–323aa, 478aa–501aa and 592aa–606aa domains of PbCPR showed no homology with MmCPRs', and all domains were exposed on the surface of the protein.ConclusionsNOS can't be found in Plasmodium berghei and other Plasmodium species. PbCPR may be a possible resistance site of antimalarial drug, and the targets of antimalarial drug and vaccine. It may be also one of the mechanisms of immune evasion. This study on Plasmodium berghei may be more suitable to Plasmodium vivax. And 137aa–200aa, 201aa–218aa, 220aa–230aa, 232aa–248, 269aa–323aa, 478aa–501aa and 592aa–606aa domains of PbCPR are more ideal targets of antimalarial drug and vaccine.

Gut Microbiota Reconstruction Following Host Infection with Bloodstage Plasmodium berghei ANKA Strain in a Murine Model

Malaria remains a global health problem.The relationship between Plasmodium spp.and the gut microbiota as well as the impact of Plasmodium spp.on the gut microbiota in vertebrate hosts is unclear.The aim of the current study was to evaluate the effect of blood-stage Plasmodium parasites on the gut microbiota of mice.The gut microbiota was analyzed by 16S rRNA sequencing and bioinformatic analyses at three stages.The gut microbiota changed during the three phases:the healthy stage,the infection stage,and the cure stage(on the 9th day after malarial elimination).Moreover,the gut microbiota of these infected animals did not recover after malaria infection.There were 254 operational taxonomic units(OTUs)across all three stages,and there were unique strains or OTUs at each stage of the experiment.The percentages of community abundance of 8 OTUs changed significantly(P<0.05).The dominant OTU in both the healthy mice and the mice with malaria was OTU265,while that in the cured mice was OTU234.In addition,the changes in OTU147 were the most noteworthy.Its percentage of community abundance varied greatly,with higher values during malaria than before malaria infection and after malaria elimination.These results indicated that the external environment influenced the gut microbiota after host C57BL/6 mice were infected with blood-stage P.berghei ANKA and that the same was true during and after elimination of blood-stage P.berghei ANKA.In addition,we could not isolate OTU147 for further study.This study identified gut microbiota components that were reconstructed after infection by and elimination of blood-stage P.berghei ANKA in host C57BL/6 mice,and this process was affected by P.berghei ANKA and the external environment of the host.

Antimalarial activity of the aqueous extract of Euphorbia cordifolia Elliot in Plasmodium berghei-infected mice

Objective:To evaluate the antimalarial activity of the aqueous extract of Euphorbia(E.)cordifolia Elliot against Plasmodium(P.)berghei-infected mice.Methods:Thirty healthy Swiss mice were intraperitoneally inoculated with 200μL of P.berghei parasitized-erythrocytes and divided into five groups,and then daily treated for 5 d with single dose of 10 mL/kg of distilled water for malaria control,10 mg/kg of chloroquine for the chloroquine control and 100,200 and 400 mg/kg of the aqueous extract of E.cordifolia for the three test groups.Parasitaemia was monitored by Giemsa-staining.At the end of the treatment,animals were sacrificed,and blood was collected for haematological and biochemical analyses.Organs were collected for biochemical and histopathological analyses.Statistical significance(P<0.05)was evaluated by analysis of variance followed by the Tukey post-test using Graphpad prism 7.0.Results:E.cordifolia extract decreased the parasite load to 2.46%,with an effective dose(ED50)of 113.07 mg/kg compared to the malaria group where the parasite load increased to(46.46±10.28)%.E.cordifolia extract prevented hypoglycaemia,anaemia,leucocytosis and thrombocytopenia,attenuated the increase of transaminases activities,bilirubin and creatinine rate,and improved catalase and superoxide dismutase activities,while reducing malondialdehyde contents in the liver and kidney.E.cordifolia extract significantly prevented histological damages observed in the malaria control group.No acute toxicity sign was observed in mice with plant extract at the dose up to 5000 mg/kg.Conclusions:E.cordifolia extract at 200 and 400 mg/kg showed significant antimalarial effects.This results support its traditional use in the treatment of malaria.

Effects of the methanolic seeds extract of Carica Papaya on plasmodium Berghei infected mice

Objective:The leaves extract of Carica Papaya（C.Papaya）papaya has been shown to possess antimalarial activity, thus this work aims at finding out if the plants antimalarial activity is present in or extended to the seeds.Methods:The seeds of C.papaya were collected from its fruit,air dried for 5 days and ground into fine powder.80.65 g of the powder was then soaked for 48hours in 300 mL of methanol.Filtration was carried out using Whatman No.1 filter paper.The filtrate was evaporated to dryness by a three-day continuous heating on a hot plate of 30℃.The dry extract yield was scraped out of the Petri dish weighed and refrigerated until required. The percentage extract yield was calculated out from the initial powder weight.A preliminary phytochemical study was done by re-dissolving the appropriate amount of the dry extract in distilled water and appropriate test reagent added.The LD50 of the seeds of C.papaya was carried out using arithmetic method.Swiss albino Mice of both sexes and of average weight of 18-25 g were used as animals for antimalarial activity.They were housed in standard animal house,fed on Rats/Mice pellets and had non restricted excess to both feed and water throughout the 60day study period.While the non pregnant female Mice were used as test animals,the male animals were used as malaria parasite donors.Precautions were taken to ensure that all animals in the study groups were free from infection with Eperythrozoon coocoides.The female animals were then divided into three main groups（A-C） of 25 animals per group.Group A was used for malaria suppressive study（early infection -day 0-3） and was further subdivided to 5 subgroups（a-e） of 5animals per group.Group B was used for malaria curative study（established malaria infection-day 3-7） and was further subdivided to 5subgroups（a-e） of 5animals per group.Group C was used for malaria prophylactic study（repository-4days treatment prior to malaria parasite infection） and was also further subdivided into 5subgroups（a-e） of 5animals per group.At the appropriate time,50 mg/kg/day,100 mg/kg/day and 200 mg/kg/day of crude extract of C.papaya were administered orally to the different subgroups（b-d） within the three main groups.One subgroup（a） in each main group also received orally,5 mg/kg/day of chloroquine phosphate as positive control while one subgroup （e） in each main group also received orally,0.2 mL/kg/day of distilled water as negative control.Malaria parasites infected red blood cells numbering 1×10 and suspended in 0.2 mL of physiological saline was inoculated intraperitoneally,to each animal of the subgroups（a-d） in each of the three main groups at the appropriate time.Blood smears were made from animals’tail,stained with Lieshman and examined microscopically at 100×for the presence of malaria parasite.Percentage malaria parastaemia was calculated as well as average percentage malaria parasitaemia suppression.Results:Extraction yield of 25.29%was obtained while the LD50 was 620 mg/kg.The phytochemistry showed the richly presence of alkaloids,as well as glycosides,carbohydrates, resins,fats and fixed oils.The suppressive study at doses of 200,100 and 50mg/kg/day showed 53.02%,43.43%and 19.83%suppressive activity against Plasmodium berghei respectively.This activity compared to that of chloroquine,a standard antimalaria drug that gave 95.95%suppressive anti-parasitaemia. The prophylactic study at doses of 200,100 and 50 mg/kg/day showed 63.85%,61.12%and 48.08%prevention to malaria parasitaemia respectively as against 94.78%showed by chloroquine.The curative study however,at doses of 200,100 and 50 mg/kg/day failed to suppress malaria parasitaemia with a mean survival range of 6-8days as against 27.2 days showed by chloroquine.The seeds extract of C.papaya showed a significant malaria parasitaemia suppressive activity（P≤0.05）.These activities are dose dependent and comparable to those of Chloroquine phosphate.Conclusion:The results above suggest that the seeds extracts of C.papaya possess antimalarial activity like the leaf extracts.The antimalarial activity may be attributable to the richly presence of alkaloids and or the presence of its proteolytic enzyme（Papain）.The present finding justifies the inclusion of the seeds of C.papaya in the treatment of malaria by local herbalists.The seeds extracts therefore,if well purified and characterized may be used in treatment of very early plasmodiasis as well as a good prophylactic drug in human.This work at the moment is limited to animals,thus clinical trials in humans is be recommended particularly,when C.papaya seeds are non harmful/non toxic.

Effect of pre-existing Schistosoma haematobium infection on Plasmodium berghei multiplications in imprinting control region mice

Objective: To investigate the effect of pre-existing Schistosoma haematobium(S. haematobium) infection on malaria disease severity.Methods: The study involved the use of twenty-i ve imprinting control region mice, i fteen of which were initially infected with S. haematobium. Five of the remaining ten schistouninfected mice together with i ve schisto-infected mice were infected with Plasmodium berghei(P. berghei) after four weeks(acute stage) of schistosoma infection. The remaining i ve schisto-uninfected mice together with i ve schisto-infected mice were also infected with P. berghei after seven weeks(chronic stage) of schistosoma infection. The last i ve schistoinfected mice were used as control group. They were then monitored for changes in P. berghei parasitaemia on Days 3, 5, 7, 9 and 11 post-infection. Records on their survivability were also taken.Results: The co-infected mice had signii cantly higher malaria parasitaemia, compared with the mono-infected mice during acute S. haematobium infection. In contrast, the co-infected mice had signii cantly lower malaria parasitaemia during chronic S. haematobium infection and a higher survival rate.Conclusions: Co-infection of mice with P. berghei during acute S. haematobium infection resulted in rapid P. berghei development and increased malaria parasitaemia. However, the co-infection resulted in slower P. berghei development and decreased malaria parasitaemia with enhanced survivability of the mice during chronic S. haematobium infection. Therefore, pre-existing chronic S. haematobium infection may provide some protection to the host by reducing parasitaemia.

Pharmacodynamic evaluation for antiplasmodial activity of Holarrhena antidysentrica(Kutaja) and Azadirachta indica(Neemb) in Plasmodium berghei infected mice model

Objective:To investigate in-vivo anti-plasmodial activity of aqueous extracts of plants selected based on the symptomology mentioned in Ayurveda.Methods:The aqueous extracts of Holarrhena antidysentrica(H.antidysentrica)(Kutaja) and Azadirachta indica(A.indica)(Neemb) for their antiplasmodial potential in Plasmodium berghei(P.berghei) infected mice was assessed using Peters four day suppressive test.Both the extracts were administered at 2 dose levels,full dose(1 000 mg/d) and minimized dose(200 mg/d).10~6 P.berghei infected RBCs were injected on day ’0’ and treated from day ’0’ till day ’3’ post-infection,Tail blood smears were collected, giemsa stained and analyzed.The mice were observed for survival and parasitemia was assessed till 50%of mice in control survived.Results:It was observed that the percentage of parasitemia increased gradually in all the groups,with maximum in control group(Day 3-35,Day 9-46.98) and minimum in Chloroquine arm(Day 3-14.06.Day 9-19.92).The percentage of parasitemia was compared using Mann-Whitney U test depicting that all test groups exhibited reduction in parasitemia as compared to control(P-value【0.002 for all groups).These groups showed similar percentage of survival as Chloroquine.Conclusions:The present investigation demonstrated the anti-plasmodial effects of H.antidysentrica and A.indica.which are two most commonly used medicinal plants in Ayurved for treatment of fever.

Antioxidant Activities of Ethanoilc Extracts of Spilanthes uliginosa, Ocimum basilicum, Hyptis spicigera and Cymbopogon citratus against Swiss Mice Exposed to Plasmodium berghei Anka 65

Malaria infection is associated with increased generation of free radicals. This study investigated the antioxidant activity of ethanolic leaf extracts of Spilanthes uliginosa, Ocimum basilicum, Hyptis spicigera and Cymbopogon citratus. Seventy two (72) swiss mice of both sexes were used. All the mice were treated intraperitoneally with 0.2 ml parasitized blood suspension and parasitemia assessed by thin blood films stained with Geimsa stain after seventy two hours. The mice were divided into six groups namely;A, B, C, D, E, and F of twelve mice each. Groups B, C, D and E were subdivided into three (3): B1, B2, B3, C1, C2, C3, D1, D2, D3, E1, E2 and E3, four in each subgroup. The subgroups were treated with the extracts of Spilanthes uliginosa (Sw), Ocimum basilicum, Hyptis spiligera and Cymbopogon citratus each for five (5) consecutive days with 200, 400 and 800 mg/kg body weight respectively via oral intubation. Two control groups, A and F were used. The negative control (A) was treated daily with 5 ml/kg normal saline while positive control group (F) was treated with 5 mg/kg body weight of chloroquine. The results indicated a general significant (P < 0.05) decrease in the lipid peroxidation concentrations of the parasitized treated mice when compared to parasitized untreated mice on the last day. A general significant dose dependent increase (P < 0.05) was observed in superoxide dismutase (SOD), catalase and glutathione perioxidase activities as well as reduced glutathione concentrations in the treated mice except at a dosage of 200 mg/kg body weight for all the plants. The effects of the extracts were significantly higher (P < 0.05) than that of chloroquine. These results suggest that the ethanolic extracts of these plants may contribute to the protection of malaria infected mice against oxidative damage by improving antioxidant status in a dose dependent manner.

Antiplasmodial Efficacy of Crude Cocoa Powder Extract on CD4+ T-Cell Counts of <i>Plasmodium berghei</i>Infected BALB/c Mice

Background: Drug resistance in malaria warrants the need for alternative therapy from plant food nutrients. The search for novel anti-malarial control spurred a great interest in cocoa which has been portrayed as immune booster against malaria. This study was geared towards estimation of CD4+ cells of P. berghei infected mice treated with cocoa powder extract (CPE) to provide substantive scientific evidence to authenticate the anecdotal report. Methods: Brine shrimp toxicity assay was done to determine LC50 of crude cocoa powder extract. The mice were infected with 1 × 107 of ANKA and NK65 strains of Plasmodium berghei intraperitoneally, while graded doses of the extract were administered by an intra-gastric intubation based on the body weight of mice. Blood samples were analyzed for microscopy and flow cytometry for CD4+ cell counts. Results: The onset of infection was delayed in the group treated before inoculations on day 3 and the level of P. berghei parasitemia was positively associated with induction of CD4+ cells while the negative control group that received normal saline had progressive increase of parasitemia. The mean survival time could not go beyond day14 in ANKA, though both strains responded to CPE in a similar way with chloroquine as a positive control. The CD4+ cells counted increased in both strains treated before and during inoculations and the episodes of malaria was suppressed compared with the control. Conclusion: This study has demonstrated that the antiplasmodial activity of CPE was associated with the level of CD4+ T-cells proliferation which initiated the protective immune response. This therefore calls for efforts to ensure adequate intake of cocoa powder to boost immunity against malaria.

Apoptosis of erythrocytic stage parasites of Plasmodium berghei chloro-quine-resistant strain

Objective: To explore the characteristics of crisis state at erythrocytic stage of Plasmodium berghei chloroquine-resistant (RC) strain. Methods: Agarose electrophoresis, optical and transmission electron microscopes were used. Patterns of genomic DNA structures and ultra-structures of the erythrocytic parasites were observed in ICA mice (infected with the RC strain) during rising and declining of parasitemia. Results: During the declining parasitemia, the erythrocytic stage parasites of the RC strain showed round or oval appearance with intact plasma membrane and shrank nuclei with no metabolic window, mitochondria or other membranaceous structures. Their DNA electrophoretogram revealed a ladder pattern which evidently differed from the parasites of the RC strain in the rising parasitemia and the chloroquine-sensitive (N) strain.Conclusion: The crisis state of the erythrocytic stage parasites of the P. berghei chloroquine-resistant (RC)strain is characterized by apoptosis.