Anti-encystment and amoebicidal activity of Lonicera japonica Thunb. and its major constituent Chlorogenic acid in vitro

Objective:To examine the acanthamoebicidal effects of ethyl acetate,aqueous and butanol fractions of dried flower buds of Lonicera japonica(L.japonica) Thunb.(Flos Lonicerae) in vitro.Methods:Acanthamoeba triangularis isolates were obtained from environmental water samples and identified by PCR.They were exposed to ethyl acetate,water and butanol fractions of L.japonica Thunb.at concentrations ranging from 0.5 mg/m L to 1.5 mg/m L.The extracts were evaluated for growth inhibition at 24,48 and 72 h,respectively.Chlorogenic acid at a concentration of 1 mg/mL was examined for inhibition of encystment.Results:Ethyl acetate fraction at a concentration of 1.5 mg/mL evoked a significant reduction of trophozoite viability by 48.9% after 24 h,49.2% after 48 h and 33.7% after 72 h Chlorogenic acid,the major active constituent of L.japonica Thunb.at the concentration of 1 mg/mL reduced the cysts/trophozoite ratio by 100% after 24 h,84.0% after 48 h and 72.3% after 72 h.This phenolic compound at concentration of 1 mg/mL concurrent with 0.6% hydrogen peroxide inhibited hydrogen peroxide-induced encystment by 92.8% at 72 h.Conclusions:Results obtained from this study show that ethyl acetate fraction at 1.5 mg/m L is the most potent fraction of L.japonica Thunb.and its major constituent chlorogenic acid showed the remarkable inhibition of encystment at a concentration of 1 mg/mL.

Recommended Medicinal Plants as Source of Natural Products:A Review

Natural product is one of the sources of drugs in pharmaceutical industry,and one of the notable origin of natural product is the medicinal plants.Medicinal plants tend to cure some certain diseases and could be a source for potential drugs.The Department of Health of the Philippines approved 10 medicinal plants namely Allium sativum(Garlic/Bawang),Blumea balsamifera(Nagal camphor/sambong),Cassia alata(Ringworm bush/akapulko),Clinopodium douglasii(Mint/yerba Buena),Ehretia microphylla(Scorpion bush/Tsaang Gubat),Momordica charantia(Bitter Melon/Ampalaya),Peperomia pellucida(Silver bush/ulasimang Bato),Psidium guajava(Guava/Bayabas),Quisqualis indica(Rangoon creeper/niyug-niyogan),and Vitex negundo(Five-leaved Chaste Tree/lagundi).The review was conducted to show that these medicinal plants are capable in treating infections and some diseases.It was found that Allium sativum for the treatmet of wounds,hypertension and tootache;Blumea balsimifera is effective in diuretic treatment for hypertension;Cassia alata for the treatment of scabies,fungal infection,athlete’s foot,tinea flava,ringworm;Clinopodium douglasii for the treatment of muscle pain,arthritis,rheumatism,cough,headache;Ehretia microphylla for the treatment of diarrhea and stomachache;Momordica charantia for the treatment of diabetes mellitus;Peperomia pellucida for the treatment of gout and rheumatism;Psidium guajava for the treatment of wounds and diarrhea;Quisqualis indica for anti-helmintic medicine;and Vitex negundo for the treatment of cough,asthma,and fever.In conclusion,these 10 medicinal plants have natural products that can be used as source for potential drugs.However,there are still many species of ethnobotanical plants that are not yet investigated thoroughly and might be a source for potential drugs.Therefore,more investigations should be done in other species of plant,most especially for the plants with practical used.

Effect of ABCB1 3435C>T transporter gene polymorphism on plasma efavirenz concentration in HIV-1 infected Thai adults

Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene encoding ABCB1 was genotyped using real-time PCR-based alleles in 149 HIV-infected Thai adults receiving efavirenz treatment.Plasma concentrations of efavirenz were measured by high-performance liquid chromatography 12 hr after administration.The relationship between plasma efavirenz concentrations and ABCB13435 C>T polymorphisms was analyzed.Results:Logistic regression analysis showed no significant predictors of high plasma efavirenz concentration in relation to age,gender,body weight,CD4 count and plasma HIV-1 RNA,blood biochemical parameters,antiretroviral duration or ABCB13435 C>T polymorphisms,except for height(OR=0.902,95%CI:0.835-0.973)(P<0.05).The minor allele frequency of ABCB13435 C>T was0.446.The frequency of the heterozygous mutant ABCB13435 C/T was 53.02%(n=79),ABCB13435 T/T homozygous mutant was 18.12%(n=21)and the wild type ABCB13435 C/C genotype was 28.86%(n=43).The overall median plasma concentration of efavirenz in 149 HIV-infected Thai cases was 2.41 mg/L[IQR:(1.46-4.12)mg/L].The plasma concentration of efavirenz was higher in cases with ABCB13435 T/T homozygous mutant[2.73 mg/L,IQR:(2.02-4.19)mg/L]and ABCB13435 C/T heterozygous mutant[2.29 mg/L,IQR:(1.41-4.28)mg/L]genotypes compared to the wild type ABCB13435 C/C homozygous[2.1 mg/L,IQR:(1.37-3.53)mg/L].However,there was no statistically significant difference in the efavirenz concentration between the different genotypes(P>0.05).Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene encoding ABCB1 was genotyped using real-time PCR-based alleles in 149 HIV-infected Thai adults receiving efavirenz treatment.Plasma concentrations of efavirenz were measured by high-performance liquid chromatography 12 hr after administration.The relationship between plasma efavirenz concentrations and ABCB13435 C>T polymorphisms was analyzed.Results:Logistic regression analysis showed no significant predictors of high plasma efavirenz concentration in relation to age,gender,body weight,CD4 count and plasma HIV-1 RNA,blood biochemical parameters,antiretroviral duration or ABCB13435 C>T polymorphisms,except for height(OR=0.902,95%CI:0.835-0.973)(P<0.05).The minor allele frequency of ABCB13435 C>T was 0.446.The frequency of the heterozygous mutant ABCB13435 C/T was 53.02%(n=79),ABCB13435 T/T homozygous mutant was 18.12%(n=27)and the wild type ABCB13435 C/C genotype was 28.86%(n=43).The overall median plasma concentration of efavirenz in 149 HIV-infected Thai cases was 2.41 mg/L[IQR:(1.46-4.12)mg/L].The plasma concentration of efavirenz was higher in cases with ABCB13435 T/T homozygous mutant[2.73 mg/L,IQR:(2.02-4.19)mg/L]and ABCB13435 C/T heterozygous mutant[2.29 mg/L,IQR:(1.41-4.28)mg/L]genotypes compared to the wild type ABCB13435 C/C homozygous[2.1 mg/L,IQR:(1.37-3.53)mg/L].However,there was no statistically significant difference in the efavirenz concentration between the different genotypes(P>0.05).Conclusions:There is no statistical significance for a tendency toward higher plasma efavirenz concentration in the ABCB13435 T/T and ABCB13435 C/T genotypes.No parameters of physiological characteristics in this study except for height were found to be predictors of high plasma efavirenz concentration in Thai HIV-1 infected cases.

Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system

Background:Access to clean and safe drinking water that is free from pathogenic protozoan parasites,especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans,is still an issue in Southeast Asia(SEA).This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA,using real-time polymerase chain reaction(qPCR)assays.Methods:A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia(53),Thailand(120),the Philippines(33),and Vietnam(15).A physicochemical analysis was conducted.The water samples were processed in accordance with the US Environmental Protection Agency’s methods 1622/1623.1,microscopically observed and subsequently screened using qPCR assays.Results:Cryptosporidium oocysts were detected in treated water samples from the Philippines(1/10),with a concentration of 0.06±0.19 oocyst/L,and untreated water samples from Thailand(25/93),Malaysia(17/44),and the Philippines(11/23),with concentrations ranging from 0.13±0.18 to 0.57±1.41 oocyst/L.Giardia cysts were found in treated water samples from the Philippines(1/10),with a concentration of 0.02±0.06 cyst/L,and in untreated water samples from Thailand(20/93),Vietnam(5/10),Malaysia(22/44),and the Philippines(16/23),with concentrations ranging from 0.12±0.3 to 8.90±19.65 cyst/L.The pathogens C.parvum and G.lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene,respectively.C.parvum was detected in untreated water samples from the Philippines(1/23)and Malaysia(2/44),whilst,G.lamblia detected was detected in treated water samples from the Philippines(1/10)and in untreated water samples from Thailand(21/93),Malaysia(12/44),and the Philippines(17/23).Nitrate concentration was found to have a high positive correlation with(oo)cyst(0.993).Conclusion:The presence of(oo)cysts in the water samples means that there is potential risk for zoonotic disease transmission in the studied countries.Detection using qPCR is feasible for quantifying both pathogenic C.parvum and G.lamblia in large water samples.