In vitro Mechanistic Assays of Tetracyclic Iridoid Compounds Isolated from Morinda lucida Benth in Leishmania species

Aims: This study investigates the activity of tetracyclic iridoid compounds against Leishmania spp. and the mechanism(s) of action. Study Design: An experimental study. Place and Duration: Department of Parasitology, Noguchi Memorial Institute for Medical Research, between September 2017 and July 2018. Methodology: The 50 % inhibitory concentration (IC50) of compounds against Leishmania donovani and L. major promastigotes were determined after 48 hours of incubation using the Alamar blue. Cytotoxicity of compounds was determined against cell lines using MTT assay. The anti-amastigote activity of compounds was further assessed by DAPI (4′,6-diamidino-2-phenylindole) staining. The mechanism of cell death induced by compounds was determined using nexin assay. Mitosis, cytokinesis and morphometry were monitored by DAPI and Kinetoplastid Membrane Protein (KMP) staining. Cell cycle arrest induced by compounds was analyzed by FACS. Results: Molucidin and ML-F52 inhibited the growth of promastigote in L. donovani (Molucidin; IC50 = 2.94±0.60 µM, ML-F52; IC50 = 0.91±0.50 µM) and L. major (Molucidin; IC50 = 1.85± 0.20 µM, ML-F52; IC50 = 1.77± 0.20 µM). ML-F52 had a 10-fold cytotoxic effect on parasites relative to normal cell lines. Against intracellular forms, Molucidin and ML-F52 inhibited intracellular amastigote replication and infectivity. Amphotericin B, Molucidin and ML-F52, induced a dose-dependent apoptotic effect on promastigotes. Although no change in KMP-11 expression was observed, iridoids inhibited cell division and morphological changes in promastigote cultures. Molucidin and ML-F52 induced apoptotic mechanism of cell death, inhibited cytokinesis and induced phenotypic changes in promastigotes. Molucidin further induced ‘’nectomonad-like’’ forms and loss of kDNA, ML-F52 induced ‘cell-rounding’ with loss of flagellum. Molucidin also induced cell growth arrest at G2-M phase (54.5 %). A significant induction of apoptosis (P = .05) was shown by an enhanced peak in the sub-G1 confirming the apoptotic inducing properties of iridoids. Conclusion: This study shows the anti-leishmania activity of tetracyclic iridoids which could be further investigated for the development of new chemotherapy against Leishmaniasis.

Syndrome of inappropriate secretion of antidiuretic hormone associated with strongyloidiasis.

We report a case of syndrome of inappropriate secretion of antidiuretic hormone (SIADH) with accompanying severe strongyloidiasis in a 52-year-old male. On admission, he showed drowsiness and emaciation with severe hyponatremia. We gave sodium (saline or salts) in an i.v. drip infusion and orally without improvement. A urinalysis and plasma osmotic pressure test indicated SIADH, therefore, treatment was changed to restrict his sodium intake. The hyponatremia gradually improved initially, but the appetite loss, nausea, and hyponatremia continued. Endoscopy revealed white patches on the stomach wall and histopathological examination revealed infestation of the mucosal epithelium with numerous Strongyloides stercoralis larvae. Ivermectin treatment was then initiated and the abdominal symptoms and hyponatremia gradually resolved. We carefully investigated the underlying cause of the SIADH, such as disease of the central nervous system, lung cancer, and other malignancies, but no abnormality or clear cause could be found. We concluded that the patient developed SIADH secondary to severe S. stercoralis infection.

Prevalence of asthma, rhinitis, and eczema among children in Vientiane city, Lao PDR.

An International Study of Asthma and Allergy in Childhood (ISAAC) revealed a wide range of prevalences of childhood asthma in the world. Lao PDR had no such epidemiological data yet. This study aimed to investigate the epidemiology of asthma, rhinitis, and eczema in children in the country. A cross-sectional study was conducted in Vientiane City, the capital of Lao PDR, in 1997 using the ISAAC questionnaire. From three primary schools and one high school, 395 children, age 6-7 years, and 468 children, age 13-14 years, were chosen. The prevalence of asthma for children aged 13-14 years in Lao PDR was 25.6%, which ranks the highest in international asthma prevalence. The prevalence in allergic-rhinoconjunctivitis of children was 24.4% and atopic eczema was 7.1%. Contrary to generally accepted risk factors, there were no associations revealed between asthma prevalence and smoking of family members (especially mothers), intake of fish/meat, and male gender.

Sensitive enzyme-linked immunosorbent assay with urine samples: a tool for surveillance of schistosomiasis japonica.

An enzyme-linked immunosorbent assay (ELISA) to detect antibodies to Schistosoma japonicum soluble egg antigens (SEA) in un-concentrated urine was developed. The urine ELISA was applied to samples collected in a schistosomiasis-endemic village in China. The levels of anti-SEA antibodies detected in urine correlated well with those obtained with paired serum samples (r = 0.694, p<0.0001). Among 129 serum ELISA positives, 112 (86.8%) were positive by urine ELISA, while all 40 serum ELISA negatives from a non-endemic area were negative. The levels of anti-SEA in urine samples were stable up to 8 weeks of storage at 37 degrees C, with sodium azide as a preservative. Therefore, ELISA with urine samples can be used for the surveillance of schistosomiasis.

EFFECT OF SCHISTOSOMA JAPONICUM INFECTION ON SERUM TESTOSTERONE LEVELS IN MICE / 中国寄生虫学与寄生虫病杂志

[Objective] To determine the effect of Schistosoma japonicum infection on the testoterone level in the sera from male C57BL/6 mice. [Methods] Radioimmunoassay was used to examine testosterone levels in sera of 9 male mice experimentally infected with Schistosoma japonicum. [Results] The serum testosterone levels reduced significantly in all experimentally infected animals 45 days after infection, as compared with the uninfected controls. [Conclusion] Infection with Schistosoma japonicum decreases testosterone levels in the mouse host.