Pion SD et al. (2011) Acta Trop "Individual host factors associated with Onchocerca volvulus microfilarial densities 15, 80 and 180 days after a first dose of ivermectin."

Nana-Djeunga H, Pion SD, Kamgno J, Boussinesq M, Grout L

[Acta Trop, 2011]

Reduction in Onchocerca volvulus skin microfilarial densities after treatment with ivermectin shows wide between-host variation. Data from two separate studies conducted in Cameroon on onchocerciasis patients treated for the first time with ivermectin were analyzed to identify host factors associated with microfilarial density at different time-points after treatment. In one site (Nkam valley), the dataset included 103 adult males for whom age, number of palpable onchocercal nodules and microfilarial densities on D0 (pre-treatment), D15, D80 and D180 were available. In the other site (Vina valley), analyses were conducted on 965 individuals of both sexes aged 5 years and over; in this dataset, available information included age, gender, exact dose of ivermectin received, onchocerciasis endemicity level in the village of residence and microfilarial densities on D0 and D180. Negative binomial regression models of microfilarial density at the different intervals post-treatment were fitted, using maximum likelihood, with the available independent variables. Gender and age were found to be associated with microfilarial density on D180. The initial microfilarial density influenced post-treatment densities at all the time-points. All other things being equal, microfilarial densities on D180 were higher in individuals harbouring a higher number of nodules or living in communities with high endemicity levels. This study demonstrates that O. volvulus microfilarial density measured after a first treatment with ivermectin, and thus probably the rate of skin repopulation by microfilariae (mf) varies according to several host factors. Should such factors also influence ivermectin efficacy after repeated treatment, then they should be taken into account to determine whether sub-optimal responses to treatment reported from various areas in Africa are actually due to parasite-related factors, particularly to the emergence of resistant populations.

Demanou M et al. (2003) Ann Trop Med Parasitol "Experimental studies on the transmission of Onchocerca volvulus by its vector in the Sanaga valley (Cameroon): Simulium squamosum B. Intake of microfilariae and their migration to the haemocoel of the vector."

Basanez MG, Demanou M, Boussinesq M, Enyong P, Pion SD

[Ann Trop Med Parasitol, 2003]

As large-scale ivermectin distribution is becoming the mainstay of onchocerciasis control in Africa, the issue of its impact on local transmission is increasing in importance. The vector competence of Simulium squamosum B in the severe focus of the Sanaga valley, Cameroon, was therefore investigated, by feeding 1320 flies on 14 carriers of Onchocerca volvulus microfilariae (mff). The results enabled the relationships between skin microfilarial load, microfilarial intake by the flies, the proportion and mean number of ingested mff that succeed in reaching the fly's haemocoel, and the frequency distribution of the ingested mff to be described, as functions of time post-engorgement (p.e.) and parasite density (while taking account of possible measurement error in the predictor variable). The proportion of flies with haemocoelic mff and the mean number of mff/fly increased up to 3 h p.e. The proportion of flies with ingested mff was non-linearly related to mean intake, via the negative-binomial distribution, with the overdispersion parameter k best described as an increasing (power) function of the mean. Approximately one in every three ingested mff escaped imprisonment by the peritrophic matrix, irrespective of the skin microfilarial load or the intake of mff. The relationship between successful and input mff is nearly linear (indicating proportionality) in S. squamosum B. These results are compared with those from O. volvulus-S. damnosum s.l. combinations in other West African foci.

Boussinesq M et al. (2001) Ann Trop Med Parasitol "Studies on the macrofilarial population of Onchocerca volvulus in hyper-endemic villages of the Central province of Cameroon."

Gardon J, Duke BO, Boussinesq M, Kamgno J, Pion SD

[Ann Trop Med Parasitol, 2001]

The population structure of Onchocerca volvulus macrofilariae was studied in villages of central Cameroon where onchocerciasis is hyper-endemic. One nodule selected at random was removed from each of 576 adult males, and examined by histology. The numbers of male and female worms/nodule, and the status of the female worms (fecund, non-fecund, or dead) were recorded. The observations were analysed to evaluate whether the mean numbers of worms of each category varied in relation to the patient's age, the level of endemicity in his village, the anatomical localization of the nodule, the weight of the nodule, and the total number of palpable nodules harboured by the patient. The results obtained were very similar to those reported from West Africa. The mean numbers of dead female worms/nodule were relatively high in the villages with the lowest levels of endemicity. The mean numbers of fecund females and of live males were significantly higher in the nodules located around the knees. These results provide information which might be useful in modelling the population dynamics of O. volvulus, and also in the context of trials of any potentially macrofilaricidal drugs.

Su C et al. (2020) Neurochem Int "Inhibitory Potency of 4- Substituted Sampangine Derivatives toward Cu2+ mediated aggregation of amyloid -peptide, Oxidative Stress, and Inflammation in Alzheimer's Disease."

Tang H, Chen K, Li W, Su C, Chen Y

[Neurochem Int, 2020]

Cu²⁺ plays a key role in the pathogenesis of Alzheimer's disease (AD). The dysregulation of Cu²⁺ can cause neuronal damage and aggravate development of AD. Moreover, a series of 4-substituted sampangine derivatives have been investigated as inhibitors of acetylcholinesterase and -amyloid (A) aggregation for the treatment of AD in our previous studies. In the present study, we reported that one of these derivatives SD-1 was able to modulate Cu²⁺-mediated multiple pathological elements in AD. The high selectivity of SD-1 for Cu²⁺ over other biologically relevant metal ions was demonstrated by ITC. Western blotting analysis, light-scattering study, DCF-DA assay and paralysis experiment indicated that SD-1 suppressed the formation of Cu²⁺-A species, alleviated the Cu²⁺-A species induced neurotoxicity and inhibited the production of ROS catalyzed by Cu²⁺-A species in SH-SY5Y cells over-expressing the Swedish mutant form of human APP (APPsw SH-SY5Y) and A42 transgenic C elegans (CL2020). Furthermore, SD-1 inhibited the expressions of NO, iNOS, TNF-, IL-1 and IL-6 induced by Cu²⁺ in BV2 microglial cells. Collectively, these findings provided valuable insights into the design and development of potent metal-chelating agents for AD treatment.

Fan PC et al. (1985) Southeast Asian J Trop Med Public Health "Experimental infection of subperiodic Brugia malayi in laboratory rats with emphasis on evaluation by four techniques."

Hsu YP, Huang CM, Wu CC, Ho CM, Fan PC

[Southeast Asian J Trop Med Public Health, 1985]

Infective larvae of subperiodic B. malayi from South Kalimantan (Borneo), Indonesia collected from laboratory-raised Ae. togoi mosquitoes after feeding on infected mongolian gerbils (Meriones unguiculatus) were inoculated subcutaneously into the groin areas of 15 SD and 36 LE rats. Blood was examined weekly by membrane filtration and thick smears starting 10 weeks post-infection. Microfilariae were found in 3 SD and 4 LE rats, the mf infection rate of 20% and 11% respectively. The prepatent period was significantly shorter in the SD rats (99-112 days) than those in the LE rats (110-153 days). The patent period was longer in the LE rats (208-703 days) than in the SD rats (236-543 days), and the mf density was similar (17.5 mf/20 c.mm blood against 16 mf/20 c.mm blood). At necropsy, 6 (3 female and 3 male) adult worms were recovered from 3 of 6 SD rats and 12 (9 female and 3 male) adult worms from 4 of 20 LE rats; all worms were found in the testes. The results of xenodiagnostic, histochemical staining and measuring spicules and protuberances, demonstrated clearly the difference between both species of Brugia. All dissected Ar. subalbatus mosquitoes exposed to B. pahangi became infected (100%), but none of those to subperiodic B. malayi were infected (0%). The mf of both species of Brugia in thick films stained with naphthol-AS-TR-phosphate showed that the excretory and anal pores of subperiodic B. malayi mf exhibited acid phosphatase activity and only a little activity was seen in other parts; while B. pahangi mf showed heavy diffuse acid phosphatase activity along the entire length of the body.(ABSTRACT TRUNCATED AT 250 WORDS)

Bourguinat C et al. (2007) PLoS Negl Trop Dis "Genetic selection of low fertile Onchocerca volvulus by ivermectin treatment."

Gardon J, Prichard RK, Bourguinat C, Pion SD, Boussinesq M, Kamgno J, Duke BO

[PLoS Negl Trop Dis, 2007]

BACKGROUND: Onchocerca volvulus is the causative agent of onchocerciasis, or "river blindness". Ivermectin has been used for mass treatment of onchocerciasis for up to 18 years, and recently there have been reports of poor parasitological responses to the drug. Should ivermectin resistance be developing, it would have a genetic basis. We monitored genetic changes in parasites obtained from the same patients before use of ivermectin and following different levels of ivermectin exposure. METHODS AND FINDINGS: O. volvulus adult worms were obtained from 73 patients before exposure to ivermectin and in the same patients following three years of annual or three-monthly treatment at 150 microg/kg or 800 microg/kg. Genotype frequencies were determined in beta-tubulin, a gene previously found to be linked to ivermectin selection and resistance in parasitic nematodes. Such frequencies were also determined in two other genes, heat shock protein 60 and acidic ribosomal protein, not known to be linked to ivermectin effects. In addition, we investigated the relationship between beta-tubulin genotype and female parasite fertility. We found a significant selection for beta-tubulin heterozygotes in female worms. There was no significant selection for the two other genes. Quarterly ivermectin treatment over three years reduced the frequency of the beta-tubulin "aa" homozygotes from 68.6% to 25.6%, while the "ab" heterozygotes increased from 20.9% to 69.2% in the female parasites. The female worms that were homozygous at the beta-tubulin locus were more fertile than the heterozygous female worms before treatment (67% versus 37%; p = 0.003) and twelve months after the last dose of ivermectin in the groups treated annually (60% versus 17%; p<0.001). Differences in fertility between heterozygous and homozygous worms were less apparent three months after the last treatment in the groups treated three-monthly. CONCLUSIONS: The results indicate that ivermectin is causing genetic selection on O. volvulus. This genetic selection is associated with a lower reproductive rate in the female parasites. We hypothesize that this genetic selection indicates that a population of O. volvulus, which is more tolerant to ivermectin, is being selected. This selection could have implications for the development of ivermectin resistance in O. volvulus and for the ongoing onchocerciasis control programmes.

Duke BO et al. (2002) Parasitology "Neoplastic change in Onchocerca volvulus and its relation to ivermectin treatment."

Pion SD, Gardo J, Boussinesq M, Kamgno J, Marty AM, Duke BO, Peett DL

[Parasitology, 2002]

A pleomorphic neoplasm (PN) is described from sections of Onchocerca volvulus worms in nodules excised from Cameroonian patients. PN is confined to older, non-fecund, female worms, and those classed as moribund/dead. It is mainly composed of small, roundish, basophilic cells of diverse sizes, often forming a 'rosette' pattern around amorphous eosinophilic centres. The cells have a high nuclear/cytoplasmic ratio and up to 2-3 mitoses/high-power field; some become grossly enlarged, highly polymorphic and contain large, irregular blocks of chromatin. The eukaryotic PN cells first appear posteriorly in the pseudocoelom, probably from ovarian cells; they spread anteriorly, invading or compressing the uteri. Ivermectin treatment increased the prevalence PN from 3.7% of 1422 female worms in 637 patients before treatment to 17.5% of 1134 worms in 511 patients after 3 years treatment. Ivermectin at 400-800 microg/kg annually, or at 150 microg/kg or 400-800 microg/kg 3-monthly, over 3 years, did not increase the PN prevalence significantly, as compared with standard doses of 150 microg/kg annually. In other small series of African patients, PN prevalence increased in worms 2, 4, 6 and 10 months after ivermectin treatment; but there was no increase after treatment with amocarzine, albendazole or diethylcarbamazine and suramin. PN may partly account for the increased macrofilaricidal action of ivermectin on female O. volvulus in patients treated for 3 years at 3-monthly intervals.

Nana-Djeunga H et al. (2012) Mol Biochem Parasitol "Single nucleotide polymorphisms in -tubulin selected in Onchocerca volvulus following repeated ivermectin treatment: possible indication of resistance selection."

Gardon J, Pion SD, Boussinesq M, Bourguinat C, Njiokou F, Nana-Djeunga H, Prichard RK, Kamgno J

[Mol Biochem Parasitol, 2012]

The control of onchocerciasis or river blindness by mass treatment of the population with ivermectin (IVM) has been a great success until now, so that in certain foci its elimination has become feasible. However, after more than 20 years of repeated IVM mass treatment, the disease still persists in many endemic countries. Sub-optimal responses and genetic changes have been reported in Onchocerca volvulus populations under high IVM pressure but more work is needed to determine whether resistance is developing. The situation needs to be urgently clarified to preserve the achievements of onchocerciasis control programs. In this study, O. volvulus adult worms were collected from the same individuals, before IVM exposure and following three years of annual or three-monthly treatments at 150 g/kg or 800 g/kg. Four single nucleotide polymorphisms (SNPs) occurring in the -tubulin gene of these parasites were investigated. We found changes in genotype frequencies in O. volvulus -tubulin gene associated with IVM treatments. The SNP at position 1545 (A/G) showed a significant increase in frequency of the less common nucleotide in the female worms following treatment. After 13 three-monthly treatments, female worm homozygotes with the less common genotype, prior to treatment, increased in frequency. The selected homozygotes, as well as heterozygotes, appeared to be less fertile (without or with very few embryonic stages in their uteri) than the wild-type homozygotes. These results provide additional evidence for genetic selection and strengthen the warning that selection for IVM resistance may be occurring in some O. volvulus populations.

Bourguinat C et al. (2006) Parasitology "Genetic polymorphism of the beta-tubulin gene of Onchocerca volvulus in ivermectin naive patients from Cameroon, and its relationship with fertility of the worms."

Pion SD, Boussinesq M, Gardon-Wendel N, Prichard RK, Kamgno J, Duke BO, Gardon J, Bourguinat C

[Parasitology, 2006]

Observations of low response of patients infected with Onchocerca volvulus to ivermectin suggest that the parasite may be under a selection process toward potential resistance. To limit the extension of this phenomenon, it is crucial to characterize the genes of O. volvulus that are involved. For this, O. volvulus adult worms collected before the introduction of ivermectin in an onchocerciasis endemic area of central Cameroon were genotyped for beta-tubulin. To derive a baseline to investigate the selective pressure of ivermectin, we analysed (1) the frequency distribution of the beta-tubulin alleles, and (2) the relationship between the different beta-tubulin related genotypes and the fertility status of the female worms. The frequency of allele b of the beta-tubulin gene was very low, as it was observed in West Africa. We observed a deficit of heterozygous female worms leading to Hardy Weinberg disequilibrium, which might be explained by a shorter life-span of these worms compared to the homozygous worms. Unexpectedly, our results also show that the heterozygous female worms were much less fertile than the homozygotes: more than two thirds of the homozygotes were fertile, whereas only 37% of the heterozygotes were fertile. These results will be further considered when analysing post-treatment data.

Yu Z et al. (2013) J Hazard Mater "Inhibitions on the behavior and growth of the nematode progeny after prenatal exposure to sulfonamides at micromolar concentrations."

Zhang J, Deng H, Yin D, Chen X, Yu Z

[J Hazard Mater, 2013]

Sulfonamides are one typical antibiotic which is an emerging hazardous material to the ecological stability due to their continuously application and biological effects to non-target organisms. The parent-progeny transgenerational effects need investigations to indicate their long-term consequences. Currently, we tested the transgenerational effects of sulfadiazine (SD), sulfapyridine (SP) and sulfamethazine (SMZ) on L3 larva of Caenorhabditis elegans. The nematodes were exposed to aqueous sulfonamides at micromolar concentrations for 96 h, and then the effects on the behavior and growth in the exposed parent and unexposed progeny were measured. Results showed that SD, SP and SMZ inhibited three behavior indicators including body bending frequency (BBF), reversal movement (RM) and Omega turn (OT), and the growth indicator (body length, BL). Behavior indicators showed higher sensitivities than the growth indicator, and BBF had the highest sensitivity among the behavior indicators. Moreover, the effects of sulfonamides were also observed in the unexposed progeny with partially rescued or more severe inhibitions on the indicators. The behavior also showed higher sensitivity than the growth in the progeny. The transgenerational effects of sulfonamides indicated that parental exposure can multiply the harmful effects of antibiotic pollution in following generations and their potential ecological risks at environmental concentrations were further raised.