Infectivity of asymptomatic Plasmodium-infected human populations to Anopheles dirus mosquitoes in western Thailand

The infectivity of Plasmodium-infected humans in western Thailand was estimated by feeding laboratory-reared Anopheles dirus Peyton and Harrison mosquitoes on venous blood placed in a membrane-feeding apparatus. Between May 2000 and November 2001, a total of 6,494 blood films collected during an active malaria surveillance program were checked by microscopy for the presence of Plasmodium parasites: 3.3, 4.5, and 0.1% of slides were P. falciparum- (Pf), P. vivax- (Pv), and P. malariae (Pm)-positive. Venous blood was collected from 70, 52, 6, and 4 individuals infected with Pf, Pv, Pm, and mixed Pf/Pv, respectively, with 167 uninfected individuals serving as negative controls. Only 10% (7/70), 13% (7/52), and 0% (0/6) of membrane feeds conducted on Pf-, Pv-, and Pm-infected blood yielded infected mosquitoes. One percent (2/167) of microscope-negative samples infected mosquitoes; however, both samples were subsequently determined to be Pf-positive by polymerase chain reaction. Gametocytes were observed in only 29% (4/14) of the infectious samples. All infections resulted in low oocyst loads (average of 1.2 oocysts per positive mosquito). Only 4.5% (10/222) of mosquitoes fed on the seven infectious Pf samples developed oocysts, whereas 2.9% (9/311) of mosquitoes fed on the seven infectious Pv samples developed oocysts. The probability of a mosquito becoming infected with Pf or Pv after a blood meal on a member of the human population in Kong Mong Tha was estimated to be 1 in 6,700 and 1 in 5,700, respectively. The implications toward malaria transmission in western Thailand are discussed.     

Sesquiterpene lactone helenalin suppresses Leydig and adrenocortical cell steroidogenesis by inhibiting expression of the steroidogenic acute regulatory protein

Helenalin is a potent anti-inflammatory and anti-neoplastic agent isolated from several plant species of the Asteracea family. Here, we have investigated the effects of helenalin on steroidogenesis activated by adrenocorticotropic hormone (ACTH) and human chorionic gonadotropin (hCG) in primary cultures of rat adrenocortical and Leydig cells. Our findings demonstrate that helenalin inhibits both ACTH- and hCG-activated steroidogenesis in these cells. This effect was already evident after 2–3 h treatment with helenalin. In contrast, steroidogenesis from 22R-OHC, a cell-permeable form of cholesterol, was not inhibited by helenalin, suggesting that the expression of the steroidogenic acute regulartory (StAR) protein might be inhibited by this compound. Indeed, helenalin attenuated StAR protein expression activated by ACTH and hCG in adrenocortical and Leydig cells as assessed by PAGE/Western analyses. This inhibitory action of helenalin on steroidogenic cell functions indicates novel mechanisms of action of this compound which may be of potential therapeutic interest. However, it also poses safety concerns relating to possible negative side-effects on anabolism and systemic stress.     

Genetic markers for the identification and characterization of Opisthorchis viverrini, a medically important food borne trematode in Southeast Asia

The liver fluke, Opisthorchis viverrini, is one of the major food borne trematodes in Southeast Asia, where infection causes hepatobiliary disease and subsequent development of cholangiocarcinoma. In Thailand, O. viverrini is most prevalent in the northeast where there is marked regional variation in the rate of infection in humans at provincial, district and village levels. To date, the roles of genetic variation of O. viverrini on this observed variability in infection, transmission and associated disease are not known. We have applied multilocus enzyme electrophoresis (MEE), specifically allozyme electrophoresis, to isolates of O. viverrini from Thailand and Laos to establish genetic markers to examine its systematics and population structure. Forty-six enzymes commonly found useful for genetic characterisation in parasitic helminths were screened, and of these, 33 enzymes gave sufficient staining and resolution to act as potential genetic markers. Sixteen enzymes were monomorphic and 17 enzymes were polymorphic in the pools of worms examined. Whether they are indicative of different enzyme loci, heterozygosity or unique genotypes within the pools of worms examined remains to be determined. Preliminary investigations examining five individual worms at enzyme loci where pools of worms showed multiple bands have confirmed the diagnostic value of the enzyme loci established as well as providing evidence of potential population sub structuring and heterozygosity. For the first time, we have established at least 17 enzymes that provide the basis to undertake comprehensive genetic analyses of the systematics and population structure of O. viverrini, a medically important food borne trematode in Southeast Asia.     

Enzyme markers to identify and characterize Opisthorchis viverrini in Thailand and Lao PDR

We conducted an allozyme electrophoretic study to explore potential enzyme markers to distinguish Opisthorchis viverrini in Thailand and Lao PDR. Twenty-eight enzymes encoding presumptive 32 loci were established. The enzymes glucose-6-phosphate dehydrogenase and pyruvate kinase were diagnostic between two geographically separate isolates from Thailand. Twelve enzymes, ie, aconitate hydratase, aldolase, creatine kinase, enolase, esterases, fumarate hydratase, aspartate aminotransferase, glucose-phosphate isomerase, alanine aminotransferase, isocitrate dehydrogenase, malic enzyme, and pyruvate kinase, also provided diagnostic markers for these two isolates from Thailand and one isolate from Lao PDR.     

Dual innervation of the brachialis muscle

A study of the innervation of the brachialis muscle was carried out on 45 male and 31 female Thai cadavers between the ages of 15 and 92 years (mean = 59 years). The dissections revealed that all brachialis muscles received innervation from the musculocutaneous nerve and that 81.6% were also innervated by a branch from the radial nerve. Among the brachialis muscles with a dual nerve supply, two patterns of branching from the radial nerve were observed: in one pattern the branch to the brachialis had a descending course (58%) and in the other pattern the nerve ascended or recurred (42%) to innervate the muscle. The radial nerve branch penetrated the inferolateral part of the brachialis muscle in 83% of cases (103/124) and its middle third in 17% of specimens (21/124). The basis for the dual innervation may result from fusion of two different embryonic muscular primordia: the ventral (flexor) and the dorsal (extensor) muscle masses. In contrast to a brachialis muscle innervated by the musculocutaneous nerve only, a muscle with dual innervation may be spared significant denervation by an anterior approach to the humerus through a longitudinally bisected muscle. In a dually innervated muscle, however, separation of the brachialis and brachioradialis muscles during surgery may put the radial nerve branch to the brachialis at risk.     

The influence of pregnancy on intestinal parasite infection in Thai women

The relationship between pregnancy and both the susceptibility and pathogenicity of parasite infections is disputed. This study compares the prevalence and intensity (as measured by density of eggs in stool samples) of intestinal helminth infections in pregnant and control groups of women from Khon Kaen Province in the northeast of Thailand. Stool samples were taken at the end of the first, second and third trimesters of pregnancy as well as 2 months after parturition and compared for the two groups. There were no significant changes in the prevalence of any of the common helminth species during the course of pregnancy or between the pregnant and control groups. Nor was there any evidence that the density of helminth eggs in the stool samples differed between sample times or between the pregnant and control groups. Our study therefore supports the hypothesis that pregnancy does not influence the course of human infection with helminths.     

Expanding clinical spectrum of non-autoimmune hyperthyroidism due to an activating germline mutation, p.M453T, in the thyrotropin receptor gene

Objective  To describe clinical and genetic features of a Thai family with non-autoimmune hyperthyroidism (NAH) caused by an activating germline mutation in the thyrotropin receptor ( TSHR ) gene.
Patients  Three affected individuals from the same family (a father and his two children) were studied. Clinical and imaging findings were reviewed and compared.
Genetic analysis  Genomic DNA was extracted from peripheral blood leukocytes and mutation analysis of the entire coding sequence of the TSHR gene was performed in both children and their parents by direct DNA sequencing.
Results  A heterozygous germline T to C transition in exon 10 of the TSHR gene (c.1358T→C) resulting in the substitution of methionine (ATG) by threonine (ACG) at codon 453 (p.M453T) was identified in the father and his two children. They presented with different clinical severity and variable age of onset. In addition to hyperthyroidism, ventriculomegaly and bilateral shortening of the fifth metacarpal bones and the middle phalanges of the fifth fingers were consistently found in all affected individuals.
Conclusions  Ventriculomegaly and bilateral shortening of the fifth metacarpal bones and the middle phalanges of the fifth fingers might be characteristic features of NAH because of an activating TSHR germline mutation. In addition, the shortening of the middle phalanges of the fifth fingers has never been previously described, expanding the phenotypic spectrum of the disease.     

Immunodiagnosis of human fascioliasis using an antigen of Fasciola gigantica adult worm with the molecular mass of 27 kDa by a dot-ELISA

Immunodominant antigens of an approximate molecular mass of 27 kDa (FG 27) were obtained from an excretory-secretory product of adult Fasciola gigantica by a simple continuous-elution method. A dot-ELISA using the FG 27 antigen was developed for the detection of specific antibodies from patients infected with F. gigantica. Control sera were obtained from patients with other parasitic infections and healthy volunteers. The accuracy, sensitivity, specificity, and positive and negative predictive values were 98.2%, 100%, 97.4%, 76.9% and 100%, respectively. This dot-ELISA is a specific, sensitive and easy to perform method for the rapid diagnosis of fascioliasis, particularly when more complex laboratory tests are unavailable.