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Biomedical subjects

F Partono

Publications and source records attributed to F Partono.

At least 19 recordsLinked to original sources

Ivermectin and diethylcarbamazine trials in leaf monkeys (Presbytis cristatus) infected with Wuchereria kalimantani.

Clinical trials of Ivermectin in single oral doses of 200, 400, and 1,000 mg/kg body weight or in multiple doses of 200 mg/kg body weight for 5 consecutive days were performed in leaf monkeys (Presbytis cristatus) infected with Wuchereria kalimantani. Optimal microfilaricidal effect occurred at 200 mg/kg body weight. The drug was less effective than diethylcarbamazine in this animal model for human filariasis but had no adverse effects.

Animals

Heat shock cognate 70 is a prominent immunogen in Brugian filariasis.

A cDNA expression library constructed from RNA derived from adult stage Brugia pahangi (mixed sexes) was screened with pooled sera from chronic, amicrofilaremic cases of human lymphatic filariasis from the Indonesian island of Tanjungpinang, where Brugia malayi is endemic. Polyclonal antisera raised to purified beta-galactosidase fusion proteins from two of the most highly reactive clones identified a protein of Mr 70,000 in all stages examined (microfilariae, L3 and adults) of both B. malayi and Brugia pahangi. Derivation of the amino acid sequence from these two overlapping cDNAs identified the encoded protein as a member of the heat shock protein 70 family, and showed the closest similarity to the constitutively expressed "heat shock cognate 70" (hsc70) protein. Hybridization of hsc70 cDNAs to RNA and DNA from B. pahangi under stringent conditions identified a major transcript of 2.4 kb and revealed the existence of a family of related genes. In vitro culture of larval stages of B. pahangi at elevated temperatures (43 degrees C) resulted in increased expression of hsc70, and a classic heat shock response in which five proteins (mr 18,500, 22,000, 62,000, 70,000, and 85,000) were exclusively synthesized in microfilariae. Analysis of cross-reactivities by Western blotting implied that antibody generated by infection with B. malayi was directed at filarial-specific determinants of Brugia hsc70. However, ELISA with recombinant fusion proteins for both Plasmodium falciparum and Schistosoma mansoni hsc70 indicated that some individuals with Brugian or Bancroftian filariasis did produce antibodies which cross-reacted with plasmodial and schistosomal homologs. Thus filarial-specific antibody responses were not generated in all individuals, indicating that this molecule would not be suitable for diagnostic purposes. ELISA with a purified beta-galactosidase fusion protein from B. pahangi showed antibody responses to hsc70 across the clinical spectrum of filariasis. Alignment of the derived amino acid sequences from B. pahangi, P. falciparum, S. mansoni and rat hsc70 homologs, and comparison of the immunologic reactivity of the products of the two cDNA clones by Western blotting and ELISA suggested that these determinants were located primarily at the C terminus of the protein.

Amino Acid Sequence

Secreted antigens of filarial nematodes: a survey and characterization of in vitro excreted/secreted products of adult Brugia malayi.

We report here a broad analysis of the excretory/secretory (E/S) products of adult Brugia malayi, collected by in-vitro cultivation of the parasite. Culture media and conditions were optimized, and non-essential amino acids were found to be crucial for efficient protein synthesis under cell- and serum-free culture conditions. A close correlation was found between total protein secretion, phosphorylcholine-bearing antigen release and lactate production on each day of culture, indicating that E/S molecules are actively secreted. Parasites cultured in vitro take 2-3 days to adjust to the new environment, and show peak levels of secretion at days 3 and 4. The active secretion of phosphorylcholine by the parasite therefore justifies the measurement of this molecule as an indication of active infection, possibly reflecting total worm burdens. By comparing metabolically labelled E/S from male and female worms, several molecules of low mol. wt, namely 10,000, 13,000, 14,000 and 22,000, together with high mol. wt components of above 12,000 were found to be female specific. Tracing the origin of the E/S products, several molecules were also found to be associated with the surface. Among these, there are at least two glycoproteins, 29,000 and 51,000 of which the 29,000 molecule is a major surface protein. The immunogenicity of the E/S was examined and antigenic cross-reactivity was found with sera from most filarial infections but not with non-filarial nematodiases such as hookworm or Trichinella. However, two molecules of low mol. wt, 15,000 and 19,000, were not recognized by anti-Onchocerca sera and appeared to be potential Brugia-specific diagnostic molecules. Possible functional roles of the adult E/S products were examined but we could find no evidence of protease activity in the E/S or glutathione S-transferase activity in either the E/S or in whole somatic extract.

Animals

Monoclonal antibody to a unique surface epitope of the human filaria Brugia malayi identifies infective larvae in mosquito vectors.

We describe properties of an IgM monoclonal antibody (NEB-D1E5) raised against the human filarial parasite Brugia malayi. The antibody reacts with a stage- and species-specific determinant located on the surface of the infective-stage larva, as determined by indirect immunofluorescence. To use this reagent in epidemiological field studies, we developed an enzyme-linked immunoassay with which B. malayi larvae can be differentiated from other filarial parasites in mosquito vectors, including the morphologically indistinguishable parasite of animals Brugia pahangi. The immunoenzyme assay was 91-94% specific and 90-97% sensitive when performed on infected mosquitoes. In the absence of mosquito tissue, the levels of specificity and sensitivity increased to 100% and 97.5-100%, respectively. Binding of antibody to the surface of living larvae was abrogated by treatment of the worms with the enzymes pronase and proteinase K and with the detergents Triton X-100, octyl beta-D-glucopyranoside, and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulphonate (CHAPS). In contrast, treatment with trypsin, endoglycosidase-F, O-Glycanase, N-Glycanase, lipase, various phospholipases, boiling, 2-mercaptoethanol at 37 degrees C, or periodate did not reduce the antigenicity of the larval surface to antibody NEB-D1E5. These results suggest that the species-specific epitope is a peptide domain attached to a hydrophobic anchoring residue.

Animals

A simple method to control Brugia timori by diethylcarbamazine administration.

The results of mass treatment using 50 mg diethylcarbamazine per kg body-weight followed one year later by short term selective re-treatment in a highly endemic area of Brugia timori are described. The criteria for selection of re-treatment are simple and practical for use in rural areas. The microfilaria rate by finger prick decreased from 24% to 0 and by Nuclepore filtration from 30% to 2.5%. The disease rates were also affected favourably. 88% of persons receiving the drug reacted to treatment, this percentage slightly exceeding the total filarial infection rate (71%). The prevalence, onset, duration and nature of side reactions are briefly discussed and related to the presence of microfilaraemia and disease manifestations among the study population.

Brugia

Experimental transmission of Wuchereria bancrofti to monkeys.

Infective larvae of Wuchereria bancrofti from laboratory-raised Culex pipiens fatigans and Aedes togoi mosquitoes fed on human volunteers in Jakarta, Indonesia (J strain) and Kinmen Island, China (K strain) were introduced into Taiwan monkeys (Macaca cyclopis) by subcutaneous inoculation, by foot puncture, or by permitting infected mosquitoes to feed weekly on the monkeys. Some animals were splenectomized and others were treated with varying regimens of immunosuppressants. Necropsy was done on monkeys that died or were killed and the entire bodies were examined for worms. A total of 78 monkeys (43 males and 35 females) were exposed to infection and parasites were found in 29% of the females and 63% of males. In infections of 38 days or less worms were recovered from the testes of males and the pelt, carcass and lymph nodes of both sexes, but after 42 days of infection most worms were in the testes of males, and a few were recovered from lymph nodes and carcasses of females. Worms recovered at 8-11 days were third-stage, those found between 14 and 38 days fourth-stage, and ones found between 42 and 103 days were young adults. After 148 days most were adults and microfilariae were seen in the uteri of female worms at 160 days and later. The parasites continued to grow in size with time. Microfilariae were detected in the blood of nine monkeys between 8 and 18 months and the patent period varied from 5-21 months. Microfilarial densities were low and erratic, and periodicity could not be determined. The effectiveness of methods of administering infections and the value of various treatment regimens seem uncertain; monkey antilymphocytic sera, however, appeared to have some influence. Parasites were found in 36% of the Taiwan monkeys given the J strain and 54% of those given the K strain. A limited number of M. mulatta (3), M.irus (fascicularis) (3) and Aotus trivirgatus (4) were also given infective larvae and adult W. bancrofti were recovered from the testes of one male M. mulatta and one male M. irus; uterine microfilariae were found in one female worm from the latter monkey. A. trivirgatus were negative. Low numbers of infective larvae recovered from mosquitoes fed on patent monkeys were introduced intermittently into seven clean monkeys and one became microfilaremic between 11 and 17 months postinoculation.

Aedes

A multidisciplinary study on bancroftian filariasis in Jakarta.

A study on bancroftian filariasis in Jakarta has indicated that one person in one year could be exposed to 223,000 bites of Culex pipens fatigans and to 1,941 infective-stage larvae of Wuchereria bancrofti. Blood surveys with 20 mm3 samples revealed a microfilaria rate of 6%. Although some cases of hydrocele (4% of 272 males examined) were found, there was little evidence of severe filarial disease in either males or females. Amongst wild-caught mosquitoes only 0.3% contained infective larvae, but much higher levels of vectorial competence were established under laboratory conditions. High daily vector mortality (30%) coupled with noticeable improvements in standards of living could have been important factors preventing an increase in endemicity.

Adolescent

Epidemiological and clinical features of Brugia timori in a newly established village. Karakuak, West Flores, Indonesia.

The epidemiological and clinical features of Brugia timori filariasis in a newly established village, Karakuak, West Flores, is described. The microfilarial rate by finger stick and Nuclepore filtration was 24% and 30%, respectively, and the disease rate 64%. Infected persons were found in every family and household with no predominant age or sex preference. Development of elephantiasis in the population was associated with residence in the new village of Karakuak, where extensive rice field cultivation was initiated soon after arrival. The irrigated fields provided excellent breeding sites for the vector, Anopheles barbirostris. People with no previous exposure to the parasite developed elephantiasis earlier and more frequently than those originating from other endemic areas.

Adolescent

The association of clinical filariasis and Wuchereria bancrofti infections in Jakarta.

In a study carried out in Jakarta, Indonesia between 1969--1977, patients with clinical manifestations of filariasis were examined for microfilaremia. Finger blood (3 X 20 microliter) were examined by conventional methods, venous blood by membrane filtration, urine and hydrocele fluid by concentration methods. A total of 1,422 patients were examined and 182 (12.8%) were positive by blood smear; however, examination by membrane filtration and concentration of urine and hydrocele fluid did not appreciably increase the association between clinical manifestations and Wuchereria bancrofti infection. Approximately 16% of the patients without patent infection were indirectly associated with filariasis, since they co-habitated with persons with microfilaremia. Results of these studies showed that more than 60% of the patients were without patent infection. Factors that may be responsible for these findings are briefly discussed.

Adolescent

Serum immunoglobulin levels in Indonesians with bancroftian filariasis.

Serum immunoglobulin (Ig) levels (IgA, IgM, IgG, IgE) were determined on 185 Indonesians mostly males with and without microfilaremia and with and without clinical disease (hydrocele and elphantiasis). No significant differences were found in IgA and IgE levels in any of the groups but significantly higher IgG levels were found in microfilariae positive persons than those without microfilaremia. Persons with microfilariae alone and symptoms alone, but not both, had higher levels of IgM. There were no correlations between disease category and Ig levels.

Adolescent

Brugia malayi in Kresek, West Java, Indonesia: the effect of environmental changes of filarial endemicity.

A survey was conducted in Kresek, West Java, Indonesia and 813 persons from six villages examined for filariasis. Only eight had microfilaremia and one male person had elephantiasis. Comparing results obtained approximately ten years earlier and the present survey, there was a marked decrease in the microfilarial rate in Kresek, especially in Penameng, during the interim. There was no drug treatment, insecticide application or any dramatic population movement. The development of a well-organized irrigation system and the conversion of the swamps into rice-fields resulted in a decrease of breeding sites for Mansonia mosquitoes and subsequently a decrease in filariasis.

Adolescent

Clinical features of timorian filariasis among immigrants to an endemic area in West Flores, Indonesia.

Fifty-nine persons, who immigrated into a Brugia timori endemic area from non-filarial areas on the island of Flores, Indonesia were examined for filariasis after residing in the village for 2 to 10 years. Six persons had B. timori microfilaremia and 31 had filarial disease. The disease seems to affect immigrants from non-filarial areas severely within a relatively short period. Among those residing in the village, for 2 years, the microfilaria rate was 5% and the elephantiasis rate 21%. Selective treatment using 50 mg diethylcarbamazine per Kg body weight was given to all microfilaria (Mf) positive persons. Approximately one year later the Mf-rate by finger stick and Nuclepore filtration was 9% and 18% respectively. There was indirect indications that the Mf-rate might increase with the passage of time. However, the total filarial disease rate remained constant during the one year period. The relationship between these findings and American servicemen exposed to filariasis during World World II is briefly discussed.

Adolescent