Glucocerebrosidase genetic variants in Malays with early and late-onset Parkinson’s disease

@#Background: Mutations in glucocerebrosidase (GBA) have been associated with the risk of developing Parkinson’s disease (PD) in different ethnic populations. The prevalence of GBA mutations among Malay PD patients is unknown. Thus, the aim of this study was to determine the frequency of GBA mutations among Malay PD patients, focusing on early (EOPD) and late-onset (LOPD) patients. Methods:EOPD (n = 50) and LOPD (n = 50) patients along with 50 ethnically and age-matched control wererecruited. The GBA exons of these patients were sequenced using the Ion Torrent PGMTM System. Results: Five heterozygous mutations exclusive to EOPD patients were identified; c.-203A>G,p.S146L, p.R159Q, p.L483P and p.L483R+c.-145G>A. In LOPD patients, c.543C>T(p.(F181=)), c.28-10C>A and p.R202Q were identified in which this p.R202Q was also present in a control subject. In addition, c.259C>A(p.(R87=)) and c.-145G>A were identified in two control subjects. In summary, we observed GBA mutations in 8% and 6% of Malay PD cases and control subject, respectively. The prevalence of GBA mutations was higher in EOPD (10%) than LOPD (6%). However, these differences were not statistically significant; [PD vs. controls: OR = 1.36, 95%CI 0.35-5.38, p = 0.752] and [EOPD vs. LOPD: OR = 1.74, 95%CI 0.39-7.71, p = 0.715]. Conclusion: We identified five exclusive heterozygous GBA mutations in EOPD patients which might predict the increase susceptibility of Malays to develop PD at young age. These findings could add knowledge into the existing evidences linking genetic alterations in GBA and PD.

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

Asthma symptoms improvement in moderate persistent asthma patients with gastroesophageal reflux disease (GERD): the role of proton-pump inhibitor.

This study aimed to evaluate effect of proton pump inhibitor (esomeprazole) on asthma symptoms, use of inhaled bronchodilator and peak expiratory flow rate (PEFR) in moderate persistent asthma with gastroesofageal refluks disease (GERD). This randomized single blind, controlled clinical trial study was conducted at Persahabatan Hospital, Jakarta from July 2004 until October 2005. Samples were moderate persistent asthma patients with GERD. GERD is diagnosed GERD symptoms and proof of oesophagitis from endoscopy and or histapatologic examination from oesophagus biopsy. Phase 1:2 week run-in period patient received inhaled budesonide 2x200 ug/day. Phase 2: patient randomised to receive inhaled budesonide 2 x 400 ug/day with esomeprazole 40 mg/day or without esomeprazole (control group) for 8 weeks. Phase 3: 4 week wash out period, patient receive inhaled budesonide 2 x 200 ug/day. Diary cards were assessed at run-in periode, after treatment 4 weeks, 8 weeks and wash out. There were 32 patients (23 female and 9 male) completed the study. Mean total asthma symptoms score daily were significantly decreased on esomeprazole vs without esomeprazole after 8 weeks (-2.29 vs -0.90; p < 0.05). Mean use of inhaled bronchodilator was significantly decreased on esomeprazole vs without esomeprazole after 8 weeks (-1.09 vs -0.42; p < 0.05). Morning and evening PEFR improved higher on esomeprazole than without esomeprazol but were not significantly difference. In conclusion, administration esomeprazole 40 mg daily improved asthma symptoms and lower the use of inhaled bronchodilator in moderate persistent asthma patients with GERD.

Testosterone undecanoate and depo medroxyprogesterone acetate induced azoospermia through increased expression of spermatogenic cell caspase 3.

The administration of a combination of testosterone undecanoate (TU, a long-acting androgen) and depo-medroxyprogesterone acetate (DMPA) were investigated in term of suppression of rat sperm concentration in vivo to azoospermia through increasing activity of spermatogenic cell caspase 3. Adult Sprague Dawley rats received TU and DMPA of 2.5 mg and 1.25 mg, respectively, a regimen known to rapidly reduce intra testicular testosterone and to produce azoospermia within 12 weeks. Caspase 3 positive sperm cells increased compared with control levels during 6 weeks post-injection and increased further through 60 weeks. Immunohistochemistry for caspase 3 revealed that spermatocytes represented the predominant caspase 3 positive germ cells. Modest immunoreactivity for caspase-3 was localized to nuclear region of the germ cells of control and treated testes. Immunohistochemistry study revealed significantly increased caspase-3 expression in nuclei of germ cells during administration of TU+DMPA to rats. Additionally, the caspase 3 content was significantly increased in germ cells during rats were administered TU+DMPA (453.90±84.88 cells/200 seminiferous tubules) and caspase 3 significant increase in immunoreactivity was localized to the nuclei of spermatogonia, spermatocytes, and spermatids. Taken together, these results indicated that azoospermia due to reduced intratesticular testosterone concentration was caspase-3 activation dependent and suggested that the increase in active caspase-3 in the nucleus may be involved in the induction of decreased sperm production.

Malaria in Jayapura District, Papua Province, Indonesia and resistance to sulfadoxine-pyrimethamine.

Malaria poses a major public health problem in Papua. The rapid spread of drug-resistant parasites and deterioration of the malaria control program following the monetary crisis and civil strife in the late 90’es have worsened the malaria situation in many areas of Papua. To re-assess the current magnitude of malaria problem, a malariometric survey was conducted in three sub-districts of Jayapura District, as well as antimalarial drug sensitivity test using sulfadoxine-pyrimethamine (SP) at the outpatient clinics of Abepura General Hospital (RSU), Hamadi Primary Health Center and Panti Asuhan Katolik Clinics, Sentani, Jayapura District, Papua Province. Screening of 543 subjects in three sub-districts revealed 185 malaria positive subjects, either with P. falciparum, P. vivax or mixed of the two species. All of the malaria cases were asymptomatic. In the SP drug sensitivity test, of the 56 subjects involved, 32 were classified as early treatment failure and two subjects were classified as late treatment failure. The remaining 22 subjects were classified as adequate clinical and parasitological response (ACPR). These results indicated that the surveyed area has a high malaria prevalence and treatment failure to SP and therefore alert to the need of deploying new antimalarial regiment that suit the remote setting.

Malaria and Anopheles spp in the villages of Salubarana and Kadaila, Mamuju District, South Sulawesi Province, Indonesia.

Malaria epidemiology in Salubarana and Kadaila villages, Mamuju District, South Sulawesi Province, Indonesia was studied from July-December 2002 to obtain baseline data related to the parasite, mosquito vector, human host, and environmental factors. Monthly malariometric surveys conducted during the six- month period revealed a relatively high prevalence of malaria in Kadaila in comparison to Salubarana. Kadaila was mainly inhabited by migrants from Java, Bali, Lombok, and from other districts of South Sulawesi. Salubarana, on the other hand, was inhabited mainly by indigenous Mandarese. Analysis of 1,113 blood smears taken from individuals randomly involved in the survey revealed 59 positive samples, consisting of Plasmodium falciparum or P. vivax. These two species predominated the samples examined and were occasionally found as mixed infection. Anopheles barbirostris was confirmed as a vector for malaria in this area whereas 7 other species An. barbumbrosus, An. parangensis, An. vagus, An. crawfordi, An.pseudobarbirostris, An. Tessellatus, and An. subpictus should be considered as potential vectors. The first mentioned species predominated, using mainly riverbeds and ground pools as breeding places with a habit of indoor or outdoor resting. These findings could be useful for establishing evidence-based malaria control program in the area.

Age structure of Anopheles subpictus (Diptera:Culicidae) collected by a light trap in Halmahela, Indonesia.

Age structure of Anopheles subpictus Grassi was studied at a Halmahela village, Indonesia. Ovary development of females reared with 2% sugar solution after emergence ceased at Stage I and their ovary size remained small (mean length x width = 0.68 x 0.18 mm). Females with such small ovaries occupied 21.2% of unfed Stage I females in cattle-bait samples. The remaining unfed Stage I or II females had cleary larger ovaries (mean 1.17 x 0.39 mm). Unfed Stage I or II females collected by the outdoor light trap all had large ovaries (mean 1.42 x 0.34 mm) irrespective of stages and parity. The parous rate of unfed stage I or II females collected by the light trap (86.6%) was significantly higher than that of cattle-bait samples (69.6%). Feasibility of using outdoor light trapping in malaria entomology was discussed.

Prevalence of intestinal helminthic infections in Kao District, north Halmahera, Indonesia.

A parasitological survey was conducted on the inhabitants of six villages of Kao District, Halmahera Island, North Maluku, Indonesia, in July 1993. A total of 422 fecal samples were examined by using Kato-Katz thick smear, modified Harada-Mori culture and formalin ether concentration techniques. Seven nematode species, ie Ascaris lumbricoides, Trichuris trichiura, Necator americanus, Ancylostoma duodenale, Strongyloides stercoralis, Enterobius vermicularis and unidentified rhabditoids of free-living nature, were detected. Trematode and cestode infection was not proven. Necator americanus was the predominant species of hookworm. Soil-transmitted nematode infections were highly prevalent. Among the young inhabitants aged less than 15, positive rates of Ascaris, Trichuris and hookworm infections were 32.7, 52.7 and 68.6%, respectively. Among the people aged 15 or more, the positive rate for hookworm (85.9%) was much higher than that for Ascaris and Trichuris (13.5 and 40.5%, respectively). Egg count revealed that more than 90% of inhabitants with Trichuris or hookworm had light infections. The latrines in the surveyed area seemed to have only limited effects on the improvement of the parasitological status because the prevalence of Trichuris infections was much higher in a village where most houses were provided with latrines. These conflicting conditions were considered to have been caused by many factors including the inadequate structure of the latrines.

Intestinal parasitic infections in Campalagian district, south Sulawesi, Indonesia.

Intestinal parasitic infections were surveyed in the inhabitants of 3 coastal and 2 inland villages of Campalagian District, South Sulawesi, Indonesia, in July 1992. A total of 398 fecal samples were examined by using Kato-Katz thick smear, Harada-Mori culture and agar-plate culture techniques. Protozoan cysts were examined by formalin ether concentration technique on 380 fecal samples. Soil-transmitted helminth infections were highly prevalent with the overall positive rates as follows: Ascaris lumbricoides 25.3%, Trichuris trichiura 59.3%, hookworm 68.3% and Strongyloides stercoralis 2.3%. Eight species of protozoan were detected with the overall prevalence as follows: Entamoeba histolytica 10.9%, E. hartmanni 16.3%, E. coli 31.9%, Endolimax nana 12.5%, Iodamoeba buetschlii 5.4%, Giardia lamblia 4.6%, Chilomastix mesnili 0.8% and Blastocystis hominis 18.0%. In the inland villages, prevalence of hookworm infection was higher than Ascaris and Trichuris infections, while in the coastal villages Trichuris infection was predominant. Egg count revealed that the infection level was light in most of the hookworm and Trichuris carriers. Prevalence of lavatories among houses appeared to be inversely proportional to the prevalence of hookworm infection. Meanwhile, the incomplete structure of the lavatories might result in contamination of environment with Ascaris and Trichuris eggs. Harada-Mori culture was the most efficient method in the detection of hookworm infection compared to other techniques. Both Necator americanus and Ancylostoma duodenale were found in all villages, but the former was the predominant species. An adult pinworm was detected by agar-plate culture of feces. Two types of pinworm males, corresponding to Enterobius vermicularis and E. gregorii, were observed.