The relationship between anthropometric indicators of nutritional status and malaria infection among youths in Khammouane Province, Lao PDR.

We assessed anthropometric indicators of the nutritional status among children and adolescents in Khammouane Province in the Lao PDR and examined the relation between malnutrition and malaria infection. The survey was conducted from July to August 1999 using a sample of 309 youths aged 2 to 18 years. Malnutrition was categorized as stunting (below -2 Z scores height-for-age) and wasting (below -2 Z scores weight-for-height). The prevalence of stunting and wasting were 45.1% and 9.2%, respectively, which were classified by WHO as "very high" prevalence. Compared with the results of previous national surveys in Lao PDR, similar prevalence was shown. The prevalence of wasting in youths with P. falciparum infection was 17%, significantly higher than those of not infected (4%). On the other hand, P. vivax infection was not associated with any indicators of malnutrition. In conclusion, this study showed that the nutritional status in youths was poor and P. falciparum infection was associated with acute malnutrition.

Possibility of false-positive detection for sporozoites in mosquitos (Diptera: Culicidae) by nested polymerase chain reaction using Plasmodium yoelii genomic DNA.

Anopheles stephensi Liston and An. saperoi Bohart and Ingram infected with the rodent malaria parasite Plasmodium yoelii nigeriense. They were examined 12 and 19 days after blood feeding for sporozoites in head with anterior thorax (HT) and oocysts in abdomen with posterior thorax (AB) by light microscopy and by the nested polymerase chain reaction (nested PCR-based on the amplification of the sequences of the small subunit ribosomal RNA gene). The detection rate of parasite DNA by nested PCR in HT samples 12 days after blood feeding was similar to that by microscopic method. However, in HT samples 19 days after blood feeding, the rate by the PCR method was higher than that by the microscopic method. The incidence of sporozoites in salivary glands of infected mosquitos for 12 days after blood sucking was examined by the PCR method. Parasite DNA in HT of Aedes albopictus Skuse (a non vector for the rodent malaria) as well as An. stephensi and An. saperoi was detected for up to 4 days after feeding on mouse with the rodent malaria parasites. The results indicate that when the PCR method is used for detection of sporozoites of human malaria in mosquitos collected in the field, there are possibilities of including false-positive data for mosquitos that have just or recently fed on human blood infected with malaria (erythrocytic form).

Aedes (Finlaya) japonicus (Diptera: Culicidae), a newly recognized mosquito in the United States: analyses of genetic variation in the United States and putative source populations.

Introduction of potential disease vectors into a new geographic area poses health risks to local human, livestock, and wildlife populations. It is therefore important to gain understanding of the dynamics of these invasions, in particular its sources, modes of spread after the introduction, and vectorial potential. We studied the population genetics of Aedes (Finlaya) japonicus japonicus (Theobald), an Asian mosquito that was recognized for the first time in the United States in 1998. We examined patterns of genetic diversity using random amplified polymorphic DNA and sequences of ND4 of mtDNA by comparing samples from populations spanning the range of this mosquito in Japan (six samples) and the United States (nine samples) as well as specimens intercepted in New Zealand in 1999. We found geographically differentiated populations in Japan, indicating limited gene flow even on small spatial scales. In the United States, we found evidence of significant genetic differentiation between samples from New York, Connecticut, and New Jersey and those from mid-Pennsylvania and Maryland. We were unable to pinpoint the source location(s) in Japan, although some of the U.S. samples are genetically close to samples from south Honshu and western Kyushu. Further studies should include samples from Korean populations. Distinct genetic signatures in U.S. populations undergoing expansion suggest the possibility of local increases in genetic diversity if and where they meet.

Identification of Culex vishnui subgroup (Diptera: Culicidae) mosquitoes from the Ryukyu Archipelago, Japan: development of a species-diagnostic polymerase chain reaction assay based on sequence variation in ribosomal DNA spacers.

The Culex vishnui subgroup includes three important vectors of Japanese encephalitis virus, Culex tritaeniorhynchus Giles, Cx. pseudovishnui Colless, and Cx. vishnui Theobald, all of which occur in the Ryukyu Archipelago, Japan. Although these three species have been shown to be vectors of JE virus in many areas of Southeast Asia, it is not yet known what role each plays in the transmission of the virus in this region. Reliable identification of adult, field-collected specimens is a critical component in epidemiological studies of virus transmission. Mosquitoes in the Cx. vishnui subgroup can be reliably identified in the larval stage. However, because females of these species are very similar, it is difficult to distinguish among them using morphology. We developed a polymerase chain reaction (PCR) assay for the identification of these species. Three species-specific primers were developed for the PCR assay based on a comparative analysis of the nucleotide sequence of the first internal transcribed spacer (ITS1) in the ribosomal DNA gene array. The primers, CT2REV, CP1REV, and CV1REV were designed to amplify a single DNA fragment each from Cx. tritaeniorhynchus, Cx. pseudovishnui, and Cx. vishnui, respectively, when paired with a single forward primer that is complementary to the highly conserved 18S rDNA gene. The amplified fragments were separated easily and identified on an agarose gel to facilitate species identification.

Effects of passive smoking on the regulation of rat aortic cholesteryl ester hydrolases by signal transduction.

The effects of exogenous oxidative stress due to passive smoking on cholesteryl ester (CE)-metabolizing enzymes and their regulatory kinases were examined by exposing rats to cigarette smoke (CS) for a 1-h period twice a day for 8, 12, or 20 wk. An oxidatively modified low density lipoprotein (Ox-LDL) with a high lipid peroxide was identified in three CS groups after all three exposure periods. The rat aortic acid and neutral CE hydrolases (ACEH and NCEH) were activated to similar extents by both cAMP-dependent protein kinase (PKA) and protein kinase C (PKC) in the presence of their respective cofactors. The aortic PKC activity in the three CS groups exhibited significant reductions of 72, 84, and 75% as compared with the respective controls, which coincided with the reductions in the ACEH activities (86, 71, and 80%, respectively), whereas the PKA activities increased to 121, 197, and 252% in the three CS groups, respectively. Reflecting the increase of the PKA activity, the NCEH activity exhibited increases of 112% at 8 wk and 140% until 12 wk of exposure and decreased by 50% of the control value at 20 wk of exposure, suggesting inactivation of NCEH itself. The activation of acyl-CoA:cholesterol O-acyltransferase activity was associated with an increase of free cholesterol in aorta. The vitamin E diet prevented the formation of Ox-LDL and the oxidative inactivation of most enzymes, especially PKC, until 12 wk, but was less effective by 20 wk. The oxidative inactivation of PKC, particularly its activated form that translocated to the membrane fraction, was confirmed in the in vitro exposure to active oxygen generators at an optimal concentration; this inactivation was prevented by catalase and superoxide dismutase. These results suggested that the formation of Ox-LDL and alterations in CE-metabolizing enzymes caused by passive smoking could contribute to a twofold increase in the aortic CE content, thereby contributing to one of the mechanisms for atherosclerosis associated with smoking.

Malaria prevalence and a brief entomological survey in a village surrounded by rice fields in Khammouan province, Lao PDR.

We surveyed Nongceng, a village in a south-eastern province of Lao PDR, for malaria and its vectors. Nongceng is situated in a basin and surrounded by rice fields. In February 1998 (dry season), 28.6% of 126 villagers were infected with malaria, and in September 1998 (rainy season), 16.3% of 147 villagers. The prevalence of malaria infection was consistently high in children under 10, and the predominant malaria species was Plasmodium falciparum. In brief surveys of the mosquitoes performed on the same day as the malaria surveys, 2007 Anopheles females from 12 species were collected by means of human bait, animal bait and resting collections. Of the vector species known to be important in transmitting malaria in neighbouring Thailand - An. minimus, An. dirus, and An. maculatus groups - only An. minimus was found. Its density was, however, very low in both seasons and it was therefore unlikely to be the vector. In fact, An. nivipes accounted for more than 65% of all mosquitoes collected and was the most common species collected from human baits. The results of this study show that endemic areas of malaria in Lao PDR are not necessarily related to forest. Rather, An. nivipes is suspected to be the most important vector.

Alterations of protein kinase C, 8-hydroxydeoxyguanosine, and K-ras oncogene in rat lungs exposed to passive smoking.

To investigate the effects of exposure to sidestream cigarette smoke (CS) on the initiation and promotion of lung cancer, two groups of 8 or 10 rats were exposed to CS for a 1 h period twice a day for 8, 12, or 20 weeks. The protein kinase C (PKC) activity of the lung exhibited significant changes of 120, 86 and 81% in the CS groups, compared with the respective control group values in the three exposure periods. The in vitro activation of PKC by the active oxygens was efficiently eliminated by hydroxyl radical scavengers, indicating that hydroxyl radicals are responsible for the PKC activation. For the alterations in the lung nucleus caused by passive smoking, the 12- and 20-week exposure CS groups showed significant increases in the accumulation of 8-hydroxydeoxyguanosine. One rat with K-ras activation by G:C transversion (GGT-->GCT) at codon 12 was found among 26 rats of the CS groups in the three exposure periods. These results show that active oxygens introduced by passive smoking may contribute to K-ras activation as an initiator of a tumor model, possibly through the oxygen-induced DNA damage, and may also contribute to an initial activation and the subsequent down-regulation of PKC as a promoter.

Mark-release-recapture experiment with adult Anopheles minimus (Diptera: Culicidae) on Ishigaki Island, Ryukyu Archipelago, Japan.

Movement of adult An. minimus Theobald between larval habitats and feeding sites was investigated by mark-release-recapture in March 1993 on Ishigaki Island, Ryukyu Archipelago, Japan. Unfed (998 females) and fed (1,485 females) cohorts from a laboratory colony were marked with different colors of fluorescent dye, and released near Nishihama stream where larvae were abundant. Overall, 42 fed (2.82%) and 13 unfed (1.30%) females were recaptured. Among these, 12 females were recaptured at a cowshed, the nearest blood meal source. Most females in the fed cohort recaptured at the release point had developing or matured ovaries, whereas 55% at the cowshed were parous. The movement of released females from larval habitat to the cowshed for blood feeding indicated a flight distance > 1 km.

Questionnaire survey and prevalence of intestinal helminthic infections in Barru, Sulawesi, Indonesia.

A questionnaire survey with parasitological study was carried out on the inhabitants of 4 villages in Barru district, Sulawesi, Indonesia from 1994 to 1995. The questionnaire dealt with life style and sanitary conditions. In 482 houses in the 4 villages, interviews for the items of the questionnaire were conducted with the owner, housekeeper and children of the same family. In Pancana and Lalolang, 37.7% and 50% respectively of man inhabitants surveyed were fishermen, while in Lompo Riaja and Pattappa, 38.6% and 65.5% respectively were farmers. The highest proportion of official workers was 33.7% in Lompo Riaja. Educational level was low; 88.4% in Pancana, 90.4% in Lalolang, 62.1% in Lompo Riaja and 91.2% in Pattappa had elementary or below elementary school education. In Lompo Riaja, 30.8% of the inhabitants graduated from senior high school or university. The percentage of families having their own latrine was 30.3% in Pancana, 13.2% in Lalolang, 31.9% in Pattapa and 60% in Lompo Riaja. The people without latrines usually defecated in rice fields, seaside or riverside. A total of 654 fecal samples was examined by the modified Kato-Katz thick smear method. Five nematode species, Ascaris lumbricoides, Trichuris trichiura, Necator americanus, Strongyloides stercoralis and unidentified Rhabditoids of free-living nature were detected. Cestode, Hymenolepis nana infection was confirmed. All the hookworms examined by the modified Harada-Mori culture technic were Necator americanus. Trichuris infection was most common, followed by hookworm and Ascaris infections, both in young (aged 4-14) and older (aged over 15) age groups. The prevalence of hookworm infection was significantly higher in males than in females of older age. Among the older age group, the prevalence of Trichuris infection was significantly lower in Lompo Riaja, while hookworm infection was the highest in Pattappa. Among all the inhabitants examined for parasite infection, 17.4% had 3 kinds of nematode, Ascaris, Trichuris and hookworm. However, egg counts revealed that most of the inhabitants with Trichuris or hookworm had light infections. The inhabitants with higher education background had significantly lower infection rates of Ascaris and Trichuris. The prevalence of hookworm infection was not significantly different between the inhabitants owning latrine and without it, but the prevalence of Ascaris and Trichuris, differed significantly.

Auto-destruction of the articular cartilage and free radical mediators.

The effect of static compression on the release of superoxide (SO) and nitric oxide (NO) from cartilage obtained from rabbit knee joints with tissue defects was studied. The rabbits were divided into two groups: (1) those that had 5 mm diameter full chondral defects (defect group) and (2) those in which chondral defects were filled with autogenous perichondrial grafts (grafted group). Histologically, cartilage was regenerated in the grafted group 3 weeks after the operation, although only a fibrous tissue filled the defects in the defect group even 16 weeks after the operation. A static pressure of 5 kg for 10 minutes applied to the cartilage chips obtained from the area surrounding the defects released significant amounts of SO and NO into the medium. Maximum increases were observed in the defect group 3 weeks after the operation.

Age-related changes in the activation of aortic cholesteryl ester hydrolases by protein kinases in rats.

Age-related changes in the activities of acid and neutral cholesteryl ester hydrolases (ACEH and NCEH) and their activation by protein kinase A (PKA) and also by protein kinase C (PKC) were examined in the aortae of 4-, 8-, 12- and 20-week-old rats in relation to their aortic lipid and lipid peroxides and lipid contents. The physiological basal activity as well as total activities of the ACEH and NCEH activated by the two kinases, which were high in the aortae of the 4- and 8-week-old rats, decreased gradually with increasing age to about 40% (ACEH) and 50% (NCEH) by 20 weeks of age. The vitamin E intake and ad libitum-diet intake of the rats each modified the age-related decline of CEH activities. The aortic PKA and PKC activities were reflected by the CEH activities to some degree. The in vitro exposure of the aortic CEH to active oxygen (AO) generators revealed the PKC-mediated activation of CEH, which was inhibited by superoxide dismutase and catalase. These results suggested that the activities of ACEH and NCEH and their regulatory enzymes may be modulated by the dual effect of endogenous AO; an activation of CEH at low doses and an inactivation at high doses, or upon a long-term exposure in aging to a low level of endogenous AO.

Description of Topomyia irianensis n. sp. and new records of To. papuensis from Maluku and Irian Jaya, Indonesia (Diptera:Culicidae).

The adults, male genitalia, pupa, and larva of Topomyia (Topomyia) irianensis n. sp. from Irian Jaya, Indonesia, are described and illustrated. The new species is compared to closely related species, Topomyia dejesusi Baisas and Feliciano, 1953, and Topomyia argyropalpis Leicester, 1908. Topomyia (Suaymyia) papuensis Marks was recorded for the first time from Maluku and Irian Jaya. This is the first record of the genus Topomyia from Maluku and Irian Jaya, Indonesia.

Breeding habitats of Aedes aegypti (L) and Aedes. albopictus (Skuse) in villages of Barru, South Sulawesi, Indonesia.

The breeding habitats of the dengue vector, Aedes aegypti and Aedes albopictus, were studied using larval collection method inside and outside houses in 6 villages of Barru, South Sulawesi, Indonesia from July 1994 to August 1995. Aedes aegypti was the dominant species, being abundant indoors especially in the coastal areas. Aedes albopictus was breeding primarily in outdoor containers in the hill and mountain areas. Earthen jar was the most common breeding habitat of Aedes aegypti in all villages surveyed. Drum can was the most common outdoor breeding habitat of Aedes albopictus in the hill and mountain areas. The high Breteau indices of Aedes aegypti and Aedes albopictus suggests that these species may play an important role in the transmission of dengue hemorrhagic fever in Barru where epidemics of the fever occur occasionally.

Collection of anopheline mosquitos in three villages endemic for malaria in Khammouane, Lao PDR.

Anopheline mosquito collections were made during the months of May and June (dry season) and August (rainy season), 1996 at villages, Nakham, Namdik and Hinboon Neua in Khammouane Province. Larval collections were also made in 3 slow running streams around Nakham villages in May 1996. Altogether, 3,549 adult females consisting 19 Anopheles species were collected by 3 nights of human and 6 animal bait traps and 3 resting collections. Among them, Anopheles vagus (19.75%), An. philippinensis (15.02%), An. nivipes (11.55%), An. pallidus (11.27%), An. peditaeniatus (8.34%), An. kochi (8.00%) and An. barbirostris (7.27%) were predominant species. The immatures of An. barbirostris, An. minimus, An. maculatus sensu lato, and An. culicifacies were found in the streams of Nakham. It is noteworthy that An. minimus, An. dirus and An. maculatus sensu lato, which are well known malaria vectors in Thailand, were recorded for the first time in this area.

[Survey of allergic diseases in 3-year old children in Okinawa City].

A questionnaire survey was conducted between September 1993 and March 1994 to ascertain the presence of allergic diseases among 3-year old children in Okinawa City of Okinawa Prefecture. Nine hundred and twenty-two questionnaires were sent to caretakers of 3-year old children, with 697 responses of which five invalid cases were deleted for a total of 692 responses which were analyzed. About 30% of these children had allergic diseases as diagnosed by the medical doctor. Among the respondents 15.1% had experience of atopic dermatitis, while 13.9% had experience of asthma. The incidence of atopic dermatitis in this survey was lower than those reported in other prefectures of Japan. Symptoms for atopic dermatitis were more severe during summer. About 15% of the children had been on a diet regimen for prevention or treatment of allergic diseases since the time of conception. Eighteen percent of the respondents used a vacuum cleaner on their bedding. A high proportion of respondents (65.9%) obtained information from newspapers or magazines about allergic diseases.

Historical review of mosquito control as a component of malaria eradication program in the Ryukyu Archipelago.

In the Ryukyu Archipelago, where malaria used to be endemic, eradication of the disease was achieved by the year 1962, as a consequential effect of a planned malaria eradication program in the area. This achievement was facilitated by concerted efforts in controlling vector mosquitos and treatment of all detected and presumptive cases of malaria infections. Anopheles minimus Theobald and An. sinensis Wiedemann were common in all areas endemic for malaria. Knowledge of the biology and bionomics of the mosquitos in malaria endemic areas formed the basis for formulating strategies for the control of vectors and subsequent surveillance activities. Insecticide residual spray, larvivorous fishes and environmental management were the basic strategies for vector control. The whole program was augmented by an active community participation in all eradication activities.

Inter- and intraspecific variation in resistance to desiccation by adult Aedes (Stegomyia) spp. (Diptera: Culicidae) from Indonesia.

Desiccation survival times of adult mosquitoes were studied for yellow fever mosquito, Aedes aegypti (L.) (6 strains), Asian tiger mosquito, Ae. albopictus (Skuse) (5 strains), and Ae. paullusi Stone & Farmer (1 strain) colonized from South Sulawesi, Indonesia. At both 90 and 70% RH, Ae. aegypti males and females outlived the other species. The forest species Ae. paullusi was least resistant to desiccation. Strains of Ae. aegypti and Ae. albopictus from urban areas were more resistant to desiccation than conspecific strains from rural villages. When water was accessible, Ae. albopictus females outlived Ae. aegypti females, indicating that death from deprivation of water was not a result of energy depletion. Ae. aegypti was largest in body size, followed by Ae. albopictus and Ae. paullusi. Among strains within species, the relationship between body size and survival time was significant only for Ae. aegypti females at 70% RH. Adult desiccation resistance may be a factor that influences distribution and abundance of Aedes (Stegomyia) species.

Occurrence of Aedes (Stegomyia) spp. mosquitoes (Diptera: Culicidae) in Halmahela villages, Indonesia.

Aedes (Stegomyia) spp. mosquitoes breeding in containers in and around human houses were surveyed in Halmahela villages, the North Moluccas, Indonesia. Adults reared from larvae found in containers and those reared from eggs collected by ovitraps revealed the dominance of Aedes scutellaris (Walker) in residential areas. Breeding of Aedes aegypti (L.) and Aedes albopictus (Skuse) was confirmed in a few coastal villages with high human densities.

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.

Mosquito (Diptera: Culicidae) and predator abundance in irrigated and rain-fed rice fields in north Sulawesi, Indonesia.

Immature mosquito species composition and abundance were studied in irrigated and rain-fed rice fields of North Sulawesi, Indonesia. Irrigated rice fields were characterized by the prevalence of aquatic macrophytes and cyprinodont larvivorous fish, Aplocheilus panchax (Hamilton), but abundance per dip of most aquatic insect predators was lower than that in rain-fed rice fields. Anopheles peditaeniatus (Leicester), Culex vishnui Theobald, and Culex tritaeniorhynchus Giles, were dominant in both irrigated and rain-fed fields, but the abundance of the Culex species was lower in irrigated fields. The effect of irrigation system introduction on regional mosquito abundance cannot be evaluated by the enlarged surface water area alone. Changes in habitat quality, expressed as the abundance per dip (index of density per unit water area), also need to be considered.