Sex- and age-related variations in the in vitro heparin-releasable lipoprotein lipase from mononuclear leukocytes in blood.

An in vitro heparin release of lipoprotein lipase (LPL) from whole blood, mainly from monocytes, was demonstrated by (1) the time-course of lipolytic activity with the presence of 10 U/ml heparin at 37 degrees C, (2) the distribution of LPL activity in monocyte and lymphocyte fractions, (3) an immuno-inactivation with anti-LPL immunoglobulin (IgG) and (4) responses to various compounds such as NaCl, protamine sulfate, heparin, and serum activator. The in vitro heparin-releasable LPL activity from blood correlated well with the LPL activity of postheparin plasma obtained from both normolipidemic and hyperlipidemic rabbits. Studies in humans revealed sex- and age-related variations in the in vitro heparin-releasable LPL from monocytes in the blood of 134 normal subjects and 24 hypertriglyceridemic subjects: The mean LPL activity was significantly higher in normal females over the age of 30, than in the corresponding males. In the hypertriglyceridemic group, the LPL activity was also higher in females than in males, but it was not significant. The in vitro heparin-releasable LPL activity from monocytes in blood was comparable to the LPL activity derived from adipose tissue and postheparin plasma, and thus it reflects lipoprotein metabolism.

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.

Effect of cyclic AMP on urokinase-type plasminogen activator receptor and fibrinolytic factors in a human osteoblast-like cell line.

We investigated the effect of cyclic AMP (cAMP) on the pericellular fibrinolytic system in NY cells. Dibutyryl cAMP (dbcAMP) or forskolin increased the level of urokinase-type plasminogen activator (u-PA) mRNA and enhanced the secretion of u-PA antigen into the conditioned medium. These agents also increased u-PA antigen on the cell surface. PA inhibitor-1 (PAI-1) antigen was inhibited by dbcAMP or forskolin. Butyrate had no effect on the production and secretion of u-PA and PAI-1. A binding assay of 125I-DFP-u-PA to NY cells revealed a single class of binding sites with a Kd of 3.85 nM and Bmax of 0.89.10(5) binding sites/cell. The Bmax was increased by dbcAMP (1 mM or 10 mM), forskolin (2 microM or 20 microM) of 1.0-, 1.4-, 1.2- and 1.8-fold, respectively. However, the Kd value was not changed. Furthermore, the level of mRNA for the u-PA receptor (u-PAR) was increased by these agents 1.2-, 1.7-, 1.8- and 2.5-fold, respectively. However, butyrate did not alter either the Bmax or the u-PAR mRNA level. These results indicated that the pericellular fibrinolytic activity induced by u-PA/u-PAR is modulated by cAMP in osteoblast-like cells.

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.

Colonization of rice fields by mosquitoes (Diptera: Culicidae) and larvivorous predators in asynchronous rice cultivation areas in the Philippines.

Colonization of rice fields by mosquitoes and larvivorous predators was studied in asynchronous rice cultivation areas in the Philippines. Dipper samples were taken from rice fields at six phases of maturity (fallow, ploughed, nursery, newly transplanted, after tillering, mature). All phases were present concurrently at each of two study sites. Abundance levels of the vishnui subgroup of Culex and of the genus Anopheles were high in ploughed fields, nurseries, and newly transplanted fields; this was primarily because of the concentration of Culex vishnui. Theobald and Anopheles vagus Doenitz in those fields with short, sparse vegetation. Dytiscidae, Anisoptera, and Zygoptera were more abundant in fallow or mature fields. The abundance of aquatic predators decreased at the onset of ploughing and then recovered slowly as rice plants grew. The abundance of surface predators showed a similar pattern, but less conspicuously. The abundance of nonpredators (other than mosquitoes) was relatively stable with regard to rice field phases. Alleviation of mortality by predation was considered to be a secondary cause of increased mosquito abundance in the fields under early phases of rice cultivation.

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.

A new species of Toxorhynchites mosquito (Diptera: Culicidae) from the Ryukyu Archipelago, Japan.

Toxorhynchites (Toxorhynchites) okinawensis Toma, Miyagi, and Tanaka, n. sp., from Okinawajima, the Ryukyu Archipelago, Japan, is described and illustrated. Its taxonomy, bionomics, and zoogeography are discussed briefly.

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.

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.

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.

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.

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.

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.

A survey of larval mosquitoes on Kume Island, Ryukyu Archipelago, Japan.

Mosquito collections were carried out during May 1991 in Kume Island, Ryukyu Archipelago. Eighteen species of mosquitoes in 8 genera were collected; 11 species were new records for Kume Island. Characteristics of the larval habitats of these species are also described.

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.

Seasonal appearance and size of egg rafts of Culex halifaxii and Culex fuscanus in Okinawajima, the Ryukyu Archipelago, Japan.

The seasonal chronology of eggs of Culex halifaxii, Cx. fuscanus and their cohabitant Cx. quinquefasciatus were observed from March 1984 to early May 1985 in Okinawa prefecture, Japan. In Okinawajima, egg rafts of Cx. halifaxii and Cx. quinquefasciatus were found throughout the year. The greatest peak in eggs of Cx. halifaxii were observed in early June. The egg rafts of Cx. fuscanus were first collected on June 15, with the greatest peak of oviposition in early November. The egg raft of Cx. fuscanus was larger and both the maximum and mean numbers of eggs per raft were approximately two times greater than those of Cx. halifaxii.

A new mutant, White larva, of the mosquito Toxorhynchites splendens: genetics and cannibalism.

A strain of a new body-color mutant, white larva (wl), was established from a field-collected wild-type strain of Toxorhynchites splendens. The mutant can be distinguished from the wild type in both the larval and pupal stages, but not in the adult. Crossing experiments confirmed its mode of inheritance to be a single recessive system. This is the first visible mutant found in Tx. splendens. Larvae of the wl phenotype seem to be recognized as prey by other individuals in mass larvae rearing.

Laboratory bionomics of Tripteroides aranoides.

Tripteroides aranoides was colonized in the laboratory. Total duration of the immature stages was ca. 3 weeks at 28 degrees C, L:D = 15.5:8.5 with an ample food supply. Retardation of 4th instar development was observed in larvae fed on insufficient food. Females were autogenous for the first clutch of eggs and required a blood meal for maturation of the second clutch. Mating was initiated in flight and copulation occurred on the cage wall. Gravid females hovered in small oblique loops above water in bamboo cups, whereupon a white egg appeared at the abdominal tip, which was propelled by the swing of the abdomen towards water surface. The females propelled eggs in the same manner into small apertures (11 x 4 mm) bored in bamboo.

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.

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.