Melanopsin as a sleep modulator: circadian gating of the direct effects of light on sleep and altered sleep homeostasis in Opn4(-/-) mice.

Light influences sleep and alertness either indirectly through a well-characterized circadian pathway or directly through yet poorly understood mechanisms. Melanopsin (Opn4) is a retinal photopigment crucial for conveying nonvisual light information to the brain. Through extensive characterization of sleep and the electrocorticogram (ECoG) in melanopsin-deficient (Opn4(-/-)) mice under various light-dark (LD) schedules, we assessed the role of melanopsin in mediating the effects of light on sleep and ECoG activity. In control mice, a light pulse given during the habitual dark period readily induced sleep, whereas a dark pulse given during the habitual light period induced waking with pronounced theta (7-10 Hz) and gamma (40-70 Hz) activity, the ECoG correlates of alertness. In contrast, light failed to induce sleep in Opn4(-/-) mice, and the dark-pulse-induced increase in theta and gamma activity was delayed. A 24-h recording under a LD 1-hratio1-h schedule revealed that the failure to respond to light in Opn4(-/-) mice was restricted to the subjective dark period. Light induced c-Fos immunoreactivity in the suprachiasmatic nuclei (SCN) and in sleep-active ventrolateral preoptic (VLPO) neurons was importantly reduced in Opn4(-/-) mice, implicating both sleep-regulatory structures in the melanopsin-mediated effects of light. In addition to these acute light effects, Opn4(-/-) mice slept 1 h less during the 12-h light period of a LD 12ratio12 schedule owing to a lengthening of waking bouts. Despite this reduction in sleep time, ECoG delta power, a marker of sleep need, was decreased in Opn4(-/-) mice for most of the (subjective) dark period. Delta power reached after a 6-h sleep deprivation was similarly reduced in Opn4(-/-) mice. In mice, melanopsin's contribution to the direct effects of light on sleep is limited to the dark or active period, suggesting that at this circadian phase, melanopsin compensates for circadian variations in the photo sensitivity of other light-encoding pathways such as rod and cones. Our study, furthermore, demonstrates that lack of melanopsin alters sleep homeostasis. These findings call for a reevaluation of the role of light on mammalian physiology and behavior.

Study on phlebotomine sand fly (Diptera: Psychodidae) fauna in Belo Horizonte, state of Minas Gerais, Brazil

A study on the phlebotomine sand fly fauna in Belo Horizonte city, state of Minas Gerais, Brazil, was carried out. From April 2001 to March 2003, monthly systematic collections were performed in three houses from each of the nine regions of the city, using CDC light traps for four consecutive days. The traps were set into the houses and in peridomestic areas totaling 54 traps. A number of 3871 sand fly specimens of the genera Lutzomyia and Brumptomyia were collected. Sixty eight percent of the specimens were L. longipalpis and 16% L. whitmani, insect vectors of visceral and American cutaneous leishmaniasis, respectively. Environmental factors such as temperature, humidity, and frequency of precipitation suggest that the number of insects increases after rainy periods. During the same period mentioned above, seasonal captures were carried out in parks and green areas of Belo Horizonte, using Shannon trap. A total of 579 phlebotomine sand flies were collected from which 398 (68.7%) were females with the predominance of L. whitmani and L. monticola. Those specimens were used for natural infection examination, by polymerase chain reaction. No Leishmania DNA was present in any of the specimens tested.

A pea plasma membrane protein exhibiting blue light-induced phosphorylation retains photosensitivity following triton solubilization.

Phosphorylation of a polypeptide of approximately 120 kD in pea (Pisum sativum L.) plasma membranes in response to blue light has been shown to be involved in phototropic curvature, but the relationship of this protein to the kinase and photoreceptor acting upon it is uncertain. Using two-phase aqueous partitioning to isolate right-side-out plasma membrane vesicles, we have obtained evidence suggesting that the photoreceptor, kinase, and substrate are localized to the plasma membrane fraction. Latent phosphorylation accessible through Triton X-100 or freeze/thaw treatments of purified plasma membrane vesicles indicates that at least the kinase moiety is present on the internal face of the plasma membrane. Effects of solubilization of vesicles on fluence-response characteristics and on phosphorylation levels provide evidence that the receptor, kinase, and protein substrate are present together in individual mixed detergent micelles, either as a stable complex or as domains of a single polypeptide. In vivo blue-light irradiation results in a small but significant decrease in mobility of the 120-kD phosphorylated protein on sodium dodecylsulfate gel electrophoresis. This mobility shift is evident on Coomassie-stained gels and on western blots probed with polyclonal antibodies raised against the 120-kD protein. Among the plasma membrane proteins bound to the reactive nucleotide analog fluorosulfonylbenzoyladenine (FSBA), a distinct protein band at 120 kD can be detected on blots probed with anti-FSBA antibodies. This band exhibits an in vivo light-dependent mobility shift identical to that observed for the protein band and antibodies specific for the 120-kD protein, implying that the 120-kD protein has an integral nucleotide binding site and consistent with the possibility that the substrate protein is also a kinase.

Light receptor action is critical for maintaining plant biomass at warm ambient temperatures.

The ability to withstand environmental temperature variation is essential for plant survival. Former studies in Arabidopsis revealed that light signalling pathways had a potentially unique role in shielding plant growth and development from seasonal and daily fluctuations in temperature. In this paper we describe the molecular circuitry through which the light receptors cry1 and phyB buffer the impact of warm ambient temperatures. We show that the light signalling component HFR1 acts to minimise the potentially devastating effects of elevated temperature on plant physiology. Light is known to stabilise levels of HFR1 protein by suppressing proteasome-mediated destruction of HFR1. We demonstrate that light-dependent accumulation and activity of HFR1 are highly temperature dependent. The increased potency of HFR1 at warmer temperatures provides an important restraint on PIF4 that drives elongation growth. We show that warm ambient temperatures promote the accumulation of phosphorylated PIF4. However, repression of PIF4 activity by phyB and cry1 (via HFR1) is critical for controlling growth and maintaining physiology as temperatures rise. Loss of this light-mediated restraint has severe consequences for adult plants which have greatly reduced biomass.

Field evaluation of effectiveness of the BG-Sentinel, a new trap for capturing adult Aedes aegypti (Diptera: Culicidae)

In recent years, the development of new tools to gather field information about vector ecological parameters has increased. This report evaluated the BG-Sentinel Trap (BGS-Trap), a promising new attempt to improve collection of the dengue vector, Aedes aegypti. The efficacy of the BGS-Trap was compared with the CDC backpack aspirator, one of the commonest used methods for capturing adult mosquitoes. BGS-Traps captured significantly more Ae. aegypti males (chi2 = 21.774, df = 1, P < 0.05) and females (chi2 = 56.007, df = 1, P < 0.05) than CDC aspirator during all days of field collection. However, CDC aspirator was significantly more efficient to capture Culex quinquefasciatus males (chi2 = 5.681, df = 1, P < 0.05) and females (chi2 = 6.553, df = 1, P < 0.05). BGS-Traps captured host-seeking females (varying between 68.75 to 89.8%) in detriment of females in other behavioral and physiological stages. BGS-Traps proved to be efficient and can be used for monitoring adult mosquito populations.

Laboratory and field evaluation of an oviposition trap for Culex quinquefasciatus(Diptera: Culicidae)

An ovitrap (BR-OVT) based on physical and chemical stimuli for attracting gravid Culex quinquefasciatus (Diptera: Culicidae) females was developed and evaluated under laboratory and field conditions. Attractants were assayed using alternative chamber bioassays prior to being used in the BR-OVT oviposition trap. A significant preference of gravid females for sites containing conspecific egg rafts was observed, as a response to the natural oviposition pheromone, as well as for sites treated with the synthetic pheromone erythro-6-acetoxy-5-hexadecanolide. Five- to 20-day old grass infusion was strongly attractive to gravid females for laying eggs. On the other hand, entomopathogenic Bacillus sphaericus (Bs) did not influence the choice of an oviposition site when used in combination with grass infusion and can therefore be used as a larvicide in ovitraps. Results from field trials showed that the BR-OVT with grass infusion and with or without Bs works as a preferred oviposition site for Cx. quinquefasciatus. The BR-OVT was more effective for egg collection when placed indoors and comparison with the number of egg rafts laid in cesspits over 40 days indicates that this very simple ovitrap may be a useful tool for monitoring populations of the most important of the vectors of bancroftian filariasis.

Therapeutic Blockade of LIGHT Interaction With Herpesvirus Entry Mediator and Lymphotoxin β Receptor Attenuates In Vivo Cytotoxic Allogeneic Responses.

BACKGROUND: Tumor necrosis factor/tumor necrosis factor receptor superfamily members conform a group of molecular interaction pathways of essential relevance during the process of T-cell activation and differentiation toward effector cells and particularly for the maintenance phase of the immune response. Specific blockade of these interacting pathways, such as CD40-CD40L, contributes to modulate the deleterious outcome of allogeneic immune responses. We postulated that antagonizing the interaction of LIGHT expression on activated T cells with its receptors, herpesvirus entry mediator and lymphotoxin β receptor, may decrease T cell-mediated allogeneic responses. METHODS: A flow cytometry competition assay was designed to identify anti-LIGHT monoclonal antibodies capable to prevent the interaction of mouse LIGHT with its receptors expressed on transfected cells. An antibody with the desired specificity was evaluated in a short-term in vivo allogeneic cytotoxic assay and tested for its ability to detect endogenous mouse LIGHT. RESULTS: We provide evidence for the first time that in mice, as previously described in humans, LIGHT protein is rapidly and transiently expressed after T-cell activation, and this expression was stronger on CD8 T cells than on CD4 T cells. Two anti-LIGHT antibodies prevented interactions of mouse LIGHT with its two known receptors, herpesvirus entry mediator and lymphotoxin β receptor. In vivo administration of anti-LIGHT antibody (clone 10F12) ameliorated host antidonor short-term cytotoxic response in wild type B6 mice, although to a lesser extent than that observed in LIGHT-deficient mice. CONCLUSION: The therapeutic targeting of LIGHT may contribute to achieve a better control of cytotoxic responses refractory to current immunosuppressive drugs in transplantation.

Phlebotomine fauna (Diptera: Psychodidae) and species abundance in an endemic area of American cutaneous leishmaniasis in southeastern Minas Gerais, Brazil

This study was undertaken to identify the phlebotomine fauna and species abundance in domiciliary and peridomiciliary (hen-house and guava-tree) environments, on a lake shore, a cultivated area of coffee and banana, and a forested area of Conceição da Aparecida municipality, southeastern the state of Minas Gerais, to provide information for the control and epidemiological surveillance of leishmaniasis in this area. The captures were carried out monthly between May 2001 and November 2002, with automatic light and Shannon traps. A total of 1444 sand flies were captured, 951 (76.5%) with automatic light traps and 493 (23.5%) with the Shannon trap. Thirteen species were captured, the most frequent being Nyssomyia whitmani (62.7%), Migonemyia migonei (21.4%), Pintomyia fischeri (6.9%), and Evandromyia lenti (3.6%). Species abundance was determined using the automatic light traps installed in the six environments. The most abundant species according to the standardized index of species abundance were Ny. whitmani (1.0) and Mg. migonei (0.82). In view of the dominance of these two species, known vectors of cutaneous leishmaniasis in other Brazilian areas, their participation in the transmission of the disease in this county is suggested. The diversity and evenness indexes in the domicile were the lowest due to the high frequency (83%) of Ny. whitmani. The capture of Lutzomyia longipalpis, rarely recorded in the south-eastern and southern regions of Minas Gerais, is also noteworthy.

Modification of Disney trap for capture of sand flies (Diptera: Psychodidae: Phlebotominae)

This paper describes the modifications made to the original model of the Disney trap, with a view to easier handling of the same, greater practicability in the collection of sand flies, protection of the animal bait and durability of the trap in the field.

Le myélome à chaînes légères ou les pièges du diagnostic du myélome [Light chain myeloma or the pitfalls of myeloma diagnosis]

The diagnosis of multiple myeloma is often suggested by disturbances found in routine laboratory tests such as sedimentation rate, electrophoresis of serum proteins and search for proteinuria. In light chain myeloma these tests are nonspecific and therefore misleading. We present 8 cases of light chain myeloma and discuss the diagnosis of multiple myeloma with its associated pitfalls.

Morphological study of the eggs and nymphs of Triatoma dimidiata (Latreille, 1811) observed by light and scanning electron microscopy (Hemiptera: Reduviidae: Triatominae)

Eggs and nymphs of Triatoma dimidiata were described using both light and scanning electron microscopy. The egg body and operculum have an exochorion formed by irregular juxtaposed polygonal cells; these cells are without sculpture and the majority of them are hexagonal in shape. The five instars of T. dimidiatacan be distinguished from each other by characteristics of the pre, meso and metanotum. The number of setiferous tubercles increases progressively among instars. The sulcus stridulatorium of 1st instar nymphs is amorphous, showing median parallel grooves; from the 2nd instar on the sulcus is, progressively, elongate, deep and posteriorly pointed with stretched parallel grooves. All instars have a trichobothrium on the apical 1/3 of segment II of the antenna. The opening of the Brindley's gland is on the mesopleura. Fifth instar nymphs have an apical ctenidium on the ventral surface of the fore tibia. Dorsal glabrous patches are found on the lateral 1/3 of abdomen. Bright oval patches are found on the ventral median line of the abdomen, from segment IV-VI; 1st instar nymphs lack these patches. Abdominal dorsal plates are present from the 1st-5th instars; the 1st instar also contains a rectangular plate in segment IX. From the 2nd instar on, variably-shaped plates are present on segments VII to IX. Morphometric data were also obtained and proved to be useful for distinguishing T. dimidiata instars.