Benefits of Coronectomy in Lower Third Molar Surgery: A Systematic Review and Meta-analysis.

PURPOSE: The purpose of this study was to measure and compare coronectomy versus extraction in patients at increased risk for inferior alveolar nerve (IAN) injuries associated with third molar removal in terms of IAN injury and other complications. METHODS: The review adhered to the Preferred Reporting Items for Systematic Reviews and Meta-analyses checklist. We conducted a comprehensive literature search across six databases and the gray literature from July 15 to August 01, 2022. We employed Rayyan software to identify and remove duplicate articles to ensure data integrity. Our research followed the strategy patient (P), intervention (I), comparison (C), outcome (O), and study (S): (P) patients needing lower third molar surgery at higher risk of IAN injury; (I) surgery options, coronectomy or complete extraction; (C) comparisons included reduced risks of nerve injuries, postoperative complications (pain, infection, alveolitis), and increased risks of reoperation, root migration, and extraction; (O) desired outcomes were preventing nerve injuries and reducing other surgical complications; and (S) observational study designs (cohort, case-control). Excluded from consideration were studies involving teeth other than lower third molars, as well as reviews, letters, conference summaries, and personal opinions. To gauge the certainty of evidence, we employed the Grading of Recommendation, Assessment, Development, and Evaluation instrument, selecting the most current papers with the highest levels of evidence for inclusion. The primary outcome variable of our study centered on evaluating the incidence of IAN injury, and secondly, the lingual nerve (LN) injury, the postoperative pain, infection, localized alveolitis, the necessity for surgical reintervention, root migration, and extraction. These assessments were carried out with respect to their chosen operative technique for managing third molars, either coronectomy or extraction, as predictor variables. We also considered covariates such as age, gender, and the presence of systemic diseases in our analysis to account for potential confounding factors. The pooled data underwent rigorous analysis utilizing an inverse variance method with both random and fixed effect models by the "metabin" function in the R program's meta-package. Additionally, we assessed the risk of bias in the selected studies by utilizing the Joanna Briggs Institute's Critical Appraisal Checklist for Studies Reporting Prevalence Data and the Critical Appraisal Checklist for Case Reports. RESULTS: Of the 1,017 articles found, after applying the inclusion and exclusion criteria, 42 were included in this study (29 cohort and 13 case-control studies), including 3,095 patients from 18 countries. The meta-analysis showed that coronectomy reduced the risk of IAN injury [OR (Odds Ratio): 0.14; 95% CI (confidence intervals): 0.06-0.30; I2 (inconsistency index) = 0%; P = .0001], postoperative pain (OR: 0.97; 95% CI: 0.33-2.86; I2 = 81%; P = .01), and alveolitis (OR: 0.38; 95% CI: 0.13-1.09; I2 = 32.2%; P = .01) when compared to complete tooth extraction. However, it also highlighted a greater risk of reintervention (OR: 5.38; 95% CI: 1.14-25.28; I2 = 0.0%; P = .01). CONCLUSIONS: This study has demonstrated that coronectomy is associated with a decreased risk for IAN injury and decreased pain and localized alveolitis when compared to complete tooth extraction. However, it is essential to acknowledge the higher likelihood of requiring reintervention with coronectomy. Therefore, clinicians should carefully consider the advantages and potential drawbacks of both techniques and tailor their choices to the unique clinical circumstances of each patient.

Deinstitutionalization and autonomy: outcomes from a Brazilian mental health policy. / Modos de autonomia em Serviços Residenciais Terapêuticos e sua relação com estratégias de desinstitucionalização.

Therapeutic Residential Services (SRT, in Portuguese) are structures implemented in the context of the Brazilian Psychiatric Reform (BPR), which aims to support psychiatric deinstitutionalization. This paper compares residents of SRTs in two cities in Brazil, analyzing the relationship among the deinstitutionalization strategies and the different types of autonomy they produce. Nineteen individuals from two cities (referred to as Paulo Delgado - PB and Franco Basaglia - FB) participated in this study. Participant observation visits were performed, and five managers and professionals who worked at the psychosocial care networks were interviewed. Narratives were created based on the field diaries and the transcription of the interviews. All the participants raised their degree of autonomy, when compared to the time in which they lived in the psychiatric hospitals. The different ways in which SRTs are organized in both cities produced different manners of exercising autonomy: in PD, home autonomy predominated, while in FB, autonomy in circulating throughout the territory predominated. In both cities, autonomy in managing financial resources was restricted. It was concluded that public policies for deinstitutionalization were effective, although their operationalization could be enhanced.

Serviços Residenciais Terapêuticos (SRTs) são dispositivos implantados no âmbito da Reforma Psiquiátrica brasileira para apoiar a desinstitucionalização psiquiátrica. Esta pesquisa objetivou comparar as formas de autonomia desenvolvidas por moradores de SRTs de dois municípios da Bahia, analisando sua relação com as estratégias de desinstitucionalização construídas em cada território. Participaram 19 indivíduos dos municípios referidos pelos pseudônimos Paulo Delgado (PD) e Franco Basaglia (FB). Foram realizadas visitas para observação participante e entrevistas com cinco gestores e profissionais das redes de atenção psicossocial. Registros em diários de campo e transcrições de entrevistas subsidiaram a construção de narrativas. Todos os participantes ampliaram o grau de autonomia em relação ao período em que saíram das instituições asilares. As formas de organização dos SRTs nos dois municípios produziram diferentes modos de exercer autonomia: em PD predominou a autonomia no habitar, enquanto em FB predominou a autonomia no transitar pelo território. Em ambos, a autonomia na administração de recursos financeiros foi limitada. Consideramos que as políticas públicas para desinstitucionalização foram efetivas, embora sua operacionalização possa ser aprimorada.

Modos de autonomia em Serviços Residenciais Terapêuticos e sua relação com estratégias de desinstitucionalização / Deinstitutionalization and autonomy: outcomes from a Brazilian mental health policy

Resumo Serviços Residenciais Terapêuticos (SRTs) são dispositivos implantados no âmbito da Reforma Psiquiátrica brasileira para apoiar a desinstitucionalização psiquiátrica. Esta pesquisa objetivou comparar as formas de autonomia desenvolvidas por moradores de SRTs de dois municípios da Bahia, analisando sua relação com as estratégias de desinstitucionalização construídas em cada território. Participaram 19 indivíduos dos municípios referidos pelos pseudônimos Paulo Delgado (PD) e Franco Basaglia (FB). Foram realizadas visitas para observação participante e entrevistas com cinco gestores e profissionais das redes de atenção psicossocial. Registros em diários de campo e transcrições de entrevistas subsidiaram a construção de narrativas. Todos os participantes ampliaram o grau de autonomia em relação ao período em que saíram das instituições asilares. As formas de organização dos SRTs nos dois municípios produziram diferentes modos de exercer autonomia: em PD predominou a autonomia no habitar, enquanto em FB predominou a autonomia no transitar pelo território. Em ambos, a autonomia na administração de recursos financeiros foi limitada. Consideramos que as políticas públicas para desinstitucionalização foram efetivas, embora sua operacionalização possa ser aprimorada.

Abstract Therapeutic Residential Services (SRT, in Portuguese) are structures implemented in the context of the Brazilian Psychiatric Reform (BPR), which aims to support psychiatric deinstitutionalization. This paper compares residents of SRTs in two cities in Brazil, analyzing the relationship among the deinstitutionalization strategies and the different types of autonomy they produce. Nineteen individuals from two cities (referred to as Paulo Delgado - PB and Franco Basaglia - FB) participated in this study. Participant observation visits were performed, and five managers and professionals who worked at the psychosocial care networks were interviewed. Narratives were created based on the field diaries and the transcription of the interviews. All the participants raised their degree of autonomy, when compared to the time in which they lived in the psychiatric hospitals. The different ways in which SRTs are organized in both cities produced different manners of exercising autonomy: in PD, home autonomy predominated, while in FB, autonomy in circulating throughout the territory predominated. In both cities, autonomy in managing financial resources was restricted. It was concluded that public policies for deinstitutionalization were effective, although their operationalization could be enhanced.

Feasibility of Using Intraoperative Neuromonitoring in the Prophylaxis of Dysesthesia in Transforaminal Endoscopic Discectomies of the Lumbar Spine.

(1) Background: Postoperative nerve root injury with dysesthesia is the most frequent sequela following lumbar endoscopic transforaminal discectomy. At times, it may be accompanied by transient and rarely by permanent motor weakness. The authors hypothesized that direct compression of the exiting nerve root and its dorsal root ganglion (DRG) by manipulating the working cannula or endoscopic instruments may play a role. (2) Objective: To assess whether intraoperative neurophysiological monitoring can help prevent nerve root injury by identifying neurophysiological events during the initial placement of the endoscopic working cannula and the directly visualized video endoscopic procedure. (3) Methods: The authors performed a retrospective chart review of 65 (35 female and 30 male) patients who underwent transforaminal endoscopic decompression for failed non-operative treatment of lumbar disc herniation from 2012 to 2020. The patients' age ranged from 22 to 86 years, with an average of 51.75 years. Patients in the experimental group (32 patients) had intraoperative neurophysiological monitoring recordings using sensory evoked (SSEP), and transcranial motor evoked potentials (TCEP), those in the control group (32 patients) did not. The SSEP and TCMEP data were analyzed and correlated to the postoperative course, including dysesthesia and clinical outcomes using modified Macnab criteria, Oswestry disability index (ODI), visual analog scale (VAS) for leg and back pain. (4) Results: The surgical levels were L4/L5 in 44.6%, L5/S1 in 23.1%, and L3/L4 in 9.2%. Of the 65 patients, 56.9% (37/65) had surgery on the left, 36.9% (24/65) on the right, and the remaining 6.2% (4/65) underwent bilateral decompression. Postoperative dysesthesia occurred in 2 patients in the experimental and six patients in the control group. In the experimental neuromonitoring group, there was electrodiagnostic evidence of compression of the exiting nerve root's DRG in 24 (72.7%) of the 32 patients after initial transforaminal placement of the working cannula. A 5% or more decrease and a 50% or more decrease in amplitude of SSEPs and TCEPs recordings of the exiting nerve root were resolved by repositioning the working cannula or by pausing the root manipulation until recovery to baseline, which typically occurred within an average of 1.15 min. In 15 of the 24 patients with such latency and amplitude changes, a foraminoplasty was performed before advancing the endoscopic working cannula via the transforaminal approach into the neuroforamen to avoid an impeding nerve root injury and postoperative dysesthesia. (5) Conclusion: Neuromonitoring enabled the intraoperative diagnosis of DRG compression during the initial transforaminal placement of the endoscopic working cannula. Future studies with more statistical power will have to investigate whether employing neuromonitoring to avoid intraoperative compression of the exiting nerve root is predictive of lower postoperative dysesthesia rates in patients undergoing videoendoscopic transforaminal discectomy.

The Summer Is Coming: nocte and timeless Genes Are Influenced by Temperature Cycles and May Affect Aedes aegypti Locomotor Activity.

Mosquitoes exhibit activity rhythms, crucial for the transmission of pathogens, under the control of a circadian clock. Aedes aegypti is one of the world's leading vectors. For decades, several studies have linked the rise in ambient temperature with the increase in their activity. Here, we identify candidate genes whose expression is influenced by temperature cycles and may affect Aedes locomotor activity. We observed that timeless completely lost its rhythmic expression in light/dark, with out-of-phase temperature cycles, and by RNAi mediated knockdown of nocte, an important gene for Drosophila circadian synchronization by temperature cycles. Thus, timeless and nocte are important genes for synchronization by temperature cycles in Aedes aegypti. To reinforce our findings, we simulated in the laboratory the gradual temperature fluctuations that were as close as possible to daily temperature variations in Brazil. We observed that the activity and the expression of the molecular circadian clock of Ae. aegypti differs significantly from that of mosquitoes subjected to constant or rectangular abrupt changes in temperature. We suggest that for understanding the circadian behavior of Aedes with possible implications for intervention strategies, the seminatural paradigm needs to replace the traditional laboratory study.

Effects of Light and Temperature on Daily Activity and Clock Gene Expression in Two Mosquito Disease Vectors.

Most organisms feature an endogenous circadian clock capable of synchronization with their environment. The most well-known synchronizing agents are light and temperature. The circadian clock of mosquitoes, vectors of many pathogens, drives important behaviors related to vectoral capacity, including oviposition, host seeking, and hematophagy. Main clock gene expression, as well as locomotor activity patterns, has been identified in Aedes aegypti and Culex quinquefasciatus under artificial light-dark cycles. Given that these mosquito species thrive in tropical areas, it is reasonable to speculate that temperature plays an important role in the circadian clock. Here, we provide data supporting a different hierarchy of light and temperature as zeitgebers of two mosquito species. We recorded their locomotor activity and quantified mRNA expression of the main clock genes in several combinations of light and temperature cycles. We observed that A. aegypti is more sensitive to temperature, while C. quinquefasciatus is more responsive to light. These variations in clock gene expression and locomotor activity may have affected the mosquito species' metabolism, energy expenditure, fitness cost, and pathogen transmission efficiency. Our findings are relevant to chronobiology studies and also have epidemiological implications.

Cryptic diversity in an Atlantic Forest malaria vector from the mountains of South-East Brazil.

BACKGROUND: Anopheles (Kerteszia) cruzii is the primary vector of human and simian malarias in Brazilian regions covered by the Atlantic Rainforest. Previous studies found that An. cruzii presents high levels of behavioural, chromosomal and molecular polymorphisms, which led to the hypothesis that it may be a complex of cryptic species. Here, An. cruzii specimens were collected in five sites in South-East Brazil located at different altitudes on the inner and coastal slopes of two mountain ranges covered by Atlantic Rainforest, known as Serra do Mar and Serra da Mantiqueria. Partial sequences for two genes (Clock and cpr) were generated and compared with previously published sequences from Florianópolis (southern Brazil). Genetic diversity was analysed with estimates of population structure (F ST ) and haplotype phylogenetic trees in order to understand how many species of the complex may occur in this biome and how populations across the species distribution are related. RESULTS: The sequences from specimens collected at sites located on the lower coastal slopes of Serra do Mar (Guapimirim, Tinguá and Sana) clustered together in the phylogenetic analysis, while the major haplotypes from sites located on higher altitude and at the continental side of the same mountains (Bocaina) clustered with those from Serra da Mantiqueira (Itatiaia), an inner mountain range. These two An. cruzii lineages showed statistically significant genetic differentiation and fixed characters, and have high F ST values typical of between species comparisons. Finally, in Bocaina, where the two lineages occur in sympatry, we found deviations from Hardy-Weinberg equilibrium due to a deficit of heterozygotes, indicating partial reproductive isolation. These results strongly suggest that at least two distinct lineages of An. cruzii (provisorily named "Group 1" and "Group 2") occur in the mountains of South-East Brazil. CONCLUSIONS: At least two genetically distinct An. cruzii lineages occur in the Atlantic Forest covered mountains of South-East Brazil. The co-occurrence of distinct lineages of An. cruzii (possibly incipient species) in those mountains is an interesting biological phenomenon and may have important implications for malaria prevalence, Plasmodium transmission dynamics and control.

Rhodnius prolixus and R. robustus (Hemiptera: Reduviidae) nymphs show different locomotor patterns on an automated recording system.

BACKGROUND: Circadian rhythms of triatomines, vectors of the etiological agent Trypanosoma cruzi responsible for Chagas disease, have been extensively studied in adults of the two most epidemiologically relevant vector species, Rhodnius prolixus and Triatoma infestans. However, little attention has been dedicated to the activity patterns in earlier developmental stages, even though triatomine nymphs are equally capable of transmitting T. cruzi to humans. Because circadian rhythms may differ even between closely related species, studies that focus on this behavioral trait can also be used to shed light on the taxonomy of controversial taxa, which becomes especially relevant regarding vector species. METHODS: We compared the daily locomotor activity patterns of second- and third-instar nymphs of Rhodnius prolixus and Rhodnius robustus in order to unveil possible behavioral differences between these cryptic species. Mitochondrial and nuclear markers were sequenced to confirm species identification. RESULTS: Nymphs of both species had a bimodal pattern of locomotion and similar daily activity patterns, but R. prolixus is more active under light/dark cycles and depicts a more pronounced activity rhythm under constant darkness conditions. CONCLUSIONS: We describe the implementation of an often-used automated method for the recording of individual locomotor activity to differentiate sibling species of Rhodnius with distinct epidemiological relevance. The higher levels of activity observed in the nymphs of R. prolixus could potentially contribute to increased vector capacity.

DNA Barcoding of Neotropical Sand Flies (Diptera, Psychodidae, Phlebotominae): Species Identification and Discovery within Brazil.

DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23-19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.

Clocks do not tick in unison: isolation of Clock and vrille shed new light on the clockwork model of the sand fly Lutzomyia longipalpis.

BACKGROUND: Behavior rhythms of insect vectors directly interfere with the dynamics of pathogen transmission to humans. The sand fly Lutzomyia longipalpis is the main vector of visceral leishmaniasis in America and concentrates its activity around dusk. Despite the accumulation of behavioral data, very little is known about the molecular bases of the clock mechanism in this species. This study aims to characterize, within an evolutionary perspective, two important circadian clock genes, Clock and vrille. FINDINGS: We have cloned and isolated the coding sequence of L. longipalpis' genes Clock and vrille. The former is structured in eight exons and encodes a protein of 696 amino acids, and the latter comprises three exons and translates to a protein of 469 amino acids. When compared to other insects' orthologues, L. longipalpis CLOCK shows a high degree of conservation in the functional domains bHLH and PAS, but a much shorter glutamine-rich (poly-Q) C-terminal region. As for L. longipalpis VRILLE, a high degree of conservation was found in the bZIP domain. To support these observations and provide an elegant view of the evolution of both genes in insects, phylogenetic analyses based on maximum-likelihood and Bayesian inferences were performed, corroborating the previously known insect systematics. CONCLUSIONS: The isolation and phylogenetic analyses of Clock and vrille orthologues in L. longipalpis bring novel and important data to characterize this species' circadian clock. Interestingly, the poly-Q shortening observed in CLOCK suggests that its transcription activity might be impaired and we speculate if this effect could be compensated by other clock factors such as CYCLE.

Phenotypic differentiation in love song traits among sibling species of the Lutzomyia longipalpis complex in Brazil.

BACKGROUND: Brazilian populations of Lutzomyia longipalpis may constitute a complex of cryptic species, and this report investigates the distribution and number of potential sibling species. One of the main differences observed among Brazilian populations is the type of acoustic signal produced by males during copulation. These copulation song differences seem to be evolving faster than neutral molecular markers and have been suggested to contribute to insemination failure observed in crosses between these sibling species. In previous studies, two main types of copulation songs were found, burst-type and pulse-type. The latter type can, in turn, be further subdivided into five different patterns. METHODS: We recorded male song from 13 new populations of the L. longipalpis complex from Brazil and compared the songs with 12 already available. RESULTS: Out of these 25 populations, 16 produce burst-type and 9 produce pulse-type songs. We performed a principal component analysis in these two main groups separately and an additional discriminant analysis in the pulse-type group. The pulse-type populations showed a clear separation between the five known patterns with a high correspondence of individuals to their correct group, confirming the differentiation between them. The distinctiveness of the burst-type subgroups was much lower than that observed among the pulse-type groups and no clear population structure was observed. This suggests that the burst-type populations represent a single species. CONCLUSION: Overall, our results are consistent with the existence in Brazil of at least six species of the L. longipalpis complex, one with a wide distribution comprising all the populations with burst-type songs, and five more closely related allopatric siblings with different pulse-type song patterns and more restricted distribution ranges.

Hybrid sterility in crosses between two Brazilian sibling species of the Anopheles albitarsis complex.

BACKGROUND: Complexes of cryptic species are common in several taxa and this is also the case in the Anopheles genus, a group including all known human malaria vectors. The Anopheles albitarsis complex comprises at least nine cryptic species, some of which are implicated as vectors of human malaria. Several different types of data have been generated for this species complex such as cytogenetics, alloenzymes, morphological and feeding behavioral, hybridization experiments, RAPD-PCR and RFLP and mitochondrial and nuclear markers. Studies focused on its postzygotic isolation are still somewhat rare in the literature despite their importance to understand the speciation process and the level of gene flow potentially occurring among the different sibling species. METHODS: Hybridization experiments between Anopheles albitarsis s.s. and Anopheles marajoara, as well as backcrosses between hybrids and Anopheles albitarsis s.s., were performed using the induced mating technique. Results were compared to intraspecific crosses. Larva-to-adult viability and sex ratio were also assessed. RESULTS: Male hybrids show very low insemination rates and nearly complete sterility, apparently due to abnormalities in their reproductive organs. Evidence of partial sterility among the hybrid females was also observed. CONCLUSIONS: Our data indicated that Anopheles albitarsis s.s. and Anopheles marajoara show a high level of postzygotic isolation with a strong hybrid male sterility. This result is consistent with the Haldane's rule which states that in interspecific crosses the heterogametic sex is the first to be affected. However, the fact that the females are not completely sterile raises the possibility of introgression between these two siblings species.

Effects of insemination and blood-feeding on locomotor activity of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) females under laboratory conditions.

BACKGROUND: Dengue is an arbovirus disease transmitted by two Aedes mosquitoes: Ae. aegypti and Ae. albopictus. Virgin females of these two species generally show a bimodal and diurnal pattern of activity, with early morning and late afternoon peaks. Although some studies on the flight activity of virgin, inseminated and blood-fed Ae. aegypti females have been carried out under laboratory conditions, little is known about the effects of such physiological states on the locomotor activity of Ae. albopictus and Ae. aegypti females. The aim of this study was to analyze, under laboratory conditions, the effects of insemination and blood-feeding on the locomotor activity of Ae. albopictus and Ae. aegypti females under LD 12:12, at 25°C. METHODS: Both Ae. albopictus and Ae. aegypti females were obtained from established laboratory colonies. Control groups were represented by virgin/unfed Ae. albopictus and Ae. aegypti females. Experiments were conducted under laboratory conditions, using an activity monitor that registers individual activity every thirty minutes. RESULTS: Virgin/unfed Ae. albopictus and Ae. aegypti females showed a diurnal and bimodal pattern of locomotor activity, with peaks at early morning and late afternoon. Insemination and blood-feeding significantly decreased the locomotor activity of Ae. aegypti females, but inseminated/blood-fed Ae. aegypti and Ae. albopictus females showed a similar significant decrease on the locomotor activity compared to virgin/unfed females. CONCLUSIONS: This study is the first demonstration of the effects of insemination and blood-feeding on the locomotor activity of Ae. albopictus and Ae. aegypti females under artificial conditions. Data suggest that Ae. albopictus and Ae. aegypti females respond in different ways to physiological status changes and such divergence between these two dengue vectors, associated with several ecological differences, could be related to the greater dengue vectorial capacity of Ae. aegypti in Americas in comparison to Ae. albopictus.

Effects of temperature and photoperiod on daily activity rhythms of Lutzomyia longipalpis (Diptera: Psychodidae).

BACKGROUND: Insect vectors have been established as models in Chronobiology for many decades, and recent studies have demonstrated a close relationship between the circadian clock machinery, daily rhythms of activity and vectorial capacity. Lutzomyia longipalpis, the primary vector of Leishmania (Leishmania) infantum in the New World, is reported to have crepuscular/nocturnal activity in the wild. However, most of these studies applied hourly CDC trap captures, which is a good indicative of L. longipalpis behaviour, but has limited accuracy due to the inability to record the daily activity of a single insect during consecutive days. In addition, very little is known about the activity pattern of L. longipalpis under seasonal variations of average temperature and day length in controlled laboratory conditions. METHODS: We recorded the locomotor activity of L. longipalpis males under different artificial regimes of temperature and photoperiod. First, in order to test the effects of temperature on the activity, sandflies were submitted to regimes of light/dark cycles similar to the equinox photoperiod (LD 12:12) combined with different constant temperatures (20°C, 25°C and 30°C). In addition, we recorded sandfly locomotor activity under a mild constant temperature (25°C with different day length regimes: 8 hours, 12 hours and 16 hours). RESULTS: L. longipalpis exhibited more activity at night, initiating dusk-related activity (onset time) at higher rather than lower temperatures. In parallel, changes of photoperiod affected anticipation as well as all the patterns of activity (onset, peak and offset time). However, under LD 16:08, sandflies presented the earliest values of maximum peak and offset times, contrary to other regimes. CONCLUSIONS: Herein, we showed that light and temperature modulate L. longipalpis behaviour under controlled laboratory conditions, suggesting that sandflies might use environmental information to sustain their crepuscular/nocturnal activity, as well as other important aspects as mating and host-seeking at appropriate times in different seasons. Our results depict previously unappreciated aspects of the L. longipalpis daily rhythms of activity that might have important epidemiological implications.

Multilocus analysis of divergence and introgression in sympatric and allopatric sibling species of the Lutzomyia longipalpis complex in Brazil.

BACKGROUND: Lutzomyia longipalpis, the main vector of visceral leishmaniasis in Latin America, is a complex of sibling species. In Brazil, a number of very closely related sibling species have been revealed by the analyses of copulation songs, sex pheromones and molecular markers. However, the level of divergence and gene flow between the sibling species remains unclear. Brazilian populations of this vector can be divided in two main groups: one producing Burst-type songs and the Cembrene-1 pheromone and a second more diverse group producing various Pulse song subtypes and different pheromones. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed 21 nuclear loci in two pairs of Brazilian populations: two sympatric populations from the Sobral locality (1S and 2S) in northeastern Brazil and two allopatric populations from the Lapinha and Pancas localities in southeastern Brazil. Pancas and Sobral 2S are populations of the Burst/Cembrene-1 species while Lapinha and Sobral 1S are two putative incipient species producing the same pheromone and similar Pulse song subtypes. The multilocus analysis strongly suggests the occurrence of gene flow during the divergence between the sibling species, with different levels of introgression between loci. Moreover, this differential introgression is asymmetrical, with estimated gene flow being higher in the direction of the Burst/Cembrene-1 species. CONCLUSIONS/SIGNIFICANCE: The results indicate that introgressive hybridization has been a crucial phenomenon in shaping the genome of the L. longipalpis complex. This has possible epidemiological implications and is particularly interesting considering the potential for increased introgression caused by man-made environmental changes and the current trend of leishmaniasis urbanization in Brazil.

Fine structure of the male reproductive system and reproductive behavior of Lutzomyia longipalpis sandflies (Diptera: Psychodidae: Phlebotominae).

BACKGROUND: The male reproductive system of insects can have several tissues responsible for the secretion of seminal fluid proteins (SFPs), such as testes, accessory glands, seminal vesicles, ejaculatory duct and ejaculatory bulb. The SFPs are transferred during mating and can induce several physiological and behavioral changes in females, such as increase in oviposition and decrease in sexual receptivity after copulation. The phlebotomine Lutzomyia longipalpis is the main vector of visceral leishmaniasis. Despite its medical importance, little is known about its reproductive biology. Here we present morphological aspects of the male L. longipalpis reproductive system by light, scanning and transmission electron microscopy, and compare the mating frequency of both virgin and previously mated females. RESULTS: The male L. longipalpis reproductive system is comprised by a pair of oval-shaped testes linked to a seminal vesicle by vasa deferentia. It follows an ejaculatory duct with an ejaculatory pump (a large bulb enveloped by muscles and associated to tracheas). The terminal endings of the vasa deferentia are inserted into the seminal vesicle by invaginations of the seminal vesicle wall, which is composed by a single layer of gland cells, with well-developed endoplasmic reticulum profiles and secretion granules. Our data suggest that the seminal vesicle acts both as a spermatozoa reservoir and as an accessory gland. Mating experiments support this hypothesis, revealing a decrease in mating frequency after copulation that indicates the effect of putative SFPs. CONCLUSION: Ultrastructural features of the L. longipalpis male seminal vesicle indicated its possible role as an accessory gland. Behavioral observations revealed a reduction in mating frequency of copulated females. Together with transcriptome analyses from male sandfly reproductive organs identifying ESTs encoding orthologs of SFPs, these data indicate the presence of putative L. longipalpis SFPs reducing sexual mating frequency of copulated females.

Evidence for the occurrence of two sympatric sibling species within the Anopheles (Kerteszia) cruzii complex in southeast Brazil and the detection of asymmetric introgression between them using a multilocus analysis.

BACKGROUND: Anopheles (Kerteszia) cruzii (Diptera: Culicidae) is a primary vector of human and simian malaria parasites in southern and southeastern Brazil. Earlier studies using chromosome inversions, isoenzymes and a number of molecular markers have suggested that An. cruzii is a species complex. RESULTS: In this study, a multilocus approach using six loci, three circadian clock genes and three encoding ribosomal proteins, was carried out to investigate in more detail the genetic differentiation between the An. cruzii populations from Florianópolis-Santa Catarina (southern Brazil) and Itatiaia-Rio de Janeiro States (southeastern Brazil). The analyses were performed first comparing Florianópolis and Itatiaia, and then comparing the two putative sympatric incipient species from Itatiaia (Itatiaia A and Itatiaia B). The analysis revealed high FST values between Florianópolis and Itatiaia (considering Itatiaia A and B together) and also between the sympatric Itatiaia A and Itatiaia B, irrespective of their function. Also, using the IM program, no strong indication of migration was found between Florianópolis and Itatiaia (considering Itatiaia A and B together) using all loci together, but between Itatiaia A and Itatiaia B, the results show evidence of migration only in the direction of Itatiaia B. CONCLUSIONS: The results of the multilocus analysis indicate that Florianópolis and Itatiaia represent different species of the An. cruzii complex that diverged around 0.6 Mya, and also that the Itatiaia sample is composed of two sympatric incipient species A and B, which diverged around 0.2 Mya. Asymmetric introgression was found between the latter two species despite strong divergence in some loci.

Assessing the effects of Aedes aegypti kdr mutations on pyrethroid resistance and its fitness cost.

Pyrethroids are the most used insecticide class worldwide. They target the voltage gated sodium channel (NaV), inducing the knockdown effect. In Aedes aegypti, the main dengue vector, the AaNaV substitutions Val1016Ile and Phe1534Cys are the most important knockdown resistance (kdr) mutations. We evaluated the fitness cost of these kdr mutations related to distinct aspects of development and reproduction, in the absence of any other major resistance mechanism. To accomplish this, we initially set up 68 crosses with mosquitoes from a natural population. Allele-specific PCR revealed that one couple, the one originating the CIT-32 strain, had both parents homozygous for both kdr mutations. However, this pyrethroid resistant strain also presented high levels of detoxifying enzymes, which synergistically account for resistance, as revealed by biological and biochemical assays. Therefore, we carried out backcrosses between CIT-32 and Rockefeller (an insecticide susceptible strain) for eight generations in order to bring the kdr mutation into a susceptible genetic background. This new strain, named Rock-kdr, was highly resistant to pyrethroid and presented reduced alteration of detoxifying activity. Fitness of the Rock-kdr was then evaluated in comparison with Rockefeller. In this strain, larval development took longer, adults had an increased locomotor activity, fewer females laid eggs, and produced a lower number of eggs. Under an inter-strain competition scenario, the Rock-kdr larvae developed even slower. Moreover, when Rockefeller and Rock-kdr were reared together in population cage experiments during 15 generations in absence of insecticide, the mutant allele decreased in frequency. These results strongly suggest that the Ae. aegypti kdr mutations have a high fitness cost. Therefore, enhanced surveillance for resistance should be priority in localities where the kdr mutation is found before new adaptive alleles can be selected for diminishing the kdr deleterious effects.

A novel classification system for evolutionary aging theories.

Theories of lifespan evolution are a source of confusion amongst aging researchers. After a century of aging research the dispute over whether the aging process is active or passive persists and a comprehensive and universally accepted theoretical model remains elusive. Evolutionary aging theories primarily dispute whether the aging process is exclusively adapted to favor the kin or exclusively non-adapted to favor the individual. Interestingly, contradictory data and theories supporting both exclusively programmed and exclusively non-programmed theories continue to grow. However, this is a false dichotomy; natural selection favors traits resulting in efficient reproduction whether they benefit the individual or the kin. Thus, to understand the evolution of aging, first we must understand the environment-dependent balance between the advantages and disadvantages of extended lifespan in the process of spreading genes. As described by distinct theories, different niches and environmental conditions confer on extended lifespan a range of fitness values varying from highly beneficial to highly detrimental. Here, we considered the range of fitness values for extended lifespan and develop a fitness-based framework for categorizing existing theories. We show that all theories can be classified into four basic types: secondary (beneficial), maladaptive (neutral), assisted death (detrimental), and senemorphic aging (varying between beneficial to detrimental). We anticipate that this classification system will assist with understanding and interpreting aging/death by providing a way of considering theories as members of one of these classes rather than consideration of their individual details.

Analysis of cycle gene expression in Aedes aegypti brains by in situ hybridization.

Even though the blood-sucking mosquito Aedes aegypti is one of the most important disease vectors, relatively little is known about the molecular mechanisms underlying processes involved in the temporal pattern of its activity and host seeking behavior. In this study, we analyzed the expression of the cycle (cyc) gene, one of the core components of the circadian clock, in Ae. aegypti brains by in situ hybridization at two different time points in light-dark conditions and compared the results with those obtained using a quantitative PCR assay (qPCR). Within the brain, differential labeling was detected according to distinct areas empirically pre-defined. Six out of seven of these areas showed significantly higher staining at ZT3 (three hours after light-on) compared to ZT11 (one before light-off), which is consistent with the qPCR data. Predominant staining was observed in three of those areas which correspond to positions of the optical and antennal lobes, as well as the region where the neurons controlling activity rhythms are presumably localized.