Transcriptome analysis reveals biosynthesis and regulation of flavonoid in common bean seeds during grain filling.

The Andean domesticated common beans (Phaseolus vulgaris) are significant sources of phenolic compounds associated with health benefits. However, the regulation of biosynthesis of these compounds during bean seed development remains unclear. To elucidate the gene expression patterns involved in the regulation of the flavonoid pathway, we conducted a transcriptome analysis of two contrasting Chilean varieties, Negro Argel (black bean) and Coscorron (white bean), at three developmental stages associated with seed color change, as well as different flavonoid compound accumulations. Our study reveals that phenolic compound synthesis initiates during seed filling, although it exhibits desynchronization between both varieties. We identified 10,153 Differentially Expressed Genes (DEGs) across all comparisons. The KEGG pathway 'Flavonoid biosynthesis' showed enrichment of induced DEGs in Negro Argel (PV172), consistent with the accumulation of delphinidin, petunidin, and malvidin hexosides in their seeds, while catechin glucoside, procyanidin and kaempferol derivatives were predominantly detected in Coscorrón (PV24). Furthermore, while the flavonoid pathway was active in both varieties, our results suggest that enzymes involved in the final steps, such as ANS and UGT, were crucial, inducing anthocyanin formation in Negro Argel. Additionally, during active anthocyanin biosynthesis, the accumulation of reserve proteins or those related to seed protection and germination was induced. These findings provide valuable insights and serve as a guide for plant breeding aimed at enhancing the health and nutritional properties of common beans.

Effectiveness of an intervention to improve ART adherence among men who have sex with men living with HIV: a randomized controlled trial in three public HIV clinics in Mexico.

We conducted a parallel-group randomized controlled trial in three HIV clinics in Mexico to evaluate a user-centred habit-formation intervention to improve ART adherence among MSM living with HIV. We randomized 74 participants to the intervention group and 77 to the control group. We measured adherence at one, four, and ten months through medication possession ratio and self-reported adherence. Additionally, we measured viral load, CD4 cell count, major depression disorder symptoms, and alcohol and substance use disorder at baseline, fourth and tenth months. We found no statistically significant effect on adherence between groups. However, the intervention demonstrated positive results in major depression disorder symptoms (21% vs. 6%, p = 0.008) and substance use disorder (11% vs. 1%, p = 0.018) in the fourth month. The latter is relevant because, in addition to its direct benefit, it might also improve the chances of maintaining adequate adherence in the long term. This trial was retrospectively registered at ClinicalTrials.gov (trial number NCT03410680) on 8 January 2018.Trial registration: ClinicalTrials.gov identifier: NCT03410680.

A Past Genetic Bottleneck from Argentine Beans and a Selective Sweep Led to the Race Chile of the Common Bean (Phaseolus vulgaris L.).

The domestication process of the common bean gave rise to six different races which come from the two ancestral genetic pools, the Mesoamerican (Durango, Jalisco, and Mesoamerica races) and the Andean (New Granada, Peru, and Chile races). In this study, a collection of 281 common bean landraces from Chile was analyzed using a 12K-SNP microarray. Additionally, 401 accessions representing the rest of the five common bean races were analyzed. A total of 2543 SNPs allowed us to differentiate a genetic group of 165 accessions that corresponds to the race Chile, 90 of which were classified as pure accessions, such as the bean types 'Tórtola', 'Sapito', 'Coscorrón', and 'Frutilla'. Our genetic analysis indicates that the race Chile has a close relationship with accessions from Argentina, suggesting that nomadic ancestral peoples introduced the bean seed to Chile. Previous archaeological and genetic studies support this hypothesis. Additionally, the low genetic diversity (π = 0.053; uHe = 0.53) and the negative value of Tajima' D (D = -1.371) indicate that the race Chile suffered a bottleneck and a selective sweep after its introduction, supporting the hypothesis that a small group of Argentine bean genotypes led to the race Chile. A total of 235 genes were identified within haplotype blocks detected exclusively in the race Chile, most of them involved in signal transduction, supporting the hypothesis that intracellular signaling pathways play a fundamental role in the adaptation of organisms to changes in the environment. To date, our findings are the most complete investigation associated with the origin of the race Chile of common bean.

Biogeographic regionalization of the Neotropical region: New map and shapefile.

We provide a map and shapefile of the 57 biogeographic provinces of the Neotropical region. Recognition of these provinces is based on their endemic species, but their delimitation on the map is based on ecoregions combining climatic, geological, and biotic criteria. These provinces belong to the Antillean, Brazilian and Chacoan subregions, and the Mexican and South American transition zones. We provide a vector file of the biogeographical regionalization by converting the map into a polygon shapefile and a raster file with all provinces.

Late Pleistocene Altitudinal Segregation and Demography Define Future Climate Change Distribution of the Peromyscus mexicanus Species Group: Conservation Implications.

Mountains harbor a significant number of the World's biodiversity, both on tropical and temperate regions. Notably, one crucial gap in conservation is the consideration of historical and contemporary patterns influencing differential distribution in small mammal mountain species and how climate change will affect their distribution and survival. The mice Peromyscus mexicanus species group is distributed across mountains in Guatemala-Chiapas and Central America, which experienced significant effects of glacial and interglacial cycles. We determined phylogeographic and demographic patterns of lowlands and highlands mountain lineages, revealing that the radiation of modern P. mexicanus lineages occurred during the Pleistocene (ca. 2.6 mya) along Nuclear Central America. In concert with climatic cycles and the distribution of habitats, lowland and highland lineages showed recent population size increase and decrease, respectively. We also estimated the current and future distribution ranges for six lineages, finding marked area size increase for two lineages for which vegetation type and distribution would facilitate migrating towards higher elevations. Contrastingly, three lineages showed range size decrease; their ecological requirements make them highly susceptible to future habitat loss. Our findings are clear evidence of the negative impacts of future climate change, while our ability to manage and conserve these vulnerable ecosystems and mountain species is contingent on our understanding of the implications of climate change on the distribution, ecology, and genetics of wildlife populations.

Water Shortage Affects Vegetative and Reproductive Stages of Common Bean (Phaseolus vulgaris) Chilean Landraces, Differentially Impacting Grain Yield Components.

Water availability for agricultural use is currently a global problem that worsens with climate change in several regions of the world. Among grain legumes, common bean (Phaseolus vulgaris) is the most cultivated in the worldwide. The Chilean germplasm of common bean is characterized by tolerance to water stress. Here, we analyzed a selection of nine ancient Chilean landraces in regard to their drought tolerance, simulating optimal (OW) and restricted watering (RW) in a Mediterranean environment. Phenological, growth, and yield traits were recorded, and correlation analysis was performed. Accordingly, leaf temperature and osmotic potential were higher under RW, while the leaf chlorophyll content decreased in all landraces. Physiological maturity days and seed-filling days were lower in RW than in OW. This similarly occurred with the grain yield. The % yield reduction was negatively correlated with the % pod reduction and the relative rate of leaf expansion (RLAE) reduction. However, the 100-seed weight value was not significantly modified by water treatment (p > 0.05). For instance, landraces that preferred to fill the grain with a lower rate of leaf expansion showed a lower loss in grain yield under drought conditions. These results suggest that the resource partitioning between growing leaves, flowers, and developing pods in Chilean landraces is variable, affecting the common bean drought tolerance.

Regulation of Anthocyanin Biosynthesis by Drought and UV-B Radiation in Wild Tomato (Solanum peruvianum) Fruit.

Anthocyanins are plant pigments derived from the phenylpropanoid pathway which are produced in many different species, contributing to defense against stresses by their antioxidant properties. Cultivated tomatoes cannot synthesize flavonoids; however, wild tomatoes such as Solanum chilense and Solanum lycopersicoides have anthocyanin pigmented skin. Other wild tomato species such as Solanum peruvianum have been poorly studied concerning anthocyanin accumulation in the fruit. This research is the first to address the regulation of anthocyanin biosynthesis mediated by drought stress and light radiation in S. peruvianum fruit. Transcript accumulation of SpAN2, encoding for a key MYB type transcription factor for the regulation of anthocyanin biosynthesis, was induced in the fruit of plants exposed to drought treatment. In addition, fruit peel accumulates a greater anthocyanin content in water deficit-treated plants. The expression of SpAN2 was also regulated according to sunlight exposure, reaching a higher expression during maximal daily UV radiation and under controlled UV-B treatments. Similar results were observed for the expression of the late flavonoid biosynthetic gene dihydroflavonol 4-reductase (SpDFR). These results suggest that SpAN2 and SpDFR are involved in anthocyanin biosynthesis under drought stress and UV radiation in S. peruvianum.

Toward a biogeographic regionalization of the Nearctic region: Area nomenclature and digital map.

We provide a preliminary nomenclatural proposal and a digital map of the Nearctic region, based on published regionalizations, especially Dice (1943), and applying the International Code of Area Nomenclature. The Nearctic region is comprised of three subregions (one of them with two dominions), one transition zone and 29 provinces. The Arctic subregion, in northern North America and Greenland, includes the Eskimoan, Hudsonian, Aleutian and Sitkan provinces. The Western subregion, in western North America, includes the Californian dominion, with the Californian and Oregonian provinces; and the Rocky Mountain dominion, including the Montanian, Saskatchewan, Palusian, Artemisian, Coloradan, Kansan, Mohavian, Navahonian, Sonoran, Chihuahuan, Comanche, and Baja California provinces. The Alleghany subregion, in eastern North America, includes the Illinoian, Canadian, Carolinian, Texan, Austroriparian, and Tamaulipan provinces. The Mexican Transition Zone, situated in the area of overlap with the Neotropical region, includes the Sierra Madre Occidental, Sierra Madre Oriental, Transmexican Volcanic Belt, Sierra Madre del Sur and Chiapas Highlands provinces.

Incidence of dengue illness in Mexican people aged 6 months to 50 years old: A prospective cohort study conducted in Jalisco.

BACKGROUND AND OBJECTIVES: The burden of dengue virus (DENV), a mosquito-borne pathogen, remains difficult to assess due to misdiagnosis and underreporting. Moreover, the large proportion of asymptomatic dengue cases impairs comprehensive assessment of its epidemiology even where effective surveillance systems are in place. We conducted a prospective community-based study to assess the incidence of symptomatic dengue cases in Zapopan and neighboring municipalities in the state of Jalisco, Mexico. METHODS: Healthy subjects aged 6 months to 50 years living in households located in the Zapopan and neighboring municipalities were enrolled for a 24-month follow-up study (NCT02766088). Serostatus was determined at enrolment and weekly contacts were conducted via phone calls and home visits. Participants had to report any febrile episode lasting for at least two days. Suspected dengue cases were tested by reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR), detection of non-structural protein 1 (NS1), anti-DENV immunoglobulin G and M (IgG and IgM) assays. RESULTS: A total of 350 individuals from 87 households were enrolled. The overall seroprevalence of anti-DENV IgG at enrolment was 19.4% (95% confidence interval [CI] 14.5-25.6) with the highest seroprevalence rate observed in the adult group. Over the 27-month study period from July 2016 to September 2018, a total of 18 suspected dengue cases were reported. Four cases were confirmed by RT-qPCR and serotyped as DENV-1. A fifth case was confirmed by the NS1 assay. The 13 remaining suspected cases were tested negative by these assays. Based on the 5 virologically confirmed cases, symptomatic dengue incidence proportion of 1.4% (95%CI 0.5-3.8) was estimated. No severe cases or hospitalizations occurred during the study. CONCLUSION: Community-based active surveillance was shown as efficient to detect symptomatic dengue cases. CLINICAL TRIAL REGISTRATION: NCT02766088.

[Acinetobacter baumannii bacteremia]. / Bacteriemia por Acinetobacter baumannii.

Acinetobacter baumannii has emerged as an important nosocomial pathogen. It is difficult to control and treat. The most seriously ill patients and those previously infected are more likely than others to be infected or colonized by A. baumannii. The epidemiology of A. baumannii infection is complex, with the coexistence of epidemic and endemic infections. The A. baumannii are the species isolated in 90% of the nosocomial infections and in 92% of the nosocomial bacteremias. The intensive care units presented the greater number of nosocomial bacteremias by A. baumannii. The most common sources of A. baumannii are respiratory tract, surgical wound, catheter, urinary tract and others. The most frequently clinical manifestation is sepsis and a fulminating course is observed when the patient presents septic shock. Progressive resistance of A. baumannii to antimicrobial limits the therapeutic options. The patients with A. baumannii multidrug-resistant present an excessive rate of attributed mortality, length of stay and costs.

Prevalence of Chagas heart disease in dilated cardiomyopathy.

OBJECTIVES: The main objective is to determine the prevalence of American trypanosomiasis in patients with dilated cardiomyopathy in a tertiary hospital in western Mexico. METHODS: From January 1991 to February 2016, 387 consecutive patients with a confirmed diagnosis of dilated cardiomyopathy were included in the study. Cases with ventricular dilatation secondary to ischemic heart disease, valvular heart disease, hypertension, lung disease, pericardial disease, or congenital heart disease were excluded from the study. Diagnosis was made detecting antibodies against Trypanosoma cruzi with two different methods or parasite in blood. RESULTS: Were included 387 patients with dilated cardiomyopathy, Chagas cardiomyopathy was confirmed in 6.9%, two patients in the acute phase (in one, suspected transfusion transmission was detected). Most patients were born in rural areas. About 96.2% showed congestive heart failure, only one patient with apical left ventricular aneurysm manifested palpitations. About 66% with right bundle branch block, left anterior fascicular block, or the association of both, in 14.8%, non-sustained ventricular tachycardia was found. CONCLUSIONS: Chagas cardiomyopathy is common in México, mainly in people who were born or lived during childhood in rural areas. It is a common cause of heart failure. Chagas' heart disease should be suspected in patients receiving a blood transfusion, even without another epidemiological history.

[Bacteremia caused by Acinetobacter baumannii among patients in critical care]. / Bacteremia por Acinetobacter baumannii en pacientes en estado crítico.

OBJECTIVE: Determine the severity and the course of Acinetobacter baumannii blood infections (AbBI). METHODS: We carried out a prospective review of all Acinetobacter baumannii (A. baumannii) isolated from the Intensive Care Unit (ICU) of a medical facility. RESULTS: During the study period, 46 patients were identified with AbBI. The rate of AbBI was 14 per 1,000 admissions. A. baumannii was ICU acquired in 85% of cases studied. The median time frame between ICU admission and acquisition of AbBI was 9 +/- 7 days. 31 patients displayed blood infection (BI) by A. baumannii alone and 15 with polymicrobial BI. The clinical manifestation of the 31 patients with AbBI included the following symptoms: 42% presented with septic shock, 42% had severe sepsis and 16% had sepsis. Two or more episodes of AbBI were observed among 13% of patients. Of A. baumannii isolates, 17% were resistant to imipenem. Patient mortality with AbBI alone was 45% and 40% for polymicrobial BI. Patient mortality for septic shock was 60%, 70% died during the first 72 hrs after AbBI. CONCLUSIONS: AbBI displayed an endemic pattern during the study period. Severe sepsis and septic shock were the most common clinical presentations of AbBI. AbBI are associated with a significant increase in the death rate of patient in the ICU studied.

Understanding the Complexity of Cold Tolerance in White Clover using Temperature Gradient Locations and a GWAS Approach.

White clover ( L.) is the most important grazing perennial forage legume in temperate climates. However, its limited capacity to survive and restore growth after low temperatures during winter constrains the productivity and wide adoption of the crop. Despite the importance of cold tolerance for white clover cultivar development, the genetic basis of this trait remains largely unknown. Hence, in this study, we performed the first genome-wide association study (GWAS) analyses in white clover to identify quantitative trait loci (QTL) for cold-tolerance-related traits. Seeds from 192 divergent genotypes from six populations in the Patagonia region of South America were collected and seed-derived plants were further clonally propagated. Clonal trials were established in three locations representing temperature gradient associated with elevation. Given the allotetraploid nature of the white clover genome, distinct genetic models (diploid and tetraploid) were tested. Only the tetraploid parameterization was able to detect the 53 loci associated with cold-tolerance traits. Out of the 53 single nucleotide polymorphism (SNP) trait associations, 17 controlled more than one trait or were stable across multiple sites. This work represents the first report of QTL for cold-tolerance-related traits, providing insights into its genetic basis and candidate genomic regions for further functional validation studies.

Stem carbohydrate dynamics and expression of genes involved in fructan accumulation and remobilization during grain growth in wheat (Triticum aestivum L.) genotypes with contrasting tolerance to water stress.

The genetic and physiological mechanisms underlying the relationship between water-soluble carbohydrates (WSC) and water stress tolerance are scarcely known. This study aimed to evaluate the main WSC in stems, and the expression of genes involved in fructan metabolism in wheat genotypes growing in a glasshouse with water stress (WS; 50% field capacity from heading) and full irrigation (FI; 100% field capacity). Eight wheat genotypes (five tolerant and three susceptible to water stress) were evaluated initially (experiment 1) and the two most contrasting genotypes in terms of WSC accumulation were evaluated in a subsequent experiment (experiment 2). Maximum accumulation of WSC occurred 10-20 days after anthesis. Under WS, the stress-tolerant genotype exhibited higher concentrations of WSC, glucose, fructose and fructan in the stems, compared to FI. In addition, the stress-tolerant genotype exhibited higher up-regulation of the fructan 1-fructosyltransferase B (1-FFTB) and fructan 1-exohydrolase w2 (1-FEHw2) genes, whereas the susceptible cultivar presented an up-regulation of the fructan 6-fructosyltransferase (6-SFT) and fructan 1-exohydrolase w3 (1-FEHw3) genes. Our results indicated clear differences in the pattern of WSC accumulation and the expression of genes regulating fructan metabolism between the tolerant and susceptible genotypes under WS.

Mexican biogeographic provinces: Map and shapefiles.

We provide a map of the 14 biogeographic provinces of Mexico based on the ecoregions recognized for the country, which combine climatic, geological and biotic criteria. These provinces belong to the Nearctic region (Californian, Baja Californian, Sonoran, Chihuahuan Desert and Tamaulipas provinces), Neotropical region (Pacific Lowlands, Balsas Basin, Veracruzan and Yucatán Peninsula provinces) and the Mexican transition zone (Sierra Madre Occidental, Sierra Madre Oriental, Transmexican Volcanic Belt, Sierra Madre del Sur and Chiapas Highlands provinces). In order to facilitate future biogeographic analyses, we provide a file of the biogeographical regionalisation of Mexico by converting the map into a polygon shapefile and a raster file with all provinces. We also separately provide each of the provinces in vector and raster format. All the maps are in geographical and Lambert Conformal Conic projections.

Large Retropharyngeal Abscesses in an Immunocompetent Adult Patient with Disseminated Tuberculosis.

BACKGROUND The retropharyngeal abscess is a rare presentation of head and neck tuberculosis. The pathogenesis of the abscess formation in the retropharyngeal space in the adult is controversial. CASE REPORT We report a case of large retropharyngeal abscesses in a 46-year-old man with disseminated tuberculosis. The patient had severe progressive dysphagia, weight loss, and a slowly enlarging bilateral cervical mass during a period of three months. His posterior pharynx wall was bulging and red, and both tonsils were enlarged and congested. The neck had an abscess of 5 cm in diameter that was firm, tender, and warm along the left sternocleidomastoid muscle. Palpable bilateral lymphadenitis was detected in the submandibular, cervical, axillary, and inguinal regions. A computed tomographic (CT) scan of the neck revealed large bilobulated retropharyngeal abscesses. A liver ultrasound showed multiple hypoechoic lesions. A Ziehl-Neelsen smear for acid-fast bacilli was positive from different abscess samples, and mycobacterial cultures subsequently yielded Mycobacterium tuberculosis. Antituberculous therapy was begun and the retropharyngeal abscesses were aspirated by external incision with complete drainage and relief of symptoms. CONCLUSIONS Large retropharyngeal abscess is a rare entity in which Mycobacterium tuberculosis etiology should be considered, especially in endemic countries, and the diagnosis may be difficult because symptoms and signs are influenced by abscess size and time of onset, or if the etiology is not suspected.

Physiological Traits Associated with Wheat Yield Potential and Performance under Water-Stress in a Mediterranean Environment.

Different physiological traits have been proposed as key traits associated with yield potential as well as performance under water stress. The aim of this paper is to examine the genotypic variability of leaf chlorophyll, stem water-soluble carbohydrate content and carbon isotope discrimination (Δ(13)C), and their relationship with grain yield (GY) and other agronomical traits, under contrasting water conditions in a Mediterranean environment. The study was performed on a large collection of 384 wheat genotypes grown under water stress (WS, rainfed), mild water stress (MWS, deficit irrigation), and full irrigation (FI). The average GY of two growing seasons was 2.4, 4.8, and 8.9 Mg ha(-1) under WS, MWS, and FI, respectively. Chlorophyll content at anthesis was positively correlated with GY (except under FI in 2011) and the agronomical components kernels per spike (KS) and thousand kernel weight (TKW). The WSC content at anthesis (WSCCa) was negatively correlated with spikes per square meter (SM2), but positively correlated with KS and TKW under WS and FI conditions. As a consequence, the relationships between WSCCa with GY were low or not significant. Therefore, selecting for high stem WSC would not necessary lead to genotypes of GY potential. The relationship between Δ(13)C and GY was positive under FI and MWS but negative under severe WS (in 2011), indicating higher water use under yield potential and MWS conditions.

Prevalence of Chagas heart disease in dilated cardiomyopathy / Prevalencia de cardiopatía chagásica en la miocardiopatía dilatada

Abstract Objectives: The main objective is to determine the prevalence of American trypanosomiasis in patients with dilated cardiomyopathy in a tertiary hospital in western Mexico. Methods: From January 1991 to February 2016, 387 consecutive patients with a confirmed diagnosis of dilated cardiomyopathy were included in the study. Cases with ventricular dilatation secondary to ischemic heart disease, valvular heart disease, hypertension, lung disease, pericardial disease, or congenital heart disease were excluded from the study. Diagnosis was made detecting antibodies against Trypanosoma cruzi with two different methods or parasite in blood. Results: Were included 387 patients with dilated cardiomyopathy, Chagas cardiomyopathy was confirmed in 6.9%, two patients in the acute phase (in one, suspected transfusion transmission was detected). Most patients were born in rural areas. About 96.2% showed congestive heart failure, only one patient with apical left ventricular aneurysm manifested palpitations. About 66% with right bundle branch block, left anterior fascicular block, or the association of both, in 14.8%, non-sustained ventricular tachycardia was found. Conclusions: Chagas cardiomyopathy is common in México, mainly in people who were born or lived during childhood in rural areas. It is a common cause of heart failure. Chagas’ heart disease should be suspected in patients receiving a blood transfusion, even without another epidemiological history.

Resumen Objetivo: El objetivo principal del estudio es conocer la prevalencia de tripanosomiasis americana en pacientes con cardiomiopatía dilatada, en un hospital de concentración en el occidente de México. Métodos: Desde enero de 1991 a febrero de 2016 se incluyeron 387 pacientes consecutivos con diagnóstico de cardiomiopatía dilatada, se excluyeron los casos con dilatación ventricular secundaria a cardiopatía isquémica, valvulopatías, hipertensión arterial sistémica, enfermedad pulmonar, enfermedad pericárdica o cardiopatías congénitas. El diagnóstico se realizó mediante la detección de anticuerpos anti-tripanosoma cruzi con 2 métodos positivos diferentes o con la detección del parásito en sangre. Resultados: Se incluyeron 387 paciente con cardiomiopatía dilatada, en el 6.9% se confirmó cardiopatía chagásica; dos pacientes en fase aguda (uno con sospecha de transmisión transfusional). La mayoría de los pacientes provenían de zonas rurales. El 96.2% de los casos presentó insuficiencia cardiaca congestiva, un paciente con aneurisma apical del ventrículo izquierdo solo manifestó palpitaciones. El 66% presentó bloqueo de la rama derecha del haz de His, hemibloqueo anterior izquierdo o la asociación de ambos, en el 14.8% se encontró taquicardia ventricular no sostenida. Conclusiones: La cardiopatía chagásica es frecuente en nuestro medio, principalmente en personas que nacieron o vivieron durante la infancia en áreas rurales. Es causa común de insuficiencia cardiaca. La cardiomiopatía chagásica debe sospecharse en pacientes que reciben transfusión sanguínea, incluso sin otros antecedentes epidemiológicos

Understanding the roles of Lys33 and Arg45 in the binding-site stability of LjLTP10, an LTP related to drought stress in Lotus japonicus.

In Lotus japonicus, as in most plants, long-chain fatty acids are important components of cuticular wax, one of the principal functions of which is to act as a barrier to water loss in response to drought stress. It is thought that lipid transfer proteins (LTPs) are involved in the process of cuticle formation. We previously described LjLTP10 as an LTP involved in cuticle formation during acclimation response to drought stress in L. japonicus. The structural model of LjLTP10 had two residues (K33 and R45) in the hydrophobic cavity, although the role of these residues was unclear. In the present work, we investigated the molecular mechanism involved in the transport of lipid precursors in L. japonicus and clarified the importance of the residues K33 and R45. First, in silico site-directed mutagenesis studies were carried out on the LjLTP10 structure. Structural analysis showed that LjLTP10 mutants possess similar structures but their hydrophobic cavities are somewhat different. Unfavorable energies for the interactions of the mutant proteins with different ligands were found by molecular docking and molecular dynamics simulations. We also examined the contributions of energetic parameters to the free energy of the protein-ligand complex using the MM-GBSA method. Results showed that the different complexes present similar, favorable van der Waals interactions, whereas electrostatic interactions were not favored in the mutant structures. Our study indicates that the residues K33 and R45 play a crucial role in maintaining the binding pocket structure required for lipid transport.

Climate-induced range shifts and possible hybridisation consequences in insects.

Many ectotherms have altered their geographic ranges in response to rising global temperatures. Current range shifts will likely increase the sympatry and hybridisation between recently diverged species. Here we predict future sympatric distributions and risk of hybridisation in seven Mediterranean ischnurid damselfly species (I. elegans, I. fountaineae, I. genei, I. graellsii, I. pumilio, I. saharensis and I. senegalensis). We used a maximum entropy modelling technique to predict future potential distribution under four different Global Circulation Models and a realistic emissions scenario of climate change. We carried out a comprehensive data compilation of reproductive isolation (habitat, temporal, sexual, mechanical and gametic) between the seven studied species. Combining the potential distribution and data of reproductive isolation at different instances (habitat, temporal, sexual, mechanical and gametic), we infer the risk of hybridisation in these insects. Our findings showed that all but I. graellsii will decrease in distributional extent and all species except I. senegalensis are predicted to have northern range shifts. Models of potential distribution predicted an increase of the likely overlapping ranges for 12 species combinations, out of a total of 42 combinations, 10 of which currently overlap. Moreover, the lack of complete reproductive isolation and the patterns of hybridisation detected between closely related ischnurids, could lead to local extinctions of native species if the hybrids or the introgressed colonising species become more successful.