Identification and characterization of the EXPA7, EXPA18 and EXT10 genes in Turbinicarpus lophophoroides (Werderm.) Buxb. & Backeb; and their expression analysis in the root under abiotic stress.

Expansin and extensin are proteins involved in resistance to various abiotic stresses by processes of cell wall modification and in the formation and elongation of the hairy root. They are located in several organs of the plant included root epidermis. Turbinicarpus lophophoroides is a cactus model to studies these genes in adventitious and transformed roots. In this study, we identified and characterized the expansin7, expansin18 and extensin10 genes in T. lophophoroides. Bioinformatic analysis indicated that the expansin sequences contained the motifs: HTFYG, HFD, YRR, VPC and YW; and certain conserved cysteine (C) residues. Regarding extensin10, the sequence contains the conserved SPPPP (SP4), YYS and YV motifs. The expression analysis in adventitious and transformed roots under osmotic stress (300 mM mannitol), heat (37 °C) and cold (4 °C); shows a higher expression of TlExpA18 in both roots, a decrease in TlExpA7 in transformed roots and a null expression in TlExt10 in both roots. In addition, a morphological comparison of the maturation/differentiation zone, meristem and cap between adventitious and transformed roots by SEM was performed, finding differences in the quantity and length of the hairy roots and the shape of the root cap. Overall, the study concluded that TlExpA18 and TlExpA7 belong to expansin family and TlExt10 belong to extensin family. The expression characteristics of TlExpA18, TlExpA7 and TlExt10 will facilitate the investigation of its function in stress response and other physiological processes in T. lophophoroides.

Repeated AAV-mediated gene transfer by serotype switching enables long-lasting therapeutic levels of hUgt1a1 enzyme in a mouse model of Crigler-Najjar Syndrome Type I.

Adeno-associated virus (AAV) -mediated gene therapy is a promising strategy to treat liver-based monogenic diseases. However, two major obstacles limit its success: first, vector dilution in actively dividing cells, such as hepatocytes in neonates/children, due to the non-integrating nature of the vector; second, development of an immune response against the transgene and/or viral vector. Crigler-Najjar Syndrome Type I is a rare monogenic disease with neonatal onset, caused by mutations in the liver-specific UGT1 gene, with toxic accumulation of unconjugated bilirubin in plasma, tissues and brain. To establish an effective and long lasting cure, we applied AAV-mediated liver gene therapy to a relevant mouse model of the disease. Repeated gene transfer to adults by AAV-serotype switching, upon neonatal administration, resulted in lifelong correction of total bilirubin (TB) levels in both genders. In contrast, vector loss over time was observed after a single neonatal administration. Adult administration resulted in lifelong TB levels correction in male, but not female Ugt1-/- mice. Our findings demonstrate that neonatal AAV-mediated gene transfer to the liver supports a second transfer of the therapeutic vector, by preventing the induction of an immune response and supporting the possibility to improve AAV-therapeutic efficacy by repeated administration.

Is cystic echinoccocosis re-emerging in western Spain?

Cystic echinococcosis (CE) remains an important health problem in many areas of the world, including the Mediterranean region. We performed a retrospective study of cases reported from 1998 to 2012 in order to review and update the epidemiology of this disease in a highly endemic area situated in western Spain. A total of 471 patients were diagnosed with hydatid disease. Of these cases, 55·8% were male, with an average age of 62·3 ± 19·5 years. More importantly, 1·5% of patients were children, and 20·5% were aged <45 years. An active therapeutic approach was implemented for 92·6% of the CE patients with primary diagnoses; however, a 'watch and wait' strategy was used in 59·3% of all secondary CE diagnoses. The incidence rate of hydatid disease was significantly higher compared to the incidence described in the Notifiable Disease System in this area. Furthermore, a significant decrease in hydatid incidence during the years included in the study was observed (ß = -0·4357, P < 0·001). CE incidence has diminished in recent years, although active transmission remains in paediatric cases. Additionally, CE incidence remains high in our region despite public health plans for its control. The documented incidence of CE disease clearly underestimates the real numbers.

Unusually high SO2 emissions and plume height from Piton de la Fournaise volcano during the April 2020 eruption.

Piton de la Fournaise volcano, La Réunion, France, erupted between the 2 and 6 April 2020, one of a series of eruptive phases which occur typically two or three times per year. Here, we use back trajectory analysis of satellite data from the TROPOMI instrument to determine that gas emissions during the June 2020 eruption were of unusually high intensity and altitude, producing 34.9 ± 17.4 kt of SO2 and plume heights up to 5 km a.s.l. The early stages of the eruption (2-4 April 2020) were characterised by relatively low SO2 emission rates despite strong low frequency tremor (LFT); the latter phase followed an increase in intensity and explosivity in the early hours of 5 April 2020. This period included lava fountaining, significantly increased SO2 emission rates, increased high frequency tremor (HFT) and decreased LFT. Using the PlumeTraj back trajectory analysis toolkit, we found the peak SO2 emission rate was 284 ± 130 kg/s on the 6 April. The plume altitude peaked at ~ 5 km a.s.l. on 5 April, in the hours following a sudden increase in explosivity, producing one of the tallest eruption columns recorded at Piton de la Fournaise. PlumeTraj allowed us to discriminate each day's SO2, which otherwise would have led to a mass overestimate due to the plumes remaining visible for more than 24 h. The eruption exhibited a remarkable decoupling and anti-correlation between the intensity of the LFT signal and that of the magma and gas emission rates. LFT intensity peaked during the first phase with low magma and SO2 emissions, but quickly decreased during the second phase, replaced by unusually strong HFT. We conclude that the observation of strong HFT is associated with higher intensity of eruption, degassing, and greater height of neutral buoyancy of the plume, which may provide an alert to the presence of greater hazards produced by higher intensity eruptive activity. This might be particularly useful when direct visual observation is prevented by meteorological conditions. This eruption shows the importance of combining multiple data sets when monitoring volcanoes. Combining gas and seismic data sets allowed for a much more accurate assessment of the eruption than either could have done alone. Supplementary Information: The online version contains supplementary material available at 10.1007/s00445-023-01628-1.

Apoptotic mechanisms are involved in the death of Strongyloides venezuelensis after triggering of nitric oxide.

Despite progress in understanding the role of nitric oxide (NO) in the pathogenesis of helminth infections, the role in strongyloidosis is unknown. Firstly, we studied the production of NO in mice infected with Strongyloides venezuelensis as well as in macrophage cultures stimulated with parasite antigens. Somatic larvae 3 (L3) and excretory-secretory female antigens stimulate specific NO production measured by Griess reaction and expression of inducible NO synthase by RT-PCR and quantitative PCR. Moreover, mice infected with S. venezuelensis produce NO in migration stages. Secondly, we analysed the effect of NO production on L3 and females of S. venezuelensis using NO donors such as diethylenetriamine and 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene. Parasites died after NO donor treatment in a dose-dependent manner. Finally, apoptotic mechanisms are involved in the death of S. venezuelensis larvae.

Evaluation of the role of angiogenic factors in the pathogenesis of schistosomiasis.

Schistosomiasis is one disease produced by helminths, which affect many people in tropical areas. Granuloma formation is the main mechanism involved in the pathogenesis of this disease. Experimental studies have demonstrated angiogenesis (blood vessels formation from pre-existing vessels) in the initial phase of granuloma formation. In the present work, VEGF (vascular endothelial growth factor) levels were analyzed in sera from people diagnosed with different helminthic infections. Patients with schistosomiasis and filariasis had significantly high VEGF levels in compared with healthy people and patients diagnosed with hookworms. In addition, the effects of angiogenesis inhibition using anti-angiogenic factors (endostatin) were evaluated in a schistosomiasis murine model. A lesion decrease was observed in mice infected with Schistosoma mansoni and treated with endostatin. Finally, mechanisms of angiogenesis induction were studied and observed that cercariae antigens stimulated the angiogenic factors by host alveolar macrophages.

Role of angiogenic factors in acute experimental Strongyloides venezuelensis infection.

This study aims to investigate the role of angiogenic factors in the pathogenesis of experimental strongyloidiasis. Two complementary approaches were used: Firstly, CD1 mice were treated with endostatin, an angiogenesis inhibitor, and infected with Strongyloides venezuelensis. Also, the mechanisms involved in this process were studied. Parasitological examination revealed a significant decrease in egg per gram of faeces, number of collected larvae from lung tissue and number of collected adult females in mice treated with endostatin. Direct mechanisms with diminution of angiogenesis factors and an indirect mechanism with increase of eosinophil perhaps produced their effect. Secondly, the effect of the antigens responsible for stimulation of angiogenic factors [vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2)] from alveolar macrophages and the mechanisms involved in their production were investigated. Alveolar macrophage cells obtained by bronchoalveolar lavage were incubated at different concentrations of somatic and excretory/secretory antigens of S. venezuelensis. Also, mRNA levels of VEGF and FGF2 in macrophage cells were detected by RT-PCR. L3-PBS larvae antigens induced angiogenic factors. The relationship between angiogenesis factors and nitric oxide has been observed using nitric oxide synthase inhibitors.

Paroxysmal eruptions tracked by variations of helium isotopes: inferences from Piton de la Fournaise (La Réunion island).

Helium (He) with its isotopes (3He, 4He) is a key tracer enabling the Earth's mantle and dynamics to be characterized. Enrichment in primordial helium (3He) has been detected in volcanic gases of numerous magmatic systems in different geodynamic settings. Despite past use to monitor volcano-tectonic unrest, temporal 3He/4He variability in volcanic emissions is still poorly constrained. Here, we investigate noble gas chemistry of Piton de la Fournaise hotspot volcano, where temporal fluctuations of 3He/4He in response to the eruptive activity have never been studied. We compare the 3He/4He signature of volcanic gases and fluid inclusions and we highlight analogous evolution of the 3He/4He signature in both during the last decades of eruptive activity (1990-2017), even during the same eruption. We show that the maximum enrichment in 3He is found in magmatic fluids that fed the most voluminous eruptions which culminated in caldera collapse events. We argue that this enrichment in 3He mostly reflects a greater contribution of magmatic fluids from a primitive component of the mantle plume. These results emphasize that He isotopes may provide warnings of increases in deep magmatic contributions that potentially herald paroxysmal eruptions, as documented here at Piton de la Fournaise (2007) and also at Kilauea (2018).

Trichinella: differential expression of angiogenic factors in macrophages stimulated with antigens from encapsulated and non-encapsulated species.

The newborn larval stage of Trichinella spiralis enters the host striated skeletal muscle cell resulting in the formation of the nurse cell. Vascular Endothelial Growth Factor (VEGF) was detected in cells in the area immediately surrounding the nurse cells. However, no data are available on the antigens involved, the role of other angiogenic factors or the relationship of angiogenesis with Nitric Oxide (NO) production. Using macrophage cell culture we study the effect of different Trichinella L1 antigens from one encapsulated (T. spiralis) and one non-encapsulated (Trichinellapseudospiralis) on the expression of VEGF and basic Fibroblast Growth Factor (FGF2). Also, we investigate the relationship between the production of NO and angiogenic mediators. The results show that encapsulated and non-encapsulated Trichinella species are different in their capacity to stimulate the expression of VEGF and FGF2 from host macrophages. Finally, there is no relationship between angiogenic factors and NO production by T. spiralis antigen.

Very- and ultra-long-period seismic signals prior to and during caldera formation on La Réunion Island.

Early detection of the onset of a caldera collapse can provide crucial information to understand their formation and thus to minimize risks for the nearby population and visitors. Here, we analyse the 2007 caldera collapse of Piton de la Fournaise on La Réunion Island recorded by a broadband seismic station. We show that this instrument recorded ultra-long period (ULP) signals with frequencies in the range (0.003-0.01 Hz) accompanied by very-long period (VLP) signals (between 0.02 and 0.50 Hz) prior to and during the caldera formation suggesting it is possible to detect the beginning of the collapse at depth and anticipate its surface formation. Interestingly, VLP wave packets with a similar duration of 20 s are identified prior to and during the caldera formation. We propose that these events could result from repeating piston-like successive collapses occurring through a ring-fault structure surrounding a magma reservoir from the following arguments: the source mechanism from the main collapse, the observations of slow source processes as well as observations from the field and the characteristic ring-fault seismicity.

Progress in loop-mediated isothermal amplification assay for detection of Schistosoma mansoni DNA: towards a ready-to-use test.

Schistosomiasis is one of the most prevalent Neglected Tropical Disease, affecting approximately 250 million people worldwide. Schistosoma mansoni is the most important species causing human intestinal schistosomiasis. Despite significant efforts in recent decades, the global disease burden of schistosomiasis remains extremely high. This could partly be attributed to the absence of accurate diagnostic tools, primarily in endemic areas. Loop-mediated isothermal amplification (LAMP) is increasingly used in molecular diagnostics as a field-friendly alternative to many other complex molecular methods and it has been proposed as an ideal candidate for revolutionizing point-of-care molecular diagnostics. In a previous work, a LAMP-based method to detect S. mansoni DNA (SmMIT-LAMP) was developed by our research group for early diagnosis of active schistosomiasis in an experimental infection murine model. The SmMIT-LAMP has been further successfully evaluated in both human stool and snail samples and, recently, in human urine samples. In this study, we developed an important improvement for SmMIT-LAMP molecular assay, transforming it into a cold maintenance dry format suitable for potentially manufacturing as kit for ready-to-use for schistosomiasis diagnosis. This procedure could be applied to create dry LAMP kits for a laboratory setting and for diagnostic applications for other neglected tropical diseases.

The addition of a new immunomodulator with the adjuvant adaptation ADAD system using fatty acid binding proteins increases the protection against Fasciola hepatica.

Fatty acid binding proteins (FABP) have shown protective immune response against Fasciola hepatica infection. We evaluated the protection induced by the Fh12 FABP from F. hepatica (Fh12) combined with the new immunomodulator the lipidic aminoalcohol OA0012 in the ADAD system in mice and sheep. In this work we introduced a lipidic aminoalcohol OA0012 as immunomodulator alone or in combination with the hydroalcoholic extract of Phlebodium pseudoaureum; PAL. Mice vaccinated with ADAD containing OA0012+Fh12 or OA0012+Qs+Fh12 had survival rates of 40-50%. Sheep ADAD-vaccinated with OA0012+Qs+Fh12 showed lower fluke recovery, less hepatic lesions and higher post-infection daily weight gain than F. hepatica infected control animals. Sheep ADAD-vaccinated with OA0012 combined PAL and Qs+Fh12 showed lower fluke recovery (42%), lower adult worms count (57%) lower faecal egg count (38%), less hepatic lesions and higher post-infection daily weight gain than F. hepatica infected control animals. Thus, the addition of a new immunomodulator of synthesis to ADAD system with FABPs increased the protection against F. hepatica.

Control of source fertility on the eruptive activity of Piton de la Fournaise volcano, La Réunion.

The eruptive activity of basaltic hotspot volcanoes displays major fluctuations on times scales of years to decades. Theses fluctuations are thought to reflect changes in the rate of mantle melt supply. However, the crustal filter generally masks the mantle processes involved. Here, we show that the cyclic and generally increasing activity of the Piton de la Fournaise volcano (La Réunion) since the mid 20th century is tightly linked to the fertility of its source, as recorded by 87Sr/86Sr and incompatible trace elements ratios of lavas. We identify a twofold control of source fertility on eruptive activity: melt extraction from fertile, incompatible element-enriched veins initiates decadal-scale eruptive sequences, so that vein distribution in the plume source directly controls the cyclic activity. Indirectly, reactive flow of enriched melts increases mantle porosity and promotes melts extraction from the peridotite matrix. This process is thought to have caused a fourfold increase in magma supply between 1998 and 2014 at Piton de la Fournaise, and could also explain magma surges at other frequently active hotspot volcanoes, such as Kilauea, Hawaii. The short-term eruptive activity of hotspot volcanoes appears to be ultimately linked to the distribution and size of lithological heterogeneities in mantle plumes.

Regulation of fibronectin EDA exon alternative splicing: possible role of RNA secondary structure for enhancer display.

The fibronectin primary transcript undergoes alternative splicing in three noncoordinated sites: the cassette-type EDA and EDB exons and the more complex IIICS region. We have shown previously that an 81-nucleotide region within the EDA exon is necessary for exon recognition and that this region contains at least two splicing-regulatory elements: a polypurinic enhancer (exonic splicing enhancer [ESE]) and a nearby silencer element (exonic splicing silencer [ESS]). Here, we have analyzed the function of both elements in different cell types. We have mapped the ESS to the nucleotide level, showing that a single base change is sufficient to abolish its function. Testing of the ESE and ESS elements in heterologous exons, individually or as part of the complete EDA regulatory region, showed that only the ESE element is active in different contexts. Functional studies coupled to secondary-structure enzymatic analysis of the EDA exon sequence variants suggest that the role of the ESS element may be exclusively to ensure the proper RNA conformation and raise the possibility that the display of the ESE element in a loop position may represent a significant feature of the exon splicing-regulatory region.

Progress in the development of Fasciola hepatica vaccine using recombinant fatty acid binding protein with the adjuvant adaptation system ADAD.

Fatty acid binding proteins (FABP) have been designed as a potential vaccine against fasciolosis. In this work, the immunoprophylaxis of the recombinant Fh15 FABP from F. hepatica (Fh15) in adjuvant/immunomodulator ADAD system was evaluated using mice and sheep challenged with F. hepatica. The ADAD system combines the Fh15 antigen with an immunomodulator (hydroalcoholic extract of Polypodium leucotomos; PAL) and/or an adjuvant (saponins of Quillaja saponaria; Qs) in a water/oil emulsion (30/70) with a non-mineral oil (Montanide). All the infected control mice died by 41-48 days post-infection. The mice vaccinated with ADAD only with PAL+Fh15 present a survival rate of 40-50% and those vaccinated with ADAD containing PAL+Qs+Fh15 had a survival rate of 50-62.5%. IgG1 antibodies were lower in surviving mice in comparison with non-surviving mice. The sheep vaccinated with ADAD PAL+Qs+Fh15 showed lower fluke recovery (43%), less hepatic lesions and higher post-infection daily weight gain than F. hepatica infected control animals. Thus, the ADAD system using recombinant fatty acid binding proteins from F. hepatica could be a good option to develop vaccines against F. hepatica.

Genetic and immunological characterization of the 14-3-3xi molecule from Schistosoma bovis.

Currently available candidate vaccines against schistosomiasis elicit only partial protection. In addition, the type of immune response that could lead to the highest level of protection against schistosomes has not yet been described. Thus, efforts should be made in both the identification of novel proteins essential for the parasite cycle and in the modulation of immune responses against these novel candidates through the combined use of immunomodulatory molecules. Several parasites have 14-3-3 proteins, and these proteins are known to play a key role in parasite biology. In the present work, we report the isolation and characterization of a new 14-3-3 gene from Schistosoma bovis and offer new information regarding the genetic structure of the gene. In addition, we have produced the corresponding recombinant protein. Finally, we describe the immune responses elicited by this protein when combined with 4 different immunomodulators in immunized mice.

The Aotus nancymaae erythrocyte proteome and its importance for biomedical research.

The Aotus nancymaae species has been of great importance in researching the biology and pathogenesis of malaria, particularly for studying Plasmodium molecules for including them in effective vaccines against such microorganism. In spite of the forgoing, there has been no report to date describing the biology of parasite target cells in primates or their biomedical importance. This study was thus designed to analyse A. nancymaae erythrocyte protein composition using MS data collected during a previous study aimed at characterising the Plasmodium vivax proteome and published in the pertinent literature. Most peptides identified were similar to those belonging to 1189 Homo sapiens molecules; >95% of them had orthologues in New World primates. GO terms revealed a correlation between categories having the greatest amount of proteins and vital cell function. Integral membrane molecules were also identified which could be possible receptors facilitating interaction with Plasmodium species. The A. nancymaae erythrocyte proteome is described here for the first time, as a starting point for more in-depth/extensive studies. The data reported represents a source of invaluable information for laboratories interested in carrying out basic and applied biomedical investigation studies which involve using this primate. SIGNIFICANCE: An understanding of the proteomics characteristics of A. nancymaae erythrocytes represents a fascinating area for research regarding the study of the pathogenesis of malaria since these are the main target for Plasmodium invasion. However, and even though Aotus is one of the non-human primate models considered most appropriate for biomedical research, knowledge of its proteome, particularly its erythrocytes, remains unknown. According to the above and bearing in mind the lack of information about the A. nancymaae species genome and transcriptome, this study involved a search for primate proteins for comparing their MS/MS spectra with the available information for Homo sapiens. The great similarity found between the primate's molecules and those for humans supported the use of the monkeys or their cells for continuing assays involved in studying malaria. Integral membrane receptors used by Plasmodium for invading cells were also found; this required timely characterisation for evaluating their therapeutic role. The list of erythrocyte protein composition reported here represents a useful source of basic knowledge for advancing biomedical investigation in this field.

[Diagnosis and treatment of imported eosinophilia in travellers and immigrants: Recommendations of the Spanish Society of Tropical Medicine and International Health (SEMTSI)]. / Diagnóstico y tratamiento de la eosinofilia importada en viajeros e inmigrantes: recomendaciones de la Sociedad Española de Medicina Tropical y Salud Internacional (SEMTSI).

According to published data, prevalence of imported eosinophilia among travellers and immigrants is set between 8% and 28.5%. Etiological diagnosis is often troublesome, and depending on the depth of the study and on the population analyzed, a parasitic cause is identified in 17% to 75.9% of the individuals. Among the difficulties encountered to compare studies are the heterogeneity of the studied populations, the type of data collection (prospective/retrospective) and different diagnostic protocols. In this document the recommendations of the expert group of the Spanish Society of Tropical Medicine and International Health (SEMTSI) for the diagnosis and treatment of imported eosinophilia are detailed.

Differential molecular identification of Taeniid spp. and Sarcocystis spp. cysts isolated from infected pigs and cattle.

In the present work, the species-specific identification of Taeniid spp. cysticerci and sarcocystis cysts isolated from infected pigs and cattle was achieved by PCR. In particular: (i) multiplex-PCR derived from HDP2 DNA fragment, specific for Taenia saginata/Taenia solium; (ii) PCRs and PCR-RFLPs of the rDNA internal transcribed spacers 1 and 2 (ITS1 and ITS2) for the differential diagnosis of taeniids; (iii) PCR derived from the 18S rRNA gene and sequencing, specific for Sarcoystis spp. The combined application of these three PCR protocols provided an unequivocally specific diagnosis of T. saginata, T. solium, T. hydatigena, Sarcocystis hominis and Sarcocystis suihominis, and may have practical application in the identification of calcified degenerating or morphologically dubious cysts, for example in the slaughter house situation or in human biopsy samples.

Caretaker's perceptions, attitudes and practices regarding childhood febrile illness and diarrhoeal diseases among riparian communities of Lake Victoria, Tanzania.

Understanding community perceptions and attitudes towards childhood illness is important in developing appropriate interventions. A cross sectional survey was therefore, conducted in a riparian community of Lake Victoria basin in Tanzania to determine caretakers' perception, attitudes and practices on childhood malaria and diarrhoeal diseases. Among 336 caretakers interviewed, 61.1% (205) reported febrile illness in children within three months and 26.0% reported a diarrhoeal episode among children within two weeks before the survey. The majority of the respondents reported high fever (98.7%) and prostration (53.7%) as major symptoms of severe malaria. Convulsions were associated with high fever by only 13.7% of the respondents. Forty percent of the respondents attributed convulsions in children to either fever or malaria, and 24.4% correctly mentioned high fever or malaria. A health facility was the first point of care for childhood malaria for the majority (73%) of the respondents. In diarrhoeal diseases, prostration-weakness (67.7%) and dysentery (20.4%) were commonly reported among the respondents. Typical symptoms of severe dehydration (sunken eyes, loss of skin turgor, dry tears) were poorly recognised as characteristics of severe diarrhoeal diseases. Over 85% of the respondents practiced appropriate dietary measures or increased fluid intake for a child who had diarrhoea. Use of anti-diarrhoea (40.8%) and antibiotic medications (34.8%) were common in the treatment of diarrhoeal diseases. It is important that health education emphasizing recognitions of danger signs/symptoms of malaria and diarrhoeal diseases and their management is strengthened among riparian communities in Lake Victoria basin.