Using C. elegans as a model for neurodegenerative diseases: Methodology and evaluation.

Caenorhabditis elegans is a nematode that has been used as an animal model for almost 50years. It has primitive and simple tissues and organs, making it an ideal model for studying neurological pathways involved in neurodegenerative diseases like Alzheimer's disease (AD) and Parkinson's disease (PD). C. elegans has conserved neurological pathways and is able to mimic human diseases, providing valuable insights into the human disease phenotype. This methodological review presents current approaches to generate neurodegenerative-like models of AD and PD in C. elegans, and evaluates the experiments commonly used to validate the diseases. These experimental approaches include assessing survival, fertility, mobility, electropharyngeogram assays, confocal mitochondrial imaging, RNA extraction for qRT-PCR or RT-PCR, and rate of defecation. This review also summarizes the current knowledge acquired on AD and PD using the aforementioned experimental approaches. Additionally, gaps in knowledge and future directions for research are also discussed in the review.

SERCA inhibition improves lifespan and healthspan in a chemical model of Parkinson disease in Caenorhabditis elegans.

Introduction: The high prevalence of neurodegenerative diseases in our population and the lack of effective treatments encourage the search for new therapeutic targets for these pathologies. We have recently described that submaximal inhibition of the Sarco-Endoplasmic Reticulum Ca2+ ATPase (SERCA), the main responsible for ER calcium storage, is able to increase lifespan in Caenorhabditis elegans worms by mechanisms involving mitochondrial metabolism and nutrient-sensitive pathways. Methods: We have studied here the effects of submaximal SERCA inhibition in a chemical model of Parkinson's disease (PD) induced in C. elegans worms by treatment with the mitochondrial complex I inhibitor rotenone. For specific SERCA inhibition, we treated worms with RNAi against sca-1, the sole orthologue of SERCA in C. elegans. Results and Discussion: Our results show that rotenone produces alterations in worms that include decreased lifespan, smaller size, reduced fertility, decreased motility, defecation and pumping rate, increased mitochondrial ROS production, reduced mitochondrial membrane potential and oxygen consumption rate, altered mitochondrial structure, and altered ethanol preference in behavioral studies. Most of these alterations were either fully or partially reversed in worms treated with sca-1 RNAi, suggesting that SERCA inhibition could be a novel pharmacological target in the prevention or treatment of neurodegeneration.

SLC25A46 promotes mitochondrial fission and mediates resistance to lipotoxic stress in INS-1E insulin-secreting cells.

Glucose sensing in pancreatic ß-cells depends on oxidative phosphorylation and mitochondria-derived signals that promote insulin secretion. Using mass spectrometry-based phosphoproteomics to search for downstream effectors of glucose-dependent signal transduction in INS-1E insulinoma cells, we identified the outer mitochondrial membrane protein SLC25A46. Under resting glucose concentrations, SLC25A46 was phosphorylated on a pair of threonine residues (T44/T45) and was dephosphorylated in response to glucose-induced Ca2+ signals. Overexpression of SLC25A46 in INS-1E cells caused complete mitochondrial fragmentation, resulting in a mild mitochondrial defect associated with lowered glucose-induced insulin secretion. In contrast, inactivation of the Slc25a46 gene resulted in dramatic mitochondrial hyperfusion, without affecting respiratory activity or insulin secretion. Consequently, SLC25A46 is not essential for metabolism-secretion coupling under normal nutrient conditions. Importantly, insulin-secreting cells lacking SLC25A46 had an exacerbated sensitivity to lipotoxic conditions, undergoing massive apoptosis when exposed to palmitate. Therefore, in addition to its role in mitochondrial dynamics, SLC25A46 plays a role in preventing mitochondria-induced apoptosis in INS-E cells exposed to nutrient stress. By protecting mitochondria, SLC25A46 might help to maintain ß-cell mass essential for blood glucose control.

Cell surface detection of vimentin, ACE2 and SARS-CoV-2 Spike proteins reveals selective colocalization at primary cilia.

The SARS-CoV-2 Spike protein mediates docking of the virus onto cells prior to viral invasion. Several cellular receptors facilitate SARS-CoV-2 Spike docking at the cell surface, of which ACE2 plays a key role in many cell types. The intermediate filament protein vimentin has been reported to be present at the surface of certain cells and act as a co-receptor for several viruses; furthermore, its potential involvement in interactions with Spike proteins has been proposed. Nevertheless, the potential colocalization of vimentin with Spike and its receptors on the cell surface has not been explored. Here we have assessed the binding of Spike protein constructs to several cell types. Incubation of cells with tagged Spike S or Spike S1 subunit led to discrete dotted patterns at the cell surface, which consistently colocalized with endogenous ACE2, but sparsely with a lipid raft marker. Vimentin immunoreactivity mostly appeared as spots or patches unevenly distributed at the surface of diverse cell types. Of note, vimentin could also be detected in extracellular particles and in the cytoplasm underlying areas of compromised plasma membrane. Interestingly, although overall colocalization of vimentin-positive spots with ACE2 or Spike was moderate, a selective enrichment of the three proteins was detected at elongated structures, positive for acetylated tubulin and ARL13B. These structures, consistent with primary cilia, concentrated Spike binding at the top of the cells. Our results suggest that a vimentin-Spike interaction could occur at selective locations of the cell surface, including ciliated structures, which can act as platforms for SARS-CoV-2 docking.

Injerto graso como rescate de herida dehiscente sobre bypass vascular. Caso clínico / Fat graft as rescue for dehiscent wound over vascular bypass. Case report

La dehiscencia de heridas quirúrgicas es un problema habitual en la práctica clínica que requiere una solución eficaz, sobre todo cuando se exponen estructuras nobles o material protésico. El injerto de grasa autóloga es una técnica segura y sencilla que puede solucionar úlceras crónicas. Presentamos el caso de un paciente diabético con una úlcera refractaria con exposición de su bypass, que alcanzó la curación completa tras realizar un injerto graso. (AU)

Surgical wound dehiscence is a common problem in clinical practice that requires an effective solution, especially when noble structures or prosthetic material are exposed. Autologous fat grafting is a safe and simple technique that can solve chronic ulcers. We present the case of a diabetic patient with a refractory ulcer with exposure of his bypass, who achieved complete healing after performing a fat graft. (AU)

Albumin internalizes and inhibits endosomal TLR signaling in leukocytes from patients with decompensated cirrhosis.

Human serum albumin (HSA) is an emerging treatment for preventing excessive systemic inflammation and organ failure(s) in patients with acutely decompensated (AD) cirrhosis. Here, we investigated the molecular mechanisms underlying the immunomodulatory properties of HSA. Administration of HSA to patients with AD cirrhosis with elevated circulating bacterial DNA rich in unmethylated cytosine-phosphate-guanine dideoxynucleotide motifs (CpG-DNA) was associated with reduced plasma cytokine concentrations. In isolated leukocytes, HSA abolished CpG-DNA-induced cytokine expression and release independently of its oncotic and scavenging properties. Similar anti-inflammatory effects were observed with recombinant human albumin. HSA exerted widespread changes on the immune cell transcriptome, specifically in genes related to cytokines and type I interferon responses. Our data revealed that HSA was taken up by leukocytes and internalized in vesicles positively stained with early endosome antigen 1 and colocalized with CpG-DNA in endosomes, where the latter binds to Toll-like receptor 9 (TLR9), its cognate receptor. Furthermore, HSA also inhibited polyinosinic:polycytidylic acid- and lipopolysaccharide-induced interferon regulatory factor 3 phosphorylation and TIR domain-containing adapter-inducing interferon-ß-mediated responses, which are exclusive of endosomal TLR3 and TLR4 signaling, respectively. The immunomodulatory actions of HSA did not compromise leukocyte defensive mechanisms such as phagocytosis, efferocytosis, and intracellular reactive oxygen species production. The in vitro immunomodulatory effects of HSA were confirmed in vivo in analbuminemic humanized neonatal Fc receptor transgenic mice. These findings indicate that HSA internalizes in immune cells and modulates their responses through interaction with endosomal TLR signaling, thus providing a mechanism for the benefits of HSA infusions in patients with cirrhosis.

The antiandrogenic vinclozolin induces differentiation delay of germ cells and changes in energy metabolism in 3D cultures of fetal ovaries.

Vinclozolin is a pesticide with antiandrogenic activity as an endocrine disruptor compound. Its effects upon the progression of primordial follicles were assessed in cultures of mouse fetal ovaries from the onset of meiotic differentiation of germ cells (13.5 days post coitum) and from both in vivo exposed mice and in vitro exposed ovaries. Exposure of ovaries to vinclozolin-at in vitro dosages ranging from 10 to 200 µM and in 3D ex vivo culture following in vivo exposure to 50 mg/kg bw/day-showed delays in meiocyte differentiation and in follicle growth, even at the lowest in vitro dose exposure. Immunofluorescent analysis showed the presence of the proteins MSY2 and NOBOX in the primary follicles but no difference in the level of protein signals or in the number of follicles in relation to treatment. However, assessing the cytological differentiation of germ cells by detecting the synaptonemal complex protein SYCP3, the exposure to vinclozolin delayed meiotic differentiation from both in vitro- and in vivo-exposed ovaries. These effects were concomitant with changes in the energy metabolism, detected as a relative increase of glycolytic metabolism in live-cell metabolic assays in exposed ovaries.

Concordance between two rapid diagnostic tests for the detection of antibodies against SARS-CoV-2 / Concordancia entre dos pruebas de detección rápida para la exploración de anticuerpos contra el SARS-CoV-2

OBJECTIVE: To assess the agreement between two rapid detection tests (RDT) for antibodies against SARS-CoV-2 infection. MATERIALS AND METHODS: This was a cross-sectional study that used a random sample of non-hospitalized patients from the primary care management division of the Healthcare Area of Leon (58 RT-PCR-positive cases and 52 RT-PCR-negative cases). Information regarding symptoms was collected and all patients were simultaneously tested using two RDTs (Combined - cRDT and Differentiated - dRDT). The results of both tests were evaluated using the chi-square test and, for degree of agreement, the kappa coefficient. RESULTS: About 52% of the participants were women (mean age: 48.2±11.0 years). A total of 58.2% were positive for d-RDT and 41.2% were positive for c-RDT. In the subjects who were RT-PCR-positive, d-RDT was positive in 72.4% and c-RDT in 55.2%; in those who were RT-PCR-negative, the percentages were 42.3% and 26.9%, respectively. The kappa coefficient observed between the two RDTs was 0.644, and was higher in patients without a fever or anosmia (0.725) and lower in those with a fever or anosmia (0.524). CONCLUSIONS: There is good agreement between the tests used in this study. Given the sensitivity observed, they can be very useful as a complement to RT-PCR

OBJETIVO: Evaluar la concordancia entre dos pruebas de detección rápida (PDR) de anticuerpos en la infección por SARS-CoV-2. MATERIALES Y MÉTODOS: Estudio transversal. Se utilizó una muestra aleatoria de pacientes no hospitalizados de la Gerencia de Atención Primaria del Área de Salud de León (58 con RT-PCR positiva y 52 con RT-PCR negativa). Se recogió información sobre síntomas y a todos se les realizó simultáneamente dos PDR (combinada: PRD-C y diferenciada: PRD-D). Los resultados de ambas pruebas fueron evaluados mediante X2 y el grado de concordancia con el índice Kappa. RESULTADOS: Un 52% de los participantes fueron mujeres (edad media: 48,2 ± 11,0 años). El 58,2% fue positivo a la PDR-D y 41,2% a la PDR-C. En los sujetos RT-PCR + la PDR-D fue positiva en el 72,4% y la PDR-C en el 55,2%; en el caso de los RT-PCR - en el 42,3% y 26,9%, respectivamente. El índice Kappa observado entre las dos PDR fue del 0,644, siendo mayor en pacientes sin fiebre ni anosmia (0,725) y menor en aquellos con fiebre o anosmia (0,524). CONCLUSIONES: Existe una buena concordancia entre los test utilizados en este estudio. Dada la sensibilidad obtenida, pueden ser de gran utilidad como complemento a las RT-PCR

Concordance between two rapid diagnostic tests for the detection of antibodies against SARS-CoV-2.

OBJECTIVE: To assess the agreement between two rapid detection tests (RDT) for antibodies against SARS-CoV-2 infection. MATERIALS AND METHODS: This was a cross-sectional study that used a random sample of non-hospitalized patients from the primary care management division of the Healthcare Area of Leon (58 RT-PCR-positive cases and 52 RT-PCR-negative cases). Information regarding symptoms was collected and all patients were simultaneously tested using two RDTs (Combined - cRDT and Differentiated - dRDT). The results of both tests were evaluated using the chi-square test and, for degree of agreement, the kappa coefficient. RESULTS: About 52% of the participants were women (mean age: 48.2±11.0 years). A total of 58.2% were positive for d-RDT and 41.2% were positive for c-RDT. In the subjects who were RT-PCR-positive, d-RDT was positive in 72.4% and c-RDT in 55.2%; in those who were RT-PCR-negative, the percentages were 42.3% and 26.9%, respectively. The kappa coefficient observed between the two RDTs was 0.644, and was higher in patients without a fever or anosmia (0.725) and lower in those with a fever or anosmia (0.524). CONCLUSIONS: There is good agreement between the tests used in this study. Given the sensitivity observed, they can be very useful as a complement to RT-PCR.

Novel Osteoinductive and Osteogenic Scaffolds of Monetite, Amorphous Calcium Phosphate, Hydroxyapatite, and Silica Gel: Influence of the Hydroxyapatite/Monetite Ratio on Their In Vivo Behavior and on Their Physical and Chemical Properties.

In this work, a new family of multiphasic materials composed of the same amount of silica gel and variable amount of three calcium phosphates with very different solubilities, monetite > amorphous calcium phosphate > hydroxyapatite (HAp), was studied. Silicon was added to calcium phosphate to increase bioactivity and osteinductivity. The influence of the HAp/monetite ratio on the material resorption and bone regeneration was investigated in critical bone defects in sheep and was related to their chemical and physical properties. It was concluded that a minimum rate of HAp/monetite is necessary to achieve an appropriate compromise between material resorption and bone regeneration. Above this minimum rate, bone regeneration and material resorbtion did not change significantly. Physical properties such as particle size, specific surface area, porosity, and granulate cohesion played a more critical role on material resorption than the solubility of their components. A huge difference between in vitro solubility and in vivo resorption was observed. It was related to the fastest cellular-mediated resorption of monetite compared to the other components. Computerized axial tomography, histology, histomorphometric, and multiple fluorochrome labeling studies showed a very advanced bone regeneration of the defects when materials with the highest HAp/monetite rate were implanted. It was also demonstrated that all materials induce bone formation and vascularization.

Protein O-Fucosyltransferase 2 Is Not Essential for Plasmodium berghei Development.

Thrombospondin type I repeat (TSR) domains are commonly O-fucosylated by protein O-fucosyltransferase 2 (PoFUT2), and this modification is required for optimal folding and secretion of TSR-containing proteins. The human malaria parasite Plasmodium falciparum expresses proteins containing TSR domains, such as the thrombospondin-related anonymous protein (TRAP) and circumsporozoite surface protein (CSP), which are O-fucosylated. TRAP and CSP are present on the surface of sporozoites and play essential roles in mosquito and human host invasion processes during the transmission stages. Here, we have generated PoFUT2 null-mutant P. falciparum and Plasmodium berghei (rodent) malaria parasites and, by phenotyping them throughout their complete life cycle, we show that PoFUT2 disruption does not affect the growth through the mosquito stages for both species. However, contrary to what has been described previously by others, P. berghei PoFUT2 null mutant sporozoites showed no deleterious motility phenotypes and successfully established blood stage infection in mice. This unexpected result indicates that the importance of O-fucosylation of TSR domains may differ between human and RODENT malaria parasites; complicating our understanding of glycosylation modifications in malaria biology.

Evaluación de competencias en Urología. Esscolap® basic, como aproximación a la mejora de las capacidades de los urólogos / Evaluation of competencies in urology. Esscolap® basic, as aproximation to urologists skills improvement

Introducción: La Urología necesita de modelos de evaluación de capacidades, a pesar de que existe una variada oferta de herramientas que no están integradas en los programas de formación. Contexto: No existe un criterio universal para medir el nivel de competencia. Los programas de formación deben proporcionar conocimientos y destrezas, y deben considerar las habilidades cognitivas, la formación basada sobre simulación y modelo animal. La validez es un concepto complejo que hace referencia a la capacidad del instrumento de evaluación, por lo que es necesario establecer varios tipos de validación para asegurar la capacidad de un método, reforzarse con distintos test de fiabilidad y cálculo de consistencia interna entre evaluadores. Objetivo: A partir de un dossier estructurado de competencias quirúrgicas, clasificadas por grupos, se planteó el sistema ESSCOLAP® Basic con 5 ejercicios sobre simulador, para la evaluación de las competencias básicas en Laparoscopia. Una vez validado, en el CCMIJU, se planteó ampliar el alcance e implementación del mismo en otras localizaciones. Resultados: Nuestro sistema no ha demostrado aún su validez en el ámbito clínico real, porque no presenta una validez predictiva con datos clínicos de resultados en salud. Existe, además, un cierto rango de subjetividad, por lo que se requiere establecer criterios claros y definidos para cualquier situación. El número de evaluadores y de los ejercicios a evaluar, va a influir en los test de fiabilidad que miden el grado de acuerdo entre evaluadores, de modo que sólo obteniendo un elevado número de casos evaluados, podremos acercarnos a una mayor fiabilidad de nuestro sistema. Por último, asumimos que la incorporación de este tipo de herramientas implica un coste añadido a cargo de las instituciones públicas y privadas responsables, que sólo se considerará rentable cuando se demuestre su trazabilidad real y positiva en resultados sanitarios. Conclusiones: ESSCOLAP® Basic, con capacidad de implementación rápida y sencilla, ha sido validado y contrastado para la evaluación de las habilidades técnicas básicas en laparoscopia (AU)

Introduction: Urology needs models of competencies assessment, although there is a wide range of tools not yet integrated into the official training programs. Context: At present, there is no universal framework for measuring surgeons ́ level of competence. Urology training programs should provide and consider knowledge, pyschomotor/cognitive skills, and simulator, cadaver or animal models-based training. Validity is a complex concept that refers to the capacity of the evaluation tool, so it is necessary to demonstrate several types of validation to assure the capacity of a method, reinforced with different reliability tests and calculation of internal consistency between evaluators. Objective: Based on a structured dossier of surgical skills, classified by groups, the ESSCOLAP® Basic system was proposed with 5 simulator tasks to evaluate basic laparoscopic skills. Once validated in the JUMISC (Spain), the tool was proposed to extend its scope and implementation in other locations. Results: Our system has not yet demonstrated a full validity in the real clinical setting because a predictive validity needs to be demonstrated on the basis of clinical data. It also suffers from a certain range of subjectivity, thus implying clear and defined criteria for any situation. Factors like the number of evaluators and tasks to assess will influence the reliability tests that measure the degree of agreement between evaluators, so that a higher number of evaluated cases would imply a greater reliability of our system. Finally, we assume that the incorporation of this type of tools implies an added cost, charged to the public and private responsible institutions, which will only be considered cost-effective when it is demonstrated its real and positive traceability in health outcomes. Conclusions: ESSCOLAP® Basic, of quick and simple implementation capacity, has been validated and calibrated for the evaluation of basic technical skills in laparoscopy (AU)

[Evaluation of competencies in urology. Esscolap® basic, as aproximation to urologists skills improvement.] / Evaluación de competencias en Urología. Esscolap® basic, como aproximación a la mejora de las capacidades de los urólogos.

Urology needs models of competencies assessment, although there is a wide range of tools not yet integrated into the official training programs. CONTEXT: At present, there is no universal framework for measuring surgeons' level of competence. Urology training programs should provide and consider knowledge, pyschomotor/cognitive skills, and simulator, cadaver or animal models-based training. Validity is a complex concept that refers to the capacity of the evaluation tool, so it is necessary to demonstrate several types of validation to assure the capacity of a method, reinforced with different reliability tests and calculation of internal consistency between evaluators. OBJECTIVE: Based on a structured dossier of surgical skills, classified by groups, the ESSCOLAP® Basic system was proposed with 5 simulator tasks to evaluate basic laparoscopic skills. Once validated in the JUMISC (Spain), the tool was proposed to extend its scope and implementation in other locations. RESULTS: Our system has not yet demonstrated a full validity in the real clinical setting because a predictive validity needs to be demonstrated on the basis of clinical data. It also suffers from a certain range of subjectivity, thus implying clear and defined criteria for any situation. Factors like the number of evaluators and tasks to assess will influence the reliability tests that measure the degree of agreement between evaluators, so that a higher number of evaluated cases would imply a greater reliability of our system. Finally, we assume that the incorporation of this type of tools implies an added cost, charged to the public and private responsible institutions, which will only be considered cost-effective when it is demonstrated its real and positive traceability in health outcomes. CONCLUSIONS: ESSCOLAP® Basic, of quick and simple implementation capacity, has been validated and calibrated for the evaluation of basic technical skills in laparoscopy.

The disruption of GDP-fucose de novo biosynthesis suggests the presence of a novel fucose-containing glycoconjugate in Plasmodium asexual blood stages.

Glycosylation is an important posttranslational protein modification in all eukaryotes. Besides glycosylphosphatidylinositol (GPI) anchors and N-glycosylation, O-fucosylation has been recently reported in key sporozoite proteins of the malaria parasite. Previous analyses showed the presence of GDP-fucose (GDP-Fuc), the precursor for all fucosylation reactions, in the blood stages of Plasmodium falciparum. The GDP-Fuc de novo pathway, which requires the action of GDP-mannose 4,6-dehydratase (GMD) and GDP-L-fucose synthase (FS), is conserved in the parasite genome, but the importance of fucose metabolism for the parasite is unknown. To functionally characterize the pathway we generated a PfGMD mutant and analyzed its phenotype. Although the labelling by the fucose-binding Ulex europaeus agglutinin I (UEA-I) was completely abrogated, GDP-Fuc was still detected in the mutant. This unexpected result suggests the presence of an alternative mechanism for maintaining GDP-Fuc in the parasite. Furthermore, PfGMD null mutant exhibited normal growth and invasion rates, revealing that the GDP-Fuc de novo metabolic pathway is not essential for the development in culture of the malaria parasite during the asexual blood stages. Nonetheless, the function of this metabolic route and the GDP-Fuc pool that is generated during this stage may be important for gametocytogenesis and sporogonic development in the mosquito.

Laparoscopic Cholecystoduodenostomy in Dogs: Canine Cadaver Feasibility Study.

OBJECTIVE: To evaluate the feasibility of laparoscopic cholecystoduodenostomy in canine cadavers using barbed self-locking sutures. STUDY DESIGN: In vivo experimental study. ANIMALS: Fresh male Beagle cadavers (n=5). METHODS: Surgery was performed by a single veterinary surgeon. Dogs were placed in dorsal recumbency and 15° reverse Trendelenburg position. The surgical procedure was performed with four 5 mm entry ports and a 5 mm 30° telescope. The cholecystoduodenostomy technique included dissection, incision of the gallbladder, and lavage, followed by gallbladder transposition over the duodenum, incision of the duodenum, and anastomosis. The latter was performed with a 4-0 barbed self-locking suture (V-Loc® 180). Subsequently, a leak test was performed by submerging the anastomosis in saline and insufflating air into the duodenum through a catheter. Total operative time and completion times for each procedural step were recorded. RESULTS: The median total operative time was 151 minutes (range, 129-159). One conversion to open surgery occurred because of vascular hemorrhage. The 3 longest intraoperative steps were posterior wall anastomosis, gallbladder dissection, and anterior wall anastomosis. Intraoperative anastomotic leakage sites were identified in 3 of 5 dogs. Leaks were managed by placement of a single reinforcing conventional intracorporeal suture, which was adequate to obtain a watertight anastomosis. CONCLUSION: This technique cannot be recommended in clinical practice until further studies are performed and the technique is further refined.

Evaluation of a chemiluminescent enzyme-linked immunosorbent assay for the diagnosis of Trypanosoma cruzi infection in a nonendemic setting.

The disappearance of lytic, protective antibodies (Abs) from the serum of patients with Chagas disease is accepted as a reliable indicator of parasitological cure. The efficiency of a chemiluminescent enzyme-linked immunosorbent assay based on a purified, trypomastigote-derived glycosylphosphatidylinositol-anchored mucin antigen for the serologic detection of lytic Abs against Trypanosoma cruzi was evaluated in a nonendemic setting using a panel of 92 positive and 58 negative human sera. The technique proved to be highly sensitive {100%; 95% confidence interval (CI) = 96-100} and specific (98.3%; 95% CI = 90.7-99.7), with a kappa score of 0.99. Therefore, this assay can be used to detect active T. cruzi infection and to monitor trypanosomicidal treatment.

Biosynthesis of GDP-fucose and other sugar nucleotides in the blood stages of Plasmodium falciparum.

Carbohydrate structures play important roles in many biological processes, including cell adhesion, cell-cell communication, and host-pathogen interactions. Sugar nucleotides are activated forms of sugars used by the cell as donors for most glycosylation reactions. Using a liquid chromatography-tandem mass spectrometry-based method, we identified and quantified the pools of UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, GDP-mannose, and GDP-fucose in Plasmodium falciparum intraerythrocytic life stages. We assembled these data with the in silico functional reconstruction of the parasite metabolic pathways obtained from the P. falciparum annotated genome, exposing new active biosynthetic routes crucial for further glycosylation reactions. Fucose is a sugar present in glycoconjugates often associated with recognition and adhesion events. Thus, the GDP-fucose precursor is essential in a wide variety of organisms. P. falciparum presents homologues of GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase enzymes that are active in vitro, indicating that most GDP-fucose is formed by a de novo pathway that involves the bioconversion of GDP-mannose. Homologues for enzymes involved in a fucose salvage pathway are apparently absent in the P. falciparum genome. This is in agreement with in vivo metabolic labeling experiments showing that fucose is not significantly incorporated by the parasite. Fluorescence microscopy of epitope-tagged versions of P. falciparum GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase expressed in transgenic 3D7 parasites shows that these enzymes localize in the cytoplasm of P. falciparum during the intraerythrocytic developmental cycle. Although the function of fucose in the parasite is not known, the presence of GDP-fucose suggests that the metabolite may be used for further fucosylation reactions.

Clinicians' assessment of mobile monitoring: a comparative study in Japan and Spain.

BACKGROUND: The gradual but steady shift toward telemedicine during the past decades is a clear response to important health problems that most industrialized countries have been facing. The growing elderly population and changing dietary habits have led to an increase in people with chronic diseases and overall health care expenditures. As more consumers use their mobile device as their preferred information and communication technology (ICT) device, mobile health monitoring has been receiving increasing attention in recent years. OBJECTIVE: This study examines clinicians' perception of factors determining mobile health monitoring acceptance in Japan and Spain. The study proposes a causal model consisting of innovation seeking, new ICT attributes (perceived value, time-place flexibility, and compatibility), and usage intention. In addition, cross-country differences are posited for the hypothesized relationships among the proposed constructs. METHODS: A questionnaire survey was performed to test our research model and hypotheses. The sample consisted of clinicians from various medical specialties. In total, 471 and 497 usable responses were obtained in Japan and Spain, respectively. RESULTS: In both countries, the collected data fit the model well with all the hypothesized paths among the constructs being supported. Furthermore, the moderating effects of psychic distance were observed in most of the paths. CONCLUSIONS: Our study demonstrates the importance of new ICT attributes, namely perceived value, time-place flexibility, and compatibility, in the clinicians' adoption of mobile health monitoring. In particular, our results clearly indicated that perceived medical value and ubiquitous nature of the tool are the two main benefits clinicians are likely to perceive (and appreciate) in both countries. This tendency will be stronger for those with a greater propensity to seek innovation in ICT. In terms of cross-country comparison, the strength of the path from innovation seeking to perceived value was greater in Japan than in Spain. Since the number of clinicians per 10,000 residents is substantially fewer in Japan compared with Spain, clinicians with a greater propensity to seek innovation in ICT may have perceived greater value in using mobile health monitoring to improve remote patient care.

Factors affecting mobile diabetes monitoring adoption among physicians: questionnaire study and path model.

BACKGROUND: Patients with type 1 and type 2 diabetes often find it difficult to control their blood glucose level on a daily basis because of distance or physical incapacity. With the increase in Internet-enabled smartphone use, this problem can be resolved by adopting a mobile diabetes monitoring system. Most existing studies have focused on patients' usability perceptions, whereas little attention has been paid to physicians' intentions to adopt this technology. OBJECTIVE: The aim of the study was to evaluate the perceptions and user acceptance of mobile diabetes monitoring among Japanese physicians. METHODS: A questionnaire survey of physicians was conducted in Japan. The structured questionnaire was prepared in a context of a mobile diabetes monitoring system that controls blood glucose, weight, physical activity, diet, insulin and medication, and blood pressure. Following a thorough description of mobile diabetes monitoring with a graphical image, questions were asked relating to system quality, information quality, service quality, health improvement, ubiquitous control, privacy and security concerns, perceived value, subjective norms, and intention to use mobile diabetes monitoring. The data were analyzed by partial least squares (PLS) path modeling. RESULTS: In total, 471 physicians participated from 47 prefectures across Japan, of whom 134 were specialized in internal and gastrointestinal medicine. Nine hypotheses were tested with both the total sample and the specialist subsample; results were similar for both samples in terms of statistical significance and the strength of path coefficients. We found that system quality, information quality, and service quality significantly affect overall quality. Overall quality determines the extent to which physicians perceive the value of mobile health monitoring. However, in contrast to our initial predictions, overall quality does not have a significant direct effect on the intention to use mobile diabetes monitoring. With regard to net benefits, both ubiquitous control and health improvement are significant predictors. Net benefits in turn significantly motivate physicians to use mobile health monitoring, and has a strong influence on perceived value. Perceived value and subjective norms are predictors of intention to use. In our sample, concerns over privacy and security risk have no significant effects on intention to use mobile diabetes monitoring. Among the 3 control variables, only age significantly affected intention to use mobile diabetes monitoring, whereas experience and gender were not significant predictors of intention. CONCLUSIONS: Physicians consider perceived value and net benefits as the most important motivators to use mobile diabetes monitoring. Overall quality assessment does affect their intention to use this technology, but only indirectly through perceived value. Net benefits seem to be a strong driver in both a direct and indirect manner, implying that physicians may perceive health improvement with ubiquitous control as a true utility by enhancing cost-effective monitoring, and simultaneously recognize it as a way to create value for their clinical practices.