Morphological and molecular data on the bat flies, Basilia silvae (Brèthes, 1913) and Trichobius parasiticus Gervais, 1844 parasitizing Chilean bats.

Among mammals, bats harbour the greatest taxonomic diversity of ectoparasitic arthropods. This is mainly due to their high mobility, wide distribution range and gregarious social behaviour. In Chile, 17 species of bats have been reported; however, their ectoparasitic arthropofauna has been little studied. There are currently 12 taxa recorded, belonging to two classes and associated with only four species of bats. This study aimed to investigate the diversity of parasitic flies associated with bats in three ecoregions: Atacama Desert, Chilean Matorral and Valdivian temperate forest. During 2021, using mist nets, bats were captured in Anzota caves (Atacama Desert ecoregion), Huelquén and Alto Jahuel (Chilean Matorral ecoregion), Cherquenco and San Patricio (Valdivian temperate forest). Each bat was carefully checked during a 15-min interval for the collection of ectoparasites. The arthropods were deposited in vials with 96% ethanol. A total of 26 bats corresponding to three species (Vespertilionidae: Histiotus magellanicus Philippi, 1866, Myotis arescens (Osgood, 1943); Phyllostomidae: Desmodus rotundus (Geoffroy, 1810)) were captured from which a total of 142 ectoparasitic arthropods were collected. Bat flies were separated/identified under a stereomicroscope. Additionally, from the fieldwork, we report the presence of other ectoparasites associated with Chilean bats. In our study, we report new host-parasite associations between Trichobius parasiticus Gervais, 1844 (Diptera: Streblidae) on D. rotundus, and Basilia silvae (Brèthes, 1913) (Diptera: Nycteribiidae) in M. arescens in Chile. Our study extended the latitudinal range of distribution for B. silvae to Araucanía region, and we report for first time T. parasiticus in the country. Additionally, partial sequences of the cytochrome c oxidase I gene were obtained from these specimens. Although there is slight morphological variation in the specimens of T. parasiticus, phylogenetic analyses suggest that they correspond to the same species. The sequences generated for B. silvae represent the first for the species. Authors recommend the use of an integrative approach in the identification of ectoparasites in poorly studied ecoregions and hosts. The integration of different markers is necessary to determine more precisely the phylogenetic relationships between South American populations and species of the genera Basilia and Trichobius.

Are Rattus rattus fleas invasive? Evaluation of flea communities in invasive and native rodents in Chile.

Co-invasion, characterized by the simultaneous introduction of hosts and parasites with the latter establishing themselves in native hosts, is a phenomenon of ecological concern. Rattus rattus, a notorious invasive species, has driven the extinction and displacement of numerous avian and mammalian species and serves as a key vector for diseases affecting both humans and wildlife. Among the parasites hosted by R. rattus are fleas, which exhibit obligate parasitic behaviour, a generalist nature and high prevalence, increasing the likelihood of flea invasion. Simultaneously, invasive species can serve as hosts for native parasites, leading to potential amplification or dilution of parasite populations in the environment. In Chile, R. rattus has been present since the 17th century because of the arrival of the Spanish colonizers through the ports and has spread throughout urban, rural and wild Chilean territories. This study aims to evaluate whether co-invasion of native fleas of invasive rats occurs on native rodents in Chile and to determine whether black rats have acquired flea native to Chile during their invasion. For this, we captured 1132 rodents from 26 localities (20° S-53° S). Rattus rattus was found coexisting with 11 native rodent species and two species of introduced rodents. Among the native rodents, Abrothrix olivacea and Oligoryzomys longicaudatus exhibited more extensive sympatry with R. rattus. We identified 14 flea species associated with R. rattus, of which only three were native to rats: Xenopsylla cheopis, Leptopsylla segnis and Nosopsyllus fasciatus. These three species presented a higher parasite load in black rats compared to native fleas. Leptopsylla segnis and N. fasciatus were also found associated with native rodent species that cohabit with R. rattus. The remaining species associated with R. rattus were fleas of native rodents, although they were less abundant compared to those associated with native rodents, except for Neotyphloceras pardinasi and Sphinctopsylla ares. Although there has been evidence of flea transmission from rats to native species, the prevalence and abundance were relatively low. Therefore, it cannot be definitively concluded that these fleas have established themselves in native rodent populations, and hence, they cannot be classified as invasive fleas. This study underscores R. rattus' adaptability to diverse environmental and geographical conditions in Chile, including its capacity to acquire fleas from native rodents. This aspect has critical implications for public health, potentially facilitating the spread of pathogens across various habitats where these rats are found.

MOTIVOS DE DESPERDICIO DE ALIMENTOS DEL PLATO PRINCIPAL DEL MENÚ DE UN COMEDOR ESCOLAR / FOOD WASTE REASONS FOR THE MAIN DISH OF A SCHOOL CAFETERIA MENU

Introducción: La pérdida y el desperdicio de alimentos es una problemática mundial, compleja y que involucra a todos los actores de la cadena de suministro de alimentos. Debido a sus características ha sido incluida en la agenda 2030 del desarrollo sostenible. Objetivo: Conocer los motivos de desperdicio de alimentos del plato principal de los alumnos de sexto grado, en el servicio de alimentación de una escuela de la ciudad de Gualeguaychú. Materiales y métodos: Se desarrolló bajo la lógica cualitativa y tuvo un alcance exploratorio. La unidad de análisis fueron los alumnos de sexto grado. Se realizaron grupos focales y como complemento se implementó una guía de observación, fotografía y escala hedónica. Resultados: Los escolares prefieren las preparaciones de sus mamás y los motivos se relacionaron, principalmente, por las características organolépticas de las comidas. Preferían almorzar en el aula antes que en el comedor porque era un lugar más tranquilo. La aceptabilidad de los menús fue heterogénea y casi el 50% de los platos poseían algún tipo de desperdicio de alimentos, con predominio del desperdicio mayor al 25% de la porción servida. Conclusión: El estudio permitió conocer en mayor profundidad los motivos de desperdicio de alimentos. El rol del licenciado en Nutrición es importante para la gestión del servicio de alimentación escolar, para la planificación de menús de calidad nutricional y aceptados por los escolares. De esta manera, se contribuiría a reducir los desperdicios y mejorar la alimentación de los escolares

Food loss and waste is a global, complex problem that involves all actors in the food supply chain. Due to its characteristics, it has been included in the 2030 agenda for sustainable development. The objective of this study was to know the reasons for wasting food from the main dish of sixth grade students, in the food service of a school in the city of Gualeguaychú. It was developed under a qualitative approach. The unit of analysis was sixth grade students. Focus groups were held and, as a complement, the following were implemented: observation guide, photography and hedonic scale. The results showed that schoolchildren prefer their mothers' preparations and the reasons were mainly related to the organoleptic characteristics of the meals. They preferred to have lunch in the classroom rather than in the dining room because it was a quieter place. The acceptability of the menus was heterogeneous and almost 50% of the dishes had some type of food waste, most of the waste was greater than 25% of the portion served.In conclusion, the study allowed us to know in greater depth the reasons for food waste. The role of the Nutritionist is important for the management of the school food service, for the planning of menus of nutritional quality and appealing to schoolchildren; In this way, it would help to reduce waste and improve schoolchildren's nutrition

Group membership, not diet, structures the composition and functional potential of the gut microbiome in a wild primate.

The gut microbiome has the potential to buffer temporal variations in resource availability and consumption, which may play a key role in the ability of animals to adapt to a broad range of habitats. We investigated the temporal composition and function of the gut microbiomes of wild common marmosets (Callithrix jacchus) exploiting a hot, dry environment-Caatinga-in northeastern Brazil. We collected fecal samples during two time periods (July-August and February-March) for 2 years from marmosets belonging to eight social groups. We used 16S rRNA gene amplicon sequencing, metagenomic sequencing, and butyrate RT-qPCR to assess changes in the composition and potential function of their gut microbiomes. Additionally, we identified the plant, invertebrate, and vertebrate components of the marmosets' diet via DNA metabarcoding. Invertebrate, but not plant or vertebrate, consumption varied across the year. However, gut microbiome composition and potential function did not markedly vary across study periods or as a function of diet composition. Instead, the gut microbiome differed markedly in both composition and potential function across marmosets residing in different social groups. We highlight the likely role of factors, such as behavior, residence, and environmental heterogeneity, in modulating the structure of the gut microbiome. IMPORTANCE: In a highly socially cohesive and cooperative primate, group membership more strongly predicts gut microbiome composition and function than diet.

Molecular survey of tick-borne infectious agents in cricetid rodents (Rodentia: Cricetidae) in Central and Southern Chile.

Tick-borne infectious agents (TBIAs) include several bacteria and protozoa that can infect vertebrates, including humans. Some of these agents can cause important diseases from both a public health perspective, such as Lyme disease, and from an animal health and production viewpoint, such as Texas fever. In Chile, several studies have assessed the presence of tick-borne disease agents in vectors and mammal hosts, mainly in the northern regions, but few studies have assessed the presence of these agents in Central and Southern Chile. This study aimed to assess the presence of three groups of TBIAs-Borrelia, Anaplasmataceae, and Piroplasmida-in cricetid rodents of Central and Southern Chile. A total of 207 specimens from 13 localities between the O'Higgins and Los Lagos regions were captured. DNA was extracted from the liver and spleen, and subsequently underwent polymerase chain reaction (PCR) amplification targeting the 16S rRNA, flaB, and 18S rRNA genes to detect DNA from Borrelia, Anaplasmataceae, and Piroplasmida, respectively. Although no DNA from these TBIAs was detected, the DNA extraction process was validated by optimal DNA purity ratios (an A260/A280 ratio within the 1.6-2.0 range) and successful internal control amplification in all samples. These results, in addition to findings from previous reports, suggest a very low prevalence of these TBIAs in the rodent population studied. Further research into the factors influencing the presence of these agents and their vectors will provide insight into the reasons underlying this low prevalence.

Occupational Therapy Practice in Schools: National Survey in Spain.

AIMS: Interdisciplinary collaboration between health and education professionals is one of the principles of inclusive education. This study aimed to describe the practices and service delivery by pediatric occupational therapists (OTs) in schools in Spain. METHODS: Data came from a convenience sample of pediatric OTs recruited through Spanish professional colleges and associations for a web-based survey, mixed-method research study. Thirty-five respondents were excluded because they didn't meet the eligibility criteria. Ninety-two responses were obtained for data analysis. Twenty-nine OTs (31.5%) work in schools (school-OTs), and sixty-three participants (68.5%) don't work there but collaborate with schools (non-school OTs). RESULTS: Participants support students with and without a medical diagnosis. Services provided by OTs external to the educational system are mainly funded by families or only partially covered. Findings show that most participants use standardized assessments and apply several intervention approaches. They believe there are barriers to the collaboration and implementation of occupational therapy services in schools. CONCLUSIONS: This preliminary study shows a need for changes in Spain's State and Regional Education legislation and provides recommendations to improve school service delivery. Pediatric OTs are encouraged to reflect on and identify opportunities for changes to enhance school-based occupational therapy practice in Spain.

A Mouse Model of the Protease Activated Receptor 4 (PAR4) Pro310Leu Variant has Reduced Platelet Reactivity.

Background: Protease activated receptor 4 (PAR4) mediates thrombin signaling on platelets and other cells. Our recent structural studies demonstrated a single nucleotide polymorphism in extracellular loop 3 (ECL3), PAR4-P310L (rs2227376) leads to a hypo-reactive receptor. Objectives: The goal of this study was to determine how the hypo-reactive PAR4 variant in ECL3 impacts platelet function in vivo using a novel knock-in mouse model (PAR4-322L). Methods: A point mutation was introduced into the PAR4 gene, F2rl3, via CRISPR/Cas9 to create PAR4-P322L, the mouse homolog to human PAR4-P310L. Platelet response to PAR4 activation peptide (AYPGKF), thrombin, ADP, and convulxin was monitored by αIIbß3 integrin activation and P-selectin translocation using flow cytometry or platelet aggregation. In vivo responses were determined by the tail bleeding assay and the ferric chloride-induced carotid artery injury model. Results: PAR4-P/L and PAR4-L/L platelets had a reduced response to AYPGKF and thrombin measured by P-selectin translocation or αIIbß3 activation. The response to ADP and convulxin was unchanged among genotypes. In addition, both PAR4-P/L and PAR4-L/L platelets showed a reduced response to thrombin in aggregation studies. There was an increase in the tail bleeding time for PAR4-L/L mice. The PAR4-P/L and PAR4-L/L mice both showed an extended time to arterial thrombosis. Conclusions: PAR4-322L significantly reduced platelet responsiveness to AYPGKF and thrombin, which is in agreement with our previous structural and cell signaling studies. In addition, PAR4-322L had prolonged arterial thrombosis time. Our mouse model provides a foundation to further evaluate the role of PAR4 in other pathophysiological contexts.

Multicellular tumor spheroid model to study the multifaceted role of tumor-associated macrophages in PDAC.

While considerable efforts have been made to develop new therapies, progress in the treatment of pancreatic cancer has so far fallen short of patients' expectations. This is due in part to the lack of predictive in vitro models capable of accounting for the heterogeneity of this tumor and its low immunogenicity. To address this point, we have established and characterized a 3D spheroid model of pancreatic cancer composed of tumor cells, cancer-associated fibroblasts, and blood-derived monocytes. The fate of the latter has been followed from their recruitment into the tumor spheroid to their polarization into a tumor-associated macrophage (TAM)-like population, providing evidence for the formation of an immunosuppressive microenvironment.This 3D model well reproduced the multiple roles of TAMs and their influence on drug sensitivity and cell migration. Furthermore, we observed that lipid-based nanosystems consisting of sphingomyelin and vitamin E could affect the phenotype of macrophages, causing a reduction of characteristic markers of TAMs. Overall, this optimized triple coculture model gives a valuable tool that could find useful application for a more comprehensive understanding of TAM plasticity as well as for more predictive drug screening. This could increase the relevance of preclinical studies and help identify effective treatments.

Keeping reptiles as pets in Brazil: keepers' motivations and husbandry practices.

BACKGROUND: Reptiles are considered one of the most popular pets in the world and are often associated with an incorrect belief that they are simple, highly adaptable, and easy to keep animals when compared with other pets, such as dogs and cats. However, keeping reptiles as pets can pose several challenges in meeting their needs in a domestic setting, requiring specific conditions and effort to maintain their health, well-being, and survival. METHODS: During 2015, using online semi-structured questionnaires applied to 719 Brazilian pet reptile keepers who participated in online groups of reptile breeders on the social network Facebook, this study aimed to identify Brazilian keepers' motivations for maintaining reptiles as pets, investigate their monthly expenses, and the husbandry practices for the maintenance, such as housing and feeding conditions, handling of the animal, health issues, and treatment provided. RESULTS: We found multiple motivations for keeping reptiles as pets (mostly snakes, lizards, and chelonians), the main motivation being emotional reasons, followed by entertainment and convenience reasons. The great majority of keepers (69%) declared to spend less than or up to US$30 per month in maintaining their reptiles. Most reptiles were kept alone in terrarium/aquarium enclosures, with basic environmental complexity in terms of physical elements. Lizards and chelonians were fed with a few insect species, cultivated fruits and vegetables, while snakes were fed mainly with domestic rodents, rabbits, or birds. Keepers declared frequent cleaning of the enclosure, but inappropriately handled their animals directly with their hands, which might result in potential threats to human and reptile health and safety. Several diseases or injuries were mentioned and 55.6% of the keepers declared taking the reptile to the vet for treatment. CONCLUSIONS: Overall, our findings revealed several challenges that reptiles face when kept in domestic environments, including issues related to housing, nutrition, and healthcare. Even though keepers demonstrated positive feelings toward their pets, suggesting a positive relationship and a willingness to provide them with proper care, it seems that without the proper knowledge and awareness, reptiles may unintentionally be kept with poor husbandry. Addressing these challenges on husbandry practices is essential for improving reptiles' welfare and promoting a responsible pet ownership.

Genomic versus phenotypic selection to improve corn borer resistance and grain yield in maize.

Introduction: The study of yield and resistance/tolerance to pest are related traits fundamental for maize breeding programs. Genomic selection (GS), which uses all marker information to calculate genomic breeding values, is presented as an emerging alternative to phenotypic and marker-assisted selections for improving complex traits controlled by many genes with small effects. Therefore, although phenotypic selection (PS) has been effective for increasing resistance and yield under high infestation with maize stem borers, higher genetic gains are expected to be obtained through GS based on the complex architecture of both traits. Our objective was to test whether GS is more effective than PS for improving resistance and/or tolerance to maize stem borers and grain yield. Methods: For this, we compared different selection programs based on phenotype and genotypic value for a single trait, resistance or yield, and for both traits together. Results and discussion: We obtained that GS achieved the highest genetic gain for yield, meanwhile phenotypic selection for yield was the program that achieved the highest reduction of tunnel length, but was ineffective for increasing yield. However, phenotypic or genomic selection for increased resistance may be more effective in improving both traits together; although the gains per cycle would be small for both traits.

Survey and Molecular Characterization of Sarcocystidae protozoa in Wild Cricetid Rodents from Central and Southern Chile.

In Chile, studies of parasites from the family Sarcocystidae (Apicomplexa) have mostly been related to domestic animals. We aimed to assess the presence of Sarcocystidae taxa in cricetid rodents from Central and Southern Chile. We studied 207 rodents, encompassing six species, from 13 localities. We isolated DNA from tissue samples, amplified the Sarcocystidae 18S rRNA gene with polymerase chain reaction, and performed phylogenetic analyses using maximum likelihood and Bayesian inferences. In addition, we examined blood smears and performed histological studies in organs from Sarcocystidae DNA-positive animals. Three specimens were DNA-positive and three genotypes were retrieved and named: Sarcocystis sp. P61, related to Sarcocystis strixi, was detected in two Abrothrix olivacea. Toxoplasmatinae gen. sp. P99 was retrieved from those same two specimens, and was related to Toxoplasma and other genera, although it branched independently. Besnoitia sp. R34 was detected in one Abrothrix hirta, and was clustered with congeneric species associated with rodents. No protozoa were found during microscopic studies; thus, it was not possible to confirm parasitic interactions rather than accidental encounters. However, the close relatedness of the retrieved genotypes to parasites of rodents supports the hypothesis of host-parasite associations. All three genotypes are suggested as potential new taxa, including a putative new genus.

PRGF Membrane with Tailored Optical Properties Preserves the Cytoprotective Effect of Plasma Rich in Growth Factors: In Vitro Model of Retinal Pigment Epithelial Cells.

The present study evaluates the ability of a novel plasma rich in growth factors (PRGF) membrane with improved optical properties to reduce oxidative stress in retinal pigment epithelial cells (ARPE-19 cells) exposed to blue light. PRGF was obtained from three healthy donors and divided into four main groups: (i) PRGF membrane (M-PRGF), (ii) PRGF supernatant (S-PRGF), (iii) platelet-poor plasma (PPP) membrane diluted 50% with S-PRGF (M-PPP 50%), and (iv) M-PPP 50% supernatant (S-PPP 50%). ARPE-19 cells were exposed to blue light and then incubated with the different PRGF-derived formulations or control for 24 and 48 h under blue light exposure. Mitochondrial and cell viability, reactive oxygen species (ROS) production, and heme oxygenase-1 (HO-1) and ZO-1 expression were evaluated. Mitochondrial viability and cell survival were significantly increased after treatment with the different PRGF-derived formulations. ROS synthesis and HO-1 expression were significantly reduced after cell treatment with any of the PRGF-derived formulations. Furthermore, the different PRGF-derived formulations significantly increased ZO-1 expression in ARPE-19 exposed to blue light. The new PRGF membrane with improved optical properties and its supernatant (M-PPP 50% and S-PPP 50%) protected and reversed blue light-induced oxidative stress in ARPE-19 cells at levels like those of a natural PRGF membrane and its supernatant.

Correction: Effectiveness of a novel gene nanotherapy based on putrescine for cancer treatment.

Correction for 'Effectiveness of a novel gene nanotherapy based on putrescine for cancer treatment' by Saínza Lores et al., Biomater. Sci., 2023, https://doi.org/10.1039/d2bm01456d.

Description of a Cohort with a New Truncating MYBPC3 Variant for Hypertrophic Cardiomyopathy in Northern Spain.

BACKGROUND: The pathogenicity of the different genetic variants causing hypertrophic cardiomyopathy (HCM) and the genotype/phenotype correlations are difficult to assess in clinical practice, as most mutations are unique or identified in non-informative families. Pathogenic variants in the sarcomeric gene MYBPC3 inherited with an autosomal dominant pattern, whereas incomplete and age-dependent penetrance are the most common causes of HCM. METHODS: We describe the clinical characteristics of a new truncating MYBPC3 variant, p.Val931Glyfs*120, in 75 subjects from 18 different families from northern Spain with the p.Val931Glyfs*120 variant. RESULTS: Our cohort allows us to estimate the penetrance and prognosis of this variant. The penetrance of the disease increases with age, whereas 50% of males in our sample developed HCM by the age of 36 years old, and 50% of women developed the disease by the time they reached 48 years of age (p = 0.104). Men have more documented arrhythmias with potential risk of sudden death (p = 0.018), requiring implantation of cardioverter defibrillators (p = 0.024). Semi-professional/competitive sport among males is related to earlier onset of HCM (p = 0.004). CONCLUSIONS: The p.Val931Glyfs*120 truncating variant in MYBPC3 is associated with a moderate phenotype of HCM, with a high penetrance, onset in middle age, and a worse outcome in males due to higher risk of sudden death due to arrhythmias.

Quantitative PET tracking of intra-articularly administered 89Zr-peptide-decorated nanoemulsions.

Intra-articular (IA) administration of drugs for the treatment of diseases such as rheumatoid arthritis, osteoarthritis and psoriatic arthritis is a common strategy; however, the rapid clearance from the synovial fluid restricts their effectivity due to the limited retention time. Drug Delivery Systems (DDS) are currently being developed to increase their joint retention time. This study compares the biodistribution and retention time of a senolytic peptide (PEP), with potential application in osteoarthritis disease, and this senolytic peptide encapsulated in a DDS based on a lipid nanoemulsion (PEPNE) by using positron emission tomography (PET) imaging. To this aim, the PEP was conjugated with a chelating agent (DFO) and radiolabeled with zirconium-89 (89Zr). Then, [89Zr]-PEP was encapsulated in a novel nanoemulsion formulation, composed by vitamin E, sphingomyelin, and a lipid-PEG. Afterward, healthy rats were administered with either the [89Zr]-PEP or the [89Zr]-PEP-NE via IA injection and underwent PET scans at 0.5-, 24-, 48-, 72-, 168-, 240- and 336 h post-injection. To assess the biodistribution of both radiotracers, several volume-of-interest were manually drawn in different organs of the rat body and the %ID/organ was calculated. The [89Zr]-PEP was successfully encapsulated in the NE and their physicochemical properties were minimally affected by the radiolabeling buffer. Adequate stability of both [89Zr]-PEP and [89Zr]-PEP-NE was found in synovial fluid over 72 h. Quantitative data from PET images revealed a significantly higher [89Zr]-PEP-NE retention in the injected knee than with [89Zr]-PEP in all follow-up PET scans. The [89Zr]-PEP %ID/organ values in the liver and kidney were significantly higher than those from [89Zr]-PEP-NE, which might indicate a faster elimination of the [89Zr]-PEP. Therefore, the study highlights the higher retention time on the target site of the [89Zr]-PEP-NE which may improve the therapeutic effects of the peptide. Thereby, the novel nanoemulsion formulation seems to be a successful DDS for IA injection. In addition, these results represent the first study that evaluates the distribution of a PET-guided DDS after its IA administration.

Efficacy of Platelet-Rich Plasma in Women with a History of Embryo Transfer Failure: A Systematic Review and Meta-Analysis with Trial Sequential Analysis.

Assisted reproductive technology (ART) is used to enhance pregnancy in infertile women. In this technique, the eggs are removed from the ovary and fertilized and injected with sperm to make embryos. Unfortunately, embryo implantation failures still occur in many of these women. Platelet-rich plasma (PRP) therapies use a patient's own platelets to promote tissue healing and growth, including endometrium. The growth factors provided by the platelets play a criterial role on the regenerative ability of PRP. In the last years, PRP treatments have been gaining a lot of popularity to treat women with repeated ART failures. In this study, we collected and summarized all information published in the scientific literature to assess the evidence of the PRP effect on pregnancy. We only considered randomized controlled trials (RCT), a type of study designed to be unbiased and considered at the highest level of evidence. Our analysis indicates that PRP therapies might be an effective treatment in cases of poor responsiveness to conventional ART. However, additional studies (well-designed) are necessary to confirm this beneficial effect of PRP.

Effectiveness of a novel gene nanotherapy based on putrescine for cancer treatment.

Gene therapy has long been proposed for cancer treatment. However, the use of therapeutic nucleic acids presents several limitations such as enzymatic degradation, rapid clearance, and poor cellular uptake and efficiency. In this work we propose the use of putrescine, a precursor for higher polyamine biosynthesis for the preparation of cationic nanosystems for cancer gene therapy. We have formulated and characterized putrescine-sphingomyelin nanosystems (PSN) and studied their endocytic pathway and intracellular trafficking in cancer cells. After loading a plasmid DNA (pDNA) encoding the apoptotic Fas Ligand (FasL), we proved their therapeutic activity by measuring the cell death rate after treatment of MDA-MB-231 cells. We have also used xenografted zebrafish embryos as a first in vivo approach to demonstrate the efficacy of the proposed PSN-pDNA formulation in a more complex model. Finally, intratumoral and intraperitoneal administration to mice-bearing MDA-MB-231 xenografts resulted in a significant decrease in tumour cell growth, highlighting the potential of the developed gene therapy nanoformulation for the treatment of triple negative breast cancer.

Development of a new plasma rich in growth factors membrane with improved optical properties.

PURPOSE: The aim of the present work was to develop a fibrin membrane using plasma rich in growth factors (PRGF) technology with improved optical properties to be used for the treatment of ocular surface diseases. BASIC PROCEDURES: Blood was drawn from three healthy donors, and the volume of PRGF obtained from each donor was divided into two main groups: i) PRGF or ii) platelet-poor plasma (PPP). Each membrane was then used pure or diluted to 90 %, 80 %, 70 %, 60 % and 50 %. The transparency of each of the different membranes was evaluated. The degradation and morphological characterization of each membrane was also performed. Finally, a stability study of the different fibrin membranes was performed. MAIN FINDINGS: The transmittance test showed that the fibrin membrane with the best optical characteristics was obtained after removal of platelets and dilution of fibrin to 50 % (50 % PPP). No significant differences (p > 0.05) were observed between the different membranes in the fibrin degradation test. The stability test showed that the membrane at 50 % PPP retains its optical and physical characteristics after storage at - 20 °C for 1 month compared to storage at 4 °C. PRINCIPAL CONCLUSIONS: The present study describes the development and characterization of a new fibrin membrane with improved optical characteristics while maintaining mechanical and biological characteristics. The physical and mechanical properties of the newly developed membrane are preserved after storage for at least 1 month at - 20 °C.

Beneficial Effects of Plasma Rich in Growth Factors (PRGF) Versus Autologous Serum and Topical Insulin in Ocular Surface Cells.

PURPOSE: In the last few decades, several blood derived products such as platelet-rich plasma (PRP), plasma rich in growth factors (PRGF) and autologous serum (AS) have been used for the treatment of ocular surface disorders. Recently, insulin has been proposed to be used as an alternative for the treatment of ocular surface diseases. The aim of this study was to evaluate the biological potential of PRGF eye drops in comparison with AS and insulin on ocular surface cells. METHODS: Blood from three healthy young donors was collected to obtain autologous serum (AS) eye drops and plasma rich in growth factors (PRGF) eye drops. Insulin (Actrapid®) was diluted at 1 and 0.2 IU/mL. The biological potential of PRGF, AS and insulin was assessed by proliferation in HCE, HK and HConF cells. Wound healing assay was performed in HCE cells after incubation with the different treatments. HConF and HK cells were differentiated to myofibroblast after treatment with 2.5 ng/mL of TGF-ß1 and then incubated with all treatments. RESULTS: PRGF eye drops induced significantly higher proliferation rate compared to AS or insulin in HConF and HK cells, but not in HCE cells. In addition, the percentage of wound healing area was significantly reduced after PRGF treatment in comparison with AS or insulin treatment. Furthermore, PRGF significantly reduced the number of myodifferentiated cells compared to AS and insulin at both concentrations analyzed. CONCLUSION: The results obtained in the present study show that PRGF increases the biological activity of the ocular surface cells and reduces the expression of fibrosis marker compared to insulin or AS. TRANSLATIONAL RELEVANCE: The present study suggests that plasma rich in growth factors eye drops are a more effective therapy than insulin and autologous serum eye drops for the treatment of ocular surface diseases.

Eco-epidemiology of rodent-associated trombiculid mites and infection with Orientia spp. in Southern Chile.

BACKGROUND: Scrub typhus is a potentially severe infection caused by bacteria of the genus Orientia, endemic in Asia-Pacific and recently discovered in southern Chile. The presented study aimed to determine the prevalence and species richness of rodent-associated trombiculid mites and their infection with Orientia spp. in different areas of two regions in southern Chile. METHODOLOGY/PRINCIPAL FINDINGS: During summer 2020, trombiculid mites were collected from rodents captured in three areas in southern Chile known to be endemic for scrub typhus (Cochamó and Chiloé Island in the Los Lagos Region and Tortel in the Aysén Region). A total of 132 rodents belonging to five species were captured using Sherman-like traps; 89.4% were infested with trombiculids. Mite specimens were morphologically identified and subsequently tested by Orientia-specific qPCR. Six mite species were identified. Among chigger-infested rodents, 33.9% carried Orientia-positive mites; this rate was higher in Tortel (63.8%) than in Cochamó (45.0%) and Chiloé Island (2.0%). The analysis of individual mites (n = 901) revealed that 31.2% of Herpetacarus antarctica samples (n = 202) were positive for Orientia DNA; the prevalence was 7.0% in Paratrombicula neuquenensis (n = 213), 6.9% in Herpetacarus eloisae (n = 144), 3.6% in Argentinacarus expansus (n = 55), and 0% in Paratrombicula goffi (n = 110) and Quadraseta chiloensis (n = 177). The southernmost site (Tortel) showed the highest rates of trombiculid infestation, trombiculid load, and Orientia infection in the captured rodents. CONCLUSIONS/SIGNIFICANCE: Our study provides new insights into the trombiculid fauna and prevalence of Orientia in mites collected from wild rodents in southern Chile. Orientia DNA was detected in four of the six mite species. Rates of infestation, mite loads, and Orientia prevalences differed geographically and were highest in the Aysén Region. Our data improve our knowledge on possible vectors of scrub typhus and their distribution in Chile.