The vacuolar iron transporter mediates iron detoxification in Toxoplasma gondii

Iron is essential to living cells, acting as a cofactor in a number of important enzymes in metabolism; however in the absence of correct storage iron forms dangerous oxygen radicals. In both yeast and plants, iron is stored in a membrane-bound vacuole through the action of a vacuolar iron transporter (VIT). This transporter is conserved in the apicomplexan family of obligate intracellular parasites, including in Toxoplasma gondii, a pathogen of medical and veterinary importance. Here, we assess the role of VIT, and iron storage, in T. gondii. We show that iron is restricted to a compartment in the parasite that does not overlap with zinc. By deleting VIT we find a slight growth defect in vitro, however the absence of VIT leads to hypersensitivity to iron, confirming its essential role in iron detoxification in the parasite. This hypersensitivity can be rescued by scavenging of oxygen radicals. In the absence of VIT, parasites store less iron and are at a growth disadvantage when moving into an iron-depleted environment. We show parasite VIT expression is regulated by iron levels at both the transcript and protein level, and by altering the distribution of VIT within the cell. In the absence of VIT, we find that T. gondii responds by altering expression of genes with a role in iron metabolism and by increasing the activity of the antioxidant protein catalase. We also show that iron detoxification has an important role both in parasite survival within macrophages and in virulence in a mouse model. Together, by demonstrating a critical role for VIT during iron detoxification in T. gondii, we reveal the importance of iron storage in the parasite and provide the first insight into the machinery involved.

Environmental and molecular approach to dye industry waste degradation by the ascomycete fungus Nectriella pironii

Textile industry effluents and landfill leachate contain chemicals such as dyes, heavy metals and aromatic amines characterized by their mutagenicity, cytotoxicity and carcinogenicity. The aim of the present study was investigation of the ascomycete fungus N. pironii isolated from urban postindustrial textile green space for its ability to grow and retain metabolic activity in the presence of the dye industry waste. Research focused mainly on dyes, heavy metals and aromatic amines, which had been detected in landfill leachate via HPLC–MS/MS analysis. Presence of all tested compounds as well as leachate in the growth medium clearly favored the growth of fungal biomass. Only slight growth limitation was observed in the presence of 50 mg L-1o-tolidine. The fungus eliminated o-tolidine as well as dyes at all tested concentrations. The presence of metals slightly influenced the decolorization of the azo dyes; however, it was still similar to 90%. During fungal growth, o-tolidine was hydroxylated and/or converted to toluidine and its derivatives. Laccase and cytochrome P450 involvement in this process has been revealed. The results presented in the paper provide a valuable background for the development of a fungus-based system for the elimination of toxic pollutants generated by the textile industry.

Estudo do padrão de melhora da alopecia androgenética em pacientes com uso de microagulhamento associado a minoxidil e/ou plasma rico em plaquetas e mesoterapia / Study of the pattern of improvement in androgenetic alopecia in patients using microneedling associated with minoxidil and/or platelet-rich plasma and mesotherapy

Introdução: a alopécia ligada a fatores hormonais androgênicos é algo que há muito tempo tem sido alvo de estudos e aplicações de conhecimento científico em todo mundo. Este padrão de perda capilar, principalmente ligada ao sexo masculino dispõe de terapias que atualmente se limitam à interrupção da queda de fios com leve progressão de crescimento em alguns casos. Neste tocante, uma nova técnica de microagulhamento, com ou sem terapias adjuvantes, associado a métodos já consagradas como minoxidil tem levado o tratamento desta alteração estética para melhores níveis de resultado. Objetivo: conduzir uma investigação sobre os resultados atuais de pesquisas que envolvam o microagulhamento associado ao uso de Minoxidil, plasma rico em plaquetas e/ou mesoterapia. Metodologia: foram feitas pesquisas no banco de dados da PubMed e Scielo em abril de 2021 buscando-se os termos “minoxidil AND microneedling”. Resultados e Discussões: os resultados obtidos têm convergido para a proposta de que a associação do método de rolagem (microagulhamento) com a aplicação tópica do vasodilatador (minoxidil) apresenta um padrão de crescimento capilar nunca alcançado pela terapia convencional disponível, e que, se acrescentar plasma rico em plaquetas e/ou soluções através da mesoterapia, os resultados podem ser ainda mais vantajosos. Conclusão: novos estudos são necessários para abranger as vertentes ainda não elucidadas para o tratamento da alopecia, contudo, os resultados até aqui obtidos são promissores para o sucesso da reposição capilar.Palavras chave: Microagulhamento, Minoxidil, Alopecia AbstractIntroduction: alopecia linked to androgenic hormonal factors is something that has long been the subject of studies and knowledge applications worldwide. This pattern of hairloss, mainly related to males, has therapies that are currently limited to interruption of hair loss with slight growth progression in some cases. In this regard, a new microneedlingtechnique, with or without adjuvant therapies, associated with well-established methods such as minoxidil has led the treatment of this esthetic alteration to better levels of results. Objective: to conduct an investigation into the current results of research involving microneedling associated with the use of Minoxidil, platelet-rich plasma and/or mesotherapy. Methodology: searches were conducted in the PubMed and Scielo database in April 2021, searching for the terms “minoxidil AND microneedling”. Results and Discussions: the results obtained have converged to the proposal that the association of the rolling method(microneedling) with the topical application of a vasodilator (minoxidil) presents a pattern of hair growth never achieved by the available conventional therapy, and that, if addedplatelet-rich plasma and/or solutions through mesotherapy, the results can be even more advantageous. Conclusion: further studies are needed to cover aspects that have notyet been elucidated for the treatment of alopecia, however, the results obtained so far are promising for the success of hair replacement.Keywords: Microneedling, Minoxidil, Alopecia

Shared and specific functions of Arfs 1–5 at the Golgi revealed by systematic knockouts

ADP-ribosylation factors (Arfs) are small GTPases regulating membrane traffic in the secretory pathway. They are closely related and appear to have overlapping functions, regulators, and effectors. The functional specificity of individual Arfs and the extent of redundancy are still largely unknown. We addressed these questions by CRISPR/Cas9-mediated genomic deletion of the human class I (Arf1/3) and class II (Arf4/5) Arfs, either individually or in combination. Most knockout cell lines were viable with slight growth defects only when lacking Arf1 or Arf4. However, Arf1+4 and Arf4+5 could not be deleted simultaneously. Class I Arfs are nonessential, and Arf4 alone is sufficient for viability. Upon Arf1 deletion, the Golgi was enlarged, and recruitment of vesicle coats decreased, confirming a major role of Arf1 in vesicle formation at the Golgi. Knockout of Arf4 caused secretion of ER-resident proteins, indicating specific defects in coatomer-dependent ER protein retrieval by KDEL receptors. The knockout cell lines will be useful tools to study other Arf-dependent processes.

The vacuolar iron transporter mediates iron detoxification in Toxoplasma gondii

Iron is essential to living cells, acting as a cofactor in a number of essential enzymes in metabolism; however, iron requires proper storage or it can be dangerous to the cell. In both yeast and plants, iron is stored in a vacuole through the action of a vacuolar iron transporter (VIT). This transporter is conserved in the apicomplexan family of obligate intracellular parasites, including in Toxoplasma gondii, a pathogen of medical and veterinary importance. Here, we assess the role of VIT in T. gondii. We show that deletion of VIT causes a slight growth defect in vitro, however leads to hypersensitivity in the presence of excess iron, confirming its essential role in iron detoxification in the parasite. In the absence of VIT, parasites contain less iron and are at a growth disadvantage when moving into an iron-depleted environment. We show parasite VIT expression is regulated by environmental iron levels at both the transcript and protein level, and by altering the distribution of VIT within the cell. In the absence of VIT, we find that the T. gondii responds by altering expression of genes with a role in iron metabolism and by increasing the activity of the antioxidant protein catalase. We also show that iron detoxification has an important role both in parasite survival within macrophages and in pathogenesis in a mouse model. Together, by demonstrating a critical role for VIT during iron detoxification in T. gondii, we reveal the importance of iron storage in the parasite and provide the first insight into the machinery involved.

Shared and specific functions of Arfs 1–5 at the Golgi revealed by systematic knockouts

The ADP-ribosylation factors (Arfs) are small GTPases regulating membrane traffic in the secretory pathway. They are closely related and appear to have overlapping functions, regulators, and effectors. The functional specificity of individual Arfs and the extent of redundancy in vivo are still largely unknown. We addressed these questions by CRISPR/Cas9-mediated genomic deletion of the human class I (Arfs 1 and 3) and class II (Arfs 4 and 5) Arfs, either individually or in combination. Cells lacking individual Arfs or certain combinations were viable with only a slight growth defect when lacking Arf1 or Arf4. However, Arf1 and 4, and Arf4 and 5 could not be deleted simultaneously. Hence, class I Arfs are not essential and Arf4 alone was found to be sufficient for cell viability. Remarkably, two single knockouts produced specific and distinct phenotypes. Upon deletion of Arf1, the Golgi complex was enlarged and recruitment of vesicle coats decreased, confirming a major role of Arf1 in coat formation at the Golgi. Cell lines deleted for Arf4 exhibited secretion of ER resident proteins, indicating a specific defect in coatomer-dependent ER protein retrieval by the KDEL receptors. The knockout cell lines will be a useful tool to study other Arf-dependent processes.

Epidemiology of Anterior Cruciate Ligament Reconstruction Surgery in Italy: A 15-Year Nationwide Registry Study

There remains little information on the epidemiology of anterior cruciate ligament reconstruction (ACL-R), therefore, we performed an epidemiological evaluation on the ACL-R procedures performed in Italy from 2001 to 2015 to highlight potential disparities in access to healthcare. The National Hospital Discharge records (SDO) maintained at the Italian Ministry of Health were analyzed from 2001 to 2015; 248,234 ACL-Rs were performed in Italy over the 15-year study period in the adult population (starting from 15 years old), and the incidence rate per year in 100,000 persons ranged from 21.70 to 33.60 over the study period. The overall male/female ratio was 4.54. The length of hospitalization ranged from four days in 2001 to two days in 2015. Italy is historically divided into north, center, and south regions, and more than half of ACL-R surgery was performed in the north (67.2%); 95.2% of ACL-Rs were underwent in public institutions. The predicted model projected a slight growth in the number of ACL-Rs in the next 10 years (2016–2025). The number of ACL-R procedures increased in the adult population from 2001 to 2015. The ACL-R procedures were concentrated in the north of Italy, suggesting that efforts on regionalization of ACL-Rs should turn toward improving quality in hospitals in the south of Italy.

Analysis of the population dynamics of the endangered American crocodile, Crocodylus acutus in Paramillo National Natural Park

Few studies have been conducted on the population dynamics of crocodiles, especially in the Neotropics. This is the first study to provide data on the population dynamics of the threatened American crocodile (Crocodylus acutus (Cuvier, 1807)) in Paramillo National Natural Park. Twenty-four survey routes of 22km each were studied, and a total of 733 sightings was recorded. During the study period (from October 2014 to March 2016), the population remained stable and experienced slight growth (λ=1.049±0.428). Offspring production by the adults II stage was the most sensitive contribution (17.05), whereas the permanence of this adult stage was the demographic process that contributed the most to population growth (0.653). However, an 18% decrease in adults II would jeopardise long-term population viability and lead to a high probability of quasi-extinction, with under 300 individuals over the next 30 years. Thus, despite the apparent stability of this population, we suggest that it be considered a key target for future conservation efforts. Additionally, we recommend that this species continue to be listed as endangered in Colombia and not be modified in the CITES Appendix I.

Epidemiology of Achilles tendon surgery in Italy: a nationwide registry study, from 2001 through 2015

Background This study aims (1) to estimate the yearly number of Achilles tendon (AT) surgeries in Italy from 2001 to 2015 based on official hospitalization records; (2) to investigate the eventual presence of geographical variation in equity in access to AT surgery between three macroregions of Italy (North, Center and South); (3) to perform statistical projections of the number of AT procedure volumes and rates based on these data. Methods We analysed the National Hospital Discharge records (SDO) maintained at the Italian Ministry of Health for a 15-year period, from 2001 through 2015. These data are anonymous and include the patient’s age (evaluated in the class of age), sex, census region, the region of hospitalization, length of the hospitalization, public or private reimbursement and diagnosis. Results During the 15-year study period, 118,652 AT repair were performed in Italy, whose peak of incidence was in 2010. More than half of AT repairs was performed in the North of Italy (52.1%), while 27.2% was performed in the South of Italy and 20.6% Center of Italy. The projection model predicted a slight growth of 2.65% in 2025 in comparison with 2015. Conclusion The current study provides detailed information about the national population-weighted incidence of AT surgery, distribution and projection. The peak of average age was 35–45 year. The majority of AT procedures was performed in the North of Italy. The projection model predicts a slight growth of AT surgery by 2025. Furthermore, this 15-year nationwide registry study shows that the age of incidence of AT injuries shifted from 30 to 40 to 35–45 years compared to the available literature. The higher prevalence of AT surgery was found in men during the working age. Moreover, a low rate of procedures in pediatric and elder age classes was observed.

Potato Virus Y (PVY) Isolates from Solanum betaceum Represent Three Novel Recombinants Within the PVYN Strain Group and Are Unable to Systemically Spread in Potato

Tamarillo, or tree tomato (Solanum betaceum), is a perennial small tree or shrub species cultivated in subtropical areas for fresh fruit and juice production. In Ecuador, tamarillo orchards are affected by several viruses, with one previously identified as potato virus Y (PVY); however, the specific strain composition of PVY in tamarillo was not determined. In 2015 and 2016, eight tamarillo plants exhibiting symptoms of leaf drop, mosaic, and mottled fruit were sampled near Tumbaco and Quito, Ecuador. These tamarillo PVY isolates were able to systemically infect tobacco, Nicotiana benthamiana, naranjilla, and tamarillo. Seven of the eight PVY isolates from tamarillo exhibited N-serotype, while one of the PVY isolates studied, Tam15, had no identifiable serotype. One isolate, Tam17, had N-serotype but produced asymptomatic systemic infection in tobacco. In tamarillo, four tamarillo isolates induced mosaic and slight growth retardation and were unable to systemically infect pepper or potato. Tamarillo, on the other hand, was unable to support systemic infection of PVY isolates belonging to the PVYO and PVYEu-N strains. The whole genomes of eight PVY isolates were sequenced from a series of overlapping RT-PCR fragments. Phylogenetically, tamarillo PVY isolates were found to belong to the large PVYN lineage, in a new tamarillo clade. Recombination analysis revealed that these tamarillo PVY isolates represent at least three novel recombinant types not reported before. The combination of the biological and molecular properties found in these eight PVY isolates suggested the existence of a new tamarillo strain of PVY that may have coevolved with S. betaceum.