Giant FAZ10 is required for flagellum attachment zone stabilization and furrow positioning in Trypanosoma brucei

The flagellum and flagellum attachment zone (FAZ) are important cytoskeletal structures in trypanosomatids, being required for motility, cell division and cell morphogenesis. Trypanosomatid cytoskeletons contain abundant high molecular mass proteins (HMMPs), but many of their biological functions are still unclear. Here, we report the characterization of the giant FAZ protein, FAZ10, in Trypanosoma brucei, which, using immunoelectron microscopy, we show localizes to the intermembrane staples in the FAZ intracellular domain. Our data show that FAZ10 is a giant cytoskeletal protein essential for normal growth and morphology in both procyclic and bloodstream parasite life cycle stages, with its depletion leading to defects in cell morphogenesis, flagellum attachment, and kinetoplast and nucleus positioning. We show that the flagellum attachment defects are probably brought about by reduced tethering of the proximal domain of the paraflagellar rod to the FAZ filament. Further, FAZ10 depletion also reduces abundance of FAZ flagellum domain protein, ClpGM6. Moreover, ablation of FAZ10 impaired the timing and placement of the cleavage furrow during cytokinesis, resulting in premature or asymmetrical cell division

National trends in the use of oral chemotherapy over 13 years

Funding Information: To the expert panel who validated the CDDs for the three indications and INFARMED I.P. for making data available. Publisher Copyright: Copyright © 2022 Moreira, Bernardo, Ramos, Aguiar and Alves da Costa.Background: Systemic cancer therapy has traditionally been administered using an intravenous (IV) route, implying patients’ frequent visits to hospitals to access to their therapy. If we consider the actual pipeline in oncology, oral chemotherapy will be the main component of cancer treatment in the next few years. This shift in the administration route requires different care models in order to guarantee treatment efficacy and safety. Objective: To analyze time trends in oral chemotherapy consumption in Portugal. Method: Oral chemotherapy consumption over a 13-year period (2008–2020) was analyzed, considering dispensed units by the administration route with respective costs, resorting to the drug regulatory agency (INFARMED I.P.) database. Oral consumption patterns were further explored using common daily doses (CDD) for three conditions, including chronic myeloid leukemia (CML), non-small-cell lung cancer (NSCLC), and breast cancer (BC), to adjust for the effect of varying doses. Data were analyzed descriptively resorting to Microsoft Office Excel 2010. Results: Overall chemotherapy consumption increased +Δ54.7%, with the highest contribution in units observed in oral forms (+Δ58.8%). The total expenditure increased +Δ96.5%, and despite the increase in oral forms (+Δ221.6%), intravenous forms continued to be the major cost driver, with an important contribution from immunotherapy. Much of the increase was led by the approval of 40 new IV and 48 new oral cancer medications with higher costs introduced in the market. Using CDD as an alternative metric to units had varying impacts by indication. The observed increases seemed to focus on specific cancer sites with varying effect; in CML, there was a 2.39-fold increase, compared to 4.41 for NSCLC and 1.86 for BC. However, for BC, two distinct sub-patterns were observed for hormone therapy (increasing 1.83) and for the novel tyrosine kinase inhibitors (increasing 40.8). Conclusion: The growing use of oral chemotherapy is obvious and calls for investments in supporting patients in managing medication adherence and adverse events. The shifts in the healthcare system are complex and need to be prioritized. Our data suggest that priority could be attributed to cancer sites driving innovation, namely, advanced breast cancer.publishersversionpublishe

SYBR Green-based Real-Time PCR targeting kinetoplast DNA can be used to discriminate between the main etiologic agents of Brazilian cutaneous and visceral leishmaniases

<p>Abstract</p> <p>Background</p> <p>Leishmaniases control has been hampered by the unavailability of rapid detection methods and the lack of suitable therapeutic and prophylactic measures. Accurate diagnosis, which can distinguish between <it>Leishmania </it>isolates, is essential for conducting appropriate prognosis, therapy and epidemiology. Molecular methods are currently being employed to detect <it>Leishmania </it>infection and categorize the parasites up to genus, complex or species level. Real-time PCR offers several advantages over traditional PCR, including faster processing time, higher sensitivity and decreased contamination risk.</p> <p>Results</p> <p>A SYBR Green real-time PCR targeting the conserved region of kinetoplast DNA minicircles was able to differentiate between <it>Leishmania </it>subgenera. A panel of reference strains representing subgenera <it>Leishmania </it>and <it>Viannia </it>was evaluated by the derivative dissociation curve analyses of the amplified fragment. Distinct values for the average melting temperature were observed, being 78.95°C ± 0.01 and 77.36°C ± 0.02 for <it>Leishmania </it>and <it>Viannia</it>, respectively (p < 0.05). Using the Neighbor-Joining method and Kimura 2-parameters, the alignment of 12 sequences from the amplified conserved minicircles segment grouped together <it>L</it>. (<it>V</it>.) <it>braziliensis </it>and <it>L</it>. (<it>V</it>.) <it>shawii </it>with a bootstrap value of 100%; while for <it>L</it>. (<it>L</it>.) <it>infantum </it>and <it>L</it>. (<it>L</it>.) <it>amazonensis</it>, two groups were formed with bootstrap values of 100% and 62%, respectively. The lower dissociation temperature observed for the subgenus <it>Viannia </it>amplicons could be due to a lower proportion of guanine/cytosine sites (43.6%) when compared to species from subgenus <it>Leishmania </it>(average of 48.4%). The method was validated with 30 clinical specimens from visceral or cutaneous leishmaniases patients living in Brazil and also with DNA samples from naturally infected <it>Lutzomyia </it>spp. captured in two Brazilian localities.</p> <p>Conclusions</p> <p>For all tested samples, a characteristic amplicon melting profile was evidenced for each <it>Leishmania </it>subgenus, corroborating the data from reference strains. Therefore, the analysis of thermal dissociation curves targeting the conserved kinetoplast DNA minicircles region is able to provide a rapid and reliable method to identify the main etiologic agents of cutaneous and visceral leishmaniases in endemic regions of Brazil.</p

Fundamentos e perspectivas do desenvolvimento sustentável e do uso de indicadores de sustentabi-lidade em propriedades leiteiras.

Ecodesenvolvimento e sustentabilidade; Desenvolvimento sustentável: origem e definição; Indicadores de sustentabilidade; Safa/FAO; RISE (Response-Inducing Sustainability Evaluation)

An unusual plank-shaped nematogen with a graphene nanoribbon core

A [12]phenacene exclusively decorated with four lateral hexylester substituents self-assembles into a nematic liquid crystal glass on cooling after melting at high temperature. This uniaxial nematic organization of a plank-shaped nanographene is unprecedented and in strong contrast to the common design rules for liquid crystals. Highly birefringent samples emitting polarized fluorescence can be obtained in homogeneously planar or twisted waveguiding configurations that are stable against crystallization at and below room temperature and up to 100 °C