Overexpression of ER Protein Processing and Apoptosis Regulator Genes in Human Embryonic Kidney 293 Cells Improves Gene Therapy Vectors Production

Gammaretroviral and lentiviral vectors (γ‐RV and LV) are among the most used vectors in gene therapy. Currently, human embryonic kidney (HEK) 293 cells, the manufacture platform of choice for these vectors, provide low transducing particle yields, challenging their therapeutic applications and commercialization. This work studies metabolic pathways, focusing on endoplasmic reticulum (ER) protein processing and anti‐apoptotic mechanisms, influencing vector productivity in HEK 293 cell substrates. To that end, four candidate genes—protein disulfide isomerase family A member 2 gene, heat shock protein family A (Hsp70) member 5 gene, X‐box binding protein 1 gene (ER protein processing), and B‐cell lymphoma 2 protein gene (anti‐apoptotic)—are individually stably expressed in the cells. How their overexpression level influence vector yields is analyzed by establishing cell populations with incremental genomic copies of each. γ‐RV volumetric productivity increases up to 97% when overexpressing ER protein processing genes. LV volumetric production increases 53% when overexpressing the anti‐apoptotic gene. Improvements are associated with higher cell specific productivities and dependent on gene overexpression level, highlighting the importance of fine‐tuning gene expression. Overall, this work discloses gene engineering targets enabling efficient gene therapy product manufacture showing that ER protein processing and anti‐apoptotic pathways are pivotal to producer cell performance.     

Phenological delay despite warming in wood frog Rana sylvatica reproductive timing: a 20-year study

Across all taxa, amphibians exhibit some of the strongest phenological shifts in response to climate change. As climates warm, amphibians and other animals are expected to breed earlier in response to temperature cues. However, if species use fixed cues such as daylight, their breeding timing might remain fixed, potentially creating disconnects between their life history and environmental conditions. Wood frogs Rana sylvatica are a cold-adapted species that reproduce in early spring, immediately after breeding ponds are free of ice. We used long-term surveys of wood frog oviposition timing in 64 breeding ponds over 20 yr to show that, despite experiencing a warming of 0.29°C per decade in annual temperature, wood frog breeding phenology has shifted later by 2.8 d since 2000 (1.4 d per decade; 4.8 d per °C). This counterintuitive pattern is likely the result of changes in the timing of snowpack accumulation and melting. Finally, we used relationships between climate and oviposition between 2000 and 2018 to hindcast oviposition dates from climate records to model longer-term trends since 1980. Our study indicates that species can respond to fine-grained seasonal climate heterogeneity within years that is not apparent or counterintuitive when related to annual trends across years.     

Genetic diversity of Puccinia kuehnii,the causal agent of orange rust of sugarcane,from Brazil

The use of resistant genotypes is the preferred method to control orange rust of sugarcane (Saccharum spp) caused by Puccinia kuehnii. This approach has been adopted in Brazil but outbreaks of the disease on previously resistant varieties showed that the efficacy of this method is limited and requires a better understanding of pathogen diversity. Nevertheless, adequate molecular markers for examining pathogen diversity at population level are not available, which limits the success of orange rust control by genetic resistance. Therefore, two independent investigations were conducted to examine genetic diversity of P. kuehnii from São Paulo state, the most important sugarcane growing state of Brazil. First, simple-sequence repeat (SSR) markers were developed in the present work and genotypic diversity of orange rust isolates from different locations investigated. Second, phenotypic diversity was examined by the single-pustule inoculation technique on P. kuehnii isolates retrieved from three susceptible commercial sugarcane cultivars. A total of 96 SSR markers were generated and tested for this species. Subsequently, 29 isolates of P. kuehnii were fingerprinted with nine SSR markers to estimate the genotypic diversity by neighbour-joining and 3D principal coordinates. The 29 isolates of the pathogen clustered into four main groups, which were identified by three SSR markers (NPRL_PK_108a, NPRL_PK_162_spka and NPRL_PK_221_spka). Phenotypic data at 21 days after the single-pustule inoculation showed that P. kuehnii from highly susceptible commercial cultivars harboured a small proportion of variants capable of causing disease on resistant cultivars. A differential reaction was demonstrated for the most virulent variant in a repeated experiment confirming the existence of races within P. kuehnii in Brazil.     

Pretreatment free of 2,4-dichlorophenoxyacetic acid improves the differentiation of sugarcane somatic embryos by affecting the hormonal balance and the accumulation of reserves

Plant Cell, Tissue and Organ Culture (PCTOC) - Several factors influence culture conditions and somatic embryogenesis responses, such as the role of plant growth regulators (PGRs) during the...     

S-adenosylmethionine induces mitochondrial dysfunction,permeability transition pore opening and redox imbalance in subcellular preparations of rat liver

Journal of Bioenergetics and Biomembranes - S-adenosylmethionine (AdoMet) predominantly accumulates in tissues and biological fluids of patients affected by liver dysmethylating diseases,...     

New conservation opportunities: Using citizen science in monitoring non-native species in Neotropical region

The combination of highly equipped smartphones, with the increased use of social media, has offered a wide database. Given this, citizen science can be used to record and monitor non-native fish fauna, target new samples and collaborate with monitoring occurrences in new areas. We aimed to demonstrate the efficiency of social media in citizen science as a tool to cooperate with monitoring studies of non-native species. Consequently, we determined the occurrence points of S. brasiliensis in the Iguaçu River basin, indicating sites of greatest occurrence and analyzing the impact of the invasion on the native fauna of the basin. Files and information available on the YouTube^® and Facebook^® media platforms were used as data, was carried out from April 2019 to April 2020. The results were 40 records, 22 videos obtained from Youtube, and seven videos and 11 photos from Facebook, the oldest record was from April 2013, while the largest number of posts was in 2016. Fish records available from online platforms can reveal the occurrence and progressive dispersion of species, in the context of biological invasions, these tools can be of great value in studies that aim to follow the progress of introduced species, contributing by helping to direct new sampling programs and corroborating the occurrence of species in new areas in conjunction with standard monitoring programs. Based on citizen science records, it was possible to update the range of occurrence of the non-native S. brasiliensis in the Iguaçu River basin, cooperating with scientific knowledge. Innovative monitoring and control measures are necessary to deal with invasive species, with citizen science proving to be competent for determining the occurrence of species and showing promise in the entire field of ichthyology.     

COVID-19-Associated Histoplasmosis in an AIDS Patient

Mycopathologia - Most reports associating fungal infections with COVID-19 have been cases of invasive aspergillosis. Here, we report a case of severe histoplasmosis and COVID-19 infections in an...     

Elicitation Improves the Leaf Area,Enzymatic Activities,Antioxidant Activity and Content of Secondary Metabolites in Achillea millefolium L. Grown in the Field

Salicylic acid (SA) is a plant hormone that stimulates the growth and metabolism of plants, also acting as an abiotic elicitor. This study aimed to evaluate the effect of SA on leaf production, leaf area and synthesis of secondary compounds in yarrow plants. The experiments were conducted under field conditions in two consecutive years and f-received SA foliar applications (T1-control; T2-1.0 mmol L⁻¹ applications at 20, 60 and 100 days after planting (DAP) and T3-1.0 mmol L⁻¹ applications at 100 DAP during 3 days). The exogenous application of SA resulted in increases in leaf area (total and specific), number of leaves and leaf mass ratio of yarrow plants, polyphenolic compounds, phenylalanine ammonia-lyase and chalcone synthase enzymes and the antioxidant activity of the plant extract. The HPLC–DAD–MS/MS analysis of phenolic compounds revealed increases in the amounts of quinic acid and rutin. The results of this research lead us to affirm that SA exerted both the hormonal effect on number of leaves and leaf area, and also acted as eliciting substance.

     

Parasitism Behavior of Tetrastichus howardi (Hymenoptera: Eulophidae) on Larvae and Pupae of Sugarcane Borers

Journal of Insect Behavior - Control of the sugarcane borers, Diatraea saccharalis and Diatraea impersonatella (= D. flavipennella) (Lepidoptera: Crambidae), in Brazil, is based on mass release of...