CRISPR-Cas greatly facilitated the integration of exogenous sequences into specific loci. However, knockin generation in multicellular animals remains challenging, partially due to the complexity of insertion screening. Here, we describe SEED/Harvest, a method to generate knockins in Drosophila, based on CRISPR-Cas and the single-strand annealing (SSA) repair pathway. In SEED (from \"scarless editing by element deletion\"), a switchable cassette is first integrated into the target locus. In a subsequent CRISPR-triggered repair event, resolved by SSA, the cassette is seamlessly removed. Germline excision of SEED cassettes allows for fast and robust knockin generation of both fluorescent proteins and short protein tags in tandem. Tissue-specific expression of Cas9 results in somatic cassette excision, conferring spatiotemporal control of protein labeling and the conditional rescue of mutants. Finally, to achieve conditional protein labeling and manipulation of short tag knockins, we developed a genetic toolbox by functionalizing the ALFA nanobody.

Background: Supporting wellbeing beyond symptom reduction is necessary in trauma care. Research suggests increased posttraumatic growth (PTG) may promote wellbeing more effectively than posttraumatic stress disorder (PTSD) symptom reduction alone. Understanding neurobiological mechanisms of PTG would support PTG intervention development. However, most PTG research to-date has been cross-sectional data self-reported through surveys or interviews.Objective: Neural evidence of PTG and its coexistence with resilience and PTSD is limited. To advance neural PTG literature and contribute translational neuroscientific knowledge necessary to develop future objectively measurable neural-based PTG interventions.Method: Alpha frequency EEG and validated psychological inventories measuring PTG, resilience, and PTSD symptoms were collected from 30 trauma-exposed healthy adults amidst the COVID-19 pandemic. EEG data were collected using custom MNE-Python software, and a wireless OpenBCI 16-channel dry electrode EEG headset. Psychological inventory scores were analysed in SPSS Statistics and used to categorise the EEG data. Power spectral density analyses, t-tests and ANOVAs were conducted within EEGLab to identify brain activity differentiating high and low PTG, resilience, and PTSD symptoms.Results: Higher PTG was significantly differentiated from low PTG by higher alpha power in the left centro-temporal brain area around EEG electrode C3. A trend differentiating high PTG from PTSD was also indicated in this same location. Whole-scalp spectral topographies revealed alpha power EEG correlates of PTG, resilience and PTSD symptoms shared limited, but potentially meaningful similarities.Conclusion: This research provides the first comparative neural topographies of PTG, resilience and PTSD symptoms in the known literature. Results provide objective neural evidence supporting existing theory depicting PTG, resilience and PTSD as independent, yet co-occurring constructs. PTG neuromarker alpha C3 significantly delineated high from low PTG and warrants further investigation for potential clinical application. Findings provide foundation for future neural-based interventions and research for enhancing PTG in trauma-exposed individuals.
Objective translational study designed to increase neural understanding of posttraumatic growth (PTG) and provide a basis for future neural-based interventions to enhance PTG.Results provide neural evidence of PTG as an independent construct that coexists, and shares limited neural relatedness with resilience and PTSD symptoms.Increased PTG was significantly related to higher alpha power in the left centro-temporal brain area around EEG electrode C3: This finding warrants further investigation for potential clinical application.

Optimization of xylanase and cellulase production by a newly isolated Aspergillus fumigatus strain grown on Stipa tenacissima (alfa grass) biomass without pretreatment was carried out using a Box-Behnken design. First, the polysaccharides of dried and ground alfa grass were characterized using chemical methods (strong and diluted acid). The effect of substrate particle size on xylanase and carboxymethylcellulase (CMCase) production by the selected and identified strain was then investigated. Thereafter, experiments were statistically planned with a Box-Behnken design to optimize initial pH, cultivation temperature, moisture content, and incubation period using alfa as sole carbon source. The effect of these parameters on the two enzyme production was evaluated using the response surface method. Analysis of variance was also carried out, and production of the enzymes was expressed using a mathematical equation depending on the influencing factors. The effects of individual, interaction, and square terms on production of both enzymes were represented using the nonlinear regression equations with significant R2 and P-values. Xylanase and CMCase production levels were enhanced by 25% and 27%, respectively. Thus, this study demonstrated for the first time the potential of alfa as a raw material to produce enzymes without any pretreatment. A set of parameter combinations was found to be effective for the production of xylanase and CMCase by A. fumigatus in an alfa-based solid-state fermentation.

BACKGROUND: This study compares the severity of SARS-CoV-2 infections caused by Alpha, Delta or Omicron variants in periods of co-circulation in Spain, and estimates the variant-specific association of vaccination with severe disease.
METHODS: SARS-CoV-2 infections notified to the national epidemiological surveillance network with information on genetic variant and vaccination status were considered cases if they required hospitalisation or controls otherwise. Alpha and Delta were compared during June-July 2021; and Delta and Omicron during December 2021-January 2022. Adjusted odds ratios (aOR) were estimated using logistic regression, comparing variant and vaccination status between cases and controls.
RESULTS: We included 5,345 Alpha and 11,974 Delta infections in June-July and 5,272 Delta and 10,578 Omicron in December-January. Unvaccinated cases of Alpha (aOR: 0.57; 95% CI: 0.46-0.69) or Omicron (0.28; 0.21-0.36) had lower probability of hospitalisation vs. Delta. Complete vaccination reduced hospitalisation, similarly for Alpha (0.16; 0.13-0.21) and Delta (June-July: 0.16; 0.14-0.19; December-January: 0.36; 0.30-0.44) but lower from Omicron (0.63; 0.53-0.75) and individuals aged 65+ years.
CONCLUSIONS: Results indicate higher intrinsic severity of the Delta variant, compared with Alpha or Omicron, with smaller differences among vaccinated individuals. Nevertheless, vaccination was associated to reduced hospitalisation in all groups.

Erythropoietin (EPO) is an exciting neurotherapeutic option. Despite its potential, concerns exist regarding the potential for thrombosis and adverse events with EPO administration in normonemic adults. Systematic review of literature using PRISMA guidelines to examine the application and risks of EPO as a treatment option for neuroprotection in normonemic adults. Independent, systematic searches were performed in July 2019. PubMed (1960-2019) and the Cochrane Controlled Trials Register (1960-2019) were screened. Search terms included erythropoietin, neuroprotection, and humans. The PubMed search resulted in the following search strategy: (\"erythropoietin\" [MeSH Terms] OR \"erythropoietin\" [All Fields] OR \"epoetin alfa\" [MeSH Terms] OR (\"epoetin\" [All Fields] AND \"alfa\" [All Fields]) OR \"epoetin alfa\" [All Fields]) AND (\"neuroprotection\" [MeSH Terms] OR \"neuroprotection\" [All Fields]) AND \"humans\" [MeSH Terms]. PubMed, Cochrane Controlled Trials Register, and articles based on prior searches yielded 388 citations. 50 studies were included, comprising of 4351 patients. There were 13 studies that noted adverse effects from EPO. Three attributed serious adverse effects to EPO and complications were statistically significant. Two of these studies related the adverse events to the co-administration of EPO with tPA. Minor adverse effects associated with the EPO group included nausea, pyrexia, headache, generalized weakness and superficial phlebitis. Most published studies focus on spinal cord injury, peri-surgical outcomes and central effects of EPO. We found no studies to date evaluating the role of EPO in post-operative pain. Future trials could evaluate this application in persistent post-surgical pain and in the peri-operative period.

Introduction: From its earliest days, the US. military has embraced the use of vaccines to fight infectious diseases. The Army Liposome Formulation (ALF) has been a pivotal innovation as a vaccine adjuvant that provides excellent safety and potency and could lead to dual-use military and civilian benefits. For protection of personnel against difficult disease threats found in many areas of the world, Army vaccine scientists have created novel liposome-based vaccine adjuvants.Areas covered: ALF consists of liposomes containing saturated phospholipids, cholesterol, and monophosphoryl lipid A (MPLA) as an immunostimulant. ALF exhibited safety and strong potency in many vaccine clinical trials. Improvements based on ALF include: ALF adsorbed to aluminum hydroxide (ALFA); ALF containing QS21 saponin (ALFQ); and ALFQ adsorbed to aluminum hydroxide (ALFQA). Preclinical safety and efficacy studies with ALF, LFA, ALFQ, and ALFQA are discussed in preparation for upcoming vaccine trials targeting malaria, HIV-1, bacterial diarrhea, and opioid addiction.Expert opinion: The introduction of ALF in the 1980s stimulated commercial interest in vaccines to infectious diseases, and therapeutic vaccines to cancer, and Alzheimer\'s disease. It is likely that ALF, ALFA, and ALFQ, will provide momentum for new types of modern vaccines with improved efficacy and safety.

One of the well-known functions of the bacterial cytoskeleton is plasmid segregation. Type II plasmid segregation systems, among the best characterized with respect to the mechanism of action, possess an actin-like cytomotive filament as the motor component. This chapter describes the essential components of the plasmid segregation machinery and their mechanism of action, concentrating on the actin-like protein family of the bacterial cytoskeleton. The structures of the actin-like filaments depend on their nucleotide state and these in turn contribute to the dynamics of the filaments. The components that link the filaments to the plasmid DNA also regulate filament dynamics. The modulation of the dynamics facilitates the cytomotive filament to function as a mitotic spindle with a minimal number of components.

BACKGROUND: Alpha-thalassemia is the most common hemoglobinopathy with a variable clinical manifestation depending on the number of allele mutations (asymptomatic/mild anemia if 1-2 allele mutations, severe disease if 3-4 allele mutations). A study was conducted from May 2011 on hemoglobinopathies found in the neonatal screening in the autonomous community of the Basque Country (CAPV).
OBJECTIVE: To analyze the impact of alpha-thalassemia in this area and the effectiveness of its neonatal screening.
METHODS: A review was made of patients with a positive gene study for alpha-thalassemia over a 2-year period (2012-2013) and an analysis was made of the age at diagnosis, ethnic group, analytical result, and treatment.
RESULTS: The genetic study was performed on 107 patients, of which 61 had some mutation, with 62% having one allele mutations and 38% with two alleles. The mean age at diagnosis was 31 years, with 28% being younger than eighteen years old. Most of the patients were European with a significant number of Africans (26%) and Arabs (13%). All patients were asymptomatic, and 28% had mild anemia. Two patients were diagnosed by neonatal screening. Most of them did not need any treatment or only required iron therapy.
CONCLUSIONS: The detection of one or two alpha gene mutations has no clinical impact, but allows genetic counseling. No patient was found with 3-4 mutations or severe symptoms in our region. Contrarily to the diagnosis of other diseases, our results does not support that routine neonatal screening for alpha-thalassemia has any clinical impact in our community.