Automated CRISPR/Cas9-based genome editing of human pluripotent stem cells using the StemCellFactory.

CRISPR/Cas9 genome editing is a rapidly advancing technology that has the potential to accelerate research and development in a variety of fields. However, manual genome editing processes suffer from limitations in scalability, efficiency, and standardization. The implementation of automated systems for genome editing addresses these challenges, allowing researchers to cover the increasing need and perform large-scale studies for disease modeling, drug development, and personalized medicine. In this study, we developed an automated CRISPR/Cas9-based genome editing process on the StemCellFactory platform. We implemented a 4D-Nucleofector with a 96-well shuttle device into the StemCellFactory, optimized several parameters for single cell culturing and established an automated workflow for CRISPR/Cas9-based genome editing. When validated with a variety of genetic backgrounds and target genes, the automated workflow showed genome editing efficiencies similar to manual methods, with indel rates of up to 98%. Monoclonal colony growth was achieved and monitored using the StemCellFactory-integrated CellCelector, which allowed the exclusion of colonies derived from multiple cells or growing too close to neighbouring colonies. In summary, we demonstrate the successful establishment of an automated CRISPR/Cas9-based genome editing process on the StemCellFactory platform. The development of such a standardized and scalable automated CRISPR/Cas9 system represents an exciting new tool in genome editing, enhancing our ability to address a wide range of scientific questions in disease modeling, drug development and personalized medicine.

Short-Term Dual Antiplatelet Therapy After Drug-Eluting Stenting in Patients With Acute Coronary Syndromes: A Systematic Review and Network Meta-Analysis.

Importance: The optimal duration of dual antiplatelet therapy (DAPT) in patients with acute coronary syndromes (ACS) undergoing percutaneous coronary intervention (PCI) remains under debate. Objectives: To analyze the efficacy and safety of DAPT strategies in patients with ACS using a bayesian network meta-analysis. Data Sources: MEDLINE, Embase, Cochrane, and LILACS databases were searched from inception to April 8, 2024. Study Selection: Randomized clinical trials (RCTs) comparing DAPT duration strategies in patients with ACS undergoing PCI were selected. Short-term strategies (1 month of DAPT followed by P2Y12 inhibitors, 3 months of DAPT followed by P2Y12 inhibitors, 3 months of DAPT followed by aspirin, and 6 months of DAPT followed by aspirin) were compared with conventional 12 months of DAPT. Data Extraction and Synthesis: This systematic review and network meta-analysis followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. The risk ratio (RR) with a 95% credible interval (CrI) was calculated within a bayesian random-effects network meta-analysis. Treatments were ranked using surface under the cumulative ranking (SUCRA). Main Outcomes and Measures: The primary efficacy end point was major adverse cardiac and cerebrovascular events (MACCE); the primary safety end point was major bleeding. Results: A total of 15 RCTs randomizing 35 326 patients (mean [SD] age, 63.1 [11.1] years; 26 954 male [76.3%]; 11 339 STEMI [32.1%]) with ACS were included. A total of 24 797 patients (70.2%) received potent P2Y12 inhibitors (ticagrelor or prasugrel). Compared with 12 months of DAPT, 1 month of DAPT followed by P2Y12 inhibitors reduced major bleeding (RR, 0.47; 95% CrI, 0.26-0.74) with no difference in MACCE (RR, 1.00; 95% CrI, 0.70-1.41). No significant differences were observed in MACCE incidence between strategies, although CrIs were wide. SUCRA ranked 1 month of DAPT followed by P2Y12 inhibitors as the best for reducing major bleeding and 3 months of DAPT followed by P2Y12 inhibitors as optimal for reducing MACCE (RR, 0.85; 95% CrI, 0.56-1.21). Conclusion and Relevance: Results of this systematic review and network meta-analysis reveal that, in patients with ACS undergoing PCI with DES, 1 month of DAPT followed by potent P2Y12 inhibitor monotherapy was associated with a reduction in major bleeding without increasing MACCE when compared with 12 months of DAPT. However, an increased risk of MACCE cannot be excluded, and 3 months of DAPT followed by potent P2Y12 inhibitor monotherapy was ranked as the best option to reduce MACCE. Because most patients receiving P2Y12 inhibitor monotherapy were taking ticagrelor, the safety of stopping aspirin in those taking clopidogrel remains unclear.

Surgical training trends in the Americas: A cross-continental assessment of minimally invasive surgery and open surgery among surgical trainees.

INTRODUCTION: Minimally invasive surgery (MIS) has become standard of care in many high-income countries, but its adoption in low- and middle-income countries (LICs/MICs) has been impeded by resource- and training-related barriers. We hypothesized that trainees in MICs perform MIS procedures less often, and that as procedure complexity increases, the rate of MIS decreases. METHODS: A 22-question survey, distributed to representative leaders across Latin America, collected country-specific graduating trainee case requirements and volumes for four index procedures (cholecystectomy, appendectomy, inguinal hernia repair, colectomy) using MIS or open surgery (OS). USA data was obtained from the Accreditation Council for Graduate Medical Education. Kruskal-Wallis and Mann-Whitney U tests were performed to determine whether the rate of MIS differed across all countries, procedure complexity classes, and high income countries (HICs)/MICs. RESULTS: Seven experts (70% response rate) completed the survey, representing: Brazil, Chile, Ecuador, Guatemala, Mexico, Panama, and Paraguay. The percentage of MIS completed by trainees varied with mean and interquartile ranges as follows: cholecystectomy (60% ± 54%), appendectomy (41% ± 69%), inguinal hernia repair (19% ± 23%), colectomy (16% ± 29%). There was a significant difference in mean MIS experience across the eight countries (H = 17.6, p = 0.014) and between most complex and least complex procedures (p = 0.039). No difference was found between MICs and HICs (p = 0.786). CONCLUSIONS: We found a significant difference of general surgery trainee exposure to MIS versus OS across the Americas, but the difference was not significantly associated with World Bank Income Groups. Different trainee experiences with MIS and OS may highlight an opportunity for international and bidirectional collaboration.

Diacerein reduces inflammasome activation and SARS-CoV-2 virus replication: a proof-of-concept translational study.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is linked to high mortality, primarily through an intense inflammatory response. Diacerein has emerged as a potential therapy for COVID-19 due to its potential impact in decreasing the inflammasome activation and coronavirus replication. This study aims to explore diacerein's influence in inhibiting both viral replication and the inflammatory response after SARS-CoV-2 infection. Methods: Human peripheral blood mononuclear cells (PBMCs) were obtained from healthy volunteers and infected in vitro with SARS-CoV-2. Additionally, we carried out a pilot randomized, double-blind, placebo-controlled study with 14 participants allocated to diacerein (n = 7) or placebo (n = 7) therapies every 12 h for 10 days. The primary endpoint was change in plasma markers of inflammasome activation (NLRP3, caspase-1, and gasdermin-D). Results: In vitro protocols have shown that rhein, diacerein's primary metabolite, decreased IL-1ß secretion caused by SARS-CoV-2 infection in human PBMCs (p < 0.05), and suppressed viral replication when administered either before or after the virus incubation (p < 0.05). This later effect was, at least partially, attributed to its inhibitory effect on 3-chymotrypsin-like protease (SARS-CoV-2 3CLpro) and papain-like protease in the SARS-CoV-2 (SARS-CoV-2 PLpro) virus and in the phosphorylation of proteins related cytoskeleton network (p < 0.05). Diacerein-treated COVID-19 patients presented a smaller area under the curve for NLRP3, caspase-1 and GSDM-D measured on days 2, 5, and 10 after hospitalization compared to those receiving a placebo (p < 0.05). Conclusion: The indicated mechanisms of action of diacerein/rhein can reduce viral replication and mitigate the inflammatory response related to SARS-CoV-2. These findings are preliminary and require confirmation in clinical trials.

A new strategy for systematically classifying HLA alleles into serological specificities: Update and refinement.

HLA antigens were historically defined according to the unique reactivity pattern of cells expressing HLA molecules with distinctive clusters of allo-antisera and/or monoclonal antibodies. Subsequently, amino acid residues determining epitopes (DEP) in the HLA molecule were correlated with reactivity patterns. In current clinical practice, the presence of allo-antibodies is assessed using Luminex-based solid phase single antigen bead (SAB) assays for transplantation. Recently, novel antigens were proposed for HLA molecules with DEP patterns that do not match any serologically defined antigens recognised by the WHO Nomenclature Committee. To validate the antigens, mean fluorescence intensity values of SABs tested on >13,000 patients' sera were extracted from clinical databases and analysed by scatter plots using a linear regression model. We found that when two proteins were considered as the same antigen in the original study, for example, HLA-A*02:01 and -A*02:06, their correlation ranked among the highest values at each locus. In contrast, discrete asymmetric outliers were observed when there were different antigens, for example, HLA-A*30:01 and -A*30:02, allowing validation and confirmation of 20 novel antigens for HLA-A, -B, -C and -DR. The outliers were confirmed to be true or false by flow cytometric crossmatches. In addition to the previously defined residues for antigen assignments, findings suggest that further distinction should be made for common antigens by including the substitutions at residue 67 of HLA-B, 67 and 74 of -DR. These serologic analyses can be applied systematically to identify and confirm novel antigens. These developments will lead to designing optimal SAB panels and further improving virtual donor-specific antibodies assessment.

Exploring maintenance rehabilitation in adults with chronic conditions: a scoping review of the literature.

OBJECTIVE: Individuals with chronic conditions have long-term needs which can be addressed by maintenance rehabilitation. The 2013 Jimmo v Sebelius settlement in the United States (US) clarified the Medicare coverage of maintenance rehabilitation as a skilled service, even when no improvement is expected. A scoping review was conducted to describe maintenance rehabilitation. MATERIALS AND METHODS: A systematic search was performed across 9 databases. Maintenance rehabilitation was defined as services (e.g. occupational, physical, or speech language therapies), intended to maintain a person's condition and prevent decline. Studies focused on medical, or non-rehabilitative, interventions (e.g. opioid cessation or chemotherapy) were excluded. Of the 734 abstracts reviewed, 90 met inclusion criteria. Two reviewers extracted data and used descriptive statistics. An exploratory thematic analysis in a convenience sample contextualized the works. RESULTS: Most works (90%) were peer-reviewed publications. The works represented 12,638 individuals, with predominant populations being cardiac (4,122 individuals) and pulmonary (2,324 individuals). Physical activity interventions were identified most frequently (87%). Outcome measures primarily focused on activities (49%) or body systems (42%). Thematic analysis suggests maintenance rehabilitation may support domains contributing to quality of life. CONCLUSIONS: While evidence exists for maintenance therapy, future research on intervention design will guide implementation by health systems.

The largest number of maintenance rehabilitation studies are focused on physical activity interventions in cardiac and pulmonary populations.Most studies measured body systems and activity outcomes rather than participation.Themes of individualized and structured interventions were identified as factors that support maintenance rehabilitation.

Modulation of Stemness and Differentiation Regulators by Valproic Acid in Medulloblastoma.

Changes in epigenetic processes such as histone acetylation are proposed as key events influencing cancer cell function and the initiation and progression of pediatric brain tumors. Valproic acid (VPA) is an antiepileptic drug that acts partially by inhibiting histone deacetylases (HDACs) and could be repurposed as an epigenetic anticancer therapy. Here, we show that VPA reduced medulloblastoma (MB) cell viability and led to cell cycle arrest. These effects were accompanied by enhanced H3K9 histone acetylation (H3K9ac) and decreased expression of the MYC oncogene. VPA impaired the expansion of MB neurospheres enriched in stemness markers, and reduced MYC while increasing TP53 expression in these spheres. In addition, VPA induced morphological changes consistent with neuronal differentiation and increased expression of differentiation marker genes TUBB3 and ENO2. Expression of stemness genes SOX2, NES, and PRTG was differentially affected by VPA in MB cells with different TP53 status. VPA increased H3K9 occupancy of the promoter region of TP53. Among genes regulated by VPA, stemness regulators MYC and NES showed association with patient survival in specific MB subgroups. Our results indicate that VPA may exert antitumor effects in MB by influencing histone acetylation, which may result in modulation of stemness, neuronal differentiation, and expression of genes associated with patient prognosis in specific molecular subgroups. Importantly, the actions of VPA in MB cells and neurospheres include a reduction in expression of MYC and increase in TP53.

Surgical stabilization of rib fractures-Does race matter?

BACKGROUND: Advances in medical technology have widened the gaps and exposed disparities in medical treatments. The prevalence of surgical stabilization for rib fractures is rising despite its controversial indications for this treatment modality. In situations of equipoise, surgeons may find themselves choosing patients for surgery, revealing potential implicit biases. We hypothesize that there exists an inequity in surgical stabilization for rib fractures performed based on race. METHODS: Data were obtained from the American College of Surgeons 2013-2021 Trauma Quality Improvement Program database. Study participants were divided into race groups according to Trauma Quality Improvement Program data registry. To assess the association between race and surgical stabilization for rib fractures, a Poisson regression model was used. Potential confounding adjusted include race, age, sex, highest abbreviated injury severity score in each region, flail chest, sternum fracture, pneumothorax, hemothorax, pulmonary contusion, and comorbidities. RESULT: Black patients were more often treated at a level 1 trauma center (74%) (P < .001). Flail chest was most common in White (3.2%) and American Indian (3.4%) patients compared with other races (P = .012). After adjusting for potential confounding in the Poisson regression analyses, Black patients were 26% less likely to undergo surgical stabilization for rib fractures (adjusted incident rate ratio [95% confidence interval]: 0.74 [0.64-0.85], P < .001) and Asian were 40% less likely to undergo surgical stabilization for rib fractures (adjusted incident rate ratio [95% confidence interval]: 0.60 [0.43-0.81], P = .001) than White patients. CONCLUSION: There is a disparity in the delivery of surgical stabilization for rib fractures in patients with rib fractures. Black and Asian patients undergo surgical stabilization for rib fractures at a significantly lower rate than their White counterparts. This discrepancy in the delivery of care is concerning and requires further study.

The "Smile Design and Space" Concept for Altering Vertical Dimension of Occlusion and Esthetic Restorative Material Selection.

OBJECTIVE: This article introduces the smile design and space (SDS) concept for managing the vertical dimension of occlusion (VDO) in full-mouth reconstructions and its influence on the selection of esthetic restorative materials. Limitations of traditional VDO adjustment methods are being addressed through a facially driven treatment approach that enhances esthetics improves function and minimizes invasiveness. CLINICAL CONSIDERATIONS: The SDS concept involves detailed evaluations of the incisal edges of the maxillary central incisors in relation to the upper lip at rest and at full smile. This includes analyzing variations in incisal exposure due to differing lip mobility, which can significantly impact gingival esthetics, particularly in cases with gingival exposure during smiling. The concept employs a strategic formula to determine the necessary alterations in VDO based on specific dental relationships and wear patterns. CONCLUSIONS: The SDS concept provides guidelines for full-mouth reconstructions, emphasizing minimal invasiveness and the integration of esthetics with functional dynamics to enhance the predictability of clinical outcomes and reduce complications associated with traditional restoration techniques. CLINICAL SIGNIFICANCE: The SDS concept enhances the precision of full-mouth reconstructions through individualized adjustment of the VDO, tailored to the esthetic and functional requirements of each patient.

Frictional Contact of Soft Polymeric Shells.

The classical Hertzian contact model establishes a monotonic correlation between contact force and area. Here, we showed that the interplay between local friction and structural instability can deliberately lead to unconventional contact behavior when a soft elastic shell comes into contact with a flat surface. The deviation from Hertzian solution first arises from bending within the contact area, followed by the second transition induced by buckling, resulting in a notable decrease in the contact area despite increased contact force. Additionally, our results invalidated a previous claim of a linear relation between friction and dissipated energy, demonstrating the suppression of both buckling and dissipation at high friction levels. Different contact regimes are discussed in terms of rolling and sliding mechanisms, providing insights for tailoring contact behaviors in soft shells.

IF1 is a cold-regulated switch of ATP synthase hydrolytic activity to support thermogenesis in brown fat.

While mechanisms controlling uncoupling protein-1 (UCP1) in thermogenic adipocytes play a pivotal role in non-shivering thermogenesis, it remains unclear whether F1Fo-ATP synthase function is also regulated in brown adipose tissue (BAT). Here, we show that inhibitory factor 1 (IF1, encoded by Atp5if1), an inhibitor of ATP synthase hydrolytic activity, is a critical negative regulator of brown adipocyte energy metabolism. In vivo, IF1 levels are diminished in BAT of cold-adapted mice compared to controls. Additionally, the capacity of ATP synthase to generate mitochondrial membrane potential (MMP) through ATP hydrolysis (the so-called "reverse mode" of ATP synthase) is increased in brown fat. In cultured brown adipocytes, IF1 overexpression results in an inability of mitochondria to sustain the MMP upon adrenergic stimulation, leading to a quiescent-like phenotype in brown adipocytes. In mice, adeno-associated virus-mediated IF1 overexpression in BAT suppresses adrenergic-stimulated thermogenesis and decreases mitochondrial respiration in BAT. Taken together, our work identifies downregulation of IF1 upon cold as a critical event for the facilitation of the reverse mode of ATP synthase as well as to enable energetic adaptation of BAT to effectively support non-shivering thermogenesis.

Influence of Fermentation Time and Inoculation of Starter Culture on the Chemical Composition of Fermented Natural Coffee Followed by Depulping.

Fermentation using starter cultures has been considered an alternative and economically viable technology for the production of specialty coffees. This type of technology promotes several benefits, such as increased sensory quality, control over the fermentation process, predictability of the final product and added value. Coffee (Coffea arabica L.) samples for this study were collected in Presidente Olegário - MG (2018/19 crop year) in the Cerrado region of Minas Gerais. The effects of natural fermentation and inoculation of the yeast Torulaspora delbrueckii and duration of fermentation (0, 24, 48, 72 and 96 hours) on the sensory and chemical quality (analysis of bioactive, volatile, and organic compounds and fatty acids) of coffee were evaluated. The objective of this study was to determine the effect of fermentation time and starter culture inoculation on the chemical composition of fermented coffees. Fermentation time significantly influenced the sensory description of the coffee beverage, with notes of honey, brown sugar and almond predominating up to 48 hours, for coffees fermented for 72 and 96 hours the notes described were and fruity, winey notes. The chemical composition was primarily influenced by fermentation time.

Numerical-experimental procedure for predicting fatigue life in SAE AMS 7475-T7351 aluminum alloy considering the effect of stress ratio.

This study addresses the prediction of fatigue life in SAE AMS 7475-T7351 aluminum alloys under variable loads, commonly used in the construction of aircraft fuselages. The main objective of the research was to develop a numerical-experimental procedure to analyze crack growth, using the Walker's approach which considers the effects of the stress ratio R on the fatigue crack growth rate d a / d N , combined with the Finite Element Method and Linear Regression of the Stress Intensity Factor. Observations showed that Walker's model effectively consolidated fatigue crack propagation data for various stress ratios when applied longitudinally to L-T rolling orientation, due to low dependence of exponent m on R -value in d a / d N equation. Simple averaging of m values effectively calculated Walker's exponent. The methodology employed experimental tests following ASTM standards for tension, fracture toughness, and fatigue, complemented by Finite Element Method (FEM) simulations. The Walker's model proved more effective, while the Paris-Erdogan model, which ignores the R effect, resulted in overly conservative service life estimates. The principle of similitude suggests that this methodology could be effective in predicting fatigue life in cases with complex geometries, where calculating the Stress Intensity Factor Fracture parameter is challenging and the Finite Element Method shows efficiency.

Enhanced antibacterial activity of starch-alginate beads by a synergistic effect between Cu2+ and Zn2+ ions with a potential wound dressing application.

The synthesis and characterization of starch/alginate composite beads, crosslinked with Cu2+, Zn2+, and Cu:Zn mixtures were investigated, focusing on their potential application in exudative wound dressings. Hydrogel beads were prepared using the external gelation method and then dried via freeze-drying to create cryogels and air-drying to create xerogels. Microstructural characterization was performed using SEM and EDS, showing the typical porous structure with a homogeneous distribution of cations across the beads. Unimetallic beads exhibited higher equilibrium water uptake compared to Cu:Zn bimetallic beads (500 % vs. 300 %). After the swelling study, the total amount of Cu2+ released was significantly below the maximum allowed level as a safeguard against copper toxicity. All beads demonstrated excellent antimicrobial activity against E. coli, S. aureus, and P. aeruginosa. Bimetallic materials, particularly cryogels with equal or greater amount of zinc relative to copper, were particularly effective against P. aeruginosa. Hence, the synthesized bimetallic starch-alginate materials presented superior water absorption capacity and significantly enhanced antibacterial response compared to unimetallic beads, due to the synergistic effect between Cu2+ and Zn2+ ions, making then suitable for use in exudative wound dressings.

Local volume concentration, packing domains, and scaling properties of chromatin.

We propose the Self Returning Excluded Volume (SR-EV) model for the structure of chromatin based on stochastic rules and physical interactions. The SR-EV rules of return generate conformationally defined domains observed by single-cell imaging techniques. From nucleosome to chromosome scales, the model captures the overall chromatin organization as a corrugated system, with dense and dilute regions alternating in a manner that resembles the mixing of two disordered bi-continuous phases. This particular organizational topology is a consequence of the multiplicity of interactions and processes occurring in the nuclei, and mimicked by the proposed return rules. Single configuration properties and ensemble averages show a robust agreement between theoretical and experimental results including chromatin volume concentration, contact probability, packing domain identification and size characterization, and packing scaling behavior. Model and experimental results suggest that there is an inherent chromatin organization regardless of the cell character and resistant to an external forcing such as RAD21 degradation.

A multi-institutional study to investigate the sparing effect after whole brain electron FLASH in mice: Reproducibility and temporal evolution of functional, electrophysiological, and neurogenic endpoints.

BACKGROUND AND PURPOSE: Ultra-high dose-rate radiotherapy (FLASH) has been shown to mitigate normal tissue toxicities associated with conventional dose rate radiotherapy (CONV) without compromising tumor killing in preclinical models. A prominent challenge in preclinical radiation research, including FLASH, is validating both the physical dosimetry and the biological effects across multiple institutions. MATERIALS AND METHODS: We previously demonstrated dosimetric reproducibility of two different electron FLASH devices at separate institutions using standardized phantoms and dosimeters. In this study, tumor-free adult female mice were given 10 Gy whole brain FLASH and CONV irradiation at both institutions and evaluated for the reproducibility and temporal evolution of multiple neurobiological endpoints. RESULTS: FLASH sparing of behavioral performance on novel object recognition (4 months post-irradiation) and of electrophysiologic long-term potentiation (LTP, 5 months post-irradiation) was reproduced between institutions. Differences between FLASH and CONV on the endpoints of hippocampal neurogenesis (Sox2, doublecortin), neuroinflammation (microglial activation), and electrophysiology (LTP) were not observed at early times (48 h to 2 weeks), but recovery of immature neurons by 3 weeks was greater with FLASH. CONCLUSION: In summary, we demonstrated reproducible FLASH sparing effects on the brain between two different beams at two different institutions with validated dosimetry. FLASH sparing effects on the endpoints evaluated manifested at later but not the earliest time points.

Correction: Virtual screening, identification and in vitro validation of small molecule GDP-mannose dehydrogenase inhibitors.

[This corrects the article DOI: 10.1039/D3CB00126A.].

CT-based comprehensive scoring system for assessing hip arthropathy in ankylosing spondylitis: feasibility study with clinical correlation.

OBJECTIVE: To develop a CT-based scoring system for assessment of hip arthropathy in AS. METHODS: All AS patients were prospectively recruited, consented, and underwent whole-body stereoradiographs and pelvis CT, which were assessed by two independent radiologists. Stereoradiographs were assessed according to Kellgreen-Lawrence and BASRI-h. For the Hip arthropathy CT score in AS (HACTSAS), joints were divided into 7 segments and scored for joint space, osteophytes, subchondral cysts/erosions. Patients were clinically assessed for range of motion (ROM), pain, and clinical scores (BASMI, BASFI, ASQol, BASDAI and ASDAS). Radiological scores correlations with clinical parameters were compared. ROM sensitivity and specificity for hip arthropathy (BASRI-h ≥ 2) were calculated. RESULTS: Sample included 112 patients, with 36/112 females and 76/112 males. Average age was 51.0 ± 11.2 years and mean duration of AS was 20.9 ± 9.6 years. ICC for HACTSAS, Kellgreen-Lawrence and BASRI-h were 0.89, 0.89 and 0.82 respectively. HACTSAS showed moderate absolute correlation with ROM (ρ=-0.41) and BASMI (ρ = 0.45), and weak with pain (ρ = 0.18) and BASFI (ρ = 0.25). BASRI-h and Kellgreen-Lawrence exhibited moderate correlation with ROM (ρ=-0.44 and ρ=-0.40, respectively), weak with pain (ρ=-.27and ρ=-0.23, respectively) and BASFI (ρ=-0.16 and ρ=-0.18, respectively), but only weak with BASMI (ρ=-0.34 and ρ=-0.36, respectively). Internal rotation <15°, abduction <31°, and intermalleolar distance <75cm were, respectively, 73%, 70% and 73% sensitivity and 81%, 65% and 68% specific for hip arthropathy. CONCLUSION: HACTSAS exhibited higher correlation with BASMI and BASFI when compared with BASRI-h, but less correlation with pain and ROM. Internal rotation was the best clinical discriminator for hip arthropathy.

Ectopic Intrasphenoidal Growth Hormone Releasing Pituitary Adenoma Associated with an Intracranial Aneurysm.

Pituitary adenomas are a type of of the most frequent intracranial tumors. These tumors can extend outside the sella, but very rarely originate ectopically to the sellar region. A 71-year-old patient presented to our institution, with prior clinical history of noncontrolled arterial hypertension and new-onset high-intensity pulsatile headache. Upon suspicion of a hypertensive emergency with probable brain compromise, a nonenhanced computed tomography of the head was performed. A mass within the sphenoid sinus was found. Endocrinological workup demonstrated a significant elevation of the growth hormone. As an incidental finding, a brain aneurysm was evidenced, which was treated endovascularly prior to the mass treatment. Subsequently, the patient successfully underwent a gross total resection through an endonasal transsphenoidal approach. Histopathological results were consistent with a pituitary ectopic adenoma. A postoperative improvement in levels of somatomedin C was documented postoperatively.

Continuous Flow Synthesis of Hexyl Laurate Using Immobilized Thermomyces Lanuginosus Lipase from Residual Babassu Mesocarp.

Brazil has one of the greatest biodiversities on the planet, where various crops play a strategic role in the country's economy. Among the highly appreciated biomasses is babassu, whose oil extraction generates residual babassu mesocarp (BM), which still needs new strategies for valorization. This work aimed to use BM as a support for the immobilization of Thermomyces lanuginosus lipase (TLL) in an 8.83 mL packed-bed reactor, followed by its application as a biocatalyst for the synthesis of hexyl laurate in an integrated process. Initially, the percolation of a solution containing 5 mg of TLL at 25 °C and flows ranging from 1.767 to 0.074 mL min-1 was investigated, where at the lowest flow rate tested (residence time of 2 h), it was possible to obtain an immobilized derivative with hydrolytic activity of 504.7 U g-1 and 31.7 % of recovered activity. Subsequent studies of treatment with n-hexane, as well as the effect of temperature on the immobilization process, were able to improve the activities of the final biocatalyst BM-TLLF, achieving a final hydrolysis activity of 7023 U g-1 and esterification activity of 430 U ⋅ g-1 against 142 U g-1 and 113.5 U g-1 respectively presented by the commercial TLIM biocatalyst. Desorption studies showed that the TL IM has 18 mg of protein per gram of support, compared to 4.92 mg presented by BM-TLL. Both biocatalysts were applied to synthesize hexyl laurate, achieving 98 % conversion at 40 °C within 2 h. Notably, BM-TLLF displayed exceptional recyclability, maintaining catalytic efficiency over 12 cycles. This reflects a productivity of 180 mg of product ⋅ h-1 U-1 of the enzyme, surpassing 46 mg h-1 U-1 obtained for TLIM. These results demonstrate the efficacy of continuous flow technology in creating a competitive and integrated process offering an exciting alternative for the valorization of residual lignocellulosic biomass.