The production of secreted therapeutic proteins, such as monoclonal antibodies, often requires cultivating CHO cell cultures in bioreactors ranging from 10 mL to 20,000 L in volume. The cells proliferate and secrete the therapeutic protein over multiple days and the bioreactor is then emptied to collect the culture broth. The first step in the purification process removes any insoluble cells and cellular debris to produce a clarified conditioned medium (CM). Manufacturing scales frequently use continuous centrifugation followed by depth filtration for clarification of CM, but this method is challenging to reproduce with small volumes generated during bench scale experiments. The typical bench scale clarification method consists of a time-consuming centrifugation step to pellet the cells and cellular debris followed by the careful removal of the supernatant which is filtered through a 0.2-μm filter. This study evaluates a robotic clarification process using the commercially available Sartorius StreamLink® CC15. Our results demonstrate that the StreamLink reduces scientist manual time and effectively clarifies material without impacting quality attributes, when compared to manual clarification. Furthermore, we provide guidance on optimizing sample and system parameters to maximize product recovery.

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Compared to batch operation, continuous bioprocessing can offer numerous advantages, including increased productivity, improved process control, reduced footprint, and increased flexibility. However, integration of traditional batch operations into a connected process can be challenging. In contrast to batch operations run at constant pressure or high flux, virus filtration in continuous processes may be operated at very low flux. This change in operating conditions may reduce the viral retention performance of the filter which has inhibited adoption of truly continuous virus filtration. To overcome this limitation, a novel approach is described that utilizes serial virus filtration, with a high area ratio between first to second stage filters, to achieve virus retention targets. In this study, virus filters were operated continuously (except for planned process interruptions) for 200 hours in a serial configuration at a first to second stage filter area ratio of 13:1 and at a first stage flux of 5 L/m 2/hr. While the minute virus of mice (MVM) retention performance of the first stage filter was about 4 log reduction value (LRV), there was no virus detected in the second stage filtrate, translating to an MVM LRV across the filtration train of ≥6.7. The second stage filter was the dominant flow resistance at the start of the run but, as it was protected from foulants by the first stage filter, it suffered minimal fouling and the life of the filter train was controlled by the first stage. A theoretical case study projected that continuous virus filtration using serial configuration at high area ratio would have about 30% longer filter changeout time, 14% higher productivity, and virus retention nearly 6 LRV greater than single stage operation. The findings of this research are expected to provide valuable insights into optimizing virus filtration in continuous bioprocessing.

TITLE: A rare case of primary vulvar diffuse large B-cell lymphoma: A case report

Introduction: Uncontrolled or severe asthma results in symptomatic usage of short-acting ß2-agonist usage (SABA). MicroRNAs (miRNAs) are post-translational regulators that can influence asthma biology. This study aims to identify miRNAs that are associated with increased SABA usage. Methods: Small RNA sequenced from blood serum of 1,132 asthmatic children aged 6 to 14 years in the Genetics of Asthma in Costa Rica Study (GACRS) was used for this analysis. Logistic regression identified miRNAs in patients who required increased SABA usage. These miRNA were validated for association with SABA induced BDR. Gene target pathway analysis was performed on validated miRNAs. Results: 21 miRNAs were significantly associated with increased SABA usage with OR ranging from 0.87 to 1.23. Two miRNAs, miR-378a-3p and miR-144-3p, had odds ratio 1.14 (1 - 1.29, p=0.05) and 1.11 (1.01-1.22, p = 0.035), respectively for increased SABA usage and were also significantly associated with bronchodilator response. Furthermore, a linear regression analysis involving these miRNA and bronchodilator response revealed that increased miR-378a-3p correlated with decreased BDR and increased expression of miR-144-3p correlated with improving pulmonary function with bronchodilators. In gene target KEGG pathway analysis, the erythroblastosis viral oncogene (ErbB) signaling pathway had among one of the highest fold enrichment and p-value. Conclusion: Increased expression miR-378a-3p and miR-144-3p was seen in this patient population who required increased SABA usage. There were different bronchodilatory effects seen in these two miRNAs, suggesting different potential mechanisms underlying increased SABA usage.

Introduction: During NICU hospitalization, children born preterm with bronchopulmonary dysplasia (BPD) are frequently prescribed diuretics for treatment of respiratory symptoms. However, less is known about diuretic use and weaning in the outpatient setting. This study sought to characterize clinical features associated with outpatient diuretic use, and timing of diuretic weaning in children with BPD. Methods: Data was obtained by chart review from registry 1224 participants born < 32 weeks gestation, discharged between 2008-2023 and recruited from outpatient BPD clinics at Johns Hopkins Children’s Center and the Children’s Hospital of Philadelphia (97.4% diagnosed with BPD). Data was analyzed using Chi-square tests, t-tests, and ANOVA tests. Results: Children with BPD prescribed diuretics as outpatients (n=737), were more likely to have lower birth weights, earlier gestational age, and severe BPD compared to those not on diuretics (n=487). Of those prescribed diuretics, most children were on a thiazide alone (46.4%) or a thiazide and a potassium sparing agent (44.8%) with a minority on loop diuretics alone (3.3%) or loop diuretic combinations (4.7%). Most children weaned off diuretics by two years of age. Public insurance, earlier gestational age, technology dependence and loop diuretics were associated with slower diuretic weaning. Conclusion: Outpatient diuretic use is common in BPD with > 75% of children being weaned by two years of age. No difference was found in weaning of home oxygen between children on one versus no diuretic. Thiazides were most commonly prescribed with slower outpatient diuretic weaning associated with public insurance, technology dependence and loop diuretic use.

Ecologists have long studied how species numbers are distributed along environmental gradients. Typically, plotting species’ abundance along a single gradient result in a polygonal point cloud, because species do not reach their maximum potential abundance at every location. The upper limit of these abundance-environment plots represents locations where the maximum abundance of the species is limited by the measured environmental variable and may be explained by the Law of the Minimum (LoM). We examine the generality of polygon-shaped plots in abundance-environment relationships and assess how often they are interpreted in the context of the LoM. Our approach combined a literature review with an analysis of observational data. Initially, we examined the prevalence of polygonal patterns in abundance-environment plots in the literature and analyzed how frequently these patterns are conceptually linked to the LoM. Additionally, we assessed the prevalence of polygon-shaped patterns in abundance-environment relationships across 186 tree and 114 bird species in North America using a novel method that discriminates polygon-shaped plots in observational data. We also developed upper quantile regressions (QR) to illustrate how to model the influence of water balance and energy on the maximum potential abundances of trees and birds. Our results revealed polygonal plots in 76% of reviewed studies and in 73% of the 300 analyzed tree and bird species. We demonstrate that polygonal shapes with upper limits are prevalent in abundance-environment plots; yet they are rarely interpreted considering the Law of the Minimum. We conclude that a wider acknowledgement of limiting relationships would improve our ecological understanding and estimation with further benefits to theoretical and applied ecology.

Background: Oral immunotherapy (OIT) has shown promise for peanut-allergic preschool children. Studies report it is safe and effective, with over 70% achieving sustained unresponsiveness. Knowledge about preschool OIT for other allergens and multiple food allergies is still limited. Objective: To assess the safety of a pragmatic OIT protocol in preschool children with single and multiple food allergies. Methods: This single-center prospective intervention study included children aged 9-24 months with proven food allergy (sensitization and a positive oral food challenge (OFC)). After a short dose escalation phase, children received one year of low-dose (300 mg protein/day) OIT, followed by an exit OFC four weeks after stopping therapy. Children with multiple food allergies received OIT for up to four allergens. Allergic dosing reactions (ADR) were recorded and classified using Sampson’s severity score (grades I-V). Results: Between May 2019 and Oct 2022, 124 children (median age 17, IQR 11-20 months) started OIT for 189 allergies. Peanut (n=52), cashew (n=46) and egg (n=38) allergies were the most common. ADR occurred in 89 children and 117 OIT trajectories, most frequently during dose escalation (65%). Reactions (95.3%) were mild (grade I-II) and mostly resolved spontaneously (55.6%) or with antihistamines (29.9%). Ten reactions were severe (grade III-IV). Reaction severity did not differ between single and multiple OIT or among the different allergens. Ten trajectories were discontinued due to side effects. Conclusions: OIT for various allergens and multiple food allergies appears safe for preschoolers. Side effects are common but typically mild. However, they may lead to therapy discontinuation.

Water-use efficiency (WUE) and the stoichiometry of plant-soil carbon (C), nitrogen (N), and phosphorus (P) are key indicators of plant growth, while stand quality is an important index for evaluating afforestation. Yet it remains unclear how WUE and stoichiometric characteristics respond to changes in stand quality in desert ecosystems. To that end, we studied the community characteristics of sand-fixing Caragana korshinskii stands differing in age (planted 10, 30, 50, and 70 years ago) in Mu Us Sandy Land, China, and measured their leaf water use efficiency (WUE) and leaf-soil C:N:P stoichiometry. The relations among the stand quality index, leaf WUE, and plant-soil stoichiometry were analyzed. After 70 years, WUE was at its lowest level, but only significantly lower than that of a 10-year-old C. korshinskii stand. Afforestation years had differential effects on C, N, and P nutrients and their stoichiometric characteristics in leaves and soil. For soil, its total P barely increased. Irrespective of stand age, the N:P ratio of leaves was > 16, which suggests P is the main factor limiting the development of C. korshinskii plantations. Notably, WUE decreased significantly as the stand quality index increased while soil stoichiometry responded more strongly than plant stoichiometry. These results can guide investigations into the role of C. korshinskii stands in plant and soil recovery effects, providing a scientific basis to evaluate the rational use of C. korshinskii sand-fixing forest in afforestation.

Despite decades of studies on the differential distribution of polyploid organisms, the causes of this pattern have yet to be elucidated. This study aimed to explore some of the possible physiological mechanisms explaining the differential northern distribution of polyploid clones of Daphnia pulex compared to the one of the diploid parental species. The critical thermal minimum (CTmin) was measured in 17 D. pulex clones of contrasted ploidy (diploid and triploid) and geographic origins (temperate and subarctic climates) reared under low and high temperatures (16 and 24 °C). Triploid clones had better cold tolerance (lower CTmin) than both sympatric and temperate diploid clones. No significant association was found between CTmin and body size nor with cell size. We suggest that triploids might express a cold shock resistant phenotype related to higher gene expression and/or fatty acid profiles. Cold tolerance can be viewed as one of the possible reasons for polyploid preponderance in subarctic climates.

Disease in crop has a great impact on agriculture resulting in a loss. Early detection of diseases through regular observing is crucial to minimize crop loss. In recent years, automatic plant disease recognition systems have been developed using image. It represents several deep learning models, including VGG16, EfficientNetB0, ResNet-50, and GoogleNet, for the identification of four rice diseases utilization. The VGG16 model with a Dense Layer achieves an accuracy of 99.747%, while the VGG16 model with 2D-CNN achieves an accuracy of 99.916%. The EfficientNetB0 model with Dense Layer achieves a validation loss of 1.3880 and a test accuracy of 27.004%, while the EfficientNetB0 model with 2D-CNN achieves a validation loss of 1.3830 and a test accuracy of 27.004%. The ResNet-50 model with Dense Layer achieves a validation loss of 0.8121 and a test accuracy of 70.211%, while the ResNet-50 model with 2D-CNN achieves a validation loss of 0.7238 and a test accuracy of 69.789%. Finally, the GoogleNet model with Dense Layer achieves an accuracy of 99.916%, while the GoogleNet model with 2D-CNN achieves an accuracy of 99.831%. The above results are Extracted on PNG image dataset. The results show that the proposed deep learning models are effective in identifying rice diseases and provide a smart agriculture solution to the problem of crop diseases, helping farmers identify and manage the diseases efficiently, leading to better crop yield and quality. Overall, it demonstrates the impact of deep learning-based approaches for the automatic identification of plant diseases and the importance of using multiple models and datasets for comprehensive evaluation. These findings can aid in the development of more accurate and robust plant disease identification systems and help address the challenges of sustainable agriculture.

A document by Natalia Galaz-Souza. Click on the document to view its contents.

Background: Respiratory Syncytial Virus (RSV) is a major health concern, particularly for infants. In France, Nirsevimab, a long-acting monoclonal antibody to prevent RSV-associated lower respiratory tract infections (LRTI) was available from September 2023. We described RSV-associated LRTI hospitalisations during the 2023-2024 season among infants younger than six months born at the Hospices Civils de Lyon (HCL), and evaluated the effectiveness of Nirsevimab against RSV-LRTI hospitalisation. Methods: This observational study included infants born and hospitalised at the HCL during the 2023-2024 season, along with pre-COVID-19 and 2022-2023 seasons. Information on Nirsevimab immunisation status, clinical and perinatal variables were collected through routine care. Infants’ characteristics and incidence risk of hospitalisation per 100 births during 2023-2024 were compared with the historical periods overall and by delay between birth and the onset of the RSV season. Nirsevimab effectiveness was computed by the screening method. Results: During the 2023-2024 season, 83 infants younger than six months were hospitalised with an RSV-associated LRTI. Compared with the historical periods these infants were older. Incidence risk for infants born during the period when immunisation was available were lower than the previous seasons; incidence risk ratios were 0.45 (95% confidence interval (CI): [0.33;0.62]) in 2023-2024 compared with pre-COVID-19 period and 0.53 (95%CI: [0.36;0.77]) compared with 2022-2023 season. Nirsevimab effectiveness was 78.3% (95%CI: [55.9 ;89.5]) with a coverage of 79.3% in the HCL maternities. Conclusions: This study revealed a change in the epidemiology of RSV-associated LRTI hospitalisations in 2023-2024. High coverage and effectiveness were estimated in real-world setting.

The advancement of technology in detecting individuals and objects is actively being pursued beyond the visible spectrum, leveraging wireless communication to identify objects in obscured areas. This approach offers the distinct advantage of not necessitating a direct line of sight, unlike traditional camera-based sensing methods. Current research efforts aim to utilize this technology in a range of applications, such as indoor positioning and activity monitoring. However, these studies primarily involve sensing within high-frequency bands such as Wi-Fi or Bluetooth, which present limitations in terms of short sensing distances and low transmittance rates. Given these challenges, our study concentrates on calculating the Power Spectral Density (PSD) in the 433MHz band to detect the presence of individuals nearby. Our findings demonstrate the real-time detection of individuals passing between transmitter and receiver nodes with a 95% probability

A document by Diego Torres-García. Click on the document to view its contents.

1. Removal of the thyroid gland often leads to vocal problems due to potential nerve damage, affecting voice quality and causing symptoms like hoarseness and vocal fatigue. 2. Vocal dysfunction post-thyroidectomy significantly impacts patients’ daily lives, affecting communication and social interactions. 3. Semi-occluded vocal tract exercises (SOVTE), such as straw phonation and lip trills, are effective in improving voice outcomes by enhancing vocal efficiency and minimizing vocal fold stress. 4. The case study presented Preliminary evidence from a single subject showed positive changes in vocal behavior, including improved intensity, frequency, and quality of voice after SOVTE therapy. 5. The study suggests that SOVTE could be beneficial in enhancing voice quality for thyroidectomy patients. However, further research with larger patient cohorts is necessary to establish the effectiveness of SOVTE as a standard therapy option in this population.

A document by Prof. Hazar Shtat. Click on the document to view its contents.

Very recently, [Wickramasinghe et al., Mann type approximation scheme for solving a new class of split inverse problems in Hilbert spaces Applicable Anal. (2023). DOI: 10.1080/00036811. 2023.2233977] introduced a new class of split inverse problems, known as split hierarchical monotone variational inclusion problems with multiple output sets. The authors proposed a Mann-type iterative method to approximate the solution of the problem in Hilbert spaces. However, to establish their convergence result, the authors required the associated single-valued operators to be co-coercive. This condition is too stringent and limits the scope of application of their results. In this study, we aim to improve their result of [Wickramasinghe et al. ] by relaxing the co-coercive condition to monotone. We also introduce a new viscosity Tseng-type iterative method for solving the problem in Hilbert spaces and obtain strong convergence results under more relaxed conditions. Furthermore, our study extends to addressing the split hierarchical variational inequality problem involving multiple output sets, the split hierarchical convex minimization problem with multiple output sets, and the generalized split hierarchical monotone variational inclusion problem. Lastly, we perform numerical experiments in both finite- and infinite-dimensional spaces and provide comparison results with existing methods discussed in the literature.

Introduction/Objectives: neuro-gastroenteritis existed since World-War-II dysen-tery outbreak, albeit mechanisms are yet ill-defined. Similarly, the post‐ COVID‐19-syndrome is a growing problem. The frequencies of neuro-gastrointestinal; gas-troesophageal-reflux (GERD), the inflammatory-bowel-syndrome (IBS), and mental health is not clear. Methods: we used descriptive analysis on stratified data obtained by online questionnaire sent nationwide and analyzed by Statistical Package for So-cial Sciences version 21 (SPSS: An IBM Company). Results: revealed the frequency and associations of different factors in the coexistence of the disorders in 2067 par-ticipants aged 18 to >60 years old. The majority were Saudis (94.7%, n= 1957), most were 18 - 29 years old (66%, n=1364) where young Saudi females were 72.4%, (n=1496). Estimated 1099 (53.2%) were students, 428 (20.7%) were unemployed, and 540 (26.1%) were employed. Psychological syndromes namely: anxiety (60.7%), stress (60.7%), and depression (60.7%) were most frequent and preceded other dis-orders in 51%, whereas IBS (48.7%) and GERD (36.3%) came 4th and 5th, respec-tively, with the latter two appeared first in 33.9%, and 24.3% respectively. In most respondents (59.2%, n=1178), the disorders first appeared recently, 33.6% (n=669) reported during adult life, and only in 7.2% (n=144) appeared in childhood (7.2%, n=144). Aggravating factors reported in family history (32.9%, n=681) of which 476 (69.9%) was IBS while 215(31.6%) and 175 (25.7%), respectively, were psycholog-ical and GERD. However, only 18.3% respondents sought treatment (n=378) with 66 (3.2%) had colectomy or a colostomy bag while 58%, (n=1201) were active outdoors. Disorders were significantly associated with age (P value =.001); GERD with old age, IBS with 40-49 years, and psychological disorders among youth. Conclusion: Thus, mental health and neuro-gastroenteritis dominated young women while the IBS and GERD occurred first in old and mid-ages, respectively. These have significant im-plications in treatment strategies and microbiome restorage. Future microbiome studies would reveal more insight into the mechanisms of disorders.

According to the Centers for Disease Control and Prevention (CDC), people living with HIV are at higher risk of smoking consequences than persons without HIV. Eswatini has put efforts in place to curb smoking in the country such as restriction of smoking in designated areas, prohibition of tobacco advertising and promotion, restrictions on tobacco packaging and labelling, but there is a dearth of literature to provide evidence on available smoking cessation programs in the country. This study aims to review and analyze the current state of smoking cessation programs in the country particularly for people living with HIV, and provide recommendations for a smoking cessation and tobacco harm reduction framework for persons living with HIV. Official documents including the National Multisectoral HIV and AIDS Strategic Framework (2018 - 2023) and the Eswatini Integrated HIV Management Guidelines (2022) provide recommendations for the screening of lifestyle factors including smoking and advice for quitting. However, sufficient light is not shed on the processes for caring for PLHIV who also smoke. The study concludes that smoking cessation and harm reduction programs for persons who smoke have not been effectively incorporated into national guidelines and programs for persons living with HIV, and thus the development of a prototype framework to guide healthcare professionals in delivering care for PLHIV who smoke across the different care models for ART delivery, including the use of Safer Nicotine products (SNPs) when necessary, can be discussed in the review of national HIV care documents.

Background: Efficacy of allergen immunotherapy is dose-dependent; however, high doses of allergen may imply a greater risk of adverse reactions. Objective: To assess the safety and tolerability of subcutaneous immunotherapy (SCIT) with mixtures of mite allergen extracts, Dermatophagoides pteronyssinus/Blomia tropicalis (Dpt/Bt) and Dermatophagoides pteronyssinus/Lepidoglyphus destructor (Dpt/Ld) at maximum concentrations, in adult patients with allergic rhinitis or rhinoconjunctivitis, and controlled allergic asthma due to a clinically relevant sensitisation to these mites. Methods: An open-label, non-controlled, non-randomised, phase IIb clinical trial was carried out in three hospitals in Spain between September 2014 and May 2018. Patients received SCIT of either Dpt/Bt (100/1000 DPP/mL) or Dpt/Ld (100/100 DPP/mL) in two phases: a rush build-up phase on the first day (0.2 mL and 0.3 mL with a 30-minute interval) and a monthly maintenance phase administration (0.5 mL) up to 48 months. Results: Forty patients were recruited for the study, seven allocated to the Dpt/Bt group and 33 to the Dpt/Ld. None experienced immediate or delayed systemic Grade ≥2 reactions (EAACI classification) (systemic reactions were mostly Grade 1) nor died during the study. Local reactions were mostly mild (0‒10 cm). Thirty-nine patients (97.5%) experienced at least one adverse event (AE). Of the 283 reported AEs, eight (2.8%) were systemic reactions experienced by six (15%) subjects and 14 (4.9%) were local reactions sustained by ten (25%) subjects. Conclusions: SCIT treatment of patients with allergic rhinitis or rhinoconjunctivitis and controlled asthma with mixtures of Dpt/Bt and Dpt/Ld allergen extracts at maximum concentrations showed a favourable safety profile. Keywords: allergic asthma, allergic rhinitis, Blomia tropicalis, Dermatophagoides pteronyssinus, House dust mites, Lepidoglyphus destructor, maximum extract concentrations. safety,

【Abstract】Promoting the expansion and sinking of high-quality medical resources and regional balanced layout is an important part of building a high-quality and efficient medical and healthcare service system, and an important healthcare reform initiative to alleviate the problem of the public’s difficulty in accessing medical care and the high cost of medical care. Over the years, Tianjin Medical University General Hospital has been actively implementing the people-centered development ideology, focusing on promoting the high-quality development of the hospital, plowing into the remote diagnosis and treatment of Internet hospitals and the construction of intelligent medical care, and exploring efficient paths and modes of expanding and sinking high-quality medical resources according to local conditions. Based on its practical exploration, this paper will analyze the macro level of horizontal comparison of healthcare reform models within and outside the province, the meso level of the overall regulation of the healthcare system in Tianjin, and the micro level of the internal healthcare reform path of the General Hospital in an attempt to contribute to the implementation of the 20th National Congress, which proposes to ”promote the expansion of high-quality healthcare resources and the balanced layout of the region”.

(±)-Elodeoidileons A-L (1-12), 12 pairs of previously undescribed filicinic acid based meroterpenoids with unique pyran-fused linear or angular 6/6/6 tricyclic skeletons, were isolated from Hypericum elodeoides. Their structures including absolute configurations were determined by modern spectroscopic techniques, nuclear magnetic resonance (NMR), high resolution electrospray ionization mass spectrometry (HR-ESI-MS), calculated electronic circular dichroism (ECD) and NMR, as well as the modified Mosher’s methods. Besides, the possible biosynthetic pathways for 1-12 were proposed. Moreover, biological activity assessments suggest that compound 1a may promote β-amyloid (Aβ) clearance by activating Retinoid X receptor-α and upregulating the expression of the ABCA1 protein, showing promise as anti-Alzheimer’s agent.

The increasing need to protect individual privacy in data releases has led to significant advancements in privacy-preserving technologies. Differential Privacy (DP) offers robust privacy guarantees but often at the expense of data utility. On the other hand, data pooling, while improving utility, lacks formal privacy assurances. Our study introduces a novel hybrid method, termed PoolDiv, which combines differential privacy with data pooling to enhance both privacy guarantees and data utility. Through extensive simulations and real data analysis, we assess the performance of synthetic datasets generated via traditional DP methods, data pooling, and our proposed PoolDiv method, demonstrating the advantages of our hybrid approach in maintaining data utility while ensuring privacy.