Search Results (98)

The development of scalable, non-invasive tools to assess tumor responsiveness to structurally active immunoformulations remains a critical unmet need in solid tumor immunotherapy. Here, we introduce a real-time, ex vivo functional system to classify tumor cell lines exposed to a phospholipoproteomic platform, without relying on cytotoxicity, co-culture systems, or molecular profiling. Tumor cells were monitored using IncuCyte^® S3 (Sartorius) real-time imaging under ex vivo neutral conditions. No dendritic cell components or immune co-cultures were used in this mode. All results are derived from direct tumor cell responses to structurally active formulations. Using eight human tumor lines, we captured proliferative behavior, cell death rates, and secretomic profiles to assign each case into stimulatory, inhibitory, or neutral categories. A structured decision-tree logic supported the classification, and a Functional Stratification Index (FSI) was computed to quantify the response magnitude. Inhibitory lines showed early divergence and high IFN-γ/IL-10 ratios; stimulatory ones exhibited a proliferative gain under balanced immune signaling. The results were reproducible across independent batches. This system enables quantitative phenotypic screening under standardized, marker-free conditions and offers an adaptable platform for functional evaluation in immuno-oncology pipelines where traditional cytotoxic endpoints are insufficient. This approach has been codified into the STIP (Structured Traceability and Immunophenotypic Platform), supporting reproducible documentation across tumor models. This platform contributes to upstream validation logic in immuno-oncology workflows and supports early-stage regulatory documentation. Full article

Background: With a high human immunodeficiency virus (HIV) adult prevalence, people living with HIV (PLHIV) in Botswana continue to experience a high burden of comorbid HIV and cancer. We sought to investigate the trends of acquired immunodeficiency syndrome (AIDS) defining cancers (ADCs), non-AIDS defining cancers (NADCs), and associated risk factors in PLHIV compared with those without HIV. Methods: We analyzed data from adults aged ≥18 years reported in Botswana National Cancer Registry and National Data Warehouse. The crude, age-standardized incidence rate (ASIR), standardized incidence ratios (SIRs) of cancers and time trends were computed. Risk factors were determined using the Cox-regression model. Results: Over a 30-year period, 27,726 cases of cancer were documented. Of these, 13,737 (49.5%) were PLHIV and 3505 (12.6%) were people without HIV and 10,484 (37.8%) had an unknown HIV status. Compared to the HIV-uninfected, the PLHIV had higher and increasing trends in the cancer incidence overall during the study period (from 44.2 to 1047.6 per 100,000; p-trend < 0.001) versus (from 1.4 to 27.2 per 100,000; p-trend < 0.001). The ASIRs also increased in PLHIV for overall ADCs, NADCs and other sub-types like cervical, lung, breast, and conjunctiva cancers (p-trend < 0.001). Further, PLHIV had elevated SIRs for cervical cancer, Kaposi sarcoma in males and some NADCs. The most common risk factors were HIV infection and female sex for ADCs incidence and advanced age and being HIV-uninfected for NADCs incidence. Conclusions: Increasing trends of ADCs and NADCs during ART expansion were observed among PLHIV compared to those without HIV highlighting a greater need for targeted effective prevention and screening strategies including the provision of access to timely HIV and cancer treatment. Full article

Yellow passion fruit provides a substrate suitable for alcoholic fermentation and holds valuable sensory and nutritional properties that support its use for producing wine. Among the different interesting aspects to consider in the winemaking process, we highlight the analysis of the proteins of the yeast or yeasts responsible for the process; in addition to providing fundamental knowledge about the biotransformations that take place, they can contribute to understanding basic aspects that affect the sensory properties of the wine obtained. This study aimed to analyze the proteomic profile of the yeast Saccharomyces cerevisiae strain E1 (ATCC: MYC-425) during passion fruit fermentation. The process was conducted in a 5 L Sartorius bioreactor with a diluted fruit puree and sucrose, making a medium with a 10% v/v alcohol potential; after 4 days, ethanol reached 8.5% v/v as sugars decreased. Subsequent LC/MS-MS analysis allowed identifying a total of 938 valid proteins: 454 from the fruit substrate and 484 attributed to yeast. Of the latter, 243 proteins were present throughout the fermentation, with GO term analysis highlighting “binding” (78%) and “catalytic activity” (66%) in molecular function, as well as “cellular” (98%) and “metabolic (90%) processes” in biological process domains. These findings may be a significant step forward in understanding the proteomic dynamics of S. cerevisiae in tropical fruit fermentations. By revealing key proteins and their roles in the unique conditions of passion fruit must, this study offers insights for optimizing the wine elaboration procedures and improving organoleptic properties and quality. Full article

This study examined the impact of different breath-holding conditions on iliopsoas and other hip flexor muscle activity in individuals with chronic ankle instability (CAI). It has been hypothesised that impaired diaphragmatic function influences iliopsoas activation, potentially contributing to motor control deficits in patients with CAI. Eleven adults with a history of chronic ankle sprain participated in this study. Maximal isometric hip flexion was assessed under three breath-holding conditions: end-expiration, end-inspiration, and the intermediate state. Surface electromyography was used to record the muscle activity of the iliopsoas, rectus femoris, sartorius, and tensor fasciae latae, while the peak hip flexion torque was measured using an isokinetic dynamometer. Under the end-inspiration condition, iliopsoas activity on the affected side was significantly lower than that on the control side (p < 0.05). However, no significant differences were observed between the affected and control sides in the activity of the other hip flexor muscles or the peak hip flexion torque across breath-holding conditions. This study highlights the association between reduced iliopsoas activity during end-inspiration and compromised diaphragmatic function in patients with CAI. Future research should explore dynamic movement tasks and larger sample sizes to elucidate neuromuscular mechanisms further and refine rehabilitation strategies. Full article

Ultrasound-guided botulinum toxin type A (BoNT-A) injections have become an essential tool in the management of lower limb spasticity. Following our previous work, which focused on upper limb muscles, this third part provides a detailed visual guide to the identification and injection of proximal lower limb muscles frequently involved in spastic gait and posture disorders. This guide presents the ultrasound anatomy, clinical relevance, and injection strategies for eleven key muscles: gluteus maximus, piriformis, psoas major, rectus femoris, sartorius, gracilis, adductor longus, adductor magnus, semimembranosus, semitendinosus, and biceps femoris. For each muscle, the Elias University Hospital (EUH) model is applied, highlighting the zones of maximum thickness and motor point density to ensure precise and effective BoNT-A delivery. Enhanced with high-resolution ultrasound images and dynamic scanning techniques, this visual guide supports clinicians in performing safe, targeted injections. It serves as both an educational and practical reference for the ultrasound-guided treatment of spasticity in the proximal lower limb, completing the series and offering a standardized framework for comprehensive BoNT-A management. By promoting accurate toxin delivery, this approach is expected to improve functional mobility, reduce spasticity-related complications, and optimize patient-centered outcomes in rehabilitation settings. Full article

Objectives: Muscles and their tendons present a considerable diversity of morphological variations. The aim of this study was to explore variants of muscles and tendons from compartments of the thigh and to raise awareness about potential problems during ultrasound examination. Materials and Methods: This comprehensive review of the literature was created on the basis of scientific articles sourced from PubMed. The search included all relevant papers related to the topic, ensuring that the most up-to-date studies were incorporated. In order to achieve these results, we created the exclusion criteria and extracted papers that did not meet the requirements of our review. Relevant papers were incorporated, and tracking of citations was fulfilled. The described method allowed for a broad yet detailed understanding, ensuring that the review of the literature covers all key aspects of the presented research. Results: Various aspects of thigh muscle anomalies were already undertaken; however, as this study has shown, current knowledge, while valuable, is insufficient to draw definitive conclusions regarding the prevalence and clinical implications of these muscle variations. A more robust body of ultrasound-based research is essential to accurately characterize these anomalies, establish their frequency, and assess their impact on clinical decision-making, including diagnostic accuracy, surgical planning, and therapeutic interventions. Conclusions: Numerous anatomical variations of the thigh muscles and tendons that were described in literature over the years might have clinical implications and could lead to mistakes during diagnosis by ultrasound imaging. Full article

Integrated serological surveillance (serosurveillance) involves testing for antibodies to multiple pathogens (or species) simultaneously and can be achieved using multiplex bead assays (MBAs). This systematic review aims to describe pathogens studied using MBAs, the operational implementation of MBAs, and how the data generated were synthesised. In November and December 2023, four databases were searched for studies utilising MBAs for the integrated serosurveillance of infectious diseases. Two reviewers independently screened and extracted data regarding the study settings and population, methodology, seroprevalence results, and operational implementation elements. Overall, 4765 studies were identified; 47 were eligible for inclusion, of which 41% (n = 19) investigated multiple malaria species, and 14% performed concurrent surveillance of malaria in combination with other infectious diseases (n = 14). Additionally, 14 studies (29%) investigated a combination of multiple infectious diseases (other than malaria), and seven studies examined a combination of vaccine-preventable diseases. Haiti (n = 8) was the most studied country, followed by Ethiopia (n = 6), Bangladesh (n = 3), Kenya (n = 3), and Tanzania (n = 3). Only seven studies were found where integrated serosurveillance was the primary objective. The synthesis of data varied and included the investigation of age-specific seroprevalence (n = 25), risk factor analysis (n = 15), and spatial analysis of disease prevalence (n = 8). This review demonstrated that the use of MBAs for integrated surveillance of multiple pathogens is gaining traction; however, more research and capabilities in lower- and middle-income countries are needed to optimise and standardise sample collection, survey implementation, and the analysis and interpretation of results. Geographical and population seroprevalence data can enable targeted public health interventions, highlighting the potential and importance of integrated serological surveillance as a public health tool. Full article

The elimination of lymphatic filariasis (LF) as a public health problem remains an ongoing challenge in the Pacific region. This study reports on antigen (Ag) and microfilaria (Mf) prevalence in Samoa in 2019, 7–9 months after the completion of the first round of triple-drug mass drug administration (MDA). It evaluates the effectiveness of the intervention for reducing Ag prevalence to below a 2% threshold, and how this differs between 5–9-year-olds and ≥10-year-olds. We surveyed 30 randomly selected and five purposefully selected primary sampling units (PSUs) in Samoa in 2018 (1–3 months post-triple-drug MDA) and, again, in 2019. In each PSU, we conducted a community survey of 15–20 households and a convenience survey of 5–9-year-old children. A finger-prick blood sample was collected from all participants to test for Ag and Mf. Demographic details were also collected. There was no significant change in adjusted Ag prevalence in the 30 randomly selected PSUs between 2018 (3.9% [95% CI: 2.7–5.6%]) and 2019 (4.1% [95% CI 2.7–5.9%]). Significantly higher Ag prevalence was observed in participants aged ≥10 years (4.6%, 95% CIs 3.0–6.7%) compared to 5–9-year-olds (1.1%, 95% CIs 0.5–2.2%), supporting existing evidence that post-MDA surveillance should not be based on Ag prevalence among 6–7-year-olds. A single round of triple-drug MDA was insufficient to break LF transmission in Samoa 7–9 months post-MDA. Full article

Immobilized enzyme reactors (IMERs) are critical tools for developing novel oligosaccharides based on the enzymatic catalysis of polysaccharides. In this paper, a novel glucuronan lyase from Peteryoungia rosettiformans was produced, purified, and then immobilized on a CIM^® CDI disk for cleaving glucuronan. The results showed that around 63.6% of glycuronan lyases (800.9 μg) were immobilized on the disk. The V_max values of immobilized glucuronan lyases did not significantly change (56.9 ± 4.7 μM∙min⁻¹), while the K_m values (0.310 ± 0.075 g∙L⁻¹) increased by 2.5 times. It is worth noting that immobilized glucuronan lyases overcame the catalytic inhibition of free enzymes observed under high glucuronan concentrations (0.5–2 g∙L⁻¹). circumscribed central composite design (CCCD) and response surface methodology (RSM) showed that glucuronan concentration, flow rate, and reaction time significantly affected the yield of oligoglucuronans. The degree of polymerization (DP) of degraded glucuronan ranged from DP 2–8 according to the results obtained by high performance anion exchange chromatography coupled with a pulsed amperometric detector (HPAEC-PAD). The IMER retained 50.9% activity after running 2373 column volumes of glucuronan. Finally, this glucuronan lyase reactor was tentatively connected to an immobilized laccase reactor to depolymerize, and gallic acid (GA) was added to glucuronan. Approximately 8.5 mg of GA was added onto 1 g of initial glucuronan, and the GA–oligoglucuronan conjugates showed notable antioxidant activity. Full article

The atmospheric environment is formed under the influence of local and distant sources as a result of horizontal and vertical transport. In the present work, microbiological analysis of 604 samples of atmospheric aerosol collected in the period from September 2020 to September 2023 at four sites differing in anthropogenic load, located in Novosibirsk and the region, was carried out. Day and night aerosol samples were collected during 12 h every two weeks by filtration using Sartorius reinforced Teflon membranes, then sown on a set of nutrient media. The taxonomic affiliation of the isolated microbial isolates was determined based on phenotypic characteristics and analysis of 16S rRNA gene nucleotide sequences. Changes in the composition and concentration of culturable microorganisms depending on the season, time of day, and site of aerosol sampling were observed. In winter, lower fungi and bacteria of the genera Bacillus, Staphylococcus, Micrococcus dominated with an average concentration from zero to 12.5 CFU/m³ of aerosol. In the warm period, the concentration and diversity of cocci, spore-forming and non-spore-forming bacteria, actinomycetes, and fungi (up to 1970 CFU/m³), among which pathogenic microorganisms were found, increased sharply in aerosols. The use of 16S metabarcoding techniques has greatly expanded the range of aerosols’ microbial diversity detectable. Full article

This study re-introduces a protein-free rapid test method for nucleic acids on paper based lateral flow assays utilizing special multichannel nitrocellulose membranes and DNA-Gold conjugates, achieving significantly enhanced sensitivity, easier protocols, reduced time of detection, reduced costs of production and advanced multiplexing possibilities. A protein-free nucleic acid-based lateral flow assay (NALFA) with a limit of detection of 1 pmol of DNA is shown for the first time. The total production duration of such an assay was successfully reduced from the currently known several days to just a few hours. The simplification and acceleration of the protocol make the method more accessible and practical for various applications. The developed method supports multiplexing, enabling the simultaneous detection of up to six DNA targets. This multiplexing capability is a significant improvement over traditional line tests and offers more comprehensive diagnostic potential in a single assay. The approach significantly reduces the run time compared to traditional line tests, which enhances the efficiency of diagnostic procedures. The protein-free aspect of this assay minimizes the prevalent complications of cross-reactivity in immunoassays especially in cases of multiplexing. It is also demonstrated that the NALFA developed in this study is amplification-free and hence does not rely on specialized technicians, nor does it involve labour-intensive steps like DNA extraction and PCR processes. Overall, this study presents a robust, efficient, and highly sensitive platform for DNA or RNA detection, addressing several limitations of current methods documented in the literature. The advancements in sensitivity, cost reduction, production time, and multiplexing capabilities mark a substantial improvement, holding great potential for various applications in diagnostics, forensics, and molecular biology. Full article

The reliable transfer of bioprocesses from single-use bioreactors (SUBs) of different scales to conventional stainless steel stirred-tank bioreactors is of steadily growing interest. In this publication, a scale-up study for SUBs with volumes of 200 L and 2000 L and the transfer to an industrial-scale conventional stainless steel stirred-tank bioreactor with a volume of 15,000 L is presented. The scale-up and transfer are based on a comparison of mixing times and the modeling of volumetric mass transfer coefficients k_La, measured in all three reactors in aqueous PBS/Kolliphor solution. The mass transfer coefficients are compared with the widely used correlation of van’t Riet at constant stirrer tip speeds. It can be shown that a van’t Riet correlation enables a robust and reliable prediction of mass transfer coefficients on each scale for a wide range of stirrer tip speeds and aeration rates. The process transfer from single-use bioreactors to conventional stainless steel stirred-tank bioreactors is proven to be uncritical concerning mass transfer performance. This provides higher flexibility with respect to bioreactor equipment considered for specific processes. Full article

The main goal of this study was to assess whether the presence of peripheral arterial disease (PAD) correlates with increased inflammatory cell infiltration. An observational, single-centre, and prospective study was conducted from January 2018 to July 2022. Clinical characteristics and anthropometric measures were registered. Consecutive PAD patients with surgical indications for a common femoral artery approach and patients with varicose veins with an indication for surgical ligation of the saphenofemoral junction were included. In both groups, samples of sartorius skeletal muscle, subcutaneous adipose tissue (SAT), and perivascular adipose tissue (PVAT) were collected from the femoral region. We analysed the characteristics of adipocytes and the presence of haemorrhage and inflammatory cells in the samples of PVAT and SAT via haematoxylin–eosin staining. We found that patients with PAD had significantly more inflammatory cells in PVAT [16 (43.24%) vs. 0 (0%) p = 0.008]. Analysing SAT histology, we observed that patients with PAD had significantly more CD45+ leucocytes upon immunohistochemical staining [32 (72.73%) vs. 3 (27.27%) p = 0.005]. Upon analysing skeletal muscle histology with haematoxylin–eosin staining, we evaluated skeletal fibre preservation, as well as the presence of trauma, haemorrhage, and inflammatory cells. We registered a significantly higher number of inflammatory cells in patients with PAD [well-preserved skeletal fibres: PAD = 26 (63.41%) vs. varicose veins = 3 (37.50%) p = 0.173; trauma: PAD = 4 (9.76%) vs. varicose veins = 2 (25.00%) p = 0.229; haemorrhage: PAD = 6 (14.63%) vs. varicose veins = 0 (0%) p = 0.248; inflammatory cells: PAD = 18 (43.90%) vs. varicose veins = 0 (0%) p = 0.018]. Patients with PAD had a higher number of inflammatory cells in skeletal muscle and adipose tissue (PVAT and SAT) when compared with those with varicose veins, emphasizing the role of inflammation in this group of patients. Full article

The current study sought to investigate the changes in surface hardness, roughness, and moisture absorption of the Vertex ThermoSens polymer (Vertex Dental, 3D Systems, The Netherlands) following immersion in artificial saliva for various periods (7, 14, and 28 days). A total of 60 rectangular specimens with dimensions of 20 mm in length, 20 mm in width, and 3 mm in thickness were made. Due to insufficient mold solidification, these specimens were made utilizing the injection molding process. A Mitutoyo Surftest 4 roughness meter (Mitutoyo, Aurora, IL, USA) was used to measure the surface roughness of the test materials. The ThermoSens polymer hardness was assessed using the Shor method and D—HSD scale, while absorption was measured with a Sartorius analytical balance. Results indicated the highest mean hardness after 28 days (M = 77.6) (Surface 1) and the lowest for the control group (M = 59) (Surface 2). The maximum surface roughness occurred in direction 2.2 pre-immersion (Ra = 2.88 μm) and 7 days post-removal (Ra = 2.95 μm). The control group exhibited the lowest absorption (Wsp = 1.524 mg/mm³), with the highest mean values over 28 days (Wsp = 1.541 mg/mm³). The elevated flask and plaster temperature slowed polymer solidification, resulting in longer macromolecules and improved mechanical properties and surface features. Full article

Adeno-associated viruses (AAV) are currently predominant viral transfer tools for gene therapy, and efforts are being made to design faster and more efficient methods and technologies for their manufacturing. The early selection of high-performing filters is essential for developing an ultrafiltration and diafiltration (UF/DF) process, especially when feed material is scarce, and timelines are short. However, few methods and technologies exist to enable process optimization with multiple variations in a single run. In this study, we explored the potential of Ambr^® Crossflow for high-throughput, automated screening of different membrane materials, pore sizes and different process conditions for the UF/DF step of AAV8. The best overall performance was obtained with a 100 kDa PES flat sheet cassette. The UF/DF process was further transferred to a larger scale to the Sartoflow^® Smart Tangential Flow Filtration (TFF) system using a 100 kDa PES Sartocon^® Slice 200 cassette and compared to a 100 kDa PES hollow fiber. Virus recovery, permeate flux and total protein removal values of the flat sheet cassette were similar to those achieved in small-scale devices, and higher than those of the hollow fiber, thus demonstrating similar performance at a larger process scale. The high-throughput, automated method described herein allowed to screen multiple materials and process parameters of a UF/DF process in a time- and resource-efficient way, making it a useful tool to accelerate early-stage downstream process development of AAV. Full article