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Abstract In this study, we aimed to establish a technique for intraprostatic implantation of prostate cancer (PCa) spheroids and to identify the impact of three-dimensional organization of PCa cells on tumor progression and metastasis in a representative in vivo model. 40,000 LNCaP cells were implanted into the prostate of immunodeficient SCID mice either as single cells (n = 8) or as preformed 3D spheroids (n = 8). For a follow up of 20 weeks, tumor growth was monitored by serum PSA and high-resolution 3D ultrasonography. Eventually, animals were sacrificed and autopsied. The organ dissects were analyzed for the presence of metastases by histology (H&E) and immunohistochemistry (AMACR, AR, Ki-67, CK5, CK8, E-Cadherin, Vimentin). Solid intraprostatic tumors developed in 50% of mice after spheroid implantation and in 50% of mice after implantation of a single cells. Primary tumors of LNCaP spheroids evolved earlier, exhibiting a shorter tumor doubling time whilst developing larger tumor volumes, which was reflected by a higher immunohistochemical expression of Ki-67 and AR, too. Spheroid tumors established lung and lymph node metastases in 75% of mice, in contrast to 50% of mice after single cell implantation. Our technique enables a variety of studies regarding the influence of the tumor microenvironment on PCa progression.

The rapid advancement of AI algorithms presents new opportunities for sensing technologies based on image recognition, such as real-time crystallization monitoring. This work investigates the use of computer vision to detect and size crystals in a lab scale draft tube baffle crystallizer (DTBC). A specially developed analytical bypass was implemented on the DTBC to enable a low-influence analysis without invasive intrusion into the process. By utilizing AI models such as YouOnlyLookOnce version 8 (YOLOv8), YOLOv8 Segmentation (YOLO8seg), and the convolutional network for biomedical image segmentation U-Net, this study assesses their effectiveness in determining crystal size distributions from photometric images. While U-Net was deemed unsuitable due to computational demands and accuracy issues, YOLOv8 and YOLO8seg performed better in terms of efficiency and precision. YOLO8seg, specifically, achieved the highest accuracy, with a mean average precision (mAP) of 82.3%, and excelling in detecting larger crystals, but underperforming with crystals smaller than 90 µm. Despite this limitation, YOLO8seg was able to compete with the manual methods and was superior to the state-of-the-art algorithm mask region convolutional neural network (Mask R-CNN) in terms of accuracy. The study suggests that specific training and adaptation of the imaging conditions could further improve the crystal detection performance.

Pharmacy compounding, the art and science of preparing customized medications to meet individual patient needs, is on the verge of transformation. Traditional methods of compounding often involve manual and time-consuming processes, presenting challenges in terms of consistency, dosage accuracy, quality control, contamination, and scalability. However, the emergence of cutting-edge technologies has paved a way for a new era for pharmacy compounding, promising to redefine the way medications are prepared and delivered as pharmacy-tailored personalized medicines. In this multi-site study, more than 30 hospitals and community pharmacies from eight countries in Europe utilized a novel automated dosing approach inspired by 3D printing for the compounding of non-sterile propranolol hydrochloride tablets. CuraBlend® excipient base, a GMP-manufactured excipient base (pharma-ink) intended for automated compounding applications, was used. A standardized study protocol to test the automated dosing of tablets with variable weights was performed in all participating pharmacies in four different iterative phases. Integrated quality control was performed with an in-process scale and NIR spectroscopy supported by HPLC content uniformity measurements. In total, 6088 propranolol tablets were produced at different locations during this study. It was shown that the dosing accuracy of the process increased from about 90% to 100% from Phase 1 to Phase 4 by making improvements to the formulation and the hardware solutions. The results indicate that through this automated and quality controlled compounding approach, extemporaneous pharmacy manufacturing can take a giant leap forward towards automation and digital manufacture of dosage forms in hospital pharmacies and compounding pharmacies.

Abstract Background Overweight and obesity represent huge concerns for children's physical and mental well-being. This study examined the relationship between body mass index (BMI) and health-related quality of life (HRQoL), somatoform complaints, and behavioral problems in children and adolescents. Additionally, the influence of sex, age, and socioeconomic status (SES) on these associations was considered. Methods In total, we studied 2350 participants between the ages of 4 and 18 years (1213 4- to 10-years-old (child sample) and 1137 11-to 18-year-olds (adolescent sample)). To assess HRQoL, somatoform complaints, and behavioral difficulties, we applied the KIDSCREEN-27, a short form of the Giessen Complaints Questionnaire, and the Strengths and Difficulties Questionnaire (SDQ). The BMI was transformed to BMI standard deviation scores (BMI-SDS), according to German gender- and age-specific reference data. Associations were investigated using linear regression analyses. Each association was checked for interaction with sex, age, and SES. Results Regarding HRQoL, we found worsening scores in physical well-being and psychological well-being with increasing BMI-SDS. Somatoform complaints were not significantly associated with BMI-SDS. Conduct problems, peer relationship problems, and emotional problems (the latter only in the adolescent sample) were positively associated with BMI-SDS. While we did not observe any significant interactions with sex, we found some significant interactions with age and/or SES. Conclusion Our findings highlight the importance of mental difficulties in children and adolescents with higher BMI and, consequently, underline the relevance of including psychological interventions in the treatment of overweight or obesity.

Arranging products in stores according to planograms, optimized product arrangement maps, is an important sales enabler and necessary for keeping up with the highly competitive modern retail market. Key benefits of planograms include increased efficiency, maximized retail store space, increased customer satisfaction, visual appeal, and increased revenue. The planograms are realized into product arrangements by humans, a process that is prone to mistakes. Therefore, for optimal merchandising performance, the planogram compliance of the arrangements needs to be evaluated from time to time. We investigate utilizing a computer vision problem setting—retail product detection—to automate planogram compliance evaluation. Retail product detection comprises product detection and classification. The detected and classified products can be compared to the planogram in order to evaluate compliance. In this paper, we propose a novel retail product detection pipeline combining a Gaussian layer network product proposal generator and domain invariant hierarchical embedding (DIHE) classifier. We utilize the detection pipeline with RANSAC pose estimation for planogram compliance evaluation. As the existing metrics for evaluating the planogram compliance evaluation performance assume unrealistically that the test image matches the planogram, we propose a novel metric, called normalized planogram compliance error (EPC), for benchmarking real-world setups. We evaluate the performance of our method with two datasets: the only open-source dataset with planogram evaluation data, GP-180, and our own dataset collected from a large Nordic retailer. Based on the evaluation, our method provides an improved planogram compliance evaluation pipeline, with accurate product location estimation when using real-life images that include entire shelves, unlike previous research that has only used images with few products. Our analysis also demonstrates that our method requires less processing time than the state-of-the-art compliance evaluation methods.

Bees (Hymenoptera) are the most important group of pollinators with about 20,000 known species worldwide. Despite the critical role of bees in providing pollination services, studies aiming at understanding which species are present in Tanzania across disturbance gradients are scarce. Limited taxononomic information for the existing and unidentified bees make their conservation strategies haphazard. Here, we present a dataset of bees obtained in surveys in the Kilimanjaro, Arusha and Manyara regions, in northern Tanzania. Our findings serve as baseline data necessary for understanding, the diversity of bees in the northern parts of the country, which is a critical step in devising robust conservation and monitoring strategies for their populations.In this paper, we present information on a total of 45 bee species in 20 genera and 4 families, sampled using a combination of sweep-netting and pantrap methods. Most species, (27 species, 60%) belong to the family Halictidae followed by 16 species (35.5%) from the family Apidae. Megachilidae and Andrenidae were least represented, each with only one species (2.2%). From a total of 953 occurrences, highest numbers were recorded in Kilimanjaro region (n= 511), followed by Arusha (n= 410), and Manyara (n= 32), but this pattern reflects the sampling efforts of the research project rather than real bias in the distributions of bees in northern Tanzania.

Originally published as ‘Michelangelo og marmoret (1876),’ Axel Sophus Guldberg ed., Fra Videnskabens Verden Almenfattelige Smaaskrifter af danske og norske Videnskabsmænd, 3rd ser., Copenhagen: Gad, 1876, Julius Lange, Billedkunst skildringer och studier fra hjemmet og udlandet, København: P. G. Philipsens forlag, 1884, pp. 68-128. Reprinted: Udvalgte skrifter af Julius Lange, udgivne af Georg Brandes og P. Købke, Tredje bind, København: Det nordiske forlag, 1903, pp. 42-77. And in German translation as ‘Michelangelo und der Marmor,’ Julius Lange’s ausgewählte Schriften (1875-1885), herausgegeben von Georg Brandes und Peter Købke, unter Mitwirkung von Alfred Wien übersetzt von Ida Anders, Strasbourg: Heitz, Erster Band, 1911, pp. 45-78.

Originally published as ‘Haenden paa Brystet,’ Tilskueren: Maanedsskrift for Literatur, Samfundsspørgsmaal og almenfattelige videnskabelige skildringer, 4th year, 1887, June-July, pp. 455-471, August pp. 571-588, reprinted: Udvalgte skrifter af Julius Lange, udgivne af Georg Brandes og P. Købke, Andet bind, København: Det nordiske forlag, 1901, pp. 10-48.

Originally published as ‘Et Udtryks Historie’, Tilskueren: Maanedsskrift for Litteratur, Samfundsspørgsmaal og Almenfattelige Videnskabelige Skildringer, vol. 12, August-September, 1895, pp. 565-583, 674-705. Reprinted: Udvalgte skrifter af Julius Lange, ed. Georg Brandes and Peter Købke, vol. 2, Copenhagen: Det nordiske forlag, 1901, pp. 89-136.

Originally published as ‘Et Motivs Historie’, Nordisk Tidskrift for vetenskap, konst och industri, Letterstedtska foereningen, 1888, pp. 475-494. Reprinted: Udvalgte skrifter af Julius Lange, udgivne af Georg Brandes og P. Købke, Andet bind, København: Det nordiske forlag, 1901, pp. 69-88.

Originally published as ‘Attiske Gravmæler,’ Nordisk Tidskrift for vetenskap, konst och industri, 1896, pp. 27-45, reprinted: Udvalgte skrifter af Julius Lange, udgivne af Georg Brandes og Peter Købke, Andet bind, København: Det nordiske forlag, 1901, pp. 385-400.

Since November 2019 the SARS-CoV-2 pandemic has caused nearly 200 million infection and more than 4 million deaths globally (Updated information from the World Health Organization, as on 2nd Aug 2021). Within only one year into the pandemic, several vaccines were designed and reached approval for the immunization of the world population. The remarkable protective effects of the manufactured vaccines are demonstrated in countries with high vaccination rates, such as Israel and UK. However, limited production capacities, poor distribution infrastructures and political hesitations still hamper the availability of vaccines in many countries. In addition, due to the emergency of SARS-CoV-2 variants with immune escape properties towards the vaccines the global numbers of new infections as well as patients developing severe COVID-19, remains high. New studies reported that about 8% of infected individuals develop long term symptoms with strong personal restrictions on private as well as professional level, which contributes to the long socioeconomic problems caused by this pandemic. Until today, emergency use-approved treatment options for COVID-19 are limited to the antiviral Remdesivir, a nucleoside analogue targeting the viral polymerase, the glucocorticosteroide Dexamethasone as well as neutralizing antibodies. The therapeutic benefits of these treatments are under ongoing debate and clinical studies assessing the efficiency of these treatments are still underway. To identify new therapeutic treatments for COVID-19, now and by the post-pandemic era, diverse experimental approaches are under scientific evaluation in companies and scientific research teams all over the world. To accelerate clinical translation of promising candidates, repurposing approaches of known approved drugs are specifically fostered but also novel technologies are being developed and are under investigation. This review summarizes the recent developments from the lab bench as well as the clinical status of emerging therapeutic candidates and discusses possible therapeutic entry points for the treatment strategies with regard to the biology of SARS-CoV-2 and the clinical course of COVID-19.

Respiratory infections with newly emerging zoonotic viruses such as SARS-CoV-2, the etiological agent of COVID-19, often lead to the perturbation of the human innate and adaptive immune responses causing severe disease with high mortality. The responsible mechanisms are commonly virus-specific and often include either over-activated or delayed local interferon responses, which facilitate efficient viral replication in the primary target organ, systemic viral spread, and rapid onset of organ-specific and harmful inflammatory responses. Despite the distinct replication strategies, human infections with SARS-CoV-2 and highly pathogenic avian influenza viruses demonstrate remarkable similarities and differences regarding the mechanisms of immune induction, disease dynamics, as well as the long-term sequelae, which will be discussed in this review. In addition, we will highlight some important lessons about the effectiveness of antiviral and immunomodulatory therapeutic strategies that this pandemic has taught us.

Background. Gastric antral vascular ectasia (GAVE) is currently recognized as an important cause of upper gastrointestinal (GI) haemorrhage, being responsible for about 4% of non-variceal upper GI haemorrhages and typically presents in middle-aged females. GAVE, also called “watermelon stomach”, is diagnosed through esophagogastroduodenoscopy and is characterized by the presence of visible columns of red tortuous enlarged vessels along the longitudinal folds of the antrum. The pathogenesis is still obscure and many hypotheses have been proposed such as mechanical stress, humoral and autoimmune factors. In the last two decades, numerous therapeutic strategies have been proposed, including surgical, endoscopic, and medical choices, yet successful treatment of GAVE continues to be a challenge. Currently, given the rapid response, safety, and efficacy, endoscopic ablative modalities have largely usurped medical treatments as first-line therapy, particularly using argon plasma coagulation. The actual GAVE prevalence in patients with end-stage renal disease (ESRD) is not clear, yet in difficult cases it should be considered as a cause of erythropoietin resistance. Case presentation. We report four clinical cases of GAVE syndrome patients diagnosed with stage 4 to 5 chronic kidney disease. All patients presented with anaemia and GI haemorrhage, the origin of which turned out to be GAVE syndrome. Conclusions. GAVE syndrome is a serious condition in ESRD patients, especially in those presenting with treatment-refractory anaemia. Realization of its aetiology and characteristics is essential to suspect, diagnose, and treat gastric ectasia. Only proper diagnosis and well-timed disease treatment can significantly improve a patient’s medical condition and future prognosis.

Background: Biomedical diagnostic and lab automation solutions built on the Lab-on-a-Disc (LoaD) platform has great potential due to their independence from specialised micro-pumps and their ease of integration, through direct pipetting, with manual or automated workflows. However, a challenge for all microfluidic chips is their cost of manufacture when each microfluidic disc must be customized for a specific application. In this paper, we present centrifugal discs with programmable fluidic networks., Results: Based on dissolvable film valves, we present two technologies. The first, based on recently introduced pulse-actuated dissolvable film valves, is a centrifugal disc which, depending on how it is loaded, is configured to perform either six sequential reagent releases through one reaction chamber or three sequential reagent releases through two reaction chambers. In the second approach, we use the previously introduced electronic Lab-on-a-Disc (eLoaD) wireless valve array, which can actuate up to 128 centrifugo-pneumatic dissolvable film valves in a pre-defined sequence. In this approach we present a disc which can deliver any one of 8 reagent washes to any one of four reaction chambers. We use identical discs to demonstrate the first four sequential washes through two reaction chambers and then two sequential washes through four reaction chambers., Significance: These programmable fluidic networks have the potential to allow a single disc architecture to be applied to multiple different assay types and so can offer a lower-cost and more integrated alternative to the standard combination of micro-titre plate and liquid handling robot. Indeed, it may even be possible to conduct multiple different assays concurrently. This can have the effect of reducing manufacturing costs and streamlining supply-chains and so results in a more accessible diagnostic platform., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jan Korvink reports financial support was provided by European Commission. Jens Ducree reports financial support was provided by European Commission. Jens Ducree reports financial support was provided by Science Foundation Ireland. Sarai Torres Delgado reports financial support was provided by National Council of Science and Technology. Jens Ducree reports financial support was provided by Fraunhofer Society for the Advancement of Applied Research., (Copyright © 2023. Published by Elsevier B.V.)

The ability of the centrifugal Lab-on-a-Disc (LoaD) platform to closely mimic the "on bench" liquid handling steps (laboratory unit operations (LUOs)) such as metering, mixing, and aliquoting supports on-disc automation of bioassay without the need for extensive biological optimization. Thus, well-established bioassays, normally conducted manually using pipettes or using liquid handling robots, can be relatively easily automated in self-contained microfluidic chips suitable for use in point-of-care or point-of-use settings. The LoaD's ease of automation is largely dependent on valves that can control liquid movement on the rotating disc. The optimum valving strategy for a true low-cost and portable device is rotationally actuated valves, which are actuated by changes in the disc spin-speed. However, due to tolerances in disc manufacturing and variations in reagent properties, most of these valving technologies have inherent variation in their actuation spin-speed. Most valves are actuated through stepped increases in disc spin-speed until the motor reaches its maximum speed (rarely more than 6000 rpm). These manufacturing tolerances combined with this "analogue" mechanism of valve actuation limits the number of LUOs that can be placed on-disc. In this work, we present a novel valving mechanism called low-high-low serial dissolvable film (DF) valves. In these valves, a DF membrane is placed in a dead-end pneumatic chamber. Below an actuation spin-speed, the trapped air prevents liquid wetting and dissolving the membrane. Above this spin-speed, the liquid will enter and wet the DF and open the valve. However, as DFs take ∼40 s to dissolve, the membrane can be wetted, and the disc spin-speed reduced before the film opens. Thus, by placing valves in a series, we can govern on which "digital pulse" in spin-speeding a reagent is released; a reservoir with one serial valve will open on the first pulse, a reservoir with two serial valves on the second, and so on. This "digital" flow control mechanism allows the automation of complex assays with high reliability. In this work, we first describe the operation of the valves, outline the theoretical basis for their operation, and support this analysis with an experiment. Next, we demonstrate how these valves can be used to automate the solid-phase extraction of DNA on on-disc LAMP amplification for applications in plant pathogen detection. The disc was successfully used to extract and detect, from a sample lysed off-disc, DNA indicating the presence of thermally inactivated Clavibacter michiganensis ssp. michiganensis (Cmm) , a bacterial pathogen on tomato leaf samples., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

By virtue of its ruggedness, portability, rapid processing times, and ease-of-use, academic and commercial interest in centrifugal microfluidic systems has soared over the last decade. A key advantage of the LoaD platform is the ability to automate laboratory unit operations (LUOs) (mixing, metering, washing etc.) to support direct translation of 'on-bench' assays to 'on-chip'. Additionally, the LoaD requires just a low-cost spindle motor rather than specialized and expensive microfluidic pumps. Furthermore, when flow control (valves) is implemented through purely rotational changes in this same spindle motor (rather than using additional support instrumentation), the LoaD offers the potential to be a truly portable, low-cost and accessible platform. Current rotationally controlled valves are typically opened by sequentially increasing the disc spin-rate to a specific opening frequency. However, due lack of manufacturing fidelity these specific opening frequencies are better described as spin frequency 'bands'. With low-cost motors typically having a maximum spin-rate of 6000 rpm (100 Hz), using this 'analogue' approach places a limitation on the number of valves, which can be serially actuated thus limiting the number of LUOs that can be automated. In this work, a novel flow control scheme is presented where the sequence of valve actuation is determined by architecture of the disc while its timing is governed by freely programmable 'digital' pulses in its spin profile. This paradigm shift to 'digital' flow control enables automation of multi-step assays with high reliability, with full temporal control, and with the number of LUOs theoretically only limited by available space on the disc. We first describe the operational principle of these valves followed by a demonstration of the capability of these valves to automate complex assays by screening tomato leaf samples against plant pathogens. Reagents and lysed sample are loaded on-disc and then, in a fully autonomous fashion using only spindle-motor control, the complete assay is automated. Amplification and fluorescent acquisition take place on a custom spin-stand enabling the generation of real-time LAMP amplification curves using custom software. To prevent environmental contamination, the entire discs are sealed from atmosphere following loading with internal venting channels permitting easy movement of liquids about the disc. The disc was successfully used to detect the presence of thermally inactivated Clavibacter michiganensis. Michiganensis (CMM) bacterial pathogen on tomato leaf samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In this paper we present a wirelessly powered array of 128 centrifugo-pneumatic valves that can be thermally actuated on demand during spinning. The valves can either be triggered by a predefined protocol, wireless signal transmission via Bluetooth, or in response to a sensor monitoring a parameter like the temperature, or homogeneity of the dispersion. Upon activation of a resistive heater, a low-melting membrane (Parafilm™) is removed to vent an entrapped gas pocket, thus letting the incoming liquid wet an intermediate dissolvable film and thereby open the valve. The proposed system allows up to 12 heaters to be activated in parallel, with a response time below 3 s, potentially resulting in 128 actuated valves in under 30 s. We demonstrate, with three examples of common and standard procedures, how the proposed technology could become a powerful tool for implementing diagnostic assays on Lab-on-a-Disc. First, we implement wireless actuation of 64 valves during rotation in a freely programmable sequence, or upon user input in real time. Then, we show a closed-loop centrifugal flow control sequence for which the state of mixing of reagents, evaluated from stroboscopically recorded images, triggers the opening of the valves. In our last experiment, valving and closed-loop control are used to facilitate centrifugal processing of whole blood., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The recent rapid growth of microfluidic applications has witnessed the emergence of several particle flow focusing techniques for analysis and/or further processing. The majority of flow focusing techniques employ an external sheath fluid to achieve sample flow focusing independent of the flow rate, in contrast to sheath-free techniques. However, the introduction of a sheath fluid to surround the sample fluid has complicated the device design and fabrication, generally involving multi-layer fabrication and bonding of multiple polydimethylsiloxane (PDMS) layers. Several promising efforts have been made to reduce the complexity of fabrication. However, most of these methods involved the use of inertial/Dean effects, which in turn demanded the use of higher sample flow rates. In this paper, we report a method of flow focusing that uses a sheath fluid to enclose the sample in a single layer of PDMS, and that possesses applicability for a wide range of sample flow rates. This method of flow focusing uses abrupt channel depth variation and a shift of one of the sample-sheath junctions (termed as 'junction-shift') against the direction of the sample flow. This configuration serves to manipulate the sample fluid with respect to the sheath fluid and achieve the desired flow focusing. This design facilitates the attainment of 3D flow focusing in two sequential steps (depth-wise and then along the lateral direction) and in distinct regions, hence enabling the regions to be used in imaging and non-imaging flow cytometric applications, respectively. Simulations were performed to characterize and determine the optimum set of design parameters. Experimental demonstrations of this technique were carried out by focusing fluorescein dye and blood cells in flow.

Typical Lab-on-a-Disc (LoaD) platforms cannot make a continuous measurement while the disc is spinning; this drawback means that the disc usually must be stopped and aligned with a sensor. This can result in measurement errors in time-dependent assays along with inaccuracies due to liquid displacement and bubble formation in the absence of a stabilising centrifugal field. This paper presents a novel concept for a wirelessly electrified-Lab-on-a-Disc (eLoaD) platform that allows continuous measurement of experimental parameters while the disc is spinning. This platform incorporates all the components needed for measurement within the rotating frame of reference, and bidirectional transmission of data outside this reference frame, thus allowing for online measurement independent of the rotation of the disc. The eLoaD platform is conceived in a modular manner whereby an interchangeable and non-disposable 'Application Disc' can be fitted to the eLoaD platform and so the system can be adapted for a range of optical, electrochemical and other measurement types. As an application example, optical readout, using the Application Disc fitted with a silicon photomultiplier, is demonstrated using a tagged chemiluminescent antibody, which is commonly used, for instance, in ELISA assays. The precision of the eLoaD platform is >94%, while its accuracy, when compared to a commercial benchtop luminometer, is higher than 96%. The modular design of this platform will permit extension of this technology to many other LoaD applications.

In this article, we present a novel approach to throughput enhancement in miniaturized microfluidic microscopy systems. Using the presented approach, we demonstrate an inexpensive yet high-throughput analytical instrument. Using the high-throughput analytical instrument, we have been able to achieve about 125,880 cells per minute (more than one hundred and twenty five thousand cells per minute), even while employing cost-effective low frame rate cameras (120 fps). The throughput achieved here is a notable progression in the field of diagnostics as it enables rapid quantitative testing and analysis. We demonstrate the applicability of the instrument to point-of-care diagnostics, by performing blood cell counting. We report a comparative analysis between the counts (in cells per μl) obtained from our instrument, with that of a commercially available hematology analyzer.