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Ashish S Shah,1,2 Brad Sobolewski,3,4 Sabina Chon,3 Bradly Cruse,5 Mike D Glisson,5 Matthew W Zackoff,3,5,6 David Davis,5 Yin Zhang,7 Daniel J Schumacher,3,4 Gary L Geis3– 5 1Department of Pediatrics, University of California – San Diego, San Diego, CA, USA; 2Division of Emergency Medicine, Rady Children’s Hospital, San Diego, CA, USA; 3Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA; 4Division of Emergency Medicine, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA; 5Center for Simulation and Research, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA; 6Division of Critical Care Medicine, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA; 7Emergency Services, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USACorrespondence: Ashish S Shah, Division of Emergency Medicine, Department of Pediatrics, University of California – San Diego, Rady Children’s Hospital, 3020 Children’s Way, San Diego, CA, 92123, USA, Tel +1 920-786-0132, Email ashah3@rchsd.org; ashah@health.ucsd.eduBackground: Early identification of shock is vital in decreasing morbidity and mortality in the pediatric population. Although residents are taught the perfusion portion of the rapid cardiopulmonary assessment at our institution, they perform it at the bedside with 8.4% completing 1 part of the assessment and 9.7% verbalizing their findings. Newer technologies, including virtual reality (VR), offer immersive training to close this clinical gap.Objective: To assess senior pediatric residents’ performance of a perfusion exam and verbalization of their perfusion assessment following VR-based Just-in-Time/Just-in-Place (JITP) training compared to video-based JITP training. We hypothesized that JITP media training was feasible, and VR JITP was more effective than video-based training.Methods: Residents were randomized to VR or video-based training during shifts in the emergency department. Clinical performance was assessed by review of a video-recorded patient encounter using a standardized assessment tool and by an in-person, two question shock assessment. Residents completed a survey assessing attitudes toward their intervention at the time of training.Results: Eighty-five senior pediatric residents were enrolled; 84 completed training. Sixty-four (76%) residents had a patient encounter available for video review (VR 33; Video 31). Fourteen residents in the VR group (42.4%, 95% CI 25.5% to 60.8%) and 13 residents in the video group (41.9%, 95% CI 24.6% to 60.9%) completed a perfusion exam AND verbalized an assessment during their next clinical encounter (X2 p-value 1.00). Fifty-one of 64 residents (79.7%) completed the two-step shock assessment; 50 (98%) agreed with supervising physician’s assessment. VR was rated more effective than reading, low-fidelity manikin, standardized patient encounters, traditional didactic teaching, and online learning. Video was rated more effective than online learning, traditional didactic teaching, and reading.Conclusion: Novel video and VR JITP perfusion exam and assessment trainings are impactful and well-received by senior pediatric residents.Keywords: resident, shock, simulation, training, virtual reality, pediatric

In this paper, the characteristics of 40 so far described virophages—parasites of giant viruses—are given, and the similarities and differences between virophages and satellite viruses, which also, like virophages, require helper viruses for replication, are described. The replication of virophages taking place at a specific site—the viral particle factory of giant viruses—and its consequences are presented, and the defence mechanisms of virophages for giant virus hosts, as a protective action for giant virus hosts—protozoa and algae—are approximated. The defence systems of giant viruses against virophages were also presented, which are similar to the CRISPR/Cas defence system found in bacteria and in Archea. These facts, and related to the very specific biological features of virophages (specific site of replication, specific mechanisms of their defensive effects for giant virus hosts, defence systems in giant viruses against virophages), indicate that virophages, and their host giant viruses, are biological objects, forming a ‘novelty’ in biology.

Abstract Chronic hepatitis C (CHC) is associated with the development of metabolic disorders, including both hepatic and extra-hepatic insulin resistance (IR). Here, we aimed at identifying liver-derived factor(s) potentially inducing peripheral IR and uncovering the mechanisms whereby HCV can regulate the action of these factors. We found ANGPTL4 (Angiopoietin Like 4) mRNA expression levels to positively correlate with HCV RNA (r = 0.46, p

Abstract Background In spite of the fact that recombinant enzymes are preferably biotechnologically obtained using recombinant clones, the purification of proteins from native microorganisms, including those encoded by bacteriophages, continues. The native bacteriophage protein isolation is often troubled by large volumes of the infected bacterial cell lysates needed to be processed, which is highly undesired in scaled-up industrial processing. A well-known ammonium sulphate fractionation is often a method of choice during purification of the native bacteriophage protein. However, this method is time-consuming and cumbersome, and requires large amounts of the relatively expensive reagent. Thus, other effective and inexpensive methods of reversible protein precipitation are highly desirable. We have previously characterized thermophilic TP-84 bacteriophage, defined a new genus TP84virus within Siphoviridae family, conducted the TP-84 genome annotation and proteomic analysis. The longest Open Reading Frame (ORF) identified in the genome is TP84_26. We have previously annotated this ORF as a hydrolytic enzyme depolymerizing the thick polysaccharides host’s capsule. Results The TP84_26 ‘capsule depolymerase’ (depolymerase) is a large, 112 kDa protein, biosynthesized by the infected Geobacillus stearothermophilus 10 (G. stearothermophilus 10) cells. The TP84_26 protein biosynthesis was confirmed by three approaches: (i) purification of the protein of the expected size; (ii) mass spectrometry (LC–MS) analysis and (iii) detection of the enzymatic activity toward G. stearothermophilus polysaccharide capsules. Streptomycin-resistant mutant of the host was generated and microbiological aspects of both the TP-84 and G. stearothermophilus 10 were determined. A new variant of polyethyleneimine (PEI)-mediated purification method was developed, using the novel TP-84 depolymerase as a model. The enzyme was characterized. Three depolymerase forms were detected: soluble, unbound proteins in the bacteriophage/cells lysate and another integrated into the TP-84 virion. Conclusions The novel TP-84 depolymerase was purified and characterized. The enzyme exists in three forms. The soluble, unbound forms are probably responsible for the weakening of the capsules of the uninfected bacterial cells. The form integrated into virion particles may generate a local passage for the invading TP-84. The developed PEI purification method appears well suited for the scaled-up or industrial production of bacteriophage proteins.

IntroductionHealth status, sickness absence, and nurses’ attrition have a direct impact on the quality of care provided and patients’ health outcomes. The Covid-19 pandemic exacerbated issues that existed within the Polish healthcare system prior to the pandemic, including staff shortages, low wages, and system inadequacies. The aim of this study was to investigate how nurses during the Covid-19 pandemic period rated the burdensomeness of job characteristics and their mental health status, as well as the correlations between factors directly caused by the Covid-19 pandemic and nurses’ subjective assessments of job characteristics and mental health.MethodThe cross-sectional study was conducted in January 2022, in Poland and involved 796 registered nurses working in hospitals.ResultsDespite the pandemic’s sweeping societal effects, this research finds limited alteration in nurses’ perceptions of job stress and self-assessed mental health. Factors such as contact with infected patients, quarantine, and isolation do not appear to substantially modify mental health perceptions among nurses. Intriguingly, nurses subjected to COVID-19 testing report heightened stress and compromised mental health.ConclusionThe interplay of diverse factors influencing the well-being of nurses is intricately complex. It is advisable to prudently execute interventions and strategies to address the pandemic, aiming to alleviate its potential adverse effects on the mental health of nurses.

This paper presents two designs of millimeter-wave single-pole single-throw switches based on AlGaN/GaN heterostructure. The switches are based on two approaches to the travelling wave concept with Schottky barrier shunt elements integrated into a coplanar waveguide. The first design utilizes a two-stage artificial transmission line topology with distributed shunt elements in Schottky diode configuration. The second one employs a single electrically large shunt element configured as a high electron mobility transistor. Both arrangements allow the switches to achieve very high operational bandwidth, exceeding 100 GHz, without the requirement for very high lithography resolution. Examined switches were fabricated in a 2- $\mu \text{m}$ GaN-on-SiC process. Measurement results demonstrate upper operating frequencies exceeding 114.5 GHz starting from 14 GHz or even DC, depending on the design. The W band on-off ratio over 17 dB and 21 dB is achieved by the first and second construction, respectively. Measurements of transient parameters show that high switching speed can also be achieved (rise/fall time as low as ≤17 ns for the first construction).

Background and Purpose Cerebral edema (CED) in ischemic stroke can worsen prognosis and about 70% of patients who develop severe CED die if treated conservatively. We aimed to describe incidence, risk factors and outcomes of CED in patients with extensive ischemia. Methods Oservational study based on Safe Implementation of Treatments in Stroke-International Stroke Treatment Registry (2003–2019). Severe hemispheric syndrome (SHS) at baseline and persistent SHS (pSHS) at 24 hours were defined as National Institutes of Health Stroke Score (NIHSS) >15. Outcomes were moderate/severe CED detected by neuroimaging, functional independence (modified Rankin Scale 0–2) and death at 90 days. Results Patients (n=8,560) presented with SHS and developed pSHS at 24 hours; 82.2% received intravenous thrombolysis (IVT), 10.5% IVT+thrombectomy, and 7.3% thrombectomy alone. Median age was 77 and NIHSS 21. Of 7,949 patients with CED data, 3,780 (47.6%) had any CED and 2,297 (28.9%) moderate/severe CED. In the multivariable analysis, age 128.5 mg/dL (RR, 1.21), and decreased level of consciousness (RR, 1.14) were associated with moderate/severe CED (for all P

Transient terahertz time-domain spectroscopy (THz-TDS) imaging has emerged as a novel non-ionizing and noninvasive biomedical imaging modality, designed for the detection and characterization of a variety of tissue malignancies due to their high signal-to-noise ratio and submillimeter resolution. We report our design of a pair of aspheric focusing lenses using a commercially available lens-design software that resulted in about 200 × 200-μm2 focal spot size corresponding to the 1-THz frequency. The lenses are made of high-density polyethylene (HDPE) obtained using a lathe fabrication and are integrated into a THz-TDS system that includes low-temperature GaAs photoconductive antennae as both a THz emitter and detector. The system is used to generate high-resolution, two-dimensional (2D) images of formalin-fixed, paraffin-embedded murine pancreas tissue blocks. The performance of these focusing lenses is compared to the older system based on a pair of short-focal-length, hemispherical polytetrafluoroethylene (TeflonTM) lenses and is characterized using THz-domain measurements, resulting in 2D maps of the tissue refractive index and absorption coefficient as imaging markers. For a quantitative evaluation of the lens effect on the image resolution, we formulated a lateral resolution parameter, R2080, defined as the distance required for a 20–80% transition of the imaging marker from the bare paraffin region to the tissue region in the same image frame. The R2080 parameter clearly demonstrates the advantage of the HDPE lenses over TeflonTM lenses. The lens-design approach presented here can be successfully implemented in other THz-TDS setups with known THz emitter and detector specifications.

Bacteriophages associated with thermophiles are gaining increased attention due to their pivotal roles in various biogeochemical and ecological processes, as well as their applications in biotechnology and bionanotechnology. Although thermophages are not suitable for controlling bacterial infections in humans or animals, their individual components, such as enzymes and capsid proteins, can be employed in molecular biology and significantly contribute to the enhancement of human and animal health. Despite their significance, thermophages still remain underrepresented in the known prokaryotic virosphere, primarily due to limited in-depth investigations. However, due to their unique properties, thermophages are currently attracting increasing interest, as evidenced by several newly discovered phages belonging to this group. This review offers an updated compilation of thermophages characterized to date, focusing on species infecting the thermophilic bacilli. Moreover, it presents experimental findings, including novel proteomic data (39 proteins) concerning the model TP-84 bacteriophage, along with the first announcement of 6 recently discovered thermophages infecting Geobacillus thermodenitrificans: PK5.2, PK2.1, NIIg10.1, NIIg2.1, NIIg2.2, and NIIg2.3. This review serves as an update to our previous publication in 2021.

The TP-84 bacteriophage, which infects Geobacillus stearothermophilus strain 10 (G. stearothermophilus), has a genome size of 47.7 kilobase pairs (kbps) and contains 81 predicted protein-coding ORFs. One of these, TP84_26 encodes a putative tail fiber protein possessing capsule depolymerase activity. In this study, we cloned the TP84_26 gene into a high-expression Escherichia coli (E. coli) system, modified its N-terminus with His-tag, expressed both the wild type gene and His-tagged variant, purified the recombinant depolymerase variants, and further evaluated their properties. We developed a direct enzymatic assay for the depolymerase activity toward G. stearothermophilus capsules. The recombinant TP84_26 protein variants effectively degraded the existing bacterial capsules and inhibited the formation of new ones. Our results provide insights into the novel TP84_26 depolymerase with specific activity against thermostable G. stearothermophilus and its role in the TP-84 life cycle. The identification and characterization of novel depolymerases, such as TP84_26, hold promise for innovative strategies to combat bacterial infections and improve various industrial processes.

The traditional laboratory investigation uses a procedure written on paper; students then record their responses on a supplied data page or laboratory notebook. In an attempt to make this process more efficient, the use of a Learning Management System (in this case D2L) was used to present the material and collect student feedback. Each student had a University supplied laptop, read the procedure from the screen and submitted answers through D2L. As anticipated there was no change in content knowledge. However surprisingly, subjects felt the paper and pencil approach was easier to use than computer entry. In a subsequent study, the same population in the same course completed an on-line only lecture and lab course. We will discuss the student's reaction to this on-line class.

Summary: Error-related potentials (ErrPs) are a prominent electroencephalogram (EEG) correlate of performance monitoring, and so crucial for learning and adapting our behavior. It is poorly understood whether ErrPs encode further information beyond error awareness. We report an experiment with sixteen participants over three sessions in which occasional visual rotations of varying magnitude occurred during a cursor reaching task. We designed a brain-computer interface (BCI) to detect ErrPs that provided real-time feedback. The individual ErrP-BCI decoders exhibited good transfer across sessions and scalability over the magnitude of errors. A non-linear relationship between the ErrP-BCI output and the magnitude of errors predicts individual perceptual thresholds to detect errors. We also reveal theta-gamma oscillatory coupling that co-varied with the magnitude of the required adjustment. Our findings open new avenues to probe and extend current theories of performance monitoring by incorporating continuous human interaction tasks and analysis of the ErrP complex rather than individual peaks.

BackgroundCatestatin (Cts) is a peptide derived from proteolytic cleavage of chromogranin A, which exhibits cardioprotective and anti-inflammatory properties. Cts has been proposed as a potential biomarker for cardiovascular (CV) disease.Objectivesexamining Cts in patients with incidentally discovered adrenocortical adenomas (AI), and its associations with CV risk factors and blood pressure (BP).Materials and methodsIn this cross-sectional study, 64 AI patients without overt CV disease other than primary hypertension were recruited along with 24 age-, sex-, and body-mass-index (BMI)-matched controls with normal adrenal morphology. Laboratory, 24-h ambulatory BP monitoring, echocardiography, and common carotid artery sonography examinations were performed.ResultsUnadjusted Cts was higher in AI patients (median 6.5, interquartile range: 4.9-37 ng/ml) versus controls (4.5 (3.5 – 28)), p=0.048, however, the difference was insignificant after adjusting for confounding variables. Cts was lower in subjects with metabolic syndrome than in those without it (5.2 (3.9- 6.9) vs. 25.7 (5.8-115) ng/ml, p

Understanding HIV-1 persistence despite antiretroviral therapy (ART) is of paramount importance. Both single-genome sequencing (SGS) and integration site analysis (ISA) provide useful information regarding the structure of persistent HIV DNA populations; however, until recently, there was no way to link integration sites to their cognate proviral sequences. Here, we used multiple-displacement amplification (MDA) of cellular DNA diluted to a proviral endpoint to obtain full-length proviral sequences and their corresponding sites of integration. We applied this method to lymph node and peripheral blood mononuclear cells from 5 ART-treated donors to determine whether groups of identical subgenomic sequences in the 2 compartments are the result of clonal expansion of infected cells or a viral genetic bottleneck. We found that identical proviral sequences can result from both cellular expansion and viral genetic bottlenecks occurring prior to ART initiation and following ART failure. We identified an expanded T cell clone carrying an intact provirus that matched a variant previously detected by viral outgrowth assays and expanded clones with wild-type and drug-resistant defective proviruses. We also found 2 clones from 1 donor that carried identical proviruses except for nonoverlapping deletions, from which we could infer the sequence of the intact parental virus. Thus, MDA-SGS can be used for "viral reconstruction" to better understand intrapatient HIV-1 evolution and to determine the clonality and structure of proviruses within expanded clones, including those with drug-resistant mutations. Importantly, we demonstrate that identical sequences observed by standard SGS are not always sufficient to establish proviral clonality.

To investigate the possibility that HIV-1 replication in lymph nodes sustains the reservoir during ART, we looked for evidence of viral replication in 5 donors after up to 13 years of viral suppression. We characterized proviral populations in lymph nodes and peripheral blood before and during ART, evaluated the levels of viral RNA expression in single lymph node and blood cells, and characterized the proviral integration sites in paired lymph node and blood samples. Proviruses with identical sequences, identical integration sites, and similar levels of RNA expression were found in lymph nodes and blood samples collected during ART, and no single sequence with significant divergence from the pretherapy population was present in either blood or lymph nodes. These findings show that all detectable persistent HIV-1 infection is consistent with maintenance in lymph nodes by clonal proliferation of cells infected before ART and not by ongoing viral replication during ART.

Objectives Hypothermia is an established method of treating severe forms of perinatal hypoxia in newborns. Some of them develop neonatal encephalopathy, which is associated with high morbidity and mortality. Therefore, prophylaxis of this pathology is important as well as determining environmental factors in mothers of newborns affected by this pathology. The aim of the study was to assess of selected environmental factors in mothers of newborns qualified for hypothermia. Material and Methods The material consisted of 102 subjects, including 51 mothers of newborns with hypoxic-ischemic encephalopathy referred for hypothermia treatment (group I) and 51 mothers of newborns without signs of hypoxia (group II). The case-control study was carried out in the third level reference centre. It is 1 of 20 centers of therapeutic hypothermia for newborn in Poland. Data was collected based on a data collection sheet. Study groups were compared in terms of demographic and environmental data. The odds ratio (OR) was determined and the logistic regression analysis of univariate and multivariate regression was used to determine the probability of the need for hypothermia in the study group. Results The groups did not differ in terms of age, BMI and place of living. The need to use hypothermia increased in pregnant women living together with their parents (OR = 6.8, 95% CI: 2.4–19.6) also in case of exposure to factors at the workplace, i.e., noise (OR = 4.1, 95% CI: 1.1–15.5). Conclusions Based on the results of our case-control study we postulate to pay attention during preconception care to proper preparation for pregnancy especially in younger women exposed to nuisance in the work environment and at home. In this area postulated activities should include education programs, in close cooperation occupational medicine practitioners and obstetricians even before the conception as a part of pre-conception counseling. Int J Occup Med Environ Health. 2023;36(1):59–68

The atmospheric electric current, “air–earth current”, flows between the low ionosphere and Earth’s surface. The source of this current is the potential difference between the global equalizing layer called the ionosphere and the ground surface. According to Wilson’s concept of the Earth’s Global Electric Circuit, in the areas of so-called fair weather, based on current measurements at the Earth’s surface, it is possible to conclude the global electrical processes in the ionosphere and higher layers. The theoretical basis for this inference is the law of continuity of electric current or the principle of conservation of electric charge. We present the results of simulations of the distribution of electric field lines for sensors with different geometries placed in a uniform electric field, representing the atmospheric electric field. The sensors are metal surfaces on which electric charges are induced or deposited. In the external measuring circuit to which the sensor is connected, an electric current [A] will flow, related to the air–earth current density [A/m2], but their relationship may be challenging to interpret. We analyze the impact of sensor geometry on the possibility of interpreting the atmospheric electric conduction and atmospheric displacement current based on the current measured in the external circuit. This present method can be used for the geometric construction of new sensors at the stage of determining the electrical characteristics of the sensor (e.g., effective collecting area). It can support the comprehensive design of a measurement system at the interface between an atmosphere, sensor, and electronic equipment.

Alcohol-related Liver Disease (ALD) is the primary cause of chronic liver disorders and hepatocellular carcinoma (HCC) development in developed countries and thus represents a major public health concern. Unfortunately, few therapeutic options are available for ALD and HCC, except liver transplantation or tumor resection for HCC. Deciphering the molecular mechanisms underlying the development of these diseases is therefore of major importance to identify early biomarkers and to design efficient therapeutic options. Increasing evidence indicate that epigenetic alterations play a central role in the development of ALD and HCC. Among them, microRNA importantly contribute to the development of this disease by controlling the expression of several genes involved in hepatic metabolism, inflammation, fibrosis, and carcinogenesis at the post-transcriptional level. In this review, we discuss the current knowledge about miRNAs’ functions in the different stages of ALD and their role in the progression toward carcinogenesis. We highlight that each stage of ALD is associated with deregulated miRNAs involved in hepatic carcinogenesis, and thus represent HCC-priming miRNAs. By using in silico approaches, we have uncovered new miRNAs potentially involved in HCC. Finally, we discuss the therapeutic potential of targeting miRNAs for the treatment of these diseases.

Objectives: The purpose of this research was to assess the extent of dyspnoea, physical activity (PA), and back pain complaints and the association of dyspnoea, PA, and back pain complaints with PA in post-COVID-19 students at medical universities in Poland. Methods: An online survey was carried out among Polish medical students (213 women and 204 men) who had had a positive test for SARS-CoV-2 within the last year. The Medical Research Council (MRC) dyspnoea scale was used to assess the degree of dyspnoea. The International Physical Activity Questionnaire (IPAQ) was used to determine the level of PA. The Oswestry Disability Index (ODI) and the Neck Disability Index (NDI) were used to assess back discomfort. Results: The study group had average levels of PA, with median total activity significantly lower in women (median total activity for women was 1189 and for men was 2044, while the standard deviation for women was 1419 and for men was 1450). More than 93% of the students reported no symptoms of dyspnoea. The following results were observed for ODI (median of 1.2 for women and 1.7 for men and standard deviation of 3.1 for women and 4.0 for men) and for NDI (median of 2.8 for women and 2.5 for men, standard deviation of 4.3 for women and 4.0 for men). Cervical spine pain was more frequent and severe. There are small, statistically significant correlations between the MRC and IPAQ measures and the ODI and NDI and IPAQ. Conclusions: In the study group of students of medicine, dyspnoea linked with a history of COVID-19 is not an issue. Post-pandemic PA levels should be increased in this group, with particular attention to female students. Urgent measures are also needed to prevent cervical pain in students at medical universities in Poland.

Chociaż tematyka oparzeń wydaje się być stosunkowo dobrze opracowana w piśmiennictwie, nadal istnieje potrzeba określenia związku patomechanizmu urazów termicznych z kluczową kwestią zakażeń w ich obrębie. Niezmiernie istotna w przypadku zakażeń ran oparzeniowych jest diagnostyka mikrobiologiczna. Prawidłowe pobranie materiału klinicznego i wykonanie badań mikrobiologicznych pozwala na ustalenie czynnika etiologicznego zakażenia i dobranie antybiotykoterapii celowanej. Jest to szczególnie istotne w dobie narastania oporności drobnoustrojów na antybiotyki i częstego izolowania szczepów wieloantybiotykoopornych. Oparzenie bowiem, zwłaszcza rozległe, stanowi unikalne środowisko, które – wraz z nieprzemyślaną, bądź niecelowaną terapią – determinuje u drobnoustrojów nabywanie oporności na antybiotyki. Fizjologiczna obrona organizmu przed urazem prowadzi do sytuacji, kiedy w ranie oparzeniowej środki przeciwdrobnoustrojowe mogą nie osiągać MIC/MBC, co drastycznie zmniejsza szanse powodzenia terapeutycznego.

Quantitative viral outgrowth assays (QVOA) use limiting dilutions of CD4+ T cells to measure the size of the latent HIV-1 reservoir, a major obstacle to curing HIV-1. Efforts to reduce the reservoir require assays that can reliably quantify its size in blood and tissues. Although QVOA is regarded as a "gold standard" for reservoir measurement, little is known about its accuracy and precision or about how cell storage conditions or laboratory-specific practices affect results. Owing to this lack of knowledge, confidence intervals around reservoir size estimates-as well as judgments of the ability of therapeutic interventions to alter the size of the replication-competent but transcriptionally inactive latent reservoir-rely on theoretical statistical assumptions about dilution assays. To address this gap, we have carried out a Bayesian statistical analysis of QVOA reliability on 75 split samples of peripheral blood mononuclear cells (PBMC) from 5 antiretroviral therapy (ART)-suppressed participants, measured using four different QVOAs at separate labs, estimating assay precision and the effect of frozen cell storage on estimated reservoir size. We found that typical assay results are expected to differ from the true value by a factor of 1.6 to 1.9 up or down. Systematic assay differences comprised a 24-fold range between the assays with highest and lowest scales, likely reflecting differences in viral outgrowth readout and input cell stimulation protocols. We also found that controlled-rate freezing and storage of samples did not cause substantial differences in QVOA compared to use of fresh cells (95% probability of < 2-fold change), supporting continued use of frozen storage to allow transport and batched analysis of samples. Finally, we simulated an early-phase clinical trial to demonstrate that batched analysis of pre- and post-therapy samples may increase power to detect a three-fold reservoir reduction by 15 to 24 percentage points.

Millimeter-wave (mm-wave) transmitters are often fabricated using advanced technology and require a sophisticated manufacturing facility. Access to such technologies is often very limited and difficult to gain particularly at the initial stage of research. Therefore, to increase the accessibility of mm-wave transmitters, this study proposes a design that can be assembled in a standard microwave laboratory from commercially available or externally ordered components. The transmitter demonstrated in this paper operates above 100 GHz and is based on a lowtemperature co-fired ceramic board in which the antenna array, microstrip lines, and power-supply lines are fabricated in a single process. Different technologies are used to assemble the module, e.g., wire-bonding, soldering, and wax adhesion. Advantages and disadvantages of the proposed design are given based on experimental evaluation of the prototype. Although the performance of the developed transmitter is not as good as that of the similar modules available in the recent literature, the results confirm the feasibility of a mm-wave transmitter that is assembled without employing advanced technologies and superior machinery.

Cystic fibrosis (CF) is characterized by chronic bacterial infections leading to progressive bronchiectasis and respiratory failure. Pseudomonas aeruginosa (Pa) is the predominant opportunistic pathogen infecting the CF airways. The guanine nucleotide exchange factor Vav3 plays a critical role in Pa adhesion to the CF airways by inducing luminal fibronectin deposition that favors bacteria trapping. Here we report that Vav3 overexpression in CF is caused by upregulation of the mRNA-stabilizing protein HuR. We found that HuR accumulates in the cytoplasm of CF airway epithelial cells and that it binds to and stabilizes Vav3 mRNA. Interestingly, disruption of the HuR-Vav3 mRNA interaction improved the CF epithelial integrity, inhibited the formation of the fibronectin-made bacterial docking platforms, and prevented Pa adhesion to the CF airway epithelium. These findings indicate that targeting HuR represents a promising antiadhesive approach in CF that can prevent initial stages of Pa infection in a context of emergence of multidrug-resistant pathogens.

This study aimed to analyze the chemotactic response of differentiated HL-60 neutrophil-like (dHL-60) cells to trans-anethole (TA)-treated Staphylococcus aureus strains. Special attention was paid to evaluate the influence of TA on the chp gene expression level, as well as molecular docking and molecular dynamics (MD) simulation studies on interactions of TA with chemotaxis inhibitory protein of S. aureus (CHIPS). The following parameters were studied: susceptibility to TA using the agar diffusion method, the chp gene detection and its expression under TA influence, and clonal diversity of S. aureus strains using molecular techniques. Furthermore, a chemotactic response of dHL-60 cells to TA-treated S. aureus using Boyden chamber assay was detected and molecular modeling using both the docking methodology and unbiased MD simulations was conducted. It was found that TA showed antibacterial activity against all strains. Three genotypes and one unique pattern were distinguished among the strains. 50% of the isolates were chp-positive. It was observed that TA reduced/inhibited chp gene expression in most S. aureus strains. Enhanced chemotactic response of dHL-60 cells to TA-treated S. aureus strains was also noted. This correlation was similar for both chp-positive and chp-negative strains. Both molecular docking and MD simulations studies confirmed that TA is preferentially bound in the complement component 5a/CHIPS interface interaction region and can interfere with any processes exploiting this binding cavity. It has been proven that dHL-60 cells exhibited a higher chemotactic response to TA-treated S. aureus strains in comparison to non-treated bacteria, regardless of the achieved expression of the chp gene or its lack. Nevertheless, further analyses are required to understand this mechanism better.

The article presents the place of 3D printing in the manufacturing and operational process. It analyzes selected incremental technologies in the product life cycle. It describes selected processes for testing the properties of materials used in 3D printing, including accelerated aging tests and simulation of operating conditions. Areas of application of 3D printing were defined, starting from design and prototype development through manufacturing of technological tools and finally finished products. Design criteria of additivelymanufactured elements in relation to the exploitation process are discussed. A methodology for the development of 3D-CAD models of manufactured elements, software processing of data and data storage format for manufacturing products and spare parts is presented. The assumptions of repair procedures based on the production of spare parts by means of 3D printing in relation to data circulation compatible with the idea of Industry 4.0 structure have been adopted.

Body fat (BF) and cardiorespiratory fitness (CRF) are important health markers that ought to be considered in screening exams. The aim of this study was to assess the value of six indicators, i.e., tri-ponderal mass index (TMI), relative fat mass (RFM), waist–BMI ratio, waist-to-height ratio (WHtR), waist-to-hip ratio (WHR) and body mass index (BMI) in predicting CRF in school-aged children. The analysis was based on the data coming from the examination of 190 children participating in school physical education (PE) classes. Their body weight (BW) and height (BH), waist and hip circumference (WC; HC) and percentage of body fat (%BF) were measured; the CRF test was performed with the use of the 20 m shuttle run test (20 mSRT); peak heart rate (HRpeak) was measured; TMI, relative fat mass pediatric (RFMp), waist–BMI ratio, WHtR, BMI and WHR were calculated. Statistical analysis was mainly conducted using regression models. The developed regression models, with respect to the sex and age of the children, revealed RFMp as the strongest CRF indicator (R2 = 51.1%) and WHR as well as waist–BMI ratio as the weakest ones (R2 = 39.2% and R2 = 40.5%, respectively). In predicting CRF in school-aged children, RFMp turned out to be comparable to body fat percentage obtained by means of the bioimpedance analysis (BIA) (R2 = 50.3%), and as such it can be used as a simple screening measure in prophylactic exams of school children. All of these models were statistically significant (p < 0.001).