Many methods exist for aligning and quantifying magnetic resonance spectroscopy (MRS) data to measure in vivo γ-aminobutyric acid (GABA). Research comparing the performance of these methods is scarce partly due to the lack of ground-truth measurements. The concentration of GABA is approximately two times higher in grey matter than in white matter. Here we use the proportion of grey matter within the MRS voxel as a proxy for ground-truth GABA concentration to compare the performance of four spectral alignment methods (i.e., retrospective frequency and phase drift correction) and six GABA signal modelling methods. We analyse a diverse dataset of 432 MEGA-PRESS scans targeting multiple brain regions and find that alignment to the creatine (Cr) signal produces GABA+ estimates that account for approximately twice as much of the variance in grey matter as the next best performing alignment method. Further, Cr alignment was the most robust, producing the fewest outliers. By contrast, all signal modelling methods, except for the single-Lorentzian model, performed similarly well. Our results suggest that variability in performance is primarily caused by differences in the zero-order phase estimated by each alignment method, rather than frequency, resulting from first-order phase offsets within subspectra. These results provide support for Cr alignment as the optimal method of processing MEGA-PRESS to quantify GABA. However, more broadly, they demonstrate a method of benchmarking quantification of in vivo metabolite concentration from other MRS sequences.

The antiretroviral (ARV) cocktailrevolved the treatment of the human immunodeficiency virus (HIV) infection. Drug combinations have been also tested to treat other infectious diseases, including the recentcoronavirus disease 2019 (COVID-19) outbreak. To simplify administration fixed-dose combinationshave been introduced, however, oral anti-HIV therapy still struggles with low oral bioavailability of many ARVs.This work investigated the co-encapsulation of two clinically relevant ARV combinations,tipranavir (TPV):efavirenz (EFV) anddarunavir (DRV):efavirenz (EFV):ritonavir (RTV),within the core of β-casein (bCN) micelles. Encapsulation efficiency in both systems was ~100%. Cryo-transmission electron microscopy and dynamic light scattering of the ARV-loaded colloidaldispersions indicatefull preservation of the spherical morphology, and x-ray diffraction confirm that the encapsulated drugs are amorphous. To prolong the physicochemical stabilitythe formulations were freeze-driedwithout cryo/lyoprotectant, and successfully redispersed, with minor changes in morphology.Then, theARV-loaded micelles were encapsulated within microparticles of Eudragit® L100, which prevented enzymatic degradation and minimized drug release under gastric-like pH conditionsin vitro. At intestinal pH, the coating polymer dissolved and released the nanocarriers and content. Overall, our results confirm the promise of this flexible and modular technology platform for oral delivery of fixed dose combinations.

Multi-scale, subject-specific quantitative methods to characterize and monitor osteoarthritis in animal models and therapeutic treatments could help reveal causal relationships in disease development and distinguish treatment strategies. In this work, we demonstrate a reproducible and sensitive quantitative image analysis to characterize bone, cartilage and joint measures describing a rat model of post-traumatic osteoarthritis. Eleven 3-month-old male Wistar rats underwent medial anterior cruciate ligament (ACL) transection and medial meniscectomy on the right knee to destabilise the right tibiofemoral joint. They were sacrificed 6 weeks post-surgery and a silicon-based micro-bead contrast agent was injected in the joint space, before scanning with micro-computed tomography (microCT). Subsequently, 3D quantitative morphometric analysis (QMA), previously developed for rabbit joints, was performed. This included cartilage, subchondral cortical and epiphyseal bone measures, as well as novel tibiofemoral joint metrics. Semi-quantitative evaluation was performed on matching two-dimensional (2D) histology and microCT images. Reproducibility of the QMA was tested on eleven age-matched additional joints. The results indicate the QMA method is accurate and reproducible and that microCT-derived cartilage measurements are valid for the analysis of rat joints. The pathologic changes caused by transection of the ACL and medial meniscectomy were reflected in measurements of bone shape, cartilage morphology, and joint alignment. Furthermore, we were able to identify model-specific predictive parameters based on morphometric parameters measured with the QMA.

Intestinal development is closely associated with inflammatory wooden breast (WB) myopathy. Vitamin E (VE) and alpha lipoic acid (ALA) with antioxidant and anti-inflammatory effects were used independently and in combination to evaluate their effects on intestinal developmental changes in ileal morphology and expression of genes related with gut nutrient transport, structure, and inflammation in broilers during the first 3 wk posthatch. A total of 160 newly hatched Ross 708 broiler chicks were randomly assigned into a control and 3 dietary treatments with 10 replicates of 4 birds each. Supplementation of VE (160 mg/kg) and ALA (500 mg/kg) independently and in combination were fed during the first 3 wk. At 1, 2, and 3 wk of age, one chick from each pen was harvested. Plasma VE concentration and ileal morphology were determined. Gene expression was measured by real-time quantitative PCR. Broilers in VE and combination of ALA and VE group had higher plasma VE concentration than the control and ALA group at 1, 2, and 3 wk of age (P < 0.01). All dietary treatments increased ileal villus height at 1 wk of age (P < 0.01) and decreased intraepithelial lymphocytes at 3 wk of age compared to the control (P ≤ 0.05). Combination of VE and ALA increased collagen type IV alpha 1 chain expression (P ≤ 0.05) and improved basement membrane structure indicating increased gut basement membrane integrity at 2 and 3 wk of age compared to the control. Expression of lipopolysaccharide-induced tumor necrosis factor-alpha factor associated with inflammation was decreased in all dietary treatments at 3 wk of age compared to the control (P < 0.01). Ileal morphology and gene expression were closely correlated with breast muscle morphology and gene expression. These results suggest that VE and ALA especially when they were combined in the diet had positive effects on mitigating intestinal inflammation and improving nutrient transport beginning at 1 wk of age, which is likely critical in reducing the severity of WB.

IL7 is a hematopoietic growth factor required for development and maintenance of lymphocytes including T cells, B cells, and natural killer cells. Recently, chicken IL7 (chIL7) has been cloned and studied in viral and parasite infection models. However, no monoclonal antibodies (mAb) that specifically detect chIL7 have been developed so far. In this study, recombinant chIL7 that expressed for immunization and mAb against chIL7 were developed and characterized to assess their immunologic properties. Five mAb exhibiting specific binding to chIL7 were generated and investigated for their applicability by Western blot, ELISA, and neutralization assays. A sandwich ELISA mAb pair that enables the measurement of chIL7 protein levels in biological samples from Eimeria-infected chickens was identified and several mAb neutralized chicken primary thymocyte proliferation mediated by chIL7. The mAb developed in this study will be valuable reagents for fundamental and applied immunological studies in poultry.

Slow-growing broilers offer differentiation in the chicken meat market for consumers who have distinct preferences based on perceived higher welfare indices and willingness to pay a higher price for the product. Although breeding for slow-growing broilers is relatively advanced in Europe and the United States, it is limited in Australia. Crossbreeding is one of the approaches taken to developing slow-growing broiler strains. Thus, the aim of this study was to compare performance, immune response, leg health, carcass characteristics, and meat quality of a novel crossbred slow-growing broiler breed (SGB) with the conventional, fast-growing Cobb 500 broiler (CB) to assess their suitability as an alternative for chicken meat production in Australia. A total of 236 one-day-old broiler chicks (116 SGB and 120 fast-growing CB) were reared on standard commercial diet in an intensive production system. Birds and feed were weighed on a weekly basis and feed intake and feed conversion ratio calculated. At 21 d of age, a 2% suspension of sheep red blood cells was injected subcutaneously into 8 broilers of each breed to compare their antibody response. Birds from both breeds were grown to a final live weight of 2.0-2.2 kg, before a latency-to-lie (LTL) test, carcass analysis and apparent metabolizable energy (AME) assay were performed. The SGB reached the target weight at 55 d of age compared with 32 d in CB. However, SGB stood for longer during LTL, had higher thigh, drumstick, and wing yields (as a percentage of carcass weight) as well as darker and redder meat in comparison with the CB. The CB had better feed conversion efficiency, higher antibody (IgM) production, higher AME, heavier breast yield, and lower meat drip loss than the SGB. Although fast-growing CB outperformed the SGB for traditional performance parameters, the crossbred in this study was comparable with other slow-growing broiler breeds and strains across different countries and is thus a suitable candidate for a slow-growing alternative in Australia.

The effects of early heat conditioning on the acute heat stress response in broilers were investigated via the growth performance, dopamine, serotonin, and corticosterone and the expression of heat shock proteins (HSP) and heat shock factors. One-day-old chicks (n = 144) were divided into 3 groups in a 35-d experiment (48 chicks per each group). Group 1 (C) was treated with an optimum temperature, group 2 (CH) was treated with 40°C ± 1°C on day 35 (5 h), and group 3 (HH) was treated with 40°C ± 1°C on day 5 (24 h) and day 35 (5 h). On day 7, the body weight gain was lower (P < 0.05) in HH than in C and CH. On day 35, the heat-treated groups (CH and HH) had lower weight gains than the C group (P < 0.05), whereas the feed conversion ratio was lower in HH (P < 0.05). Serum corticosterone was higher in CH than in C, but HH and C did not differ (P < 0.05). Liver HSP70 protein expression was higher in CH than HH and C (P < 0.05), which did not differ, and HSP40 protein expression was higher in CH than C (P < 0.05). These results suggest that early heat conditioning may reduce acute heat stress on broiler.

Besides being a common food component broadly consumed worldwide, egg yolk immunoglobulin Y (IgY) has essential therapeutic potentials. In fact, in a time of ever-increasing risk of antibiotic resistance, it is crucial to find new ways to battle infection, and oral administration of preformed specific antibodies represents one of the most attractive approaches against infection. Infectious diseases of bacterial and viral origin in humans and animals can be controlled and passively cured by orally applied IgYs isolated from chicken egg yolks. Despite multiple obvious advantages of oral administration of IgY, harvesting IgY from egg yolk in a pure form is a challenging task. In this study, we developed a fast, simple, cost-effective, and efficient protocol for IgY isolation from chicken egg yolks. First, egg yolk was collected and diluted with 5 volumes of cold distilled water, homogenized, pH adjusted, and centrifuged. Next, the supernatant was collected, to which caprylic acid at concentration of 2% v/v was added, followed by pH adjustment to pH 5.0, centrifugation at 4°C, and collection of the resulting supernatant. This step was repeated twice, with adding 2% v/v of caprylic acid each time. The final supernatant was concentrated using ultrafiltration, and the IgY purity and activities were checked by SDS-PAGE, western blotting, and ELISA. The sequential (2, 2, 2%) addition of caprylic acid yielded IgY with a purity of 63.5, 90.6, and 95.8%, respectively, and reached 97.9% after ultrafiltration at pH 9.0. The IgY activity increased exponentially to reach 99% after the ultrafiltration step. The proposed caprylic-acid-based protocol of IgY purification from the yolk of chicken eggs seems to be simple, fast, direct, and very cheap. This indicates that this protocol has great potential for scale-up processing.

Aroma is an important quality parameter of fresh culinary herbs that may be highly affected after postharvest treatments. The innovative technology of vapor essential oil (EO) application under vacuum conditions may recover aroma lost during the postharvest processing of plant products like aromatic herbs. Hence, this study assessed the aroma recovery effect of vapor EOs applied during vacuum cooling on curly parsley and dill. The volatile organic compounds (VOCs) profiles of these aromatic herbs were studied by static headspace solid-phase microextraction (SPME), and the VOCs sorption kinetics onto the SPME stir-bar coating were modeled by the Baranyi model. At the pilot plant scale, the total VOCs contents of parsley and dill (whose extractability was increased by 10-20% after a single vacuum process) were enhanced by 4.5- and 2-fold, respectively, when vapor EOs were applied. In particular, 1,3,8-p-menthatriene and carvone (parsley) increased by 18.7- and 7.3-fold, respectively, while dill ether (the characteristic VOC of dill) augmented by 2.4-fold after vapor EOs were applied under vacuum conditions. The aroma recovery of culinary herbs was successfully validated at an industrial level in an installation developed by our group to apply vapor EOs within a vacuum cooling system, reaching total VOC recoveries of 4.9- and 2.3-fold in parsley and dill, respectively.

Genetic interpretation of rare variants associated with arrhythmogenic cardiomyopathy (ACM) is essential due to their diagnostic implications. New data may relabel previous variant classifications, but how often reanalysis is necessary remains undefined. Five years ago, 39 rare ACM-related variants were identified in patients with features of cardiomyopathy. These variants were classified following the American College of Medical Genetics and Genomics' guidelines. In the present study, we reevaluated these rare variants including novel available data. All cases carried one rare variant classified as being of ambiguous significance (82.05%) or likely pathogenic (17.95%) in 2016. In our comprehensive reanalysis, the classification of 30.77% of these variants changed, mainly due to updated global frequencies. As in 2016, nowadays most variants were classified as having an uncertain role (64.1%), but the proportion of variants with an uncertain role was significantly decreased (17.95%). The percentage of rare variants classified as potentially deleterious increased from 17.95% to 23.07%. Moreover, 83.33% of reclassified variants gained certainty. We propose that periodic genetic reanalysis of all rare variants associated with arrhythmogenic cardiomyopathy should be undertaken at least once every five years. Defining the roles of rare variants may help clinicians obtain a definite diagnosis.

In recent years, transcriptome profiling studies have identified changes in host splicing patterns caused by viral invasion, yet the functional consequences of the vast majority of these splicing events remain uncharacterized. We recently showed that the host splicing landscape changes during Rift Valley fever virus MP-12 strain (RVFV MP-12) infection of mammalian cells. Of particular interest, we observed that the host mRNA for Rio Kinase 3 (RIOK3) was alternatively spliced during infection. This kinase has been shown to be involved in pattern recognition receptor (PRR) signaling mediated by RIG-I like receptors to produce type-I interferon. Here, we characterize RIOK3 as an important component of the interferon signaling pathway during RVFV infection and demonstrate that RIOK3 mRNA expression is skewed shortly after infection to produce alternatively spliced variants that encode premature termination codons. This splicing event plays a critical role in regulation of the antiviral response. Interestingly, infection with other RNA viruses and transfection with nucleic acid-based RIG-I agonists also stimulated RIOK3 alternative splicing. Finally, we show that specifically stimulating alternative splicing of the RIOK3 transcript using a morpholino oligonucleotide reduced interferon expression. Collectively, these results indicate that RIOK3 is an important component of the mammalian interferon signaling cascade and its splicing is a potent regulatory mechanism capable of fine-tuning the host interferon response.

The main purpose of the research was to analyze the influence of surface modification of the cobalt-based alloy used in dental prosthetics by applying zirconium oxide (ZrO2) layers using the ALD (Atomic Layer Deposition) method. The samples were made using the DMLS (Direct Metal Laser Sintering) technique, and their surfaces were prepared in accordance with the principles of removable partial dentures (RPDs). A 50 nm-thick zirconium oxide coating was applied to the prepared substrates. This paper deals with the issues of prosthetic stomatopathy, which is a complex of pathological changes occurring in approx. 40% of the Polish population using removable dentures. Often, these changes, occurring on the mucosa, are related to improper performance, allergic reactions or the multiplication of bacteria on the surface of partial dentures. An innovative method of surface modification was proposed, together with the analysis of its influence on the physicochemical properties of the alloy and the adhesion of bacteria to the surface.

Bladder cancer (BC) is the tenth most frequently detected cancer in both sexes. Type-I luteinizing hormone-releasing hormone (LHRH) receptor (LHRH-R-I) is expressed not only in the pituitary, but also in several types of cancer disease. There are few data about LHRH-R-I expression in human BC. This study aimed to investigate the expression of LHRH and LHRH-R-I in the transitional cell carcinoma (TCC) type of human BC. RNA was extracted from 24 human bladder tumor specimens and three BC cell lines. RT-PCR was performed to detect mRNA for LHRH and LHRH-R-I. The protein of LHRH-R-I was further studied by immunohistochemistry (IHC), ligand competition assay, and Western Blot. PCR products of LHRH were found in 19 of 24 (79%) specimens and mRNA of LHRH-R-I was detected in 20 of 24 specimens (83%). Positive immunostaining for LHRH-R-I with different expression intensity was found in all samples examined, showing negative correlation with TCC grade. Radioligand binding studies also showed the presence of specific LHRH-R-I and high affinity binding of LHRH analogs. The high incidence of LHRH-R in BC suggests that it could serve as a molecular target for therapy of human BC with cytotoxic LHRH analogs or modern powerful antagonistic analogs of LHRH.

The main aims of the research were to produce efficient nanofibrous filters with long-term antibacterial properties and to confirm the functionality of samples under real filtration conditions. A polyurethane solution was modified by micro- or nanoparticles of copper oxide in order to juxtapose the aggregation tendency of particles depending on their size. Modified solutions were electrospun by the Nanospider technique. The roller spinning electrode with a needle surface and static wire electrode were used for the production of functionalized nanofibers. The antibacterial properties of the modified nanofibrous layers were studied under simulated conditions of water and air filtration. Particular attention was paid to the fixation mechanism of modifiers in the structure of filters. It was determined that the rotating electrode with the needle surface is more efficient for the spinning of composite solutions due to the continuous mixing and the avoidance of particle precipitation at the bottom of the bath with modified polyurethane. Moreover, it was possible to state that microparticles of copper oxide are more appropriate antimicrobial additives due to their weaker aggregation tendency but stronger fixation in the fibrous structure than nanoparticles. From the results, it is possible to conclude that nanofibers with well-studied durable antibacterial properties may be recommended as excellent materials for water and air filtration applications.

Circulating tumour cells (CTCs) are the precursor cells for the formation of metastatic disease. With a simple blood draw, liquid biopsies enable the non-invasive sampling of CTCs from the blood, which have the potential to provide important insights into cancer detection and monitoring. Since gaining FDA approval in 2004, the CellSearch system has been used to determine the prognosis of patients with metastatic breast, prostate and colorectal cancers. This utilises the cell surface marker Epithelial Cell Adhesion Molecule (EpCAM), to enrich CTCs, and many other technologies have adopted this approach. More recently, the role of mesenchymal-like CTCs in metastasis formation has come to light. It has been suggested that these cells are more aggressive metastatic precursors than their epithelial counterparts; however, mesenchymal CTCs remain undetected by EpCAM-based enrichment methods. This has prompted the development of a variety of 'label free' enrichment technologies, which exploit the unique physical properties of CTCs (such as size and deformability) compared to other blood components. Here, we review a wide range of both immunocapture and label free CTC enrichment technologies, summarising the most significant advantages and disadvantages of each. We also highlight the important characteristics that technologies should possess for routine clinical use, since future developments could have important clinical implications, with the potential to direct personalised therapies for patients with cancer.

Underlying mechanisms for resistance to cisplatin-based chemotherapy in bladder cancer patients are largely unknown, although androgen receptor (AR) activity, as well as extracellular signal-regulated kinase (ERK) signaling, has been indicated to correlate with chemosensitivity. We also previously showed ERK activation by androgen treatment in AR-positive bladder cancer cells. Because our DNA microarray analysis in control vs. AR-knockdown bladder cancer lines identified BXDC2 as a potential downstream target of AR, we herein assessed its functional role in cisplatin sensitivity, using bladder cancer lines and surgical specimens. BXDC2 protein expression was considerably downregulated in AR-positive or cisplatin-resistant cells. BXDC2-knockdown sublines were significantly more resistant to cisplatin, compared with respective controls. Without cisplatin treatment, BXDC2-knockdown resulted in significant increases/decreases in cell proliferation/apoptosis, respectively. An ERK activator was also found to reduce BXDC2 expression. Immunohistochemistry showed downregulation of BXDC2 expression in tumor (vs. non-neoplastic urothelium), higher grade/stage tumor (vs. lower grade/stage), and AR-positive tumor (vs. AR-negative). Patients with BXDC2-positive/AR-negative muscle-invasive bladder cancer had a significantly lower risk of disease-specific mortality, compared to those with a BXDC2-negative/AR-positive tumor. Additionally, in those undergoing cisplatin-based chemotherapy, BXDC2 positivity alone (p = 0.083) or together with AR negativity (p = 0.047) was associated with favorable response. We identified BXDC2 as a key molecule in enhancing cisplatin sensitivity. AR-ERK activation may thus be associated with chemoresistance via downregulating BXDC2 expression in bladder cancer.

In recent years, textile industries have focused their attention on the development of functional finishing that presents durability and, consequently, controlled release. However, in the case of methyl salicylate microcapsules supported on a textile matrix, studies indicate only the interactions between substrate and microcapsules and the drug delivery system, not applying the release equations. This study reports the mechanism and kinetics of controlled release of microcapsules of gelatin and gum Arabic containing methyl salicylate as active ingredient incorporated into textile matrices. According to the results presented, it was possible to verify that the wall materials participated in the coacervation process, resulting in microcapsules with well-defined geometry, besides promoting the increase of the thermal stability of the active principle. The samples (100% cotton, CO, and 100% polyamide, PA) functionalized with microcapsules released methyl salicylate in a controlled manner, based on the adjustment made by the Korsmeyer-Peppas model, indicating a Fickian mechanism. The influence of temperature was noticeable when the samples were subjected to washing, since with higher temperature (50 °C), the release was more pronounced than when subjected to lower temperature (37 °C). The results presented in this study indicate that the mechanism of backbone release is influenced by the textile matrix and by the durability of the microcapsule during the wash cycles.

The rapid and uncontrolled proliferation of cancer cells is supported by metabolic reprogramming. Altered glucose metabolism supports cancer growth and progression. Compared with normal cells, cancer cells show increased glucose uptake, aerobic glycolysis and lactate production. Byproducts of adjusted glucose metabolism provide additional benefits supporting hallmark capabilities of cancer cells. Long non-coding RNAs (lncRNAs) are a heterogeneous group of transcripts of more than 200 nucleotides in length. They regulate numerous cellular processes, primarily through physical interaction with other molecules. Dysregulated lncRNAs are involved in all hallmarks of cancer including metabolic alterations. They may upregulate metabolic enzymes, modulate the expression of oncogenic or tumor-suppressive genes and disturb metabolic signaling pathways favoring cancer progression. Thus, lncRNAs are not only potential clinical biomarkers for cancer diagnostics and prediction but also possible therapeutic targets. This review summarizes the lncRNAs involved in cancer glucose metabolism and highlights their underlying molecular mechanisms.

MACC1 is a prognostic and predictive metastasis biomarker for more than 20 solid cancer entities. However, its role in cancer metabolism is not sufficiently explored. Here, we report on how MACC1 impacts the use of glucose, glutamine, lactate, pyruvate and fatty acids and show the comprehensive analysis of MACC1-driven metabolic networks. We analyzed concentration-dependent changes in nutrient use, nutrient depletion, metabolic tracing employing 13C-labeled substrates, and in vivo studies. We found that MACC1 permits numerous effects on cancer metabolism. Most of those effects increased nutrient uptake. Furthermore, MACC1 alters metabolic pathways by affecting metabolite production or turnover from metabolic substrates. MACC1 supports use of glucose, glutamine and pyruvate via their increased depletion or altered distribution within metabolic pathways. In summary, we demonstrate that MACC1 is an important regulator of metabolism in cancer cells.

An increasing number of studies suggests that a novel neuromodulation technique targeting the spinal circuitry enhances gait rehabilitation, but research on its application to stroke survivors is limited. Therefore, we investigated the characteristics of spinal motor-evoked responses (sMERs) from lower-limb muscles obtained by transcutaneous spinal cord stimulation (tSCS) after stroke compared to age-matched and younger controls without stroke. Thirty participants (ten stroke survivors, ten age-matched controls, and ten younger controls) completed the study. By using tSCS applied between the L1 and L2 vertebral levels, we compared sMER characteristics (resting motor threshold (RMT), slope of the recruitment curve, and latency) of the tibialis anterior (TA) and medial gastrocnemius (MG) muscles among groups. A single pulse of stimulation was delivered in 5 mA increments, increasing from 5 mA to 250 mA or until the subjects reached their maximum tolerance. The stroke group had an increased RMT (27-51%) compared to both age-matched (TA: p = 0.032; MG: p = 0.005) and younger controls (TA: p < 0.001; MG: p<0.001). For the TA muscle, the paretic side demonstrated a 13% increased latency compared to the non-paretic side in the stroke group (p = 0.010). Age-matched controls also exhibited an increased RMT compared to younger controls (TA: p = 0.002; MG: p = 0.007), suggesting that altered sMER characteristics present in stroke survivors may result from both stroke and normal aging. This observation may provide implications for altered spinal motor output after stroke and demonstrates the feasibility of using sMER characteristics as an assessment after stroke.