Search Results (38)

β-glucuronidase is an important hydrolase, which plays an important role in drug metabolism, clinical diagnostics, and biotransformation. This study focuses on the heterologous expression, isolation, purification, and its enzymatic properties of β-glucuronidase CpGUS from Clostridium perfringens, as well as its application in the whole-cell transformation of unconjugated bilirubin from pig bile. A recombinant E. coli BL21(DE3)/pET-28a-CpGUS was constructed for the heterologous expression of CpGUS, with the majority of the expressed enzyme being soluble. Enzymatic analysis showed that CpGUS displayed optimal activity at pH 5.0 and 45 °C, and it rapidly lost activity at pH < 4.5. Metal ions, such as Mg²⁺ and Fe²⁺, enhanced CpGUS catalysis, while Zn²⁺, K⁺, Fe³⁺, Mn²⁺, Cu²⁺, and Na⁺ inhibited it. Notably, Cu²⁺ and Fe³⁺ can significantly inhibit β-glucuronidase, resulting in the complete loss of its activity. The results of the whole-cell transformation experiment show that when E.coli BL21(DE3)/ pET-28a-CpGUS at an OD₆₀₀ of 10 was incubated at pH 5.0, a temperature of 45 °C, and a rotation speed of 200 rpm for 12 h, the hydrolysis rate of the conjugated bilirubin in pig bile reached 81.1%, the yield of unconjugated bilirubin was 76.8%, and the purity of unconjugated bilirubin was 98.2%. The three-dimensional structure of CpGUS was predicted using AlphaFold2 (AlphaFold v2.0, DeepMind Technologise Limited, London, UK), and p-Nitrophenyl-β-D-Glucuronide (pNPG) and conjugated bilirubin were then docked to the CpGUS protein model using SWISSDOCK. The best docked conformations of the CpGUS–pNPG and CpGUS–conjugated bilirubin complex systems were simulated by independent 500 ns molecular dynamics (MD) runs with the RSFF2C force field, and the binding dynamic and catalytic mechanism of each system were obtained. The results indicated that π-π stacking, hydrogen bonding, and hydrophobic interactions between the key residue Tyr472 and the benzene ring of pNPG molecules are crucial for its catalytic process. Similarly, for the binding and catalysis of conjugated bilirubin by CpGUS, the π-π stacking and hydrogen bonding and hydrophobic interactions between the sidechains of residues Phe368 and Tyr472 and the benzene ring of conjugated bilirubin play a synergistic role during its catalytic process. Their total binding free energy (∆G_bind) values were calculated to be as high as −65.05 ± 12.66 and −86.70 ± 17.18 kJ/mol, respectively. These results suggest that CpGUS possesses high binding and catalytic hydrolysis properties for both pNPG and conjugated bilirubin. Full article

tet(X4)-positive IncHI1 plasmids are widely prevalent in various bacteria. To understand their transmission characteristics, we analyzed two extensively drug-resistant (XDR) Escherichia coli strains isolated from pet dog feces in Henan Province, China. Strain T28R harbored tet(X4)-positive IncHI1, IncF18:A-:B-, and mcr-1-positive IncI2 plasmids, while T16R carried tet(X4)-positive IncHI1, F16:A-:B-, and mcr-1-positive IncX4 plasmids. Four representative fusion plasmids, pT28R-F1, pT28R-F2, pT28R-F3, and pT16R-F1, in transconjugants were analyzed using WGS and PCR mapping. The results showed that IS26 from the IncF18:A-:B--plasmid attacked the conjugative transfer-associated genes trhc and rsp on the IncHI1 plasmid, generating pT28R-F1 and pT28R-F2. pT28R-F3 was generated through ISCro1- and ISCR2-mediated homologous recombination, deleting the Tra1 region of the IncHI1 plasmid. T16R-F1 emerged from ISCR2- and IS1B-mediated homologous recombination, losing transfer regions of parental plasmids. Notably, fusion plasmids lost the temperature sensitivity of the IncHI1 plasmid, with conjugation frequencies between 1.57 × 10⁻⁴ and 3.84 × 10⁻⁵ at 28 °C and 37 °C. The findings suggest that tet(X4)-positive IncHI1 plasmids could be mobilized with the assistance of conjugative helper plasmids and that fusion events enhance the adaptability of these plasmids, thus facilitating the spread of antibiotic resistance, posing a growing public health threat. Full article

In addition to the mechanical properties and barrier performance, one of the key properties of flexible films used in food packaging is the resistance of their gas barrier layer to buckling and bending. Testing the gas barrier before and after mechanical stress is time-consuming and resource-intensive, but important to assure a certain gas barrier during the whole life time of the package until food consumption. The aim of this study was, on the one hand, to identify the most significant influencing factors of a composite lacquer formulation and coating on its buckling resistance and, on the other hand, to show a fast and efficient method to identify defects occurring during buckling. The influence of mechanical stress was simulated via Gelbo-Flex treatment, and the samples were examined and evaluated before and after using light microscopy. The evaluation was verified with scanning electron microscopy (SEM) and helium barrier measurement. Polyethylene terephthalate (PET) and polyethylene (PE) films were coated with composite barrier lacquers made of polyvinyl alcohol (PVA) and montmorillonite (MMT) and the wet coating layer thickness (20, 40, 80, and 130 μm) and the composition of the coating were changed. It was found that thin coatings are more resistant to buckling than thick coatings. It was also shown that a higher proportion of MMT in the coating layer leads to a better gas barrier, but poorer buckling resistance. Additionally, it was found that soft PE films are already subjected to high stresses during the coating process, which means that barrier coatings do not build up ideally. However, the barrier-coated soft film withstood mechanical stress better and lost less barrier by a lower factor than the counterpart on the basis of PET. To conclude, the evaluation of the buckling resistance with microscopy offers an efficient method during lacquer development; however, the final decision on the right lacquer composition is dependent on many factors. Full article

The global trade of non-native pet birds has increased in recent decades, and this has accelerated the introduction of invasive birds in the wild. This study employed ensemble species distribution modelling (eSDM) to assess potential habitat suitability and environmental predictor variables influencing the potential distribution of non-native pet bird species reported lost and sighted in South Africa. We used data and information on lost and found pet birds from previous studies to establish and describe scenarios of how pet birds may transition from captivity to the wild. Our study revealed that models fitted and performed well in predicting the suitability for African grey (Psittacus erithacus), Budgerigar (Melopsittacus undulatus), Cockatiel (Nymphicus hollandicus), Green-cheeked conure (Pyrrhura molinae), Monk parakeet (Myiopsitta monachus), and Rose-ringed parakeet (Psittacula krameri), with the mean weighted AUC and TSS values greater than 0.765. The predicted habitat suitability differed among species, with the suitability threshold indicating that between 61% and 87% of areas were predicted as suitable. Species with greater suitability included the African grey, Cockatiel, and Rose-ringed parakeet, which demonstrated significant overlap between their habitat suitability and reported lost cases. Human footprint, bioclimatic variables, and vegetation indices largely influenced predictive habitat suitability. The pathway scenario showed the key mechanisms driving the transition of pet birds from captivity to the wild, including the role of pet owners, animal rescues, adoption practices, and environmental suitability. Our study found that urban landscapes, which are heavily populated, are at high risk of potential invasion by pet birds. Thus, implementing a thorough surveillance survey is crucial for monitoring and evaluating the establishment potential of pet species not yet reported in the wild. Full article

The global increase in the pet trade and ownership of pet birds has heightened the introduction of emerging invasive vertebrate species. We analyzed online databases of lost, found, and sighted non-native pet bird reports in South Africa to evaluate non-native pet bird statuses, investigate geographic patterns, assess species trends, and determine the factors associated with lost pet birds. We identified a total of 1467 case reports representing 77 species across nine families from websites (n = 3) and Facebook pages (n = 13). Most reports of lost birds were within large cities, in populated provinces, including Gauteng, KwaZulu-Natal, and Western Cape. Psittacidae, Psittaculidae, and Cacatuidae were the most dominant families, with African grey (Psittacus erithacus), Cockatiel (Nymphicus hollandicus), and Rose-ringed parakeet (Psittacula krameri) among the top species reported as lost. Lower-priced species were commonly reported as lost, and there was no association between the species’ price and the likelihood of being found. In addition, we found a positive relationship between species reported as lost and the number of pet shops, human population size, species size, and docility. There was a sharp increase in lost cases from 2019 onwards; however, males were more frequently lost. Our findings highlight challenges in regulating and monitoring the pet ownership and trade of non-native pet birds and the need to address commonly kept species in conservation efforts. Online resources can be effective tools for passive surveillance of non-native pet bird species, especially potentially invasive ones. Full article

Synaptic transmission is essential for nervous system function and the loss of synapses is a known major contributor to dementia. Alzheimer’s disease dementia (ADD) is characterized by synaptic loss in the mesial temporal lobe and cerebral neocortex, both of which are brain areas associated with memory and cognition. The association of synaptic loss and ADD was established in the late 1980s, and it has been estimated that 30–50% of neocortical synaptic protein is lost in ADD, but there has not yet been a quantitative profiling of different synaptic proteins in different brain regions in ADD from the same individuals. Very recently, positron emission tomography (PET) imaging of synapses is being developed, accelerating the focus on the role of synaptic loss in ADD and other conditions. In this study, we quantified the densities of two synaptic proteins, the presynaptic protein Synaptosome Associated Protein 25 (SNAP25) and the postsynaptic protein postsynaptic density protein 95 (PSD95) in the human brain, using enzyme-linked immunosorbent assays (ELISA). Protein was extracted from the cingulate gyrus, hippocampus, frontal, primary visual, and entorhinal cortex from cognitively unimpaired controls, subjects with mild cognitive impairment (MCI), and subjects with dementia that have different levels of Alzheimer’s pathology. SNAP25 is significantly reduced in ADD when compared to controls in the frontal cortex, visual cortex, and cingulate, while the hippocampus showed a smaller, non-significant reduction, and entorhinal cortex concentrations were not different. In contrast, all brain areas showed lower PSD95 concentrations in ADD when compared to controls without dementia, although in the hippocampus, this failed to reach significance. Interestingly, cognitively unimpaired cases with high levels of AD pathology had higher levels of both synaptic proteins in all brain regions. SNAP25 and PSD95 concentrations significantly correlated with densities of neurofibrillary tangles, amyloid plaques, and Mini Mental State Examination (MMSE) scores. Our results suggest that synaptic transmission is affected by ADD in multiple brain regions. The differences were less marked in the entorhinal cortex and the hippocampus, most likely due to a ceiling effect imposed by the very early development of neurofibrillary tangles in older people in these brain regions. Full article

In positron emission tomography (PET), the original points of emission are unknown, and the scanners record pairs of photons emitting from those origins and creating lines of response (LORs) in random directions. This presents a latent variable problem, since at least one dimension of relevant information is lost. This can be solved by a statistical approach to image reconstruction—modeling the image as a Gaussian mixture model (GMM). This allows us to obtain a high-quality continuous model that is not computationally demanding and does not require postprocessing. In this paper, we propose a novel method of GMM estimation in the PET setting, directly from lines of response. This approach utilizes some well-known and convenient properties of the Gaussian distribution and the fact that the random slopes of the lines are independent from the points of origin. The expectation–maximization (EM) algorithm that is most commonly used to estimate GMMs in the traditional setting here is adapted to lower-dimensional data. The proposed estimation method is unbiased, and simulations and experiments show that accurate reconstruction on synthetic data is possible from relatively small samples. Full article

Overweight and obesity is one of the most significant health and welfare issues affecting companion animals and are linked to several serious medical conditions, reduced welfare, and shortened lifespan. The number of overweight and obese pets increases every year. Overweight and obesity are associated with multiple chronic diseases. Underlying causes include human-related and animal-related factors. Veterinarians encounter overweight and obese canine patients in daily practice and they play an important role in weight management. This research examines the level of veterinary engagement and the success of veterinary management plans for overweight and obese canines over a five-year period. Electronic medical records (EMRs) were collected for 500 canine patients assessed as either overweight or obese and statistically analyzed for level of veterinary engagement (VE), weight loss success, prescription weight loss diet (RX) use, and comorbidities. The average age at the beginning of the study was 61.5 months. A starting Body Condition Score (BCS) of 6 or 7 was most common (87.2%). Twelve different small and large dog breeds were most highly represented (61.1%). Average weight loss rates were low and more dogs gained rather than lost weight (56.5% vs. 43.5%). While VE is important, this study suggests current VE levels are not adequate to successfully combat overweight and obesity or result in improved animal welfare. Full article

The escape of pet birds into the wild raises concerns about the introduction of invasive avian species. This study investigated the impact of escaped pet birds on the introduction of non-native species in Japan. Data sourced from four lost-and-found pet websites between January 2018 and December 2021 revealed 12,125 recorded escapes exhibiting both daily occurrences and seasonal fluctuations. Statistical modeling identified the monthly average temperature (positively correlated) and maximum electricity demand (negatively correlated) as influential factors. Text analysis revealed “window” and “open” as frequently cited reasons for escapes. Budgerigars (Melopsittacus undulatus) and Cockatiels (Nymphicus hollandicus) accounted for 76% of the total escape, suggesting a low perceived risk of establishment in nonnative environments. Interestingly, two globally established invasive species, the Rose-ringed Parakeet (Psittacula krameri) and Monk Parakeet (Myiopsitta monachus), were among the escaped birds. While the Rose-ringed Parakeet is locally naturalized in Tokyo and its adjacent prefectures, the Monk Parakeet failed to establish itself in Japan. Despite the limited number of escaped Monk Parakeets, ongoing efforts are crucial for preventing the potential re-establishment of species with such capabilities. Full article

Kisspeptins (KPs, KISS1) and their receptor (KISS1R) play a pivotal role as metastasis suppressor for many cancers. Low or lost KP expression is associated with higher tumor grade, increased metastatic potential, and poor prognosis. Therefore, KP expression has prognostic relevance and correlates with invasiveness in cancers. Furthermore, KISS1R represents a very promising target for molecular imaging and therapy for KISS1R-expressing tumors. The goal of this study was to evaluate the developed KISS1-54 derivative, [⁶⁸Ga]KISS1-54, as a PET-imaging probe for KISS1R-expressing tumors. The NODAGA-KISS1-54 peptide was labeled by Gallium-68, and the stability of the resulting [⁶⁸Ga]KISS1-54 evaluated in injection solution and human serum, followed by an examination in different KISS1R-expressing tumor cell lines, including HepG2, HeLa, MDA-MB-231, MCF7, LNCap, SK-BR-3, and HCT116. Finally, [⁶⁸Ga]KISS1-54 was tested in LNCap- and MDA-MB-231-bearing mice, using µ-PET, assessing its potential as an imaging probe for PET. [⁶⁸Ga]KISS1-54 was obtained in a 77 ± 7% radiochemical yield and at a >99% purity. The [⁶⁸Ga]KISS1-54 cell uptake amounted to 0.6–4.4% per 100,000 cells. Moreover, the accumulation of [⁶⁸Ga]KISS1-54 was effectively inhibited by nonradioactive KISS1-54. In [⁶⁸Ga]KISS1-54-PET, KISS1R-positive LNCap-tumors were clearly visualized as compared to MDA-MB-231-tumor implant with predominantly intracellular KISS1R expression. Our first results suggest that [⁶⁸Ga]KISS1-54 is a promising candidate for a radiotracer for targeting KISS1R-expressing tumors via PET. Full article

Multimodal neuroimaging has gained traction in Alzheimer’s Disease (AD) diagnosis by integrating information from multiple imaging modalities to enhance classification accuracy. However, effectively handling heterogeneous data sources and overcoming the challenges posed by multiscale transform methods remains a significant hurdle. This article proposes a novel approach to address these challenges. To harness the power of diverse neuroimaging data, we employ a strategy that leverages optimized convolution techniques. These optimizations include varying kernel sizes and the incorporation of instance normalization, both of which play crucial roles in feature extraction from magnetic resonance imaging (MRI) and positron emission tomography (PET) images. Specifically, varying kernel sizes allow us to adapt the receptive field to different image characteristics, enhancing the model’s ability to capture relevant information. Furthermore, we employ transposed convolution, which increases spatial resolution of feature maps, and it is optimized with varying kernel sizes and instance normalization. This heightened resolution facilitates the alignment and integration of data from disparate MRI and PET data. The use of larger kernels and strides in transposed convolution expands the receptive field, enabling the model to capture essential cross-modal relationships. Instance normalization, applied to each modality during the fusion process, mitigates potential biases stemming from differences in intensity, contrast, or scale between modalities. This enhancement contributes to improved model performance by reducing complexity and ensuring robust fusion. The performance of the proposed fusion method is assessed on three distinct neuroimaging datasets, which include: Alzheimer’s Disease Neuroimaging Initiative (ADNI), consisting of 50 participants each at various stages of AD for both MRI and PET (Cognitive Normal, AD, and Early Mild Cognitive); Open Access Series of Imaging Studies (OASIS), consisting of 50 participants each at various stages of AD for both MRI and PET (Cognitive Normal, Mild Dementia, Very Mild Dementia); and whole-brain atlas neuroimaging (AANLIB) (consisting of 50 participants each at various stages of AD for both MRI and PET (Cognitive Normal, AD). To evaluate the quality of the fused images generated via our method, we employ a comprehensive set of evaluation metrics, including Structural Similarity Index Measurement (SSIM), which assesses the structural similarity between two images; Peak Signal-to-Noise Ratio (PSNR), which measures how closely the generated image resembles the ground truth; Entropy (E), which assesses the amount of information preserved or lost during fusion; the Feature Similarity Indexing Method (FSIM), which assesses the structural and feature similarities between two images; and Edge-Based Similarity (EBS), which measures the similarity of edges between the fused and ground truth images. The obtained fused image is further evaluated using a Mobile Vision Transformer. In the classification of AD vs. Cognitive Normal, the model achieved an accuracy of 99.00%, specificity of 99.00%, and sensitivity of 98.44% on the AANLIB dataset. Full article

Fill product residues in packagings are equivalent to product losses. They are washed out after sorting and before commencing recycling processes. Not much data have been published about how much fill product is still present in packagings dedicated for recycling. Results are often from laboratory trials. Therefore, several hundred packagings from a sorting plant of a dual system in Germany were analysed to determine the amount of fill product residues. Approximately 10 wt. % of highly viscous fill products in tubes were lost as residue. In the case of packagings that were easy to empty, such as cups, and in the case of low-viscosity fill products, such as water, less than 1 wt. % of the fill products remained in the packagings. The mean amount of residue in relation to clean packaging was 0.9 g residue in 1 g of packaging material (without residue) in tubes and 0.07 g in PET bottles. These values were significantly lower for low-viscosity fill products compared to high-viscosity fill products, as expected. Full article

Representations of play abound in Stephen King’s 1983 novel Pet Sematary and its 1989 and 2019 subsequent film adaptations. However, play in Pet Sematary is not representative of the innocent actions designed to create functioning adults who meaningfully contribute to society. In the 1989 film, for example, “play” for a newly resurrected Gage is a version of hide-and-go-seek resulting in the death of neighbour Jud. Meanwhile, the 2019 adaptation sees a newly resurrected Ellie “playing” in her dirt-stained white funereal dress. These dirt stains become markers of lost innocence and transform her dance into an uncanny performance. Since Gage and Ellie are both somewhat monstrous child figures, their play, like their bodies, is transformed into something unsettling and ventures into the realm of the uncanny. However, play itself is also performed differently between the adaptations because the central child figure also changes. In the 1989 film, it is a male toddler, and in the 2019 film, it is a pre-pubescent female. Both adaptations focus on ideal, socially acceptable forms of play according to the time in which the film was made as well as how children diverge from these behaviours. Play is often rendered dangerous when not performed properly according to the paradigms of age and gender, resulting in what I call ‘uncanny play’. When children engage with ‘uncanny play’, the adults in the narrative are permitted to execute the children for the sake of preserving the memory of them as innocent beings, or what I call the ‘Save the Child’ discourse. Linda Hutcheon argues that ‘when we adapt […] we actualize or concretize ideas’, so that the socially acceptable play put forth in King’s novel becomes more realised and thus more at risk to transgression in each successive filmic adaptation. Full article

Aerides Lour. (Orchidaceae, Aeridinae) is a group of epiphytic orchids with high ornamental value, mainly distributed in tropical and subtropical forests, that comprises approximately 20 species. The species are of great value in floriculture and garden designing because of their beautiful flower shapes and colors. Although the morphological boundaries of Aerides are clearly defined, the relationship between Aerides and other closely related genera is still ambiguous in terms of phylogeny. To better understand their phylogenetic relationships, this study used next-generation sequencing technology to investigate the phylogeny and DNA barcoding of this taxonomic unit using genetic information from six Aerides plastid genomes. The quadripartite-structure plastomes ranged from 147,244 bp to 148,391 bp and included 120 genes. Among them, 74 were protein coding genes, 38 were tRNA genes and 8 were rRNA genes, while the ndh genes were pseudogenized or lost. Four non-coding mutational hotspots (rpl20–rpl33, psbM, petB, rpoB–trnC^GCA, Pi > 0.06) were identified. A total of 71–77 SSRs and 19–46 long repeats (>30 bp) were recognized in Aerides plastomes, which were mostly located in the large single-copy region. Phylogenetic analysis indicated that Aerides was monophylic and sister to Renanthera. Moreover, our results confirmed that six Aerides species can be divided into three major clades. These findings provide assistance for species identification and DNA barcoding investigation in Aerides, as well as contributes to future research on the phylogenomics of Orchidaceae. Full article

¹⁸F-FDG PET-CT is routinely performed as part of the initial staging of numerous cancers. Other than having descriptive, predictive and prognostic values for tumors, ¹⁸F-FDG PET-CT provides full-body data, which could inform on concurrent pathophysiological processes such as malnutrition. To test this hypothesis, we measured the ¹⁸F-FDG uptake in several organs and evaluated their association with weight loss in patients at diagnosis of esophageal cancer. Forty-eight patients were included in this retrospective monocentric study. ¹⁸F-FDG uptake quantification was performed in the brain, the liver, the spleen, bone marrow, muscle and the esophageal tumor itself and was compared between patients with different amounts of weight loss. We found that Total Lesion Glycolysis (TLG) and peak Standardized Uptake Values (SUV_peak) measured in the brain correlated with the amount of weight loss: TLG was, on average, higher in patients who had lost more than 5% of their usual weight, whereas brain SUV_peak were, on average, lower in patients who had lost more than 10% of their weight. Higher TLG and lower brain SUV_peak were associated with worse OS in the univariate analysis. This study reports a new and significant association between ¹⁸F-FDG uptake in the brain and initial weight loss in patients with esophageal cancer. Full article