Hairy root systems have proven to be a viable alternative for recombinant protein production. For recalcitrant proteins, maximizing the productivity of hairy root cultures is essential. The aim of this study was to optimize a Brassica rapa rapa hairy root process for secretion of α-L-iduronidase (IDUA), a biologic of medical value. The process was first optimized with hairy roots expressing eGFP. For the biomass optimization, the highest biomass yields were achieved in modified Gamborg B5 culture medium. For the secretion induction, the optimized secretion media was obtained with additives (1.5g/l PVP + 1mg/l 2,4-D + 20.5g/l KNO 3) resulting in 3.4 fold eGFP secretion when compared to the non-induced control. These optimized conditions were applied to the IDUA-expressing hairy root clone, confirming that the highest yields of secreted IDUA occurred when using the already defined additive combination. The functionality of the IDUA protein, secreted and intracellular, was confirmed with an enzymatic activity assay. A >150-fold increase of the IDUA activity was observed using an optimized secretion medium, compared with a non-induced medium. We have proven that our B. rapa rapa hairy root system can be harnessed to secrete recalcitrant proteins, illustrating the high potential of hairy roots in plant molecular farming.

Eosinophilic gastritis (EG) is characterized by eosinophilic infiltration of any gastric layers. We report a 65- year-female presenting with abdominal pain and vomiting for two months. Chronic gastritis not responding to empirical treatment interrogated further investigations. In the absence of atopy and peripheral eosin-ophilia, successful treatment of a large solitary antral ulcer with steroids upheldthe diagnosis of EG.

Coronary artery disease (CAD) is a common comorbidity of gastro-esophageal reflux disease (GERD). Since the distal esophagus and heart share a common afferent vagal supply, the delivery of radiofrequency in the distal esophagus can stimulate the coronary artery and induce acute myocardial infarction (AMI) in a patient with CAD.

A 75-year-old man with an aortic bioprosthesis was admitted with polyarthritis in a non-febrile setting. Blood cultures were positive for Listeria monocytogenes. The diagnosis of Listeria endocarditis and spondylodiscitis was evoked. These are two unusual forms of listeriosis.The evolution was favourable after antibiotic therapy.

Saccharomyces cerevisiae produces a multicellular phenotype, known as a mat, on a semi-solid medium. This biofilm phenotype was first described in the lab strain 1278b and has been analyzed mostly in this same background. Yeast cells form a mat by spreading across the medium and adhering to each other and the surface, in part through the variegated expression of the cell adhesin, FLO11. This process creates a characteristic floral pattern and generates pH and glucose gradients outward from the center of the mat. Mats are encapsulated in a liquid which may aid in surface spreading and diffusion. Here, we examine thirteen environmental isolates that vary visually in the phenotype. We predicted that mat properties were universal and increased morphological complexity would be associated with more extreme trait values. Our results showed that pH varied significantly among strains, but was not correlated to mat complexity. Only two isolates generated significant liquid boundaries and neither produced visually complex mats. In five isolates, we tracked the initiation of FLO11 using GFP under the control of the endogenous promoter. Strains varied in when and how much GFP was detected, with increased signal associated with increased morphological complexity. Generally, the signal was strongest in the center of the mat and absent at the expanding edge. Our results show that traits discovered in one background vary and exist independently of mat complexity in natural isolates. The environment may favor different sets of traits, which could have implications for how this yeast adapts to its many ecological niches.

Consider the nonautonomous semilinear evolution equation of type: $(\star) \; u’(t)=A(t)u(t)+f(t,u(t)), \; t \in \mathbb{R},$ where $ A(t), \ t\in \mathbb{R} $ is a family of closed linear operators on a Banach space $X$, the nonlinear term $f$, acting on some real interpolation spaces, is assumed to be almost periodic only in a weak sense (i.e. in Stepanov sense) with respect to $t$ and Lipschitzian in bounded sets with respect to the second variable. We prove the existence and uniqueness of positive almost periodic solutions in the strong sense (Bohr sense) for equation $ (\star) $ using the exponential dichotomy approach. Then, we establish a new composition result of Stepanov almost periodic functions by assuming only the continuity of $f$ in the second variable. Moreover, we provide an application to a nonautonomous system of reaction–diffusion equations describing a Lotka–Volterra predator–prey model with diffusion and time–dependent parameters in a generalized almost periodic environment.

Objective: The aim of the study is to assess the therapeutic effect and applicability of pectoralis major muscle turnover flap reconstruction for treatment of deep sternal wound infection after cardiac surgery in infants and children. Methods: From march 2013 to october 2021, 23 patients with deep sternal wound infection after cardiac surgery underwent pectoralis major muscle turnover flap reconstruction.The data and outcomes of the patients were retrospectively analyzed. Results: 20 patients were treated with unilateral pectoralis major muscle turnover flap reconstruction,3 patients were treated by bilateral pectoralis major muscle turnover flap. All of the sternal wounds healed successfully. All patients survived and were discharged without evidence of infection. In a follow-up period, ranging from 15 to 83 months (mean 32.6 months), all patients demonstrated normal development with no limitations to limb movements. There were no signs of chronic sternal infection in all of them. Conclusion:Pectoralis major muscle turnover flap reconstruction is a simple,feasible and effective treatment of deep sternal wound infection after cardiac surgery in infants and children,with minimal developmental problems.

Radiofrequency (RF) ablation has been the most widely employed energy source for catheter ablation to date. However, most of conventional RF ablation energy dissipates into the bloodstream before reaching the target tissue. Technology that conveys RF energy exclusively toward target tissue may potentially improve the quality, safety, and outcome of the RF ablation procedures. RF ablation using a novel insulated-tip catheter (SMT, Sirona Medical Technologies, Windsor, CT) may refine RF ablation in the future to minimize the risk of iatrogenic complications. Although it is still unclear whether the results of the SMT catheter can be translated to a human beating heart, the data for SMT catheter of this study are very promising.

Actinobacteria are important cave inhabitants, but knowledge of how anthropization and anthropization-related visual marks affect this community on cave walls is lacking. We compared Actinobacteria communities among four French limestone caves (Mouflon, Reille, Rouffignac, Lascaux) ranging from pristine to anthropized, and within Lascaux Cave between marked (wall visual marks) and unmarked areas in different rooms (Sas-1, Passage, Apse, Diaclase). In addition to the 16S rRNA gene marker, 441 bp fragments of the hsp65 gene were used and an hsp65-related taxonomic database was constructed for identification of Actinobacteria to the species level by Illumina-MiSeq analysis. The hsp65 marker revealed higher resolution for species and higher richness (99% OTU cutoff) versus 16S rRNA gene; however, more taxa were identified at higher taxonomic ranks. Actinobacteria communities varied between Mouflon and Reille caves (both pristine), and Rouffignac and Lascaux (both anthropized). Rouffignac displayed high diversity of Nocardia, suggesting human inputs, and Lascaux exhibited high Mycobacterium relative abundance, whereas Gaiellales were typical in pristine caves and the Diaclase (least affected area of Lascaux Cave). Within Lascaux, Pseudonocardiaceae dominated on unmarked walls and Streptomycetaceae (especially Streptomyces mirabilis) on marked walls, indicating a possible role in mark formation. A new taxonomic database (https://zenodo.org/record/5576074) was developed. Although not all Actinobacteria species were represented, the use of the hsp65 marker enabled species-level variations of the Actinobacteria community to be documented based on the extent of anthropogenic pressure. This approach proved effective when comparing different limestone caves or specific conditions within one cave.

Human disturbance directly affects animal populations but indirect effects of disturbance on species behaviors are less well understood. Camera traps provide an opportunity to investigate variation in animal behaviors across gradients of disturbance. We used camera trap data to test predictions about predator-sensitive behavior in three ungulate species (caribou Rangifer tarandus; white-tailed deer, Odocoileus virginianus; moose, Alces alces) across two boreal forest landscapes varying in disturbance. We quantified behavior as the number of camera trap photos per detection event and tested its relationship to predation risk between a landscape with greater industrial disturbance and predator abundance (Algar) and a “control” landscape with lower human and predator activity (Richardson). We also assessed the influence of predation risk and habitat on behavior across camera sites within the disturbed Algar landscape. We predicted that animals in areas with greater predation risk (more wolf activity, less cover) would travel faster and generate fewer photos per event, while animals in areas with less predation risk would linger (rest, forage), generating more photos per event. Consistent with predictions, caribou and moose had more photos per event in the landscape where predation risk was reduced. Within the disturbed landscape, no prey species showed a significant behavioral response to wolf activity, but the number of photos per event decreased for white-tailed deer with increasing line of sight (m) along seismic lines (i.e. decreasing visual cover), consistent with a predator-sensitive response. The presence of juveniles was associated with shorter behavioral events for caribou and moose, suggesting greater predator sensitivity for females with calves. Only moose demonstrated a positive association with vegetation productivity (NDVI), suggesting that for other species influences of forage availability were generally weaker than those from predation risk. Behavioral insights can be gleaned from camera trap surveys and provide information about animal responses to predation risk and the indirect impacts of human disturbances.

Humans inhale, ingest and touch thousands of fungi each day. The ubiquity and diversity of the fungal kingdom are in sharp contrast with their complex and relatively blurred taxonomy and scarce knowledge about their distribution, pathogenic effects, and effective interventions at the environmental and individual levels. Here, we present an overview of salient features of fungi as permanent players of the human exposome and key determinants of human health. Improved understanding of the fungal exposome sheds new light on the epidemiology of fungal-related diseases, their immunological substratum, the currently available methods, and biomarkers for environmental and medical fungi. Unmet needs are described and potential approaches are highlighted as perspectives.

Species distributions are one of the fundamental factors needed for understanding and conserving wildlife. While the IUCN Red List of Threatened Species is the primary applied reference for biodiversity conservation, limitations in data availability and analyses of the distributions of some species may limit accurate threat classification assessments and conservation recommendations. Improving the accuracy of species distributions in light of growing data and analytical methods is a key step to increasing the efficacy of the Red List. In this study, we reassessed the distribution of takin (Budorcas taxicolor tibetana), a large ungulate in Sichuan Province, southwest China , classified by the IUCN as vulnerable. Using species distribution models and reported habitat requirements, we updated the takin distribution map. Our updated distribution range in the study area (79,449km2) was 61.31% of current distribution range (CDR) on the IUCN red list. This reduction was in large part due to the inclusion in the CDR of substantial areas of lowland plains, high human disturbance, and non-forest habitat, which provide no suitable habitat for takin. According to our results, suitable takin habitat covered 18.97% of the CDR, suggesting a substantially over-estimated distribution. However, there are high proportions of habitat (40%) still covered by the nature reserve network, indicating the importance of protected areas (PAs) in conservation threaten species. We recommend that experts apply the basic approach presented herein to update the Red List distributions for more species to increase the accuracy of assessments and resulting conservation applications.

We describe a case of pancreatic tumor associated with a giant type IV hiatal hernia that had prolapsed into the posterior mediastinum. We initially performed hiatal hernia repair, followed by laparoscopic distal pancreatectomy, because the hernia repair enables performing pancreatectomy safety and in the normal anatomical position.

High-dose methotrexate (HD-MTX) with rigorous supportive care is essential to the treatment of pediatric non-Hodgkin lymphomas (NHL). We describe the safety and tolerability of HD-MTX in patients with NHL treated at our center. In our cohort of 46 patients, the majority had at least one course of delayed clearance and/or creatinine elevation. Additionally, more than one-third of patients experienced an episode of grade ≥3 mucositis. Creatinine elevations and delayed clearance were independently associated with subsequent grade ≥3 mucositis. We advocate for greater availability of methotrexate monitoring to allow dose escalation of this essential modality around the world.

The use of an adequate protocol that accurately extracts microbial DNA from bovine milk samples is of importance for downstream analysis such as 16S rRNA gene sequencing. Although sequencing platforms such as Illumina are very common, there are reservations concerning reproducibility in challenging samples that combine low bacterial loads with high amounts of host DNA. The objective of this study was to evaluate six different DNA extraction protocols applied to four different prototype milk samples (low/high level of colony-forming units (cfu) and somatic cells). DNA extracts were sequenced on Illumina MiSeq with primers for the hypervariable regions V1V2 and V3V4. The different protocols were evaluated by analyzing the yield and purity of DNA extracts and the number of clean reads after sequencing. Three protocols with the highest median number of clean reads were selected. To assess reproducibility, these extraction replicates were re-sequenced in triplicates (n=120). The most reproducible results for alpha- and beta-diversity were obtained with the modified DNeasy Blood & Tissue kit after a chemical pre-treatment plus resuspension of the cream fraction. The unmodified QIAamp DNA Mini kit performed particularly weak in the sample representing unspecific mastitis. These results suggest that pre-treatment in combination with the modified DNeasy Blood & Tissue kit is useful in extracting microbial DNA from challenging milk samples. To increase reproducibility, we recommend that duplicates, if not triplicates, should be sequenced. We showed that high counts of somatic cells challenged DNA extraction, which shapes the need to apply modified extraction protocols.

The complex boundary of the elliptical inclusion rendered it difficult to solve the problem of wave scattering. In this study, the steady-state response was analyzed using the wave function expansion method. Subsequently, the Ricker wavelet was employed as the transient disturbance and Fourier transform was used to determine the distribution of transient dynamic stress concentration around the elliptical inclusion. The effects of wave number, elliptical axial ratio and difference in material properties on the distribution of the dynamic stress concentration around the elliptical inclusion were evaluated. The numerical results revealed that the dynamic stress concentration always appeared at both ends of the major axis and minor axis of the elliptical inclusion, and the difference in material properties between the inclusion and medium influenced the variations in the dynamic stress concentration factor with the wave number and elliptical axial ratio.

Subdural empyema is a rare intracranial infection with an accumulation of purulent material between the dura and arachnoid matter. We report a case of 17 years old presented with altered conscious level .CSF analysis showed increased WBCS. His situation has improved after treating by acyclovir, ceftriaxone, vancomycin and dexamethasone.

Submerged macrophytes play a key role in maintaining a clear-water phase and promoting biodiversity in shallow aquatic ecosystems. Since their abundance has declined globally due to anthropogenic activities, it is important to include them in aquatic ecosystem restoration programs. That macrophytes establish in early spring is crucial for maintaining the macrophyte communities for the remainder of the year. However, factors affecting this early establishment of submerged macrophytes have not been fully explored yet. Here, we conducted an outdoor experiment from winter to early spring using the submerged macrophytes Potamogeton crispus and Vallisneria spinulosa to study the effects of shading, nutrient loading, snail herbivory (Radix swinhoei) and their interactions on the early growth and stoichiometric characteristics of macrophytes. The results show that the effects strongly depend on macrophyte species. Biomass and number of shoots of P. crispus decreased, and internode length increased during low light conditions, but were not affected by nutrient loading. P. crispus shoot biomass and number showed hump-shaped responses to increased snail biomass under full light. In contrast, the biomass of the plant linearly decreased with snail biomass under low light. This indicates an interaction of light with snail herbivory. Since snails prefer grazing on periphyton over macrophytes, a low density of snails promoted growth of P. crispus by removing periphyton competition, while herbivory on the macrophyte increased during a high density of snails. The growth of V. spinulosa was not affected by any of the factors, probably because of growth limitation by low temperature. Our study demonstrates that the interaction of light with snail herbivory may affect establishment and growth of submerged macrophytes in early spring. Macrophyte restoration projects may thus benefit from lowering water levels to increase light availability and making smart use of cold-adapted herbivores to reduce light competition with periphyton.