In this study, we evaluated how gene expression differs in mature Pseudomonas aeruginosa biofilms as opposed to planktonic cells by the use of RNA sequencing technology that gives rise to both ...quantitative and qualitative information on the transcriptome. Although a large proportion of genes were consistently regulated in both the stationary phase and biofilm cultures as opposed to the late exponential growth phase cultures, the global biofilm gene expression pattern was clearly distinct indicating that biofilms are not just surface attached cells in stationary phase. A large amount of the genes found to be biofilm specific were involved in adaptation to microaerophilic growth conditions, repression of type three secretion and production of extracellular matrix components. Additionally, we found many small RNAs to be differentially regulated most of them similarly in stationary phase cultures and biofilms. A qualitative analysis of the RNA-seq data revealed more than 3000 putative transcriptional start sites (TSS). By the use of rapid amplification of cDNA ends (5'-RACE) we confirmed the presence of three different TSS associated with the pqsABCDE operon, two in the promoter of pqsA and one upstream of the second gene, pqsB. Taken together, this study reports the first transcriptome study on P. aeruginosa that employs RNA sequencing technology and provides insights into the quantitative and qualitative transcriptome including the expression of small RNAs in P. aeruginosa biofilms.
Using case examples and other common scenarios, the authors outline several types of moral distress that clinicians may experience when providing pediatric gender-affirming care. The authors also ...offer strategies for managing moral distress and issue a call to action for professional organizations to enhance support for gender-affirming clinicians.
The dramatic rise in the diagnosis of autism spectrum disorders (ASDs) has resulted in a significant explosion in visibility and a substantial increase in the understanding of this complex group of ...disorders. Over the last decade, what was once defined only as “autism” has become a spectrum of neurodevelopmental disorders, with variable presentations and levels of impairment that requires an equally broad continuum of care. A wealth of research has not discovered a single “cause” for ASDs, but rather has found a range of genetic/genomic variations that likely play a significant role in the etiology of these disorders. Routine screening for autism is becoming more common, allowing for more timely diagnosis, and hopefully for earlier entry into appropriate and effective treatment. Although there remains no single ‘‘cure’’, there are treatments available that can improve overall functioning and decrease problematic or interfering symptoms across the full spectrum of this disorder. The wide spectrum of ASD presentations complicates treatment planning, with overall ASD severity a factor in determining type, intensity, and duration of interventions and services. This article presents a two-dimensional model of “ASD severity” that considers both the level of specific ASD symptoms/deficits and the level of cognitive resources/limitations as a framework for understanding needs, challenges, and potentially effective interventions for individuals across the ASD spectrum.
Pseudomonas aeruginosa is a gram-negative bacterium that is ubiquitously present in the aerobic biosphere. As an antibiotic-resistant facultative pathogen, it is a major cause of hospital-acquired ...infections. Its rapid and accurate identification is crucial in clinical and therapeutic environments.
In a large-scale MALDI-TOF mass spectrometry-based screen of the Harvard transposon insertion mutant library of P. aeruginosa strain PA14, intact-cell proteome profile spectra of 5547 PA14 transposon mutants exhibiting a plethora of different phenotypes were acquired and analyzed.
Of all P. aeruginosa PA14 mutant profiles 99.7% were correctly identified as P. aeruginosa with the Biotyper software on the species level. On the strain level, 99.99% of the profiles were mapped to five different individual P. aeruginosa Biotyper database entries. A principal component analysis-based approach was used to determine the most important discriminatory mass features between these Biotyper groups. Although technical replicas were consistently categorized to specific Biotyper groups in 94.2% of the mutant profiles, biological replicas were not, indicating that the distinct proteotypes are affected by growth conditions.
The PA14 mutant profile collection presented here constitutes the largest coherent P. aeruginosa MALDI-TOF spectral dataset publicly available today. Transposon insertions in thousands of different P. aeruginosa genes did not affect species identification from MALDI-TOF mass spectra, clearly demonstrating the robustness of the approach. However, the assignment of the individual spectra to sub-groups proved to be non-consistent in biological replicas, indicating that the differentiation between biotyper groups in this nosocomial pathogen is unassured.
Abstract
The objective was to evaluate a novel brewer’s yeast product on ruminal fiber fermentation in vitro. The experiment was a completely randomized design and included 4 treatments. Treatments ...included a negative control (NCON), a commercial brewer’s yeast product (Emprove MX; EMX), and a prototype concentrated brewer’s yeast (T1R8) fed at low and high inclusions (T1R8-L and T1R8-H, respectively). Inclusions for low and high treatments were 0.0008 and 0.0015 g, respectively, to correspond with 7 and 14 g inclusion in a lactating cow diet. Treatments were evaluated in duplicate flasks in 2 replicated runs for a total of four observations per treatment. Flasks contained 2.5 grams dry matter (DM) of substrate to represent a lactating cow diet (30% haylage, 25% corn silage, 20% corn, 15% canola meal, and 10% soybean hulls), and 150mL of rumen fluid mixture (2 McDougall’s buffer:1 rumen fluid ratio). Four F57 sealed Ankom bags were filled with 0.25 grams DM of substrate each to facilitate measuring in vitro DM and neutral detergent fiber disappearance (IVDMD and IVNDFD, respectively). Flasks were incubated at 39°C for 30 hours with sample aliquots collected at hour 0, 8, 16, and 30 to determine pH and volatile fatty acids (VFA). Data were analyzed using the MIXED procedure of SAS 9.4 with IVDMD and IVNDFD considered a main effect of treatment while pH and VFA were analyzed with effects of treatment, time, and their interaction. No differences in IVDMD were observed (P = 0.31) among treatments. Treatment affected (P = 0.01) IVNDFD with EMX (50.6%), T1R8-L (48.6%), and T1R8-H (50.3%) being greater than NCON (45.7%). A treatment by hour interaction (P < 0.01) was observed for total VFA concentration; EMX (71.8 mM), T1R8-L (72.6 mM), and T1R8-H (66.0 mM) were greater than NCON (47.8 mM) at h 8, but all treatments were not different at h 16. A treatment by hour interaction (P < 0.01) was also observed for acetate and propionate; EMX, T1R8-H, and T1R8-L were greater than NCON at h 8, but all treatments were not different at h 16. At h 30, greater propionate was observed for T1R8-H compared with all treatments. No effects of treatment or treatment by hour (P ≥ 0.89) were observed for acetate:propionate ratio. Although a treatment by hour interaction was not observed for pH (P = 0.12), a treatment effect (P = 0.03) indicated T1R8-L was the greatest with EMX being intermediate, and NCON and T1R8-H being the least. Novel brewer’s yeast product T1R8 increased the rate of substrate fermentation in the initial 8 h and increased the extent of fiber degradation after a 30 h in vitro fermentation.
Background Non-well-differentiated cutaneous squamous cell carcinomas may display a more aggressive behavior. It is important to better define prognostic criteria for these tumors. Methods This was a ...retrospective case-control analysis of a squamous cell carcinoma database. Patients with non-well-differentiated and well-differentiated tumors were matched based on site of tumor, age, and immunocompromised status. Comparisons included demographics, histology, immunohistochemical protein expressions (Ki-67, p53, E-cadherin, cyclin D1), and clinical outcomes. Results Demographic features were similar between cases ( n = 30) and controls ( n = 30). Non-well-differentiated tumors were larger (1.8 cm versus 1.3 cm, P = 0.08), deeper (0.81 cm versus 0.32 cm, P < 0.0001), and had greater recurrence ( P = 0.003). Non-well-differentiated tumors showed increased proliferation rate, Ki-67 index (77% versus 61%, P = 0.001); no significant difference in activity of p53, E-cadherin, and cyclin D1 between the two groups. Conclusions Tumor differentiation and depth are important pathologic and prognostic criteria for cutaneous squamous cell carcinoma. Immunohistochemistry helps describe patterns of biomarker protein expression and may exemplify aggressive subtypes.
Comprehension of dental implant components and ideal placement, as well as a foundation of intraoral radiography technique is critical to a successful intraoral dental implant radiograph. Because of ...the precision required in radiographic evaluation of dental implants, the application of proper technique and the ability to troubleshoot radiographic imaging errors is essential. ...expose the radiograph, assess outcome, and troubleshoot if needed. Intraoral radiography involves placement of an imaging plate or sensor in the oral cavity and an extraoral x-ray exposure by a certified or licensed professional.4 Imaging receptors capture the projected image of focus, therefore receptor size and placement should be taken into consideration, to ensure the radiograph is diagnostic.5 The standard diagnostic criteria for periapical intraoral radiographs include optimal exposure settings, anatomically accurate images with minimal distortion, inclusion of entire tooth structure as well as the capture of 2-3 mm beyond the root apices.3 There are two techniques for intraoral radiography, paralleling technique and bisecting angle technique (see figures 1 and 2). Due to potential risk of elongation, foreshortening, and distortion with bisecting angle technique, the preferred intraoral imaging technique is paralleling.4 Intraoral Dental Implant Radiography Intraoral radiographs of natural teeth and surrounding structures are utilized to assess many conditions, including but not limited to dental caries, dental infection, and bone loss.4 Intraoral radiography, however, has the capacity to be applied to diagnosis and evaluation of dental implants, among other orofacial conditions.6 While similar, intraoral dental implant radiography has different criteria for success than natural dentition and surrounding structures, so traditional methods utilized in imaging of natural teeth may need to be modified for dental implants.
Enterobacter cloacae infections are common among burn victims, immunocompromised patients, and patients with malignancy. Most commonly these infections are manifested as nosocomial urinary tract or ...pulmonary infections. Nosocomial outbreaks have also been associated with colonization of certain surgical equipment and operative cleaning solutions. Infections of an aortobifemoral prosthesis, an aortic graft, and arteriovenous fistulae are noted in the literature. To our knowledge, this is the first isolated account of an E. cloacae infection of a femoral-popliteal expanded polytetrafluoroethylene bypass graft.
A 68-year-old Caucasian man presented with fever and rest pain in the right lower extremity five months after the placement of a vascular expanded polytetrafluoroethylene graft for femoral-popliteal bypass. Computed tomography angiography demonstrated peri-graft fluid that was aspirated percutaneously with image guidance and cultured to reveal E. cloacae. The graft was revised and then removed. The patient completed a six-week course of ceftazidime and is currently without signs of infection. There were no other reports of E. cloacae graft infections in any patients receiving treatment in the same surgical suite within a month of this report.
Isolated cases of E. cloacae infection of surgical bypass grafts are rare (unique in this setting). Clinicians should have a high index of suspicion for device contamination in such cases and should consider testing for possible microbial reservoirs. Graft removal is required due to the formation of biofilm and the recent emergence of Enterobacteriaceae producing extended-spectrum beta-lactamase in community acquired infections.
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