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Ferrocyanide-reduced osmium tetroxide has been introduced by de Bruyn and Kamovsky primarily as an ultrastructural stain and by Karnovsky secondarily as a possible method to increase contrast in existing cytochemical methods requiring conventional osmication. de Bruyn's staining medium consists of 1.65% potassium ferricyanide [K3Fe(CN)6] in 1% osmium tetroxide (OSO4) while Kamovsky's consists of 1.5% K3Fe(CN)6 in 1% osmium tetroxide. The staining complex has been proposed by de Bruyn (3) to be:
We report here our experience in comparing this staining medium with OSO4. Various rat tissues, e.g., adrenal, epididymus, heart, kidney, liver, prostate and testis were fixed in 3% glutaraldehyde-O.1 M phosphate (pH 7.4) and postfixed in potassium-osmium-cyanide complex (POCC) staining medium (1.5% K3Fe(CN)6 in 1% osmium tetroxide). In addition, liver and kidney were fixed in freshly de- polymerized 2% paraformaldehyde-0.1 M phosphate (pH 7.4) and postfixed in POCC.
Our studies of mammary tumor virus have included the application of the unlabeled antibody enzyme method of Sternberger to mammary tumor derived mouse cells in culture and observation with an electron microscope. The method avoids the extravagance of covalent binding of indicator molecules (horseradish peroxidase) with precious antibody locator molecules by relying instead upon specific antibody-antigen linkages. Our reagents included: Primary Antibody, rabbit anti-murine mammary tumor virus (MuMTV) which was antiserum 113 AV-2; Secondary Antibody, goat anti-rabbit IgG gamma chain (Cappel Laboratories); andthe Indicator, rabbit anti-horseradish peroxidase - horseradish peroxidase complex (PAP) (Cappel Labs.). Dilutions and washes were made in 0.05 M Tris 0.15 M saline buffered to pH 7.4. Cell monolayers, after light fixation in glutaraldehyde, were incubated in place by a protocol adapted from Sternberger and Graham and Karnovsky, then embedded by our usual method for monolayers. Reagents were confined to specific areas by neoprene 0-rings (Parker Seal Co.) reducing the amount of reagent needed to 50 microliters, 1/6th of that required to wet a 35 mm petri dish.
During the design and synthesis of new chemotherapeutic agents for prostatic carcinoma based on phosphorylated agents which might be enzyme-activated to cytotoxicity, phosphorylcholine, [(CH3)3+NCH2CH2OPO3Ca]Cl-, has been indicated to be a very specific substrate for prostatic acid phosphatase (PAP). This phenomenon has led to the development of specific histochemical and ultracytochemical methods for PAP using modifications of the Gomori lead method for acid phosphatase. Comparative histochemical results in prostate and kidney of the rat have been published earlier with phosphorylcholine (PC) and β-glycerophosphate (βGP). We now report the ultracytochemical results.
Minced tissues were fixed in 3% glutaraldehyde-0.1 M phosphate buffered (pH 7.4) for 1.5 hr and rinsed overnight in several changes of 0.05 M phosphate buffer (pH 7.0) containing 7.5% sucrose. Tissues were incubated 30 min to 2 hr in Gomori acid phosphatase medium (2) containing 0.1 M substrate, either PC or βGP.
The following position statement from the Union of the European Phoniatricians, updated on 25th May 2020 (superseding the previous statement issued on 21st April 2020), contains a series of recommendations for phoniatricians and ENT surgeons who provide and/or run voice, swallowing, speech and language, or paediatric audiology services.
This material specifically aims to inform clinical practices in countries where clinics and operating theatres are reopening for elective work. It endeavours to present a current European view in relation to common procedures, many of which fall under the aegis of aerosol generating procedures.
As evidence continues to build, some of the recommended practices will undoubtedly evolve, but it is hoped that the updated position statement will offer clinicians precepts on safe clinical practice.
Spot-spraying POST herbicides is an effective approach to reduce herbicide input and weed control cost. Machine vision detection of grass or grass-like weeds in turfgrass systems is a challenging task due to the similarity in plant morphology. In this work, we explored the feasibility of using image classification with deep convolutional neural networks (DCNN), including AlexNet, GoogLeNet, and VGGNet, for detection of crabgrass species (Digitaria spp.), doveweed [Murdannia nudiflora (L.) Brenan], dallisgrass (Paspalum dilatatum Poir.), and tropical signalgrass [Urochloa distachya (L.) T.Q. Nguyen] in bermudagrass [Cynodon dactylon (L.) Pers.]. VGGNet generally out-performed AlexNet and GoogLeNet in detecting selected grassy weeds. For detection of P. dilatatum, VGGNet achieved high F1 scores (≥0.97) and recall values (≥0.99). A single VGGNet model exhibited high F1 scores (≥0.93) and recall values (1.00) that reliably detected Digitaria spp., M. nudiflora, P. dilatatum, and U. distachya. Low weed density reduced the recall values of AlexNet at detecting all weed species and GoogLeNet at detecting Digitaria spp. In comparison, VGGNet achieved excellent performances (overall accuracy = 1.00) at detecting all weed species in both high and low weed density scenarios. These results demonstrated the feasibility of using DCNN for detection of grass or grass-like weeds in turfgrass systems.
The new compound (4R)-methyl-3-(1-(4-chlorophenyl)-1H-1,2,3-triazole-4-carbonyl)thiazolidin-4-carboxylate was synthesized by the 1,3-dipolar cycloaddition reaction between (4R)-methyl-3-propionyl-thiazolidin-4-carboxylate (1) and 4-chlorophenylazide using the click chemistry approach. Molecular characterization was carried out by infrared spectroscopy and mass spectrometry. The X-ray powder diffraction study determined that the title compound crystallized in an orthorhombic system with unit-cell parameters a = 20.876 (2) Å, b = 12.111 (1) Å, and c = 6.288 (9) Å. The volume of the unit cell is V = 1589.7 (2) Å3. All measured diffraction maxima were indexed and are consistent with the P2221 space group (No. 17). No detectable impurities were observed.
Background: There is an unmet need for blood-based biomarkers that can reliably detect MS disease activity. Serum Biomarkers of interest includ Neurofilament-light-chain (NfL), Glial-fibrillary-strocyte-protein(GFAP) and Tau. Bone Marrow Transplantation (BMT) is reserved for aggressive forms of MS and has been shown to halt detectable CNS inflammatory activity for prolonged periods. Significant pre-treatment tissue damage at followed by inflammatory disease abeyance should be reflected longitudinal sera collected from these patients. Methods: Sera were collected from 23 MS patients pre-treatment, and following BMT at 3, 6, 9 and 12-months in addition from 33 non-inflammatory neurological controls. Biomarker quantification was performed with SiMoA. Results: Pre-AHSCT levels of serum NfL and GFAP but not Tau were elevated compared to controls (p=0.0001), and NfL correlated with lesion-based disease activity (6-month-relapse, MRI-T2 and Gadolinium-enhancement). 3-months post-treatment, while NfL levels remained elevated, Tau/GFAP paradoxically increased (p=0.0023/0.0017). These increases at 3m correlated with MRI ‘pseudoatrophy’ at 6-months. NfL/Tau levels dropped to that of controls by 6-months (p=0.0036/0.0159). GFAP levels dropped progressively after 6-months although even at 12-months remained higher than controls (p=0.004). Conclusions: NfL was the closest correlate of MS disease activity and treatment response. Chemotherapy-related toxicity may account for transient increases in NfL, Tau and MRI brain atrophy post-BMT.
In this paper, we will report about the implementation of the self-consistent Kröner–Eshelby model for the calculation of X-ray elastic constants for general, triclinic crystal symmetry. With applying appropriate symmetry relations, the point groups of higher crystal symmetries are covered as well. This simplifies the implementation effort to cover the calculations for any crystal symmetry. In the literature, several models can be found to estimate the polycrystalline elastic properties from single crystal elastic constants. In general, this is an intermediate step toward the calculation of the polycrystalline response to different techniques using X-rays, neutrons, or ultrasonic waves. In the case of X-ray residual stress analysis, the final goal is the calculation of X-ray Elastic constants. Contrary to the models of Reuss, Voigt, and Hill, the Kröner–Eshelby model has the benefit that, because of the implementation of the Eshelby inclusion model, it can be expanded to cover more complicated systems that exhibit multiple phases, inclusions or pores and that these can be optionally combined with a polycrystalline matrix that is anisotropic, i.e., contains texture. We will discuss a recent theoretical development where the approaches of calculating bounds of Reuss and Voigt, the tighter bounds of Hashin–Shtrikman and Dederichs–Zeller are brought together in one unifying model that converges to the self-consistent solution of Kröner–Eshelby. For the implementation of the Kröner–Eshelby model the well-known Voigt notation is adopted. The 4-rank tensor operations have been rewritten into 2-rank matrix operations. The practical difficulties of the Voigt notation, as usually concealed in the scientific literature, will be discussed. Last, we will show a practical X-ray example in which the various models are applied and compared.
Identifying Research Domain Criteria (RDoC) constructs in early childhood is essential for understanding etiological pathways of psychopathology. Our central goal was to identify early emotion knowledge and self-regulation difficulties across different RDoC domains and examine how they relate to typical versus atypical symptom trajectories between ages 3 and 10. Particularly, we assessed potential contributions of children's gender, executive control, delay of gratification, and regulation of frustration, emotion recognition, and emotion understanding at age 3 to co-occurring patterns of internalizing and externalizing across development. A total of 238 3-year-old boys and girls were assessed using behavioral tasks and parent reports and reassessed at ages 5 and 10 years. Results indicated that very few children developed “pure” internalizing or externalizing symptoms relative to various levels of co-occurring symptoms across development. Four classes of co-occurring internalizing and externalizing problems were identified: low, low-moderate, rising, and severe-decreasing trajectories. Three-year-old children with poor executive control but high emotion understanding were far more likely to show severe-decreasing than low/low-moderate class co-occurring internalizing and externalizing symptom patterns. Child gender and poor executive control differentiated children in rising versus low trajectories. Implications for early intervention targeting self-regulation of executive control are discussed.
Weed interference during crop establishment is a serious concern for Florida strawberry [Fragaria×ananassa (Weston) Duchesne ex Rozier (pro sp.) [chiloensis×virginiana]] producers. In situ remote detection for precision herbicide application reduces both the risk of crop injury and herbicide inputs. Carolina geranium (Geranium carolinianum L.) is a widespread broadleaf weed within Florida strawberry production with sensitivity to clopyralid, the only available POST broadleaf herbicide. Geranium carolinianum leaf structure is distinct from that of the strawberry plant, which makes it an ideal candidate for pattern recognition in digital images via convolutional neural networks (CNNs). The study objective was to assess the precision of three CNNs in detecting G. carolinianum. Images of G. carolinianum growing in competition with strawberry were gathered at four sites in Hillsborough County, FL. Three CNNs were compared, including object detection–based DetectNet, image classification–based VGGNet, and GoogLeNet. Two DetectNet networks were trained to detect either leaves or canopies of G. carolinianum. Image classification using GoogLeNet and VGGNet was largely unsuccessful during validation with whole images (Fscore<0.02). CNN training using cropped images increased G. carolinianum detection during validation for VGGNet (Fscore=0.77) and GoogLeNet (Fscore=0.62). The G. carolinianum leaf–trained DetectNet achieved the highest Fscore (0.94) for plant detection during validation. Leaf-based detection led to more consistent detection of G. carolinianum within the strawberry canopy and reduced recall-related errors encountered in canopy-based training. The smaller target of leaf-based DetectNet did increase false positives, but such errors can be overcome with additional training images for network desensitization training. DetectNet was the most viable CNN tested for image-based remote sensing of G. carolinianum in competition with strawberry. Future research will identify the optimal approach for in situ detection and integrate the detection technology with a precision sprayer.
Parameters such as pretreatment method, enzyme type and concentration, determine the conversion efficiency of biomass’ cellulose and hemicellulose to glucose and mainly xylose in biomass-based fuel production. Chemical quantification of these processes offers no information on the effect of enzymatic hydrolysis (EH) on particle morphology. We report on the development of a microscopy method for imaging pretreated biomass particles at different EH stages. The method was based on acquiring large field of view images, typically 20×10 mm2 containing thousands of particles. Morphology of particles with lengths between 2 μm and 5 mm could be visualized and analyzed. The particle length distribution of corn stover samples, pretreated with increasing amounts of sulfuric acid at different EH stages, was measured. Particle size was shown to be dependent on pretreatment severity and EH time. The methodology developed could offer an alternative method for characterization of EH of biomass for second generation biofuels and visualization of recalcitrant structures.
This article describes a CDI outbreak in a long-term care (LTC) facility that used molecular typing techniques and whole-genome sequencing to identify widespread dissemination of the clonal strain in the environment which was successfully removed after terminal cleaning.
This study was conducted in a long-term care facility in Texas.
A recently hospitalized LTC patient was diagnosed with CDI followed shortly thereafter by 7 subsequent CDI cases. A stool specimen was obtained from each patient for culturing and typing. An environmental point-prevalence study of the facility was conducted before and after terminal cleaning of the facility to assess environmental contamination. Cultured isolates were typed using ribotyping, multilocus variant analysis, and whole-genome sequencing.
Stool samples were available for 5 of 8 patients; of these specimens, 4 grew toxigenic C. difficile ribotype 027. Of 50 environmental swab samples collected throughout the facility prior to the facility-wide terminal cleaning, 19 (38%) grew toxigenic C. difficile (most commonly ribotype 027, 79%). The terminal cleaning was effective at reducing C. difficile spores in the environment and at eradicating the ribotype 027 strain (P<.001). Using multilocus variance analysis and whole-genome sequencing, clinical and environmental strains were highly related and, in some cases, were identical.
Using molecular typing techniques, we demonstrated reduced environmental contamination with toxigenic C. difficile and the eradication of a ribotype 027 clone. These techniques may help direct infection control efforts and decrease the burden of CDI in the healthcare system.
The development of laser wakefield accelerators (LWFA) over the past several years has led to an interest in very compact sources of X-ray radiation – such as “table-top” free electron lasers. However, the use of conventional undulators using permanent magnets also implies system sizes which are large. In this work, we assess the possibilities for the use of novel mini-undulators in conjunction with a LWFA so that the dimensions of the undulator become comparable with the acceleration distances for LWFA experiments (i.e., centimeters). The use of a prototype undulator using laser machining of permanent magnets for this application is described and the emission characteristics and limitations of such a system are determined. Preliminary electron propagation and X-ray emission measurements are taken with a LWFA electron beam at the University of Michigan.
The stellar ultraviolet radiation (UVR) has been studied in the last decade and has been found to be an important factor to determine the habitability of planetary surfaces. It is known that UVR can be a constraint for life. However, most of the studies of UVR and habitability have missed some fundamental aspects: i) Accurate estimation of the planetary atmospheric attenuation, ii) The biological inferences used to represent the impact of the stellar UVR on life are theoretical and based on the action spectrum (for DNA or microorganisms) or considering parameters as the “lethal dose” obtained from non-astrobiological experiments. Therefore, the conclusions reached by previous studies about the UVR habitability of planetary bodies may be inaccurate. In this work, we propose how to address these studies in a more accurate way through an interdisciplinary approach that combines astrophysics, microbiology, and photobiology and by the use of specially designed laboratory experiments.
We present a catalog of the observed properties of Mira-type variable stars detected with the Kilodegree Extremely Little Telescope (KELT). Asymptotic giant branch (AGB) candidates were identified in KELT using a combination of photometric data from KELT and 2MASS colors. Of the 4 million objects with KELT photometry, 3332 Mira-like variables were identified. Here, we present their observed periods and luminosities which will place important constraints on future theoretical work on the effect convection has on pulsation periods and mode stability.
A fundamental understanding of the filament thinning of viscoelastic fluids is important in practical applications such as spraying and printing of complex materials. Here, we present direct numerical simulations of the two-phase axisymmetric momentum equations using the volume-of-fluid technique for interface tracking and the log-conformation transformation to solve the viscoelastic constitutive equation. The numerical results for the filament thinning are in excellent agreement with the theoretical description developed with a slender body approximation. We show that the off-diagonal stress component of the polymeric stress tensor is important and should not be neglected when investigating the later stages of filament thinning. This demonstrates that such numerical methods can be used to study details not captured by the one-dimensional slender body approximation, and pave the way for numerical studies of viscoelastic fluid flows.
OBJECTIVES/SPECIFIC AIMS: Increased retinal protein O-GlcNAcylation occurs in response to hyperglycemia and contributes to diabetic retinopathy. Renin-angiotensin system (RAS) blockers reduce the incidence of diabetic retinopathy. Beneficial effects of RAS blockers are often attributed to production of angiotensin-(1-7) (Ang1-7). The objective here is to determine the impact of Ang1-7 on retinal protein O-GlcNAcylation. METHODS/STUDY POPULATION: C57/BL6 mice were fed a high-fat diet for 8 weeks and then treated for 3 weeks with either a vehicle control, the RAS blocker captopril, or captopril and the Ang1-7 receptor antagonist A779. R28 cells were used to assess levels of O-GlcNAcylated proteins in response to Ang1-7, and the role of cAMP was investigated with addition of forskolin, 6-Bnz-cAMP-AM, and 8-pCPT-2-O-Me-cAMP-AM to cell culture medium. RESULTS/ANTICIPATED RESULTS: Captopril attenuated retinal protein O-GlcNAcylation in mice fed a high-fat diet. This effect was reversed by A779. Ang1-7 attenuated protein O-GlcNAcylation and increased cAMP levels. Forskolin and the EPAC selective cAMP analog 8-pCPT-2-O-Me-cAMP-AM, but not the PKA selective cAMP analog 6-Bnz-cAMP-AM, attenuated O-GlcNAcylation. Inhibiting EPAC blocked the effect of forskolin, whereas inhibiting PKA did not. DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrates a novel role for Ang1-7 in the retina and identifies a potential EPAC-dependent mechanism that regulates protein O-GlcNAcylation. Thus, future therapeutics targeted at an Ang1-7/EPAC axis in retina may be used to address DR.
There is little debate that the health workforce is a key component of the health care system. Since the training of doctors and nurses takes several years, and the building of new schools even longer, projections are needed to allow for the development of health workforce policies. Our work develops a projection model for the demand of doctors and nurses by Organisation for Economic Co-operation and Development (OECD) countries in the year 2030. The model is based on a country’s demand for health services, which includes the following factors: per capita income, out-of-pocket health expenditures and the ageing of its population. The supply of doctors and nurses is projected using country-specific autoregressive integrated moving average models. Our work shows how dramatic imbalances in the number of doctors and nurses will be in OECD countries should current trends continue. For each country in the OECD with sufficient data, we report its demand, supply and shortage or surplus of doctors and nurses for 2030. We project a shortage of nearly 400,000 doctors across 32 OECD countries and shortage of nearly 2.5 million nurses across 23 OECD countries in 2030. We discuss the results and suggest policies that address the shortages.