Small cell carcinoma of the prostate (SCPC) is morphologically similar to small cell carcinoma of the lung (SCLC) and maybe misinterpreted as Gleason pattern 5b prostate adenocarcinoma (HGPC). Recognition of SCPC is important because of its different clinical behavior. This study aims to characterize the immunophenotype of histologically classic SCPC using a comprehensive panel of markers, to better understand its histogenesis, aid in its classification, and evaluate potential therapeutic targets. Using the World Health Organization morphologic criteria for SCLC, 18 SCPC cases were identified; and studied for the following tumor marker groups: prostate specific/related, neuroendocrine, sex steroid hormone receptors, and prognostic/treatment target-related. Ten cases of UPC were used as controls. PSA was positive in 17% of SCPC and neuroendocrine markers were expressed in HGPC. PSA, TTF-1 and CD56 were the most helpful markers in differentiating between SCPC and HGPC (P<0.01), whereas bombesin/GRP, c-kit, bcl-2, and EGFR expression was more frequent in SCPC. SCPC is best diagnosed by following the World Health Organization diagnostic criteria for SCLC. Immunohistochemical markers can help separate SCPC from HGPC and may be useful in histologically borderline cases. Potential therapeutic targets are identified immunohistochemically in SCPC (Bombesin/GRP, c-kit, bcl-2, and EGFR).

Biosynthetic regulation of renal glomerular heparan sulfate-proteoglycans by various aldohexoses (mannose, glucose, and galactose) and laminin. Cellular ATP levels were dramatically reduced in all groups, and the maximal depletion was caused by mannose. Addition of ATP (0.1-1.0 mM) to the perfusion medium resulted in the normalization of the de novo synthesis and of the biochemical characteristics of heparan sulfate-proteoglycans. The relevance of decreased de novo synthesis of proteoglycans due to the depletion of ATP in hyperglycemic states is discussed in terms of increased glomerular permeability to plasma proteins, as seen in diabetes mellitus.During ultrafiltration, the glomerular basement membrane (GBM) restricts the passage of circulating plasma proteins into the urinary space (1). Thus, a compromise in the integrity of the GBM would be expected to lead to the leakage of proteins into the urine, as observed in a variety of renal diseases, including diabetic nephropathy (2, 3). In diabetes, there is a remarkable thickening of the GBM, and an increase in the mesangial matrices associated with proteinuria (4, 5). The GBM alterations may be due to the metabolic derangements in its various components-i.e., type IV collagen, laminin, entactin, and proteoglycans (PGs) (3, 6, 7). In this context, several studies have indicated a loss or a decrease in the synthesis of PGs in diabetes (3, 6-10). The loss of PGs has been regarded as one of the key events responsible for the proteinuric response seen in diabetes (4, 5). Although the deficiency of PGs/glycosaminoglycans (GAGs) is well documented, the mechanism(s) responsible for their decreased synthesis remains to be elucidated. This investigation relates to the study of one of the possible mechanisms-i.e., ATP depletion caused by high aldohexose levels-that may be responsible for the decreased de novo synthesis of heparan sulfate (HS)-PGs in diabetes. METHODSRadiolabeling of Glomerular PGs. An ex vivo organ perfusion system was utilized for radiolabeling of PGs with [35S]sulfate (100 ,uCi/ml; 1 ,uCi = 37 kBq) (11). The hexoses used in various experiments were mannose, glucose, and galactose. They were individually added (10-50 mM) to the perfusion medium. Mannitol (25 mM) was included in the medium as a control. After 5 hr of radiolabeling of the rat kidney, a small cortical piece was processed for tissue autoradiography and the remaining kidney was perfused with Hanks' balanced salts solution containing a mixture of protease inhibitors (11). The kidney was then bisected, the cortex was dissected out, and glomeruli were isolated. Both the isolated glomeruli and the perfusion medium were processed for isolation and characterization of PGs/GAGs.Isolation and Characterization of PGs/GAGs. The radiolabeled PGs from isolated glomeruli were extracted with 4 M guanidinium chloride containing a mixture of protease inhibitors (11). An aliquot of the extract was passed through a Sephadex G-25 column equilibrated with 4 M guanidinium chloride/0.05 M sodium acetate/0...

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Antimicrobial resistance has become an immediate threat to modern healthcare systems as it continues to spread across the globe. As development of novel antibiotics stalls, preserving the effectiveness of existing agents has become a priority. One of the major driving forces behind antimicrobial resistance is the misuse and overuse of antibiotics, often a result of data on the susceptibility of pathogens not being obtained in a convenient and timely manner, a need that conventional antimicrobial susceptibility testing struggles to meet. Here, a hydrogel microfluidic platform is reported for antimicrobial susceptibility testing purposes, capable of handling real samples and yielding results within 2.5 h of culture. By using a multiplayer design with channels crossing overhead of each other, multiple experiments, either one‐ or two‐dimensional, can be staged on the same device. Bacteria grown on the surface of the hydrogel can be easily visualized with standard Gram staining after being transferred onto a glass slide. Coupled with software‐based image analysis, the system can yield a variety of useful information on bacterial susceptibility and the effects of drugs, such as minimum inhibitory concentration and morphological changes in bacteria, either individually or in combination. Compared to conventional testing methods, this system requires less labor, reagents, and equipment to operate, and has significantly higher speed and efficiency.

Antimicrobial resistance (AMR) is a rapidly increasing threat to the effective treatment of infectious diseases worldwide. The two major remedies include: (1) using narrow-spectrum antibiotics based on rapid diagnosis; and (2) developing new antibiotics. A key part of both remedies is the antimicrobial susceptibility test (AST). However, the current standard ASTs that monitor colony formation are costly and time-consuming and the new strategies proposed are not yet practical to be implemented. Herein, we report a strategy to fabricate whole-hydrogel microfluidic chips using alginate-doped agar. This agar-based microfabrication makes it possible to prepare inexpensive hydrogel devices, and allows a seamless link between microfluidics and conventional agar-based cell culture. Different from common microfluidic systems, in our system the cells are cultured on top of the device, similar to normal agar plate culture; on the other hand, the microfluidic channels inside the hydrogel allow precise generation of linear gradient of drugs, thus giving a better performance than the conventional disk diffusion method. Cells in this system are not exposed to any shear flow, which allows the reliable tracking of individual cells and AST results to be obtained within 2-3 hours. Furthermore, our system could test the synergistic effect of drugs through two-dimensional gradient generation. Finally, the platform could be directly implemented to new drug discovery and other applications wherein a fast, cost-efficient method for studying the response of microorganisms upon drug administration is desirable.

Cosmic ray muon-computed tomography (μCT) is a new imaging modality with unique characteristics that could be particularly important for diverse applications including nuclear nonproliferation, spent nuclear fuel monitoring, cargo scanning, and volcano imaging. The strong scattering dependence of muons on atomic number Z in combination with high penetration range could offer a significant advantage over existing techniques when dense, shielded containers must be imaged. However, μCT reconstruction using conventional filtered back-projection is limited due to the overly simple assumptions that do not take into account the curved path caused by multiple Coulomb scattering prompting the need for more sophisticated approaches to be developed. In this paper, we argue that the use of improved muon tracing and scattering angle projection algorithms as well as an algebraic reconstruction technique should produce muon tomographic images with improved quality - or require fewer muons to produce the same image quality - compared to the case where conventional methods are used. We report on the development and assessment of three novel muon tracing methods and two scattering angle projection methods for μCT. Simulated dry storage casks with single and partial missing fuel assemblies were used as numerical examples to assess and compare the proposed methods. The reconstructed images showed an expected improvement in image quality when compared with conventional techniques, even without muon momentum information, which should lead to improved detection capability, even for partial defects.

This work presents a generalized muon trajectory estimation (GMTE) algorithm to estimate the path of a muon in either uniform or nonuniform media. The use of cosmic ray muons in nuclear nonproliferation and safeguards verification applications has recently gained attention due to the non-intrusive and passive nature of the inspection, penetrating capabilities, as well as recent advances in detectors that measure position and direction of the individual muons before and after traversing the imaged object. However, muon image reconstruction techniques are limited in resolution due to low muon flux and the effects of multiple Coulomb scattering (MCS). Current reconstruction algorithms, e.g., point of closest approach (PoCA) or straight-line path (SLP), rely on overly simple assumptions for muon path estimation through the imaged object. For robust muon tomography, efficient and flexible physics-based algorithms are needed to model the MCS process and accurately estimate the most probable trajectory of a muon as it traverses an object. In the present work, the use of a Bayesian framework and a Gaussian approximation of MCS are explored for estimation of the most likely path of a cosmic ray muon traversing uniform or nonuniform media and undergoing MCS. The algorithm's precision is compared to Monte Carlo simulated muon trajectories. It was found that the algorithm is expected to be able to predict muon tracks to less than 1.5 mm RMS for 0.5 GeV muons and 0.25 mm RMS for 3 GeV muons, a 50% improvement compared to SLP and 15% improvement when compared to PoCA. Further, a 30% increase in useful muon flux was observed relative to PoCA. Muon track prediction improved for higher muon energies or smaller penetration depth where energy loss is not significant. The effect of energy loss due to ionization is investigated, and a linear energy loss relation that is easy to use is proposed.

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