Development and Application of a Virtual Imaging Trial Framework for Longitudinal Quantification of Emphysema in CT.

Pulmonary emphysema is a progressive lung disease that requires accurate evaluation for optimal management. This task, possible using quantitative CT, is particularly challenging as scanner and patient attributes change over time, negatively impacting the CT-derived quantitative measures. Efforts to minimize such variations have been limited by the absence of ground truth in clinical data, thus necessitating reliance on clinical surrogates, which may not have one-to-one correspondence to CT-based findings. This study aimed to develop the first suite of human models with emphysema at multiple time points, enabling longitudinal assessment of disease progression with access to ground truth. A total of 14 virtual subjects were modeled across three time points. Each human model was virtually imaged using a validated imaging simulator (DukeSim), modeling an energy-integrating CT scanner. The models were scanned at two dose levels and reconstructed with two reconstruction kernels, slice thicknesses, and pixel sizes. The developed longitudinal models were further utilized to demonstrate utility in algorithm testing and development. Two previously developed image processing algorithms (CT-HARMONICA, EmphysemaSeg) were evaluated. The results demonstrated the efficacy of both algorithms in improving the accuracy and precision of longitudinal quantifications, from 6.1±6.3% to 1.1±1.1% and 1.6±2.2% across years 0-5. Further investigation in EmphysemaSeg identified that baseline emphysema severity, defined as >5% emphysema at year 0, contributed to its reduced performance. This finding highlights the value of virtual imaging trials in enhancing the explainability of algorithms. Overall, the developed longitudinal human models enabled ground-truth based assessment of image processing algorithms for lung quantifications.

Radiographic Imaging of Community-Acquired Pneumonia: A Case-Based Review.

The chest radiograph is the most common imaging examination performed in most radiology departments, and one of the more common indications for these studies is suspected infection. Radiologists must therefore be aware of less common radiographic patterns of pulmonary infection if they are to add value in the interpretation of chest radiographs for this indication. This review uses a case-based format to illustrate a range of imaging findings that can be associated with acute pulmonary infection and highlight findings that should prompt investigation for diseases other than community-acquired pneumonia to prevent misdiagnosis and delays in appropriate management.

Image quality of photon counting and energy integrating chest CT - Prospective head-to-head comparison on same patients.

PURPOSE: To prospectively compare the image quality of high-resolution, low-dose photon-counting detector CT (PCD-CT) with standard energy-integrating-detector CT (EID) on the same patients. METHOD: IRB-approved, prospective study; patients received same-day non-contrast CT on EID and PCD-CT (NAEOTOM Alpha, blinded) with clinical protocols. Four blinded radiologists evaluated subsegmental bronchial wall definition, noise, and overall image quality in randomized order (0 = worst; 100 = best). Cases were quantitatively compared using the average Global-Noise-Index (GNI), Noise-Power-Spectrum average frequency (fav), NPS frequency-peak (fpeak), Task-Transfer-Function-10%-frequency (f10) an adjusted detectability index (d'adj), and applied output radiation doses (CTDIvol). RESULTS: Sixty patients were prospectively imaged (27 men, mean age 67 ± 10 years, mean BMI 27.9 ± 6.5, 15.9-49.4 kg/m2). Subsegmental wall definition was rated significantly better for PCD-CT than EID (mean 71 [56-87] vs 60 [45-76]; P < 0.001), noise was rated higher for PCD-CT (48 [26-69] vs 34 [13-56]; P < 0.001). Overall image quality was rated significantly higher for PCD-CT than EID (66 [48-85] vs 61 [42-79], P = 0.008). Automated image quality measures showed similar differences for PCD-CT vs EID (mean GNI 70 ± 19 HU vs 26 ± 8 HU, fav 0.35 ± 0.02 vs 0.25 ± 0.02 mm-1, fpeak 0.07 ± 0.01 vs 0.09 ± 0.03 mm-1, f10 0.7 ± 0.08 vs 0.6 ± 0.1 mm-1, all p-values < 0.001). PCD-CT showed a 10% average d'adj increase (-49% min, 233% max). PCD-CT studies were acquired at significantly lower radiation doses than EID (mean CTDIvol 4.5 ± 2.1 vs 7.7 ± 3.2 mGy, P < 0.01). CONCLUSION: Though PCD-CT had higher measured and perceived noise, it offered equivalent or better diagnostic quality compared to EID at lower radiation doses, due to its improved resolution.

Inducible CRISPRi-Based Operon Silencing and Selective in Trans Gene Complementation in Borrelia burgdorferi.

To accelerate genetic studies on the Lyme disease pathogen Borrelia burgdorferi, we developed an enhanced CRISPR interference (CRISPRi) approach for isopropyl-ß-d-thiogalactopyranoside (IPTG)-inducible repression of specific B. burgdorferi genes. The entire system is encoded on a compact 11-kb shuttle vector plasmid that allows for inducible expression of both the sgRNA module and a nontoxic codon-optimized dCas9 protein. We validated this CRISPRi system by targeting the genes encoding OspA and OspB, abundant surface lipoproteins coexpressed by a single operon, and FlaB, the major subunit forming the periplasmic flagella. As in other systems, single guide RNAs (sgRNAs) complementary to the nontemplate strand were consistently effective in gene repression, with 4- to 994-fold reductions in targeted transcript levels and concomitant reductions of protein levels. Furthermore, we showed that ospAB knockdowns could be selectively complemented in trans for OspA expression via the insertion of CRISPRi-resistant, synonymously or nonsynonymously mutated protospacer adjacent motif (PAM*) ospA alleles into a unique site within the CRISPRi plasmid. Together, this establishes CRISPRi PAM* as a robust new genetic tool to simplify the study of B. burgdorferi genes, bypassing the need for gene disruptions by allelic exchange and avoiding rare codon toxicity from the heterologous expression of dCas9. IMPORTANCE Borrelia burgdorferi, the spirochetal bacterium causing Lyme disease, is a tick-borne pathogen of global importance. Here, we expand the genetic toolbox for studying B. burgdorferi physiology and pathogenesis by establishing a single plasmid-based, fully inducible, and nontoxic CRISPR interference (CRISPRi) system for transcriptional silencing of B. burgdorferi genes and operons. We also show that alleles of CRISPRi-targeted genes with mutated protospacer-adjacent motif (PAM*) sites are CRISPRi resistant and can be used for simultaneous in trans gene complementation. The CRISPRi PAM* system will streamline the study of essential Borrelia proteins and accelerate investigations into their structure-function relationships.

MR Imaging for the Evaluation of Diffuse Lung Disease: Where Are We?

Patients with diffuse lung diseases require thorough medical and social history and physical examinations, coupled with a multitude of laboratory tests, pulmonary function tests, and radiologic imaging to discern and manage the specific disease. This review summarizes the current state of imaging of various diffuse lung diseases by hyperpolarized MR imaging. The potential of hyperpolarized MR imaging as a clinical tool is outlined as a novel imaging approach that enables further understanding of the cause of diffuse lung diseases, permits earlier detection of disease progression before that found with pulmonary function tests, and can delineate physiologic response to lung therapies.

Radiographic Imaging of Community-Acquired Pneumonia: A Case-Based Review.

The chest radiograph is the most common imaging examination performed in most radiology departments, and one of the more common indications for these studies is suspected infection. Radiologists must therefore be aware of less common radiographic patterns of pulmonary infection if they are to add value in the interpretation of chest radiographs for this indication. This review uses a case-based format to illustrate a range of imaging findings that can be associated with acute pulmonary infection and highlight findings that should prompt investigation for diseases other than community-acquired pneumonia to prevent misdiagnosis and delays in appropriate management.

Noninvasive diagnosis of pulmonary hypertension with hyperpolarised 129Xe magnetic resonance imaging and spectroscopy.

Background: The diagnosis of pulmonary hypertension (PH) remains challenging. Pre- and post-capillary PH have different signatures on noninvasive 129Xe gas-exchange magnetic resonance imaging (MRI) and dynamic MR spectroscopy (MRS). We tested the accuracy of 129Xe MRI/MRS to diagnose PH status compared to right heart catheterisation (RHC). Methods: 129Xe MRI/MRS from 93 subjects was used to develop a diagnostic algorithm, which was tested in 32 patients undergoing RHC on the same day (n=20) or within 5 months (42±40 days) (n=12). Three expert readers, blinded to RHC, used 129Xe MRI/MRS to classify subjects as pre-capillary PH, post-capillary PH, no PH and no interstitial lung disease (ILD), or ILD. Results: For pre-capillary PH, 129Xe MRI/MRS diagnostic accuracy was 75% (95% CI 66-84) with a sensitivity of 67% (95% CI 54-79) and a specificity of 86% (95% CI 75-96); for post-capillary PH accuracy was 69% (95% CI 59-78) with sensitivity of 54% (95% CI 34-74) and specificity of 74% (95% CI 63-84). The model performed well in straightforward cases of pre-capillary PH but was less accurate in its diagnosis in the presence of mixed disease, particularly in the presence of ILD or combined post- and pre-capillary PH. Conclusion: This study demonstrates the potential to develop 129Xe MRI/MRS into a modality with good accuracy in detecting pre- and post-capillary PH. Furthermore, the combination of 129Xe dynamic MRS and gas-exchange MRI uniquely provide concurrent, noninvasive assessment of both haemodynamics and gas-exchange impairment that may aid in the detection of PH.

Role of ultrasound in the evaluation of first-trimester pregnancies in the acute setting.

In patients presenting for an evaluation of pregnancy in the first trimester, transvaginal ultrasound is the modality of choice for establishing the presence of an intrauterine pregnancy; evaluating pregnancy viability, gestational age, and multiplicity; detecting pregnancy-related complications; and diagnosing ectopic pregnancy. In this pictorial review article, the sonographic appearance of a normal intrauterine gestation and the most common complications of pregnancy in the first trimester in the acute setting are discussed.

Pancreaticobiliary Trauma: A Multimodality Imaging Update.

Pancreaticobiliary injury is an uncommon entity which more often occurs in the setting of blunt than penetrating trauma. We present cases of pancreaticobiliary traumatic injuries from our Level 1 trauma center to illustrate an imaging update on the spectrum of injuries and correlation with current grading systems.

Mechanistic Insights into Oxidative Oligomerization of p-Phenylenediamine and Resorcinol.

An efficient synthesis of a green dye from oxidative coupling of p-phenylenediamine (PPD) and resorcinol (in a 2:1 ratio) has been developed. Reactivity studies of this dye molecule with a variety of reagents (PPD, resorcinol, the oxidized form of the green dye itself, and a dinuclear indo dye) demonstrate that it cannot be the key reactive intermediate in reported oxidative oligomerization of PPD and resorcinol. However, the trinuclear species does form large aggregates. At least one viable pathway of oligomerization has been demonstrated with the dinuclear indo dye.

Bullous Pemphigoid Arising in Lower Leg Vein Graft Incision Site.

Bullous pemphigoid is a blistering disorder which can be idiopathic or arise secondary to drugs or trauma; however, blistering arising within surgical scars is rare. We present a patient with no prior skin history who developed blistering in his left calf vein harvesting scar soon after coronary artery bypass surgery.

Comparison of damage to human hair fibers caused by monoethanolamine- and ammonia-based hair colorants.

The number of Level 3 hair color products that substitute 2-aminoethanol [monoethanolamine (MEA)] for ammonia is increasing. There is some anecdotal evidence that higher levels of MEA can be more damaging to hair and more irritating than a corresponding equivalent level of the typical alkalizer, ammonia (in the form of ammonium hydroxide). Our interest was to understand in more quantitative terms the relative hair damage from the two alkalizers, particularly at the upper limits of MEA on-head use. Limiting investigations of oxidative hair damage to increases in cysteic acid content (from cystine oxidation) can underreport the extent of total damage. Hence, we complemented Fourier transform infrared spectroscopy (FTIR) cysteic acid level measurement with scanning electron microscopy (SEM) photomicrographs to visualize cuticle damage, and protein loss to understand not only the oxidative damage but also the damage caused by other damage pathways, e.g., reaction of the more nucleophilic (than ammonia) MEA with hair protein. In fact, all methods show an increase in damage from MEA-based formulations, up to 85% versus ammonia in the most extreme case. Hence, if the odor of ammonia is a concern, a better approach may be to minimize the volatility of ammonia in specific chassis rather than replacing it with high levels of a potentially more damaging alkalizer such as MEA.

Pathological endoplasmic reticulum stress mediated by the IRE1 pathway contributes to pre-insulitic beta cell apoptosis in a virus-induced rat model of type 1 diabetes.

AIMS/HYPOTHESIS: We hypothesised that pathological endoplasmic reticulum (ER) stress contributes to beta cell death during development of type 1 diabetes. In this study, we investigated the occurrence of beta cell ER stress and the signalling pathways involved during discrete stages of autoimmune diabetes progression. The virus-inducible BBDR rat model was used to systematically interrogate the three main ER stress signalling pathways (IRE1 [inositol-requiring protein-1], PERK [double-stranded RNA-dependent protein kinase (PKR)-like ER kinase] and ATF6 [activating transcription factor 6]) in pancreatic beta cells during type 1 diabetes development. METHODS: ER stress and apoptotic markers were assessed by immunoblot analyses of isolated pancreatic islets and immunofluorescence staining of pancreas sections from control and virus-induced rats. Various time points were analysed: (1) early stages preceding the development of insulitis and (2) a late stage during onset and progression of insulitis, which precedes overt hyperglycaemia. RESULTS: The IRE1 pathway, including its downstream component X-box-binding protein 1, was specifically activated in pancreatic beta cells of virus-induced rats at early stages preceding the development of insulitis. Furthermore, ER stress-specific pro-apoptotic caspase 12 and effector caspase 3 were also activated at this stage. Activation of PERK and its downstream effector pro-apoptotic CHOP (CCAAT/-enhancer-binding-protein homologous protein), only occurred during late stages of diabetes induction concurrent with insulitis, whereas ATF6 activation in pancreatic beta cells was similar in control and virus-induced rats. CONCLUSIONS/INTERPRETATION: Activation of the IRE1 pathway and ER stress-specific pro-apoptotic caspase 12, before the development of insulitis, are indicative of ER stress-mediated beta cell damage. The early occurrence of pathological ER stress and death in pancreatic beta cells may contribute to the initiation and/or progression of virus-induced autoimmune diabetes.

KLF15 is a molecular link between endoplasmic reticulum stress and insulin resistance.

Obesity places major demands on the protein folding capacity of the endoplasmic reticulum (ER), resulting in ER stress, a condition that promotes hepatic insulin resistance and steatosis. Here we identify the transcription factor, Kruppel-like factor 15 (KLF15), as an essential mediator of ER stress-induced insulin resistance in the liver. Mice with a targeted deletion of KLF15 exhibit increased hepatic ER stress, inflammation, and JNK activation compared to WT mice; however, KLF15 (-/-) mice are protected against hepatic insulin resistance and fatty liver under high-fat feeding conditions and in response to pharmacological induction of ER stress. The mammalian target of rapamycin complex 1 (mTORC1), a key regulator of cellular energy homeostasis, has been shown to cooperate with ER stress signaling pathways to promote hepatic insulin resistance and lipid accumulation. We find that the uncoupling of ER stress and insulin resistance in KLF15 (-/-) liver is associated with the maintenance of a low energy state characterized by decreased mTORC1 activity, increased AMPK phosphorylation and PGC-1α expression and activation of autophagy, an intracellular degradation process that enhances hepatic insulin sensitivity. Furthermore, in primary hepatocytes, KLF15 deficiency markedly inhibits activation of mTORC1 by amino acids and insulin, suggesting a mechanism by which KLF15 controls mTORC1-mediated insulin resistance. This study establishes KLF15 as an important molecular link between ER stress and insulin action.