Abstract. Significance: Peripheral pitting edema is a clinician-administered measure for grading edema. Peripheral edema is graded 0, 1  +  , 2  +  , 3  +  , or 4  +  , but subjectivity is a major limitation of this technique. A pilot clinical study for short-wave infrared (SWIR) molecular chemical imaging (MCI) effectiveness as an objective, non-contact quantitative peripheral edema measure is underway. Aim: We explore if SWIR MCI can differentiate populations with and without peripheral edema. Further, we evaluate the technology for correctly stratifying subjects with peripheral edema. Approach: SWIR MCI of shins from healthy subjects and heart failure (HF) patients was performed. Partial least squares discriminant analysis (PLS-DA) was used to discriminate the two populations. PLS regression (PLSR) was applied to assess the ability of MCI to grade edema. Results: Average spectra from edema exhibited higher water absorption than non-edema spectra. SWIR MCI differentiated healthy volunteers from a population representing all pitting edema grades with 97.1% accuracy (N  =  103 shins). Additionally, SWIR MCI correctly classified shin pitting edema levels in patients with 81.6% accuracy. Conclusions: Our study successfully achieved the two primary endpoints. Application of SWIR MCI to monitor patients while actively receiving HF treatment is necessary to validate SWIR MCI as an HF monitoring technology.

Heart failure (HF) has a significant impact on patient outcomes and health care costs. Objective monitoring of pitting edema level of a HF patient may help clinicians reduce the volume of patient re-admissions. To address this, ChemImage is developing a Molecular Chemical Imaging (MCI) device for noninvasive measurement of peripheral edema level in HF patients. In an initial clinical study, edema grade was predicted in HF patients with 86% accuracy. Results from a follow-up clinical trial demonstrating the capability of MCI to monitor changes in a HF patient’s peripheral edema over time during the course of treatment will be presented.

Heart failure (HF) has a large impact on patient outcomes and health care costs. Objective monitoring of the pitting edema level of a HF patient may help clinicians reduce the amount of readmissions. ChemImage is developing a Molecular Chemical Imaging (MCI) device for monitoring HF patients that will non-invasively quantify peripheral edema. Results from a completed in-human clinical trial will be presented demonstrating ability to discriminate between healthy volunteers and HF patients with all levels of pitting edema and correct prediction of peripheral edema grade across the patient population. Follow-on clinical trials will address monitoring patients during treatment.

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I (MP16)1 Apr 2020MP16-02 PROSTATIC BIOFILMS: A POTENTIAL BACTERIAL BIOMARKER FOR HIGH-GRADE PROSTATE CANCER Rachael Melton-Kreft, Arash Samiei*, Shifeng Mao, Ralph Miller, Laura Nistico, John Lyne, Nick Giannoukakis, Shweta Patel, Christopher Post, and Jeffrey Cohen Rachael Melton-KreftRachael Melton-Kreft More articles by this author , Arash Samiei*Arash Samiei* More articles by this author , Shifeng MaoShifeng Mao More articles by this author , Ralph MillerRalph Miller More articles by this author , Laura NisticoLaura Nistico More articles by this author , John LyneJohn Lyne More articles by this author , Nick GiannoukakisNick Giannoukakis More articles by this author , Shweta PatelShweta Patel More articles by this author , Christopher PostChristopher Post More articles by this author , and Jeffrey CohenJeffrey Cohen More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000841.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The link between biofilm and the risk of prostate cancer has become the center of attention recently. Understanding the role that microbiome play in the pathogenesis of prostate cancer (PCa) and its progression to metastatic disease remains largely underexplored. We hypothesize that specific bacterial biofilms in the prostate govern the behavior of PCa in regards to indolent or progressive disease in relationship to inflammatory immune markers. METHODS: A total of 25 patients with PCa who underwent radical prostatectomy were enrolled in this retrospective study. Eleven subjects had a Gleason score (GS) of 6 or 3+4, six subjects had a GS of 4+3, and eight subjects had a GS of ≥8. DNA was extracted from tissue blocks and 16S ribosomal RNA gene sequencing was completed to identify the bacterial communities within the tissue. Additionally, fluorescent in situ hybridization (FISH) was used to identify biofilms within the tissue. RESULTS: A total of 512 bacterial signatures were detected across the 25 subjects. Most of the genera detected originating from within the gut. Bacterial diversity narrowed in relationship to higher grade disease. Three hundred eighty-one detections occur in the 6 or 3+4 GS group, seventy-one detections in the 4+3 group, and sixty detections in the ≥8 group. Of the bacteria detected, 39 distinct bacterial genera were identified with 28 exhibiting significant differences in prevalence between the three GS groups. Four bacterial species, Bacteroides vulgatus, Bacteroides xylanisolvens, Bifidobacterium bifidum, and Bifidobacterium longum displayed loss of expression with statistically significant differences (p-value <0.05) between the 6 or 3+4 GS group and the 4+3 group. Additionally, bacterial biofilms were discovered within the prostate tissue by using FISH and confocal microscopy. CONCLUSIONS: The bacterial species in prostate specimens showed reduced microbial diversity in higher GS groups, suggesting a possible pathophysiological correlation between the composition of the biofilm and PCa aggressiveness. Additionally, the discovery of bacterial biofilms within the PCa tissue identifies a potential source of inflammation. The identification of Bacteroides and Bifidobacterium species differences between the GS 3+4 and 4+3 groups provides a potential bacterial biomarker for PCa and altering the microbiome may be indicated for reducing the chance of progression. Source of Funding: The Western Pennsylvania Prostate Cancer Foundation © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e214-e215 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rachael Melton-Kreft More articles by this author Arash Samiei* More articles by this author Shifeng Mao More articles by this author Ralph Miller More articles by this author Laura Nistico More articles by this author John Lyne More articles by this author Nick Giannoukakis More articles by this author Shweta Patel More articles by this author Christopher Post More articles by this author Jeffrey Cohen More articles by this author Expand All Advertisement PDF downloadLoading ...

SIGNIFICANCE:A key risk faced by oncological surgeons continues to be complete removal of tumor. Currently, there is no intraoperative imaging device to detect kidney tumors during excision.AIM:We are evaluating molecular chemical imaging (MCI) as a technology for real-time tumor detection and margin assessment during tumor removal surgeries.APPROACH:In exploratory studies, we evaluate visible near infrared (Vis-NIR) MCI for differentiating tumor from adjacent tissue in ex vivo human kidney specimens, and in anaesthetized mice with breast or lung tumor xenografts. Differentiation of tumor from nontumor tissues is made possible with diffuse reflectance spectroscopic signatures and hyperspectral imaging technology. Tumor detection is achieved by score image generation to localize the tumor, followed by application of computer vision algorithms to define tumor border.RESULTS:Performance of a partial least squares discriminant analysis (PLS-DA) model for kidney tumor in a 22-patient study is 0.96 for area under the receiver operating characteristic curve. A PLS-DA model for in vivo breast and lung tumor xenografts performs with 100% sensitivity, 83% specificity, and 89% accuracy.CONCLUSION:Detection of cancer in surgically resected human kidney tissues is demonstrated ex vivo with Vis-NIR MCI, and in vivo on mice with breast or lung xenografts.

Heart failure (HF) occurs when the heart is unable to pump enough blood to meet blood and oxygen requirements and is among the most common causes for hospitalization in the United States. A retrospective analysis determined that 22% of HF patients are readmitted within 30 days of release from the hospital, and the costs for readmission are substantial. Measuring the severity of peripheral edema is one method for monitoring the treatment of a HF patient. Pitting peripheral edema is a subjective measure administered by clinicians who create an indentation mid-tibia and observe depth and time to resolve the indentation. The results are graded 0, 1, 2, 3 or 4, and this information is used in the patient treatment plan. ChemImage is engaged in a clinical study to determine whether Molecular Chemical Imaging (MCI) in the short wave infrared (SWIR) spectral region can provide an objective measure of peripheral edema in HF patients. In this paper, the performance of SWIR MCI for discriminating between healthy volunteers and HF patients with high grade pitting edema will be presented. This technology may provide a non-invasive methodology for quantitative peripheral edema measurement. As the technology matures, it is envisioned patient self-monitoring, with wireless transmission of edema levels while at home, can aid clinicians in monitoring HF patients for necessary treatment changes remotely, to improve patient outcomes, and ultimately, reduce HF hospital readmission rates.

Background Recurrent Respiratory Papillomatosis (RRP) is a rare disease characterized by the growth of papillomas in the airway and especially the larynx. The clinical course is highly variable among individuals and there is poor understanding of the factors that drive an aggressive vs an indolent course. Methods A convenience cohort of 339 affected subjects with papillomas positive for only HPV6 or HPV11 and clinical course data available for 1 year or more, from a large multicenter international study were included. Exploratory data analysis was conducted followed by inferential analyses with frequentist and Bayesian statistics. Results We examined 339 subjects: 82% were diagnosed prior to the age of 18 years, 65% were infected with HPV6, and 69% had an aggressive clinical course. When comparing age at diagnosis with clinical course, the probability of aggressiveness is high for children under five years of age then drops rapidly. For patients diagnosed after the age of 10 years, an indolent course is more common. After accounting for confounding between HPV11 and young age, HPV type was minimally associated with aggressiveness. Fast and Frugal Trees (FFTs) were utilized to determine which algorithms yield the highest accuracy to classify patients as having an indolent or aggressive clinical course and consistently created a branch for diagnostic age at ~5 years old. There was no reliable strong association between clinical course and socioeconomic or parental factors. Conclusion In the largest cohort of its type, we have identified a critical age at diagnosis which demarcates a more aggressive from less aggressive clinical course.

Improved patient outcomes highlight the need to avoid errors in surgery. Molecular Chemical Imaging (MCI) aids in identifying critical structures without reagents and in vivo results are presented for a range of surgical applications.

Accurate identification, precise dissection, and careful preservation of critical structures, such as nerves and blood vessels, are key to successful surgical outcomes. Unintended and/or unrecognized injuries to critical structures result in debilitating short- and long-term morbidity, avoidable mortality, and considerable socioeconomic and healthcare burdens. ChemImage has developed a Molecular Chemical Imaging endoscope (MCI-E) to be deployed as an intraoperative imaging device for real-time detection of key anatomical structures. MCI-E does not require the use of contrast agents, and employs visible-near infrared (vis-NIR) reflectance hyperspectral imaging. We tested the in vivo performance of MCI-E by collecting high quality vis-NIR signatures from several anatomical structures, including ureters, arteries, and veins in live pigs under general anesthesia. In this paper, we will present successful MCI-E detection of lymph node, ureter, vessels, nerve, bowel, and thyroid in background tissues under relevant in vivo conditions. If successful, integration of MCI-E into surgical procedures will enable real-time automated detection of anatomical structures during surgeries. The benefits of this capability include the opportunity of reduced surgery time, decreased patient risk, fewer repeat surgeries, and enhancement of surgeon training.

Visualization and identification of critical structures such as blood vessels and tumor margins during surgery can often be difficult. The impact of misidentification of such structures can range from internal bleeding to inadequate excision of a malignant tumor and the subsequent need for additional surgery. We are developing an intraoperative device which will provide surgeons with the ability to visualize specified anatomic structures in real-time and without the use of labels (i.e. reagents). This imaging tool employs diffuse reflectance Molecular Chemical Imaging (MCI), a technology combining molecular spectroscopy and digital imaging for non-invasive, non-contact and reagentless evaluation of human tissues. In order to implement real-time sensing, we have developed a new approach to MCI, based on the principles of compressive sensing, and involving a novel, multivariate liquid crystal tunable filter technology. This technology can facilitate real-time detection of biological materials versus complex backgrounds with equal performance to that achieved by conventional MCI instruments. In this paper, we will present results demonstrating the capabilities and performance of a proof-of-concept intraoperative MCI surgical device.

Colorectal cancer (CRC) is the third most common cancer in men and women in the United States. Raman Molecular Imaging (RMI) is an effective technique to evaluate human tissue, cells and bodily fluids, including blood serum for disease diagnosis. ChemImage Corporation, in collaboration with clinicians, has been engaged in development of an in vitro diagnostic Raman assay focused on CRC detection. The Raman Assay for Colorectal Cancer (RACC) exploits the high specificity of Raman imaging to distinguish diseased from normal dried blood serum droplets without additional reagents. Pilot Study results from testing of hundreds of biobank patient samples have demonstrated that RACC detects CRC with high sensitivity and specificity. However, expanded clinical trials, which are ongoing, are revealing a host of important preanalytical considerations associated with sample collection, sample storage and stability, sample shipping, sample preparation and sample interferents, which impact detection performance. Results from recent clinical studies will be presented.

Haemophilus influenzae is an opportunistic pathogen. The emergence of virulent, non-typeable strains (NTHi) emphasizes the importance of developing new interventional targets. We screened the NTHi supragenome for genes encoding surface-exposed proteins suggestive of immune evasion, identifying a large family containing Sel1-like repeats (SLRs). Clustering identified ten SLR-containing gene subfamilies, each with various numbers of SLRs per gene. Individual strains also had varying numbers of SLR-containing genes from one or more of the subfamilies. Statistical genetic analyses of gene possession among 210 NTHi strains typed as either disease or carriage found a significant association between possession of the SlrVA subfamily (which we have termed, macrophage survival factor, msf) and the disease isolates. The PittII strain contains four chromosomally contiguous msf genes. Deleting all four of these genes (msfA1-4) (KO) resulted in a highly significant decrease in phagocytosis and survival in macrophages; which was fully complemented by a single copy of the msfA1 gene. Using the chinchilla model of otitis media and invasive disease, the KO strain displayed a significant decrease in fitness compared to the WT in co-infections; and in single infections, the KO lost its ability to invade the brain. The singly complemented strain showed only a partial ability to compete with the WT suggesting gene dosage is important in vivo. The transcriptional profiles of the KO and WT in planktonic growth were compared using the NTHi supragenome array, which revealed highly significant changes in the expression of operons involved in virulence and anaerobiosis. These findings demonstrate that the msfA1-4 genes are virulence factors for phagocytosis, persistence, and trafficking to non-mucosal sites.

Novel microbial detection technologies have revealed that chronic bacterial biofilms, which are recalcitrant to antibiotic treatment, are common in failed orthopedic procedures.

OBJECTIVES/HYPOTHESIS:Bacteriologic studies of otitis media with effusion (OME) using highly sensitive techniques of molecular biology such as the polymerase chain reaction have demonstrated that traditional culturing methods are inadequate to detect many viable bacteria present in OME. The presence of pathogens attached to the middle-ear mucosa as a bacterial biofilm, rather than as free-floating organisms in a middle-ear effusion, has previously been suggested to explain these observations. The suggestion has been speculative, however, because no visual evidence of such biofilms on middle-ear mucosa has heretofore been collected. The hypotheses motivating the current study were: 1) biofilms of nontypable Hemophilus influenzae will form on the middle-ear mucosa of chinchillas in an experimental model of OME, 2) these biofilms will exhibit changes in density or structure over time, and 3) biofilms are also present on tympanostomy tubes in children with refractory post-tympanostomy otorrhea. The objective of this study was to collect visual evidence of the formation of bacterial biofilms in these situations.STUDY DESIGN:Laboratory study of bacteriology in an animal model and on medical devices removed from pediatric patients.METHODS:Experimental otitis media was induced in chinchillas by transbullar injection of nontypable H. influenzae. Animals were killed in a time series and the surface of the middle-ear mucosa was examined by scanning electron microscopy (SEM) for the presence of bacterial biofilms. Adult and fetal chinchilla uninfected controls were similarly examined for comparison. In addition, tympanostomy tubes that had been placed in children's ears to treat OME and removed after onset of refractory otorrhea or other problems were examined by SEM and by confocal scanning laser microscopy for bacterial biofilms, and compared with unused control tubes.RESULTS:Bacterial biofilms were visually detected by SEM on the middle-ear mucosa of multiple chinchillas in which H. influenzae otitis media had been induced. Qualitative evaluation indicated that the density and thickness of the biofilm might increase until at least 96 hours after injection. The appearance of the middle-ear mucosa of experimental animals contrasted with that of uninjected control animals. Robust bacterial biofilms were also visually detected on tympanostomy tubes removed from children's ears for clinical reasons, in contrast with unused control tubes.CONCLUSIONS:Bacterial biofilms form on the middle-ear mucosa of chinchillas in experimentally induced H. influenzae otitis media and can form on tympanostomy tubes placed in children's ears. Such biofilms can be directly observed by microscopy. These results reinforce the hypothesis that the bacterial aggregates called biofilms, resistant to treatment by antibiotics and to detection by standard culture techniques, may play a major etiologic role in OME and in one of its frequent complications, post-tympanostomy otorrhea.

Clinicalmicrobiology ispoised tomove into adynamic ageby embracing molecular diagnostic techniques, such as polymerase chain reaction–based amplification techniques and whole-genome sequencing (WGS). These technologies will revolutionize data acquisition for theranostics and the tracking of outbreaks. Whole-genome sequencing originally proved its worth through investigations of infectious agents.1 In this issue of JAMA Internal Medicine, Reuter et al2 now advocate WGS for routine diagnostics and public health microbiology. These ideas arenot as far-fetchedas they may initially sound. Although the cost and time to conductWGSwouldhaveoncebeenprohibitive,abacterialWGS can be performed for less than $100 in 1 day’s time.Moreover, the computational systems forperforming theanalytics on the WGSdata allow for rapid turnaround times.Collectively, these metrics make WGS competitive with any technology, including traditional methods of culturing bacteria. Although there arenocommercialWGSanalytics thatcould integratewithstandard laboratory informationsystemsnow,suchtoolswill bedeveloped that canaccess cloud-basedcomputing resources containingglobalstraindatabasesannotatedwithrespect toclinical origins, spread, and associated symptoms.

Background Haemophilus influenzae colonizes the human nasopharynx as a commensal, and is etiologically associated with numerous opportunistic infections of the airway; it is also less commonly associated with invasive disease. Clinical isolates of H. influenzae display extensive genomic diversity and plasticity. The development of strategies to successfully prevent, diagnose and treat H. influenzae infections depends on tools to ascertain the gene content of individual isolates. Results We describe and validate a Haemophilus influenzae supragenome hybridization (SGH) array that can be used to characterize the full genic complement of any strain within the species, as well as strains from several highly related species. The array contains 31,307 probes that collectively cover essentially all alleles of the 2890 gene clusters identified from the whole genome sequencing of 24 clinical H. influenzae strains. The finite supragenome model predicts that these data include greater than 85% of all non-rare genes (where rare genes are defined as those present in less than 10% of sequenced strains). The veracity of the array was tested by comparing the whole genome sequences of eight strains with their hybridization data obtained using the supragenome array. The array predictions were correct and reproducible for ~ 98% of the gene content of all of the sequenced strains. This technology was then applied to an investigation of the gene content of 193 geographically and clinically diverse H. influenzae clinical strains. These strains came from multiple locations from five different continents and Papua New Guinea and include isolates from: the middle ears of persons with otitis media and otorrhea; lung aspirates and sputum samples from pneumonia and COPD patients, blood specimens from patients with sepsis; cerebrospinal fluid from patients with meningitis, as well as from pharyngeal specimens from healthy persons. Conclusions These analyses provided the most comprehensive and detailed genomic/phylogenetic look at this species to date, and identified a subset of highly divergent strains that form a separate lineage within the species. This array provides a cost-effective and high-throughput tool to determine the gene content of any H. influenzae isolate or lineage. Furthermore, the method for probe selection can be applied to any species, given a group of available whole genome sequences.

Lactobacillus forms a thick, protective biofilm in the female urogenital system that provides protection against bacterial infection. Standard obstetric and gynecologic procedures can disrupt this biofilm, increasing the patient risk for developing infectious complications. The role of biofilms in the female reproductive system was evaluated by performing Medline/PubMed literature searches using the key words biofilm, Lactobacillus, urethral, and vagina. Additional information was evaluated from previously presented doctoral theses and scientific meeting abstracts. The important protective role of Lactobacillus biofilm in the female reproductive tract is supported by research with bacterial vaginosis and sexually transmitted diseases, with mixed data for female urinary-tract infections. Lactobacillus biofilm, therefore, may be a target for treatments when developing treatments to reduce reproductive system infections. Promoting Lactobacillus biofilm growth may also be important for reducing complications after gynecologic surgery and obstetrical procedures that typically disrupt the healthy, protective vaginal biofilm.