Impact of a new hospital with close to 100% single-occupancy rooms on environmental contamination and incidence of vancomycin-resistant Enterococcus faecium colonization or infection: a genomic surveillance study.

BACKGROUND: Vancomycin-resistant Enterococcus faecium (VRE) is a leading cause of nosocomial infection, driven by its ability to spread between patients and persist in the hospital environment. AIM: To investigate the impact of a long-established cardiothoracic hospital moving to new premises with close to 100% single-occupancy rooms on the rates of environmental contamination and infection or colonization by VRE. METHODS: Prospective environmental surveillance for VRE was conducted at five time-points between April and November 2019, once in the original building, and four times in the new building. Incidence rate ratios (IRRs) of VRE infection/colonization were determined for the one-year period before and after the hospital move, and compared to a nearby hospital. FINDINGS: In the original location, the first environmental screen found 29% VRE positivity. The following four screens in the new location showed a significant reduction in positivity (1-6%; P<0.0001). The VRE infection/colonization rates were halved in the new location (IRR: 0.56; 95% confidence interval: 0.38-0.84), compared to the original location, contrasting with an increase in a nearby hospital (1.62; 1.17-2.27) over the same time-period. Genomic analysis of the environmental isolates was consistent with reduced transmission in the new hospital. CONCLUSION: The use of single-occupancy rooms was associated with reduced environmental contamination with VRE, and lower transmission and isolation of VRE from clinical samples. The cost-effectiveness of single-occupancy room hospitals in reducing healthcare-associated infections should be reassessed in the context of operational costs of emerging pandemic and increasing antimicrobial resistance threats.

Multifaceted Effects of Kinase Inhibitors on Pancreatic Cancer Cells Reveals Pivotal Entities with Therapeutic Implications.

Pancreatic cancer is one of the most aggressive forms of cancer and is the seventh leading cause of cancer deaths worldwide. Pancreatic ductal adenocarcinoma (PDAC) accounts for over 90% of pancreatic cancers. Most pancreatic cancers are recalcitrant to radiation, chemotherapy, and immunotherapy, highlighting the urgent need for novel treatment options for this deadly disease. To this end, we screened a library of kinase inhibitors in the PDAC cell lines PANC-1 and BxPC-3 and identified two highly potent molecules: Aurora kinase inhibitor AT 9283 (AT) and EGFR kinase inhibitor WZ 3146 (WZ). Both AT and WZ exhibited a dose-dependent inhibition of viability in both cell lines. Thus, we conducted an in-depth multilevel (cellular, molecular, and proteomic) analysis with AT and WZ in PANC-1 cells, which harbor KRAS mutation and exhibit quasimesenchymal properties representing pancreatic cancer cells as having intrinsic chemoresistance and the potential for differential response to therapy. Elucidation of the molecular mechanism of action of AT and WZ revealed an impact on the programmed cell death pathway with an increase in apoptotic, multicaspase, and caspase 3/7 positive cells. Additionally, the key survival molecule Bcl-2 was impacted. Moreover, cell cycle arrest was observed with both kinase inhibitors. Additionally, an increase in superoxide radicals was observed in the AT-treated group. Importantly, proteomic profiling revealed differentially regulated key entities with multifaceted effects, which could have a deleterious impact on PDAC. These findings suggest potential targets for efficacious treatment, including a possible increase in the efficacy of immunotherapy using PD-L1 antibody due to the upregulation of lactoferrin and radixin. Furthermore, combination therapy outcomes with gemcitabine/platinum drugs may also be more effective due to an increase in the NADH dehydrogenase complex. Notably, protein-protein interaction analysis (STRING) revealed possible enrichment of reactome pathway entities. Additionally, novel therapy options, such as vimentin-antibody--drug conjugates, could be explored. Therefore, future studies with the two kinases as monotherapy/combination therapy are warranted.

Disruption of Glioblastoma Multiforme Cell Circuits with Cinnamaldehyde Highlights Potential Targets with Implications for Novel Therapeutic Strategies.

Glioblastoma multiforme (GBM) is a major aggressive primary brain tumor with dismal survival outcome and few therapeutic options. Although Temozolomide (TMZ) is a part of the standard therapy, over time, it can cause DNA damage leading to deleterious effects, necessitating the discovery of drugs with minimal side effects. To this end, we investigated the effect of cinnamaldehyde (CA), a highly purified, single ingredient from cinnamon, on the GBM cell lines U87 and U251 and the neuroglioma cell line H4. On observing similar impact on the viability in all the three cell lines, detailed studies were conducted with CA and its isomer/analog, trans-CA (TCA), and methoxy-CA (MCA) on U87 cells. The compounds exhibited equal potency when assessed at the cellular level in inhibiting U87 cells as well as at the molecular level, resulting in an increase in reactive oxygen species (ROS) and an increase in the apoptotic and multicaspase cell populations. To further characterize the key entities, protein profiling was performed with CA. The studies revealed differential regulation of entities that could be key to glioblastoma cell circuits such as downregulation of pyruvate kinase-PKM2, the key enzyme of the glycolytic pathway that is central to the Warburg effect. This allows for monitoring the levels of PKM2 after therapy using recently developed noninvasive technology employing PET [18F] DASA-23. Additionally, the observation of downregulation of phosphomevalonate kinase is significant as the brain tumor initiating cells (BTIC) are maintained by the metabolism occurring via the mevalonate pathway. Results from the current study, if translated in vivo, could provide additional efficacious treatment options for glioblastoma with minimal side effects.

Casticin Impacts Key Signaling Pathways in Colorectal Cancer Cells Leading to Cell Death with Therapeutic Implications.

Colorectal cancer is the third most frequently encountered cancer worldwide. While current chemotherapeutics help to manage the disease to some extent, they have eluded achieving complete remission and are limited by their severe side effects. This warrants exploration of novel agents that are efficacious with anticipation of minimal adverse effects. In the current study, casticin, a tetramethoxyflavone, was tested for its ability to inhibit the viability of three human colorectal cancer cells: adenocarcinoma (DLD-1, Caco-2 cell lines) and human colorectal carcinoma cells (HCT116 cell line). Casticin showed potent inhibition of viability of DLD-1 and HCT116 cells. Clonogenic assay performed in DLD-1 cells revealed that casticin impeded the colony-forming efficiency of the cells, suggesting its impact on the proliferation of these cells. Further, a sustained effect of the inhibitory action upon withdrawal of the treatment was observed. Elucidation of the mechanism of action revealed that casticin impacted the extrinsic programmed cell death pathway, leading to an increase in apoptotic cells. Further, Bcl-2, the key moiety of cell survival, was affected. Notably, a significant number of cells were arrested in the G2/M phase of the cell cycle in DLD-1 cells. Due to the multifaceted action of casticin, we envision that treatment with casticin could provide an efficacious treatment option for colorectal adenocarcinomas with minimal side effects.

Disruption of Colorectal Cancer Network by Polyphyllins Reveals Pivotal Entities with Implications for Chemoimmunotherapy.

The prevalence of colorectal cancer has increased world-wide with high rates of mortality and morbidity. In the absence of efficacious drugs to treat this neoplasia, there is an imminent need to discover molecules with multifaceted effects. To this end, we opted to study the effect of steroidal saponins such as Polyphyllins. We performed anticancer activity studies with three analogs of Polyphyllins: Polyphyllin D (PD), Polyphyllin II (PII) and Polyphyllin G (PG). Here we show the potent effect of PD, PII (IC50 of 0.5-1 µM) and PG (IC50 of 3 µM) in inhibiting the viability of colorectal adenocarcinoma cells (DLD-1) and colorectal carcinoma cells (HCT116). PD and PII also showed inhibition of cell proliferation and sustained response upon withdrawal of the compounds when assessed by clonogenic assays in both the cell lines. Elucidation of the molecular mode of action revealed impact on the programmed cell death pathway. Additionally, proteomic profiling of DLD-1 revealed pivotal proteins differentially regulated by PD and PII, including a downregulated peroxiredoxin-1 which is considered as one of the novel targets to combat colorectal cancers and an upregulated elongation factor 2 (EF2), one of the key molecules considered as a tumor associated antigen (TAA) in colon cancer. Entities of cell metabolic pathways including downregulation of the key enzyme Phosphoglycerate kinase 1 of the glycolytic pathway was also observed. Importantly, the fold changes per se of the key components has led to the loss of viability of the colorectal cancer cells. We envision that the multifaceted function of PD and PII against the proliferation of colorectal carcinoma cells could have potential for novel treatments such as chemoimmunotherapy for colorectal adenocarcinomas. Future studies to develop these compounds as potent anti-colorectal cancer agents are warranted.

Automated Segmentation of Spinal Muscles From Upright Open MRI Using a Multiscale Pyramid 2D Convolutional Neural Network.

STUDY DESIGN: Randomized trial. OBJECTIVE: To implement an algorithm enabling the automated segmentation of spinal muscles from open magnetic resonance images in healthy volunteers and patients with adult spinal deformity (ASD). SUMMARY OF BACKGROUND DATA: Understanding spinal muscle anatomy is critical to diagnosing and treating spinal deformity.Muscle boundaries can be extrapolated from medical images using segmentation, which is usually done manually by clinical experts and remains complicated and time-consuming. METHODS: Three groups were examined: two healthy volunteer groups (N = 6 for each group) and one ASD group (N = 8 patients) were imaged at the lumbar and thoracic regions of the spine in an upright open magnetic resonance imaging scanner while maintaining different postures (various seated, standing, and supine). For each group and region, a selection of regions of interest (ROIs) was manually segmented. A multiscale pyramid two-dimensional convolutional neural network was implemented to automatically segment all defined ROIs. A five-fold crossvalidation method was applied and distinct models were trained for each resulting set and group and evaluated using Dice coefficients calculated between the model output and the manually segmented target. RESULTS: Good to excellent results were found across all ROIs for the ASD (Dice coefficient >0.76) and healthy (dice coefficient > 0.86) groups. CONCLUSION: This study represents a fundamental step toward the development of an automated spinal muscle properties extraction pipeline, which will ultimately allow clinicians to have easier access to patient-specific simulations, diagnosis, and treatment.

Computational Identification of Stearic Acid as a Potential PDK1 Inhibitor and In Vitro Validation of Stearic Acid as Colon Cancer Therapeutic in Combination with 5-Fluorouracil.

BACKGROUND: Colorectal cancer is the third largest cause of cancer-related mortality worldwide. Although current treatments with chemotherapeutics have allowed for management of colorectal cancer, additional novel treatments are essential. Intervening with the metabolic reprogramming observed in cancers called "Warburg Effect," is one of the novel strategies considered to combat cancers. In the metabolic reprogramming pathway, pyruvate dehydrogenase kinase (PDK1) plays a pivotal role. Identification and characterization of a PDK1 inhibitor is of paramount importance. Further, for efficacious treatment of colorectal cancers, combinatorial regimens are essential. To this end, we opted to identify a PDK1 inhibitor using computational structure-based drug design FINDSITEcomb and perform combinatorial studies with 5-FU for efficacious treatment of colorectal cancers. METHODS: Using computational structure-based drug design FINDSITEcomb, stearic acid (SA) was identified as a possible PDK1 inhibitor. Elucidation of the mechanism of action of SA was performed using flow cytometry, clonogenic assays. RESULTS: When the growth inhibitory potential of SA was tested on colorectal adenocarcinoma (DLD-1) cells, a 50% inhibitory concentration (IC50) of 60 µM was recorded. Moreover, SA inhibited the proliferation potential of DLD-1 cells as shown by the clonogenic assay and there was a sustained response even after withdrawal of the compound. Elucidation of the mechanism of action revealed, that the inhibitory effect of SA was through the programmed cell death pathway. There was increase in the number of apoptotic and multicaspase positive cells. SA also impacted the levels of the cell survival protein Bcl-2. With the aim of achieving improved treatment for colorectal cancer, we opted to combine 5-fluorouracil (5-FU), the currently used drug in the clinic, with SA. Combining SA with 5-FU, revealed a synergistic effect in which the IC50 of 5-FU decreased from 25 to 6 µM upon combination with 60 µM SA. Further, SA did not inhibit non-tumorigenic NIH-3T3 proliferation. CONCLUSIONS: We envision that this significant decrease in the IC50 of 5-FU could translate into less side effects of 5-FU and increase the efficacy of the treatment due to the multifaceted action of SA. The data generated from the current studies on the inhibition of colorectal adenocarcinoma by SA discovered by the use of the computational program as well as synergistic action with 5-FU should open up novel therapeutic options for the management of colorectal adenocarcinomas.

Interception of Signaling Circuits of Esophageal Adenocarcinoma Cells by Resveratrol Reveals Molecular and Immunomodulatory Signatures.

Deregulation of signaling pathways due to mutations sets the cell on a path to neoplasia. Therefore, recent reports of increased mutations observed in esophageal tissue reflects the enhanced risk of tumor formation. In fact, adenocarcinoma of the esophagus has been on the rise lately. Increase in mortality due to a paucity of efficacious drugs for this cancer prompted us to discover molecular signatures to combat this malady. To this end, we chose resveratrol-a polyphenol with anticancer property-and studied its impact on three esophageal adenocarcinoma cell lines (OE33, OE19 and FLO-1) by multilevel profiling. Here, we show the impact of resveratrol on the viability of the three adenocarcinoma esophageal cell systems studied, at the cellular level. Furthermore, an analysis at the molecular level revealed that the action was through the programmed cell death pathway, resulting in an increase in apoptotic and caspase-positive cells. The impact on reactive oxygen species (ROS) and a decrease in Bcl2 levels were also observed. Moreover, proteomic profiling highlighted pivotal differentially regulated signaling molecules. The phenotypic effect observed in resveratrol-treated esophageal cells could be due to the stoichiometry per se of the fold changes observed in entities of key signaling pathways. Notably, the downregulation of Ku80 and other pivotal entities by resveratrol could be harnessed for chemo-radiation therapy to prevent DNA break repair after radiation therapy. Additionally, multilevel profiling has shed light on molecular and immune-modulatory signatures with implications for discovering novel treatments, including chemo-immunotherapy, for esophageal adenocarcinomas which are known to be aggressive cancers.

Estimation and assessment of sagittal spinal curvature and thoracic muscle morphometry in different postures.

Spine models are typically developed from supine clinical imaging data, and hence clearly do not fully reflect postures that replicate subjects' clinical symptoms. Our objectives were to develop a method to: (i) estimate the subject-specific sagittal curvature of the whole spine in different postures from limited imaging data, (ii) obtain muscle lines-of-action in different postures and analyze the effect of posture on muscle fascicle length, and (iii) correct for cosine between the magnetic resonance imaging (MRI) scan plane and dominant fiber line-of-action for muscle parameters (cross-sectional area (CSA) and position). The thoracic spines of six healthy volunteers were scanned in four postures (supine, standing, flexion, and sitting) in an upright MRI. Geometry of the sagittal spine was approximated with a circular spline. A pipeline was developed to estimate spine geometry in different postures and was validated. The lines-of-action for two muscles, erector spinae (ES) and transversospinalis (TS) were obtained for every posture and hence muscle fascicle lengths were computed. A correction factor based on published literature was then computed and applied to the muscle parameters. The maximum registration error between the estimated spine geometry and MRI data was small (average RMSE∼1.2%). The muscle fascicle length increased (up to 20%) in flexion when compared to erect postures. The correction factor reduced muscle parameters (∼5% for ES and ∼25% for TS) when compared to raw MRI data. The proposed pipeline is a preliminary step in subject-specific modeling. Direction cosines of muscles could be used while improving the inputs of spine models.

Preliminary investigation of spinal level and postural effects on thoracic muscle morphology with upright open MRI.

OBJECTIVE: Spinal-muscle morphological differences between weight-bearing and supine postures have potential diagnostic, prognostic, and therapeutic applications. While the focus to date has been on cervical and lumbar regions, recent findings have associated spinal deformity with smaller paraspinal musculature in the thoracic region. We aim to quantitatively investigate the morphology of trapezius (TZ), erector spinae (ES) and transversospinalis (TS) muscles in upright postures with open upright MRI and also determine the effect of level and posture on the morphological measures. METHODS: Six healthy volunteers (age 26 ± 6 years) were imaged (0.5 T MROpen, Paramed, Genoa, Italy) in four postures (supine, standing, standing with 30° flexion, and sitting). Two regions of the thorax, middle (T4-T5), and lower (T8-T9), were scanned separately for each posture. 2D muscle parameters such as cross-sectional area (CSA) and position (radius and angle) with respect to the vertebral body centroid were measured for the three muscles. Effect of spinal level and posture on muscle parameters was examined using 2-way repeated measures ANOVA separately for T4-T5 and T8-T9 regions. RESULTS: The TZ CSA was smaller (40%, P = .0027) at T9 than at T8. The ES CSA was larger at T5 than at T4 (12%, P = .0048) and at T9 than at T8 (10%, P = .0018). TS CSA showed opposite trends at the two spinal regions with it being smaller (16%, P = .0047) at T5 than at T4 and larger (11%, P = .0009) at T9 than at T8. At T4-T5, the TZ CSA increased (up to 23%), and the ES and TS CSA decreased (up to 10%) in upright postures compared to supine. CONCLUSION: Geometrical parameters that describe muscle morphology in the thorax change with level and posture. The increase in TZ CSA in upright postures could result from greater activation while upright. The decrease in ES CSA in flexed positions likely represents passive stretching compared to neutral posture.

Cellular molecular and proteomic profiling deciphers the SIRT1 controlled cell death pathways in esophageal adenocarcinoma cells.

Worldwide prevalence of esophageal adenocarcinomas with high rates of mortality coupled with increased mutations in esophageal cells warrants investigation to understand deregulation of cell signaling pathways leading to cancer. To this end, the current study was undertaken to unravel the cell death signatures using the model human esophageal adenocarcinoma cell line-OE33. The strategy involved targeting the key epigenetic modulator SIRT1, a histone deacetylase by a small molecule inhibitor - sirtinol. Sirtinol induced a dose-dependent inhibition of cell viability under both normoxic and hypoxic conditions with long term impact on proliferation as shown by clonogenic assays. Signature apoptotic signaling pathways including caspase activation and decreased Bcl-2 were observed. Proteomic analysis highlighted an array of entities affected including molecules involved in replication, transcription, protein synthesis, cell division control, stress-related proteins, spliceosome components, protein processing and cell detoxification/degradation systems. Importantly, the stoichiometry of the fold changes of the affected proteins per se could govern the cell death phenotype by sirtinol. Sirtinol could also potentially curb resistant and recurrent tumors that reside in hypoxic environments. Overall, in addition to unraveling the cellular, molecular and proteomics basis of SIRT1 inhibition, the findings open up avenues for designing novel strategies against esophageal adenocarcinoma.

Quantitative identification and segmentation repeatability of thoracic spinal muscle morphology.

OBJECTIVE: MRI derived spinal-muscle morphology measurements have potential diagnostic, prognostic, and therapeutic applications in spinal health. Muscle morphology in the thoracic spine is an important determinant of kyphosis severity in older adults. However, the literature on quantification of spinal muscles to date has been limited to cervical and lumbar regions. Hence, we aim to propose a method to quantitatively identify regions of interest of thoracic spinal muscle in axial MR images and investigate the repeatability of their measurements. METHODS: Middle (T4-T5) and lower (T8-T9) thoracic levels of six healthy volunteers (age 26 ± 6 years) were imaged in an upright open scanner (0.5T MROpen, Paramed, Genoa, Italy). A descriptive methodology for defining the regions of interest of trapezius, erector spinae, and transversospinalis in axial MR images was developed. The guidelines for segmentation are laid out based on the points of origin and insertion, probable size, shape, and the position of the muscle groups relative to other recognizable anatomical landmarks as seen from typical axial MR images. 2D parameters such as muscle cross-sectional area (CSA) and muscle position (radius and angle) with respect to the vertebral body centroid were computed and 3D muscle geometries were generated. Intra and inter-rater segmentation repeatability was assessed with intraclass correlation coefficient (ICC (3,1)) for 2D parameters and with dice coefficient (DC) for 3D parameters. RESULTS: Intra and inter-rater repeatability for 2D and 3D parameters for all muscles was generally good/excellent (average ICC (3,1) = 0.9 with ranges of 0.56-0.98; average DC = 0.92 with ranges from 0.85-0.95). CONCLUSION: The guidelines proposed are important for reliable MRI-based measurements and allow meaningful comparisons of muscle morphometry in the thoracic spine across different studies globally. Good segmentation repeatability suggests we can further investigate the effect of posture and spinal curvature on muscle morphology in the thoracic spine.

Inhibition of Prostate Cancer Cells by 4,5-Dicaffeoylquinic Acid through Cell Cycle Arrest.

Prostate cancer is a major cause of cancer-related mortality in men. Even though current therapeutic management has contributed to reducing mortality, additional intervention strategies are warranted to further improve the outcomes. To this end, we have investigated the efficacy of dicaffeoylquinic acids, ingredients in Yerba Mate (Ilex paraguariensis), an evergreen cultivated in South America, the leaves of which are used to prepare a tea/coffee-like drink. Of the various analogs tested, 4,5-dicaffeoylquinic acid (4,5-diCQA) was the most active molecule against DU-145 prostate cancer cells with a 50% inhibitory concentration (IC50) of 5 µM. 4,5-diCQA was active both under normoxic and hypoxic conditions. The effect of 72-hour treatment on DU-145 cells persisted for an extended time period as assessed by clonogenic assay. Mechanistic studies revealed that the toxic effect was not due to induction of programmed cell death but through cell cycle arrest at S phase. Additionally, 4,5-diCQA did not impact PI3K/MAPK signaling pathway nor did it affect the depolarization of the mitochondrial membrane. 4,5-diCQA-induced accumulation of cells in the S-phase also seems to negatively impact Bcl-2 expression. 4,5-diCQA also exhibited inhibitory activity on LNCaP and PC-3 prostate cancer cells suggesting that it has therapeutic potential on a broad range of prostate cancers. Taken together, the novel inhibitory activity and mechanism of action of 4,5-diCQA opens up potential therapeutic options for using this molecule as monotherapy as well as in combinatorial therapies for the clinical management of prostate cancer.

Giant Encephalocoele: A Rare Case Report and Review of Literature.

Giant encephalocoeles are rare entities with only one case series and few case reports reported in medical literature. Encephalocoeles, which reach a size larger than the head size, are be called Giant encephalocoeles. We report a case of a 6 month old child who had giant encephalocoele with delayed motor milestones in the form of inability to hold neck. Anesthetic implications include difficulty in securing air way due without undue pressure on the sac. She underwent VP shunt followed by excision of the encephalocele sac. Patient is doing well at 1 year of follow up. Preoperative neurological status and amount of brain tissue herniating into the sac are the most important factors determining the long term prognosis.

Engineering Controlled Peritumoral Inflammation to Constrain Brain Tumor Growth.

Brain tumors remain a great clinical challenge, in part due to their capacity to invade into eloquent, inoperable regions of the brain. In contrast, inflammation in the central nervous system (CNS) due to injuries activates microglia and astrocytes culminating in an astroglial scar that typically "walls-off" the injury site. Here, the hypothesis is tested that targeting peritumoral cells surrounding tumors to activate them via an inflammatory stimulus that recapitulates the sequelae of a traumatic CNS injury, could generate an environment that would wall-off and contain invasive tumors in the brain. Gold nanoparticles coated with inflammatory polypeptides to target stromal cells in close vicinity to glioblastoma (GBM) tumors, in order to activate these cells and stimulate stromal CNS inflammation, are engineered. It is reported that this approach significantly contains tumors in rodent models of GBM relative to control treatments (reduction in tumor volume by over 300% in comparison to controls), by the activation of the innate and adaptive immune response, and by triggering pathways related to cell clustering. Overall, this report outlines an approach to contain invasive tumors that can complement adjuvant interventions for invasive GBM such as radiation and chemotherapy.