Ex vivo drug sensitivity screening predicts response to temozolomide in glioblastoma patients and identifies candidate biomarkers.

BACKGROUND: Patient-derived glioma stem-like cells (GSCs) have become the gold-standard in neuro-oncological research; however, it remains to be established whether loss of in situ microenvironment affects the clinically-predictive value of this model. We implemented a GSC monolayer system to investigate in situ-in vitro molecular correspondence and the relationship between in vitro and patient response to temozolomide (TMZ). METHODS: DNA/RNA-sequencing was performed on 56 glioblastoma tissues and 19 derived GSC cultures. Sensitivity to TMZ was screened across 66 GSC cultures. Viability readouts were related to clinical parameters of corresponding patients and whole-transcriptome data. RESULTS: Tumour DNA and RNA sequences revealed strong similarity to corresponding GSCs despite loss of neuronal and immune interactions. In vitro TMZ screening yielded three response categories which significantly correlated with patient survival, therewith providing more specific prediction than the binary MGMT marker. Transcriptome analysis identified 121 genes related to TMZ sensitivity of which 21were validated in external datasets. CONCLUSION: GSCs retain patient-unique hallmark gene expressions despite loss of their natural environment. Drug screening using GSCs predicted patient response to TMZ more specifically than MGMT status, while transcriptome analysis identified potential biomarkers for this response. GSC drug screening therefore provides a tool to improve drug development and precision medicine for glioblastoma.

Can in vitro studies aid in the development and use of antiseizure therapies? A report of the ILAE/AES Joint Translational Task Force.

In vitro preparations (defined here as cultured cells, brain slices, and isolated whole brains) offer a variety of approaches to modeling various aspects of seizures and epilepsy. Such models are particularly amenable to the application of anti-seizure compounds, and consequently are a valuable tool to screen the mechanisms of epileptiform activity, mode of action of known anti-seizure medications (ASMs), and the potential efficacy of putative new anti-seizure compounds. Despite these applications, all disease models are a simplification of reality and are therefore subject to limitations. In this review, we summarize the main types of in vitro models that can be used in epilepsy research, describing key methodologies as well as notable advantages and disadvantages of each. We argue that a well-designed battery of in vitro models can form an effective and potentially high-throughput screening platform to predict the clinical usefulness of ASMs, and that in vitro models are particularly useful for interrogating mechanisms of ASMs. To conclude, we offer several key recommendations that maximize the potential value of in vitro models in ASM screening. This includes the use of multiple in vitro tests that can complement each other, carefully combined with in vivo studies, the use of tissues from chronically epileptic (rather than naïve wild-type) animals, and the integration of human cell/tissue-derived preparations.

Comparative analysis of deeply phenotyped GBM cohorts of 'short-term' and 'long-term' survivors.

BACKGROUND: Glioblastoma (GBM) is an aggressive brain cancer that typically results in death in the first 15 months after diagnosis. There have been limited advances in finding new treatments for GBM. In this study, we investigated molecular differences between patients with extremely short (≤ 9 months, Short term survivors, STS) and long survival (≥ 36 months, Long term survivors, LTS). METHODS: Patients were selected from an in-house cohort (GLIOTRAIN-cohort), using defined inclusion criteria (Karnofsky score > 70; age < 70 years old; Stupp protocol as first line treatment, IDH wild type), and a multi-omic analysis of LTS and STS GBM samples was performed. RESULTS: Transcriptomic analysis of tumour samples identified cilium gene signatures as enriched in LTS. Moreover, Immunohistochemical analysis confirmed the presence of cilia in the tumours of LTS. Notably, reverse phase protein array analysis (RPPA) demonstrated increased phosphorylated GAB1 (Y627), SRC (Y527), BCL2 (S70) and RAF (S338) protein expression in STS compared to LTS. Next, we identified 25 unique master regulators (MR) and 13 transcription factors (TFs) belonging to ontologies of integrin signalling and cell cycle to be upregulated in STS. CONCLUSION: Overall, comparison of STS and LTS GBM patients, identifies novel biomarkers and potential actionable therapeutic targets for the management of GBM.

Genomic Exploration of Distinct Molecular Phenotypes Steering Temozolomide Resistance Development in Patient-Derived Glioblastoma Cells.

Chemotherapy using temozolomide is the standard treatment for patients with glioblastoma. Despite treatment, prognosis is still poor largely due to the emergence of temozolomide resistance. This resistance is closely linked to the widely recognized inter- and intra-tumoral heterogeneity in glioblastoma, although the underlying mechanisms are not yet fully understood. To induce temozolomide resistance, we subjected 21 patient-derived glioblastoma cell cultures to Temozolomide treatment for a period of up to 90 days. Prior to treatment, the cells' molecular characteristics were analyzed using bulk RNA sequencing. Additionally, we performed single-cell RNA sequencing on four of the cell cultures to track the evolution of temozolomide resistance. The induced temozolomide resistance was associated with two distinct phenotypic behaviors, classified as "adaptive" (ADA) or "non-adaptive" (N-ADA) to temozolomide. The ADA phenotype displayed neurodevelopmental and metabolic gene signatures, whereas the N-ADA phenotype expressed genes related to cell cycle regulation, DNA repair, and protein synthesis. Single-cell RNA sequencing revealed that in ADA cell cultures, one or more subpopulations emerged as dominant in the resistant samples, whereas N-ADA cell cultures remained relatively stable. The adaptability and heterogeneity of glioblastoma cells play pivotal roles in temozolomide treatment and contribute to the tumor's ability to survive. Depending on the tumor's adaptability potential, subpopulations with acquired resistance mechanisms may arise.

Isolated Atrial Neutrophilic Myocarditis: A Rare Cause of Death and Potential "Blind Spot" for Postmortem Computed Tomography and Postmortem Examination.

ABSTRACT: Myocarditis can focally or diffusely involve the myocardium and is associated with arrhythmia, cardiac failure, and sudden death. The majority of cases of myocarditis are lymphocytic in etiology, and neutrophilic myocarditis is relatively uncommon. Although reported in lymphocytic myocarditis, isolated/focal neutrophilic myocarditis of the atrium causing death is not reported in the postmortem literature, and its postmortem computed tomography correlation is not documented. We report a fatal case of isolated neutrophilic atrial myocarditis affecting only the left atrium in a 33-year-old woman presenting as a sudden unexpected death. Microscopy of the left atrium, a not commonly sampled area of the heart, showed florid neutrophilic myocarditis with the remaining right atrium, conduction system, and both ventricles uninvolved. Similar to previously reported fatal isolated atrial lymphocytic myocarditis, postmortem computed tomography was unable to detect any atrial abnormalities. This case highlights a potential "blind spot" for both routine postmortem imaging and examination.

Cervical Epidural Abscess: A Blind Spot for Postmortem Computed Tomography.

ABSTRACT: Spinal epidural abscess is an uncommon condition, which may have serious complications, including neurological sequelae and death. Classical symptoms include spinal pain, fever, and neurological deficit; however, diagnosis is difficult and requires a high degree of clinical suspicion. Antemortem magnetic resonance imaging (MRI) scanning is the gold-standard diagnostic tool. Computed tomography (CT) is less sensitive and as such, postmortem CT may miss the diagnosis. We report a case of Staphylococcus aureus cervical epidural abscess presenting as neck pain and causing flaccid paralysis and subsequently death. Antemortem MRI showed a small epidural collection, spinal cord edema, and fluid in the adjacent cervical disc and facet joints; however, these findings could not be appreciated on postmortem CT. Postmortem examination, guided by the antemortem imaging, was able to confirm a cervical spinal epidural abscess; however, similar deaths may occur before medical presentation. This case demonstrates a limitation of postmortem imaging in diagnosing spinal epidural abscess and highlights that such cases may be missed.

Phlegmonous Psoas Muscle Infection Causing Sepsis and Death With Missing Postmortem Computed Tomography Scan Correlation.

ABSTRACT: Infection of the psoas muscle is a rare pathology, which carries a high risk of sepsis and is a potential cause of death. Classic symptoms include back pain and fever and it may be diagnosed premortem on computed tomography or magnetic resonance imaging, where abscess formation may be identified as a discrete rim enhancing and low-attenuation lesion. Infections without abscess formation, such as phlegmonous infection, may be more difficult to identify however, particularly if there is absence of other nonspecific findings, such as gas bubbles. We report a case of Staphylococcus aureus phlegmonous psoas muscle causing sepsis and death with no postmortem computed tomography scan correlation, where clinical history of back pain and an unknown source of sepsis was the only prompt for psoas dissection. This case highlights a potential postmortem computed tomography blind spot in abdominal pathology and we recommend dissecting the psoas muscle if sepsis is suspected but a definitive septic focus is unable to be identified.

Titanium Fan Plate Reconstruction for Lateral Orbital Traumatic Defects.

ABSTRACT: Fractures of the orbit often require operative repair with materials ranging from autologous tissue to titanium implants. Reconstructive techniques of the lateral orbital rim and wall offer a unique challenge due to the structural and aesthetic demands with regard to its natural contour. The authors present a case in which a patient sustained a ballistic injury involving the lateral orbital rim and wall requiring reconstruction. The authors employed the use of a single titanium orbital mesh fan plate (DePuy Synthes, Warsaw IN) in an injury-specific and cost-effective technique to rapidly restore the contour and structure of the lateral orbit rivaling the result of a 3D printed custom plate.

Patching the Pathway and Widening the Pipeline: Models for Developing a Diverse Early Childhood Workforce in Chicago.

With the growing appreciation of the importance of early learning experiences for children's healthy development, attention to the cultivation and maintenance of a qualified workforce has steadily increased. Such a workforce must have not just the knowledge and skills related to child development and early learning, but also be linguistically and culturally prepared to meet the needs of an increasingly diverse child and family population. To ensure a highly qualified workforce, programs and policymakers must attend to both the "pipeline" through which new early childhood educators (ECEs) enter the workforce and the "pathways" by which ECEs work toward and obtain the necessary education and credentials for different roles within the field. In line with the aims of this special issue, this paper leverages the first-person account style to describe barriers to and creative solutions for the development of practitioners in low-resourced communities in Chicago, with the goal of informing practice and policy. We describe three prior and ongoing partnership programs between community-based organizations and institutions of higher education, each tailored to support a unique population in the ECE pipeline on the pathway for increased educational attainment and credentialing. Each program is grounded in a specific community of Chicago, a diverse city with a sizable population of children raised in non-English speaking homes. Each program addresses specific needs of the communities they serve, especially around the recruitment, retention, and promotion of bilingual ECEs. Program administrators and community members describe each programs' goals, development, and key components unique to their target population as well as key takeaways. We conclude with an overview of critical components that we identified across these programs in order to create pathways for change within the workforce and the communities they serve.

Disproportionate Sterilization of Latinos Under California's Eugenic Sterilization Program, 1920-1945.

OBJECTIVES: To compare population-based sterilization rates between Latinas/os and non-Latinas/os sterilized under California's eugenics law. METHODS: We used data from 17 362 forms recommending institutionalized patients for sterilization between 1920 and 1945. We abstracted patient gender, age, and institution of residence into a data set. We extracted data on institution populations from US Census microdata from 1920, 1930, and 1940 and interpolated between census years. We used Spanish surnames to identify Latinas/os in the absence of data on race/ethnicity. We used Poisson regression with a random effect for each patient's institution of residence to estimate incidence rate ratios (IRRs) and compare sterilization rates between Latinas/os and non-Latinas/os, stratifying on gender and adjusting for differences in age and year of sterilization. RESULTS: Latino men were more likely to be sterilized than were non-Latino men (IRR = 1.23; 95% confidence interval [CI] = 1.15, 1.31), and Latina women experienced an even more disproportionate risk of sterilization relative to non-Latinas (IRR = 1.59; 95% CI = 1.48, 1.70). CONCLUSIONS: Eugenic sterilization laws were disproportionately applied to Latina/o patients, particularly Latina women and girls. Understanding historical injustices in public health can inform contemporary public health practice.

The National Kidney Foundation of Illinois KidneyMobile: a mobile resource for community based screenings of chronic kidney disease and its risk factors.

BACKGROUND: Early detection and treatment of chronic kidney disease (CKD) and its risk factors improves outcomes; however, many high-risk individuals lack access to healthcare. The National Kidney Foundation of Illinois (NKFI) developed the KidneyMobile (KM) to conduct community-based screenings, provide disease education, and facilitate follow-up appointments for diabetes, hypertension, and CKD. METHODS: Cross-sectional design. Adults > = 18 years of age participated in NKFI KM screenings across Illinois between 2005 and 2011. Sociodemographic and medical history were self-reported using structured interviews; laboratory data and blood pressure were assessed using standard procedures. RESULTS: Among 20,770 participants, mean age was 53.5 years, 68% were female, 49% were African-American or Hispanic, 21% primarily spoke Spanish, and at least 27% lacked health insurance. Seventy-eight percent of participants with elevated blood pressure (≥ 140/90 mmHg) were aware of having hypertension, 93% of participants with abnormal blood glucose (fasting glucose > 126 mg/dl or a random glucose of > 200 mg/dL) were aware of having diabetes, and 19% of participants with albuminuria (> 30 mg/gm) were aware of having CKD. In participants reporting hypertension, 47% had blood pressure ≥ 140/90 mmHg, and in those reporting diabetes, 56% had blood glucose ≥ 130 mg/dl (fasting) or ≥ 180 mg/dl (random). Among 4937 participants with abnormal screening findings that participated in follow-up interviews, 69% reported having further medical evaluation. CONCLUSIONS: A high-risk disadvantaged population is being reached by the NKFI KidneyMobile and connected with healthcare services. A significant proportion of participants were newly informed of having abnormal results suggestive of diabetes, hypertension, and/or CKD or that their diabetes and hypertension were inadequately controlled.

A novel immune biomarker IFI27 discriminates between influenza and bacteria in patients with suspected respiratory infection.

Host response biomarkers can accurately distinguish between influenza and bacterial infection. However, published biomarkers require the measurement of many genes, thereby making it difficult to implement them in clinical practice. This study aims to identify a single-gene biomarker with a high diagnostic accuracy equivalent to multi-gene biomarkers.In this study, we combined an integrated genomic analysis of 1071 individuals with in vitro experiments using well-established infection models.We identified a single-gene biomarker, IFI27, which had a high prediction accuracy (91%) equivalent to that obtained by multi-gene biomarkers. In vitro studies showed that IFI27 was upregulated by TLR7 in plasmacytoid dendritic cells, antigen-presenting cells that responded to influenza virus rather than bacteria. In vivo studies confirmed that IFI27 was expressed in influenza patients but not in bacterial infection, as demonstrated in multiple patient cohorts (n=521). In a large prospective study (n=439) of patients presented with undifferentiated respiratory illness (aetiologies included viral, bacterial and non-infectious conditions), IFI27 displayed 88% diagnostic accuracy (AUC) and 90% specificity in discriminating between influenza and bacterial infections.IFI27 represents a significant step forward in overcoming a translational barrier in applying genomic assay in clinical setting; its implementation may improve the diagnosis and management of respiratory infection.

California's Sterilization Survivors: An Estimate and Call for Redress.

From 1919 to 1952, approximately 20 000 individuals were sterilized in California's state institutions on the basis of eugenic laws that sought to control the reproductive capacity of people labeled unfit and defective. Using data from more than 19 000 sterilization recommendations processed by state institutions over this 33-year period, we provide the most accurate estimate of living sterilization survivors. As of 2016, we estimate that as many as 831 individuals, with an average age of 87.9 years, are alive. We suggest that California emulate North Carolina and Virginia, states that maintained similar sterilization programs and recently have approved monetary compensation for victims. We discuss the societal obligation for redress of this historical injustice and recommend that California seriously consider reparations and full accountability.

The CYP27B1 variant associated with an increased risk of autoimmune disease is underexpressed in tolerizing dendritic cells.

Genome-wide association studies have identified a linkage disequilibrium (LD) block on chromosome 12 associated with multiple sclerosis (MS), type 1 diabetes and other autoimmune diseases. This block contains CYP27B1, which catalyzes the conversion of 25 vitamin D3 (VitD3) to 1,25VitD3. Fine-mapping analysis has failed to identify which of the 17 genes in this block is most associated with MS. We have previously used a functional approach to identify the causal gene. We showed that the expression of several genes in this block in whole blood is highly associated with the MS risk allele, but not CYP27B1. Here, we show that CYP27B1 is predominantly expressed in dendritic cells (DCs). Its expression in these cells is necessary for their response to VitD, which is known to upregulate pathways involved in generating a tolerogenic DC phenotype. Here, we utilize a differentiation protocol to generate inflammatory (DC1) and tolerogenic (DC2) DCs, and show that for the MS risk allele CYP27B1 is underexpressed in DCs, especially DC2s. Of the other Chr12 LD block genes expressed in these cells, only METT21B expression was as affected by the genotype. Another gene associated with autoimmune diseases, CYP24A1, catabolizes 1,25 VitD3, and is predominantly expressed in DCs, but equally between DC1s and DC2s. Overall, these data are consistent with the hypothesis that reduced VitD pathway gene upregulation in DC2s of carriers of the risk haplotype of CYP27B1 contributes to autoimmune diseases. These data support therapeutic approaches aimed at targeting VitD effects on DCs.

Modeling problem behaviors in a nationally representative sample of adolescents.

Research on multiple problem behaviors has focused on the concept of Problem Behavior Syndrome (PBS). Problem Behavior Theory (PBT) is a complex and comprehensive social-psychological framework designed to explain the development of a range of problem behaviors. This study examines the structure of PBS and the applicability of PBT in adolescents. Participants were 6062 adolescents; aged 12-19 (51.3% female) who took part in the My World Survey-Second Level (MWS-SL). Regarding PBS, Confirmatory Factor Analysis established that problem behaviors, such as alcohol and drug use loaded significantly onto a single, latent construct for males and females. Using Structural Equation Modeling, the PBT framework was found to be a good fit for males and females. Socio-demographic, perceived environment system and personality accounted for over 40% of the variance in problem behaviors for males and females. Our findings have important implications for understanding how differences in engaging in problem behaviors vary by gender.

A Website Intervention to Increase Knowledge About Living Kidney Donation and Transplantation Among Hispanic/Latino Dialysis Patients.

BACKGROUND: Hispanic dialysis patients often encounter barriers to learning about living kidney donation and transplantation. Effective culturally targeted interventions to increase knowledge are lacking. We developed a culturally targeted educational website to enhance informed treatment decision making for end-stage kidney disease. METHODS: A pretest/posttest intervention study was conducted among adult Hispanic patients undergoing dialysis at 5 dialysis centers in Chicago, Illinois. Surveys included a 31-item, multiple-choice pretest/posttest of knowledge about kidney transplantation and living donation, attitudes about the website, Internet use, and demographics. The intervention entailed viewing 3 of 6 website sections for a total of 30 minutes. The pretest/posttest was administered immediately before and after the intervention. Participants completed a second posttest via telephone 3 weeks thereafter to assess knowledge retention, attitudes, and use of the website. RESULTS: Sixty-three patients participated (96% participation rate). Website exposure was associated with a mean 17.1% same day knowledge score increase between pretest and posttest (P < .001). At 3 weeks, participants' knowledge scores remained 11.7% above pretest (P < .001). The greatest knowledge gain from pretest to 3-week follow-up occurred in the Treatment Options (P < .0001) and Cultural Beliefs and Myths (P < .0001) website sections. Most participants (95%) "agreed" or "strongly agreed" that they would recommend the website to other Hispanics. CONCLUSIONS: Web-based education for patients undergoing dialysis can effectively increase Hispanics' knowledge about transplantation and living kidney donation. Study limitations include small sample size and single geographic region study. Dialysis facilities could enable website access as a method of satisfying policy requirements to provide education about kidney transplantation.

Peroxiredoxin-1 protects estrogen receptor α from oxidative stress-induced suppression and is a protein biomarker of favorable prognosis in breast cancer.

INTRODUCTION: Peroxiredoxin-1 (PRDX1) is a multifunctional protein, acting as a hydrogen peroxide (H2O2) scavenger, molecular chaperone and immune modulator. Although differential PRDX1 expression has been described in many tumors, the potential role of PRDX1 in breast cancer remains highly ambiguous. Using a comprehensive antibody-based proteomics approach, we interrogated PRDX1 protein as a putative biomarker in estrogen receptor (ER)-positive breast cancer. METHODS: An anti-PRDX1 antibody was validated in breast cancer cell lines using immunoblotting, immunohistochemistry and reverse phase protein array (RPPA) technology. PRDX1 protein expression was evaluated in two independent breast cancer cohorts, represented on a screening RPPA (n = 712) and a validation tissue microarray (n = 498). In vitro assays were performed exploring the functional contribution of PRDX1, with oxidative stress conditions mimicked via treatment with H2O2, peroxynitrite, or adenanthin, a PRDX1/2 inhibitor. RESULTS: In ER-positive cases, high PRDX1 protein expression is a biomarker of improved prognosis across both cohorts. In the validation cohort, high PRDX1 expression was an independent predictor of improved relapse-free survival (hazard ratio (HR) = 0.62, 95% confidence interval (CI) = 0.40 to 0.96, P = 0.032), breast cancer-specific survival (HR = 0.44, 95% CI = 0.24 to 0.79, P = 0.006) and overall survival (HR = 0.61, 95% CI = 0.44 to 0.85, P = 0.004). RPPA screening of cancer signaling proteins showed that ERα protein was upregulated in PRDX1 high tumors. Exogenous H2O2 treatment decreased ERα protein levels in ER-positive cells. PRDX1 knockdown further sensitized cells to H2O2- and peroxynitrite-mediated effects, whilst PRDX1 overexpression protected against this response. Inhibition of PRDX1/2 antioxidant activity with adenanthin dramatically reduced ERα levels in breast cancer cells. CONCLUSIONS: PRDX1 is shown to be an independent predictor of improved outcomes in ER-positive breast cancer. Through its antioxidant function, PRDX1 may prevent oxidative stress-mediated ERα loss, thereby potentially contributing to maintenance of an ER-positive phenotype in mammary tumors. These results for the first time imply a close connection between biological activity of PRDX1 and regulation of estrogen-mediated signaling in breast cancer.

Combined hepatocellular-cholangiocarcinoma (cHCC-CC): a distinct entity.

Patients with hepatocellular carcinoma (HCC), the fifth most common cancer worldwide, display a highly variable clinical course, suggesting that HCC encompasses several biologically distinct subtypes. This heterogeneity has the potential to impede both treatment decisions and prognostic predictions for patients with HCC. One distinct, albeit rare, subtype of HCC is combined hepatocellular-cholangiocarcinoma (cHCC-CC), which overall carries a poorer prognosis than HCC and cholangiocarcinoma (CC) alone. This review discusses predominantly the histopathologic and pathogenetic intricacies of this tumor and highlights the need for an accurate diagnosis of this specific HCC subtype.

Hispanic/Latino concerns about living kidney donation: a focus group study.

CONTEXT: Given the shortage of kidneys for transplant, living kidney donation (LKD) is increasingly used to expand the organ donor pool. Although Hispanics/Latinos need disproportionately more kidney transplants, they receive a smaller proportion of living donor kidney transplants than other ethnic/racial groups. OBJECTIVE: To assess Hispanics' awareness, perceptions, misconceptions, cultural beliefs, and values about and barriers to LKD. DESIGN: Nine focus groups were conducted with 76 adult Hispanics in Chicago, Illinois, between January and March 2012. PARTICIPANTS: Focus groups included kidney transplant recipients, living kidney donors, dialysis patients, and the general Hispanic public. RESULTS: Several themes emerged as perceived barriers to LKD. Many participants identified knowledge deficits about LKD, expressing uncertainty about the differences between LKD and deceased donation, and whether kidney disease simultaneously afflicts both kidneys. Many believed that donors experience dramatically shorter life expectancies, are unable to have children, and are more susceptible to kidney disease after donating. Recipients and donors reported that family members were involved in discussions about the donor's decision to donate, with some family members discouraging donation. Financial barriers cited included fear of becoming unable to work, losing one's job, or being unable to pay household bills while recovering. Participants also identified logistic barriers for undocumented immigrants (eg, the inability to obtain government insurance for transplant candidates and uncertainty about their eligibility to donate). Donors desired information about optimizing self-care to promote their remaining kidney's health. Culturally competent interventions are needed to redress Hispanics' knowledge deficits and misconceptions and reduce LKD disparities among Hispanics.