Determination Of Phenotype Research Articles

Expression of transferrin receptor 1 and β1-integrins correlates with estrogen receptor status and immune infiltration in breast cancer

To study of transferrin receptor 1 (CD71) expression by primary breast cancer cells in correlation with tumor cell phenotype. Determination of BC phenotype: immunohistochemical staining method (immunofluorescence). Antibodies to ER (estrogen receptors), KL-1 (pancytokeratin), CD71 (transferrin receptor), CD29 (β1-integrins). CD45, CD3, CD4, CD8, CD20 infiltration was also evaluated. ZEISS microscope (AXIOSKOP; Germany), method of G.J. Hammerling et al. Statistical processing: IBM-SPSS Statistics v.21. 63% of BC cases had CD71+ phenotype. CD71-mosaic tumors were observed in 14.4%. β1-integrin expression was monomorphic in 51.6% of cases and mosaic in 38.7%. 85% of ER-positive tumors were CD71-positive with a monomorphic type of reaction; p=0.014. Among ER-negative tumors, CD71-negative reactions were 2-fold more frequent and the monomorphic type was less frequent. ER-positive tumors were CD29-positive in 73%; p=0.031. 45.5% of ER+ tumors were CD29-monomorphic. Among ER-negative tumors, the frequency of CD29-monomorphic tumors was 55%. Significant infiltration by CD3+ cells was predominant in CD71-positive tumors; p=0.016. In the CD29-monomorphic phenotype, CD45+ infiltration was 31.3%, and in the mosaic phenotype, 67.1%. BC aberrantly expresses transferrin receptors, β1-integrins. CD71 expression is associated with ER expression. ER-positive tumors are often monomorphic for CD71. Prominent CD3+ infiltration was present in CD71+ tumors. Expression of β1-integrins correlated with ER+ status and weak immune infiltration.

A study on phenotypic characterization and genotypic determination of drug resistance of Helicobacter pylori from cases of acid peptic disease in a tertiary care center

Background: Helicobacter pylori infection is one of the major risk factors for gastroduodenal disease, and invasive or non-invasive tests are available to detect it. Aims and Objectives: The study aims to identify H. pylori phenotypically and genotypically to identify drug-resistant H. pylori in suspected cases of peptic acid disorders at a tertiary care center. Materials and Methods: This 6-month prospective and observational study was performed at Saveetha Medical College and Hospitals. Patients’ gastric biopsy samples were obtained from 105 individuals with gastroduodenal disorders who had upper gastrointestinal endoscopies at the Hospital. A fiber optic endoscope and biopsy forceps were used for the endoscopy. Biopsy samples were collected from the antrum and placed in Eppendorf tubes for fast urease, culture, gram stain, polymerase chain reaction (PCR), and histological analysis. Results: A total of 78 (74%) males and 27 (26%) females were among the 105 patients. Endoscopic findings imply gastritis, gastric ulcer, and duodenal ulcer in 74 patients (70.48%). Rapid urease test (RUT), gram stain, and histology revealed positive results in 20 cases. RUT showed that 30.47% of patients tested positive. Gram stain detected 25.1% of H. pylori infections, and 12 cases were histopathologically positive. By PCR, 21.62% of gastritis patients were positive for H. pylori infection. RUT and PCR give comparable results for detecting H. pylori infection, with a strong association between H. pylori infection and peptic ulcer disease. Conclusion: A combination of RUT and PCR is an effective diagnostic approach for H. pylori infection, allowing doctors to select the most appropriate medications for specific patients.

Deciphering Collagen Phenotype Dynamics Regulators: Insights from In-Silico Analysis.

Collagen (Col) types I and III are integral components in wound healing and tissue regeneration, influencing tissue development, homeostasis, and related pathologies. Col I and Col III expression changes during different stages of wound healing and understanding the regulation of collagen phenotype determination is crucial for unraveling the complexities of these processes. Transcription factors and microRNAs, directly and indirectly, play a critical role in regulating collagen expression, however, a comprehensive understanding of the factors regulating Col I and III phenotypes remains elusive. This critically analyzed published reports with focuses on various factors regulating the expression of Col I and Col III at the transcriptional and translational levels. We performed bioinformatics analysis with an input of proinflammatory mediators, growth factors, elastases, and matrix metalloproteinases and predicted transcription factors and microRNAs involved in the regulation of collagen expression. Network analysis revealed an interaction between genes, transcription factors, and microRNAs and provided a holistic view of the regulatory landscape governing collagen expression and unveils intricate interconnections. This analysis lays a founda-tional framework for guiding future research and therapeutic interventions to promote extracellular matrix remodeling, wound healing, and tissue regeneration after an injury by modulating collagen expression. In essence, this scientific groundwork offers a comprehensive exploration of the regulatory dynamics in collagen synthesis, serving as a valuable resource for advancing both basic research and clinical interventions in tissue repair.

Identification of Hypertension in Electronic Health Records Through Computable Phenotype Development and Validation for Use in Public Health Surveillance: Retrospective Study.

Electronic health record (EHR) systems are widely used in the United States to document care delivery and outcomes. Health information exchange (HIE) networks, which integrate EHR data from the various health care providers treating patients, are increasingly used to analyze population-level data. Existing methods for population health surveillance of essential hypertension by public health authorities may be complemented using EHR data from HIE networks to characterize disease burden at the community level. We aimed to derive and validate computable phenotypes (CPs) to estimate hypertension prevalence for population-based surveillance using an HIE network. Using existing data available from an HIE network, we developed 6 candidate CPs for essential (primary) hypertension in an adult population from a medium-sized Midwestern metropolitan area in the United States. A total of 2 independent clinician reviewers validated the phenotypes through a manual chart review of 150 randomly selected patient records. We assessed the precision of CPs by calculating sensitivity, specificity, positive predictive value (PPV), F1-score, and validity of chart reviews using prevalence-adjusted bias-adjusted κ. We further used the most balanced CP to estimate the prevalence of hypertension in the population. Among a cohort of 548,232 adults, 6 CPs produced PPVs ranging from 71% (95% CI 64.3%-76.9%) to 95.7% (95% CI 84.9%-98.9%). The F1-score ranged from 0.40 to 0.91. The prevalence-adjusted bias-adjusted κ revealed a high percentage agreement of 0.88 for hypertension. Similarly, interrater agreement for individual phenotype determination demonstrated substantial agreement (range 0.70-0.88) for all 6 phenotypes examined. A phenotype based solely on diagnostic codes possessed reasonable performance (F1-score=0.63; PPV=95.1%) but was imbalanced with low sensitivity (47.6%). The most balanced phenotype (F1-score=0.91; PPV=83.5%) included diagnosis, blood pressure measurements, and medications and identified 210,764 (38.4%) individuals with hypertension during the study period (2014-2015). We identified several high-performing phenotypes to identify essential hypertension prevalence for local public health surveillance using EHR data. Given the increasing availability of EHR systems in the United States and other nations, leveraging EHR data has the potential to enhance surveillance of chronic disease in health systems and communities. Yet given variability in performance, public health authorities will need to decide whether to seek optimal balance or declare a preference for algorithms that lean toward sensitivity or specificity to estimate population prevalence of disease.

The Role of Cancer Stem Cell Markers in Ovarian Cancer.

Ovarian cancer is the most lethal gynaecological cancer and the eighth most common female cancer. The early diagnosis of ovarian cancer remains a clinical problem despite the significant development of technology. Nearly 70% of patients with ovarian cancer are diagnosed with stages III-IV metastatic disease. Reliable diagnostic and prognostic biomarkers are currently lacking. Ovarian cancer recurrence and resistance to chemotherapy pose vital problems and translate into poor outcomes. Cancer stem cells appear to be responsible for tumour recurrence resulting from chemotherapeutic resistance. These cells are also crucial for tumour initiation due to the ability to self-renew, differentiate, avoid immune destruction, and promote inflammation and angiogenesis. Studies have confirmed an association between CSC occurrence and resistance to chemotherapy, subsequent metastases, and cancer relapses. Therefore, the elimination of CSCs appears important for overcoming drug resistance and improving prognoses. This review focuses on the expression of selected ovarian CSC markers, including CD133, CD44, CD24, CD117, and aldehyde dehydrogenase 1, which show potential prognostic significance. Some markers expressed on the surface of CSCs correlate with clinical features and can be used for the diagnosis and prognosis of ovarian cancer. However, due to the heterogeneity and plasticity of CSCs, the determination of specific CSC phenotypes is difficult.

The possibility of phenotyping patients with sepsis in a chronic critical condition

The main obstacle to understanding sepsis is an excessively broad definition of this disease, which covers an extensive set of clinical and pathophysiological signs. Different combinations of these traits can naturally combine into phenotypes that have different degrees of risk of an adverse outcome, and may respond differently to treatment. Scientific work on the determination of phenotypes has focused mainly on patients in the intensive care unit. In addition, there have been no prospective or retrospective studies on the classification and phenotyping of sepsis patients in a chronic critical condition. Most likely, this is due to the fact that the term «chronic critical patient» is quite novel and has recently been widely covered in both Russian and foreign scientific literature. The purpose of this work is the theoretical determination of phenotypic groups for chronic critical patients. Methods and materials: The search for Russian publications was carried out in the database on the RSCI website, and foreign publications were searched in the PubMed and Google Scholar databases in the period of 1998–2022. When analyzing the PubMed database, the query «sepsis phenotype» resulted in 62,371 links. The works on the keywords «chronic critical illness» were also studied. The publications describing the sepsis phenotypes, the diagnosis of sepsis and septic shock, as well as the clinical picture of a chronic critical condition (illness) were analyzed, with a total of 45 scientific articles. Discussion: It is also worth noting that the study by C. Seymour, latent class analysis, and other works devoted to the treatment of sepsis consider a large number of mainly non-surgical patients, without dividing them by the main nosology and foci of infection. Meanwhile, the main axis of neurohumoral immunity, i.e. the brain — gastrointestinal tract, is disrupted in chronic critical patients. These patients, as well as patients undergoing repeated sepsis caused by a nosocomial infection, are not considered separately in any of the studies. Furthermore, neither the study by C. Seymour, nor the latent class analysis examines any instrumental method for assessing the infection focus (radiography or computed tomography of the lungs). Much attention is paid to the acid-base state of patients, but the state of the main buffer systems is not described in terms of the presence of concomitant or competing diseases, and an indication of an increase in creatinine and blood urea nitrogen can only indirectly indicate kidney dysfunction and, as a consequence, a disorder in the bicarbonate bufer system. Conclusions: Based on the above, the basic classifications for patients with sepsis in an acute state should be updated for chronic critical patients with host response options, taking into account the peculiarities of the immune system. Moreover, it is recommendable to phenotype chronic critical patients separately, considering the localization of brain damage, since patients with vertebrobasilar lesions are more prone to aspiration pneumonia and severe sepsis, while this type of pneumonia is extremely rare in patients with frontal lobe lesions. Accordingly, hypothalamic lesion leads to a change in the neurohumoral immune response to various pathogens. Thus, chronic critical patients with sepsis should be classified not only according to already known phenotyping systems, but also based on the localization of brain damage and the functionality of the gastrointestinal tract.

Determinants of dementia phenotype in North African & Mid‐eastern Israelis

AbstractBackgroundDiverse ethnic ancestry and adverse social health determinants are increasingly recognized to impact dementia on multiple dimensions: genetically and environmentally determined risk, clinical presentation, detection, and the utility of cognitive testing and management. Most evidence originates from ethnically diverse cohorts in North America, with little representation of other world regions. We aimed to assess dementia clinical and neuroimaging phenotype in individuals of Middle Eastern and Northern African (MENA) descent, a population severely underrepresented in brain health research.MethodsThe Alzheimer’s and atypical dementias in Israelis of diverse ancestry and disparate social health determinants (ADIDAS) project started on January 1st, 2022, and will prospectively recruit 200 individuals aged 40‐80 years reporting a new cognitive decline. The ADIDAS protocol includes APOE genotyping, structured assessment of early‐to‐late life social determinants, Israeli neighbourhood disadvantage index, medical history, vascular risk factors, cognitive testing, plasma Alzheimer’s biomarkers (Aβ40, Aβ42, total‐tau, p‐tau181, and NfL), and brain MRI. Ancestry was defined using participants and paternal country of birth. Countries were grouped into MENA, European, or neither, as defined by the World Bank. The project aims to assess the relationship between ethnic ancestry and associated social health determinants as predictors of brain neurodegeneration evident on neuroimaging and plasma biomarkers.Preliminary ResultsAs of January 1st, 2023, the ADIDAS cohort recruited 51 participants (55% female, mean age 65.8±8.4 years) of MENA (n = 24), European (n = 23), and neither (n = 3) ancestry. A preliminary comparison of MENA and European groups reveals MENA in our cohort are on average younger (63.87±8.9 vs. 68.3±8.2 years), have fewer years of schooling(11.7±2.5 vs. 16.2±3.4 years), and more likely to obtain only elementary or vocational education (75% vs. 13%). MENA were more likely to score poor results on cognitive testing‐ MMSE (21.6±4.9 vs. 24.8±5.5), receive a higher clinical dementia rating sum of boxes (3.4±2.5 vs. 1.45±1.4), and a lower carriage rate of apoE4 allele (11% vs. 30%).ConclusionPreliminary non‐matched comparison suggests MENA with early‐stage cognitive impairment have a different social, cognitive, and genetic risk profile than individuals of European ancestry. Results of plasma biomarkers profile and MRI‐related structural changes will be shared at AAIC2023.

Identifying Phenotypic Determinants of Infliximab Target Concentrations in Inflammatory Bowel Disease: Toward Personalized Evidence-Based Dosing Regimens.

Identifying Phenotypic Determinants of Infliximab Target Concentrations in Inflammatory Bowel Disease: Toward Personalized Evidence-Based Dosing Regimens.

Genome-wide association analysis of left ventricular imaging-derived phenotypes identifies 72 risk loci and yields genetic insights into hypertrophic cardiomyopathy

Left ventricular regional wall thickness (LVRWT) is an independent predictor of morbidity and mortality in cardiovascular diseases (CVDs). To identify specific genetic influences on individual LVRWT, we established a novel deep learning algorithm to calculate 12 LVRWTs accurately in 42,194 individuals from the UK Biobank with cardiac magnetic resonance (CMR) imaging. Genome-wide association studies of CMR-derived 12 LVRWTs identified 72 significant genetic loci associated with at least one LVRWT phenotype (P < 5 × 10−8), which were revealed to actively participate in heart development and contraction pathways. Significant causal relationships were observed between the LVRWT traits and hypertrophic cardiomyopathy (HCM) using genetic correlation and Mendelian randomization analyses (P < 0.01). The polygenic risk score of inferoseptal LVRWT at end systole exhibited a notable association with incident HCM, facilitating the identification of high-risk individuals. The findings yield insights into the genetic determinants of LVRWT phenotypes and shed light on the biological basis for HCM etiology.

Spatial and Temporal Control of 3D Hydrogel Viscoelasticity through Phototuning.

The mechanical properties of the extracellular environment can regulate a variety of cellular functions, such as spreading, migration, proliferation, and even differentiation and phenotypic determination. Much effort has been directed at understanding the effects of the extracellular matrix (ECM) elastic modulus and, more recently, stress relaxation on cellular processes. In physiological contexts such as development, wound healing, and fibrotic disease progression, ECM mechanical properties change substantially over time or space. Dynamically tunable hydrogel platforms have been developed to spatiotemporally modulate a gel's elastic modulus. However, dynamically altering the stress relaxation rate of a hydrogel remains a challenge. Here, we present a strategy to tune hydrogel stress relaxation rates in time or space using a light-triggered tethering of poly(ethylene glycol) to alginate. We show that the stress relaxation rate can be tuned without altering the elastic modulus of the hydrogel. We found that cells are capable of sensing and responding to dynamic stress relaxation rate changes, both morphologically and through differences in proliferation rates. We also exploited the light-based technique to generate spatial patterns of stress relaxation rates in 3D hydrogels. We anticipate that user-directed control of the 3D hydrogel stress relaxation rate will be a powerful tool that enables studies that mimic dynamic ECM contexts or as a means to guide cell fate in space and time for tissue engineering applications.

Characterization and Quantification of Von Willebrand Factor Proteoforms in Von Willebrand Disease Patients and Healthy Controls: Insights from the Win-Study

Background: Von Willebrand disease (VWD) is a commonly inherited bleeding disorder which is characterized by either low (type 1) or absent (type 3) levels of Von Willebrand factor (VWF) or by the occurrence of dysfunctional VWF proteoforms (type 2) in plasma. The complexity of diagnosing and managing VWD arises from limited understanding of the diverse VWF proteoforms present in both healthy individuals as well as in VWD patients. Objective: The aim of this study was to investigate the plasma proteome and to characterize VWF proteoforms in Von Willebrand disease (VWD) patients, that were enrolled in the Willebrand in the Netherlands (WiN) 1 study, compared with healthy controls. Method: We selected type 1, type 2A, type 2M, type 2N and type 3 patients from the WiN cohort for which extensive laboratory- and genotype data were available. Plasma from 64 VWD patients and ten healthy controls was analyzed using a combination of discovery- and targeted mass spectrometry-based approaches. Results: Both discovery-based as well as targeted quantitative proteomics analysis revealed significant variation in VWF levels across the selected VWD patient group, which was in line with reported clinical laboratory data. Elevated VWF-propeptide/VWF ratios were observed in plasma samples from a range of VWD patients, with notable increases observed in VWD type 2A, B, and M, indicating accelerated clearance of these VWF proteoforms. Discovery-based proteomics revealed a strong VWF-F8 relationship in all VWD subtypes and healthy controls, except for type 2N VWD. We were able to identify peptides corresponding with common polymorphisms ( e.g. p.Thr798Ala, p.Gln852Arg, p.Thr1381Ala), for which the observed peptide distribution was in line with the minor allele frequency reported by the Broad institute. In addition, we identified peptides corresponding with 16 pathogenic VWF variants (e.g. p.Arg1306Trp, p. Phe1293Leu, p.Cys1190Arg, p.Arg1374His or p.Tyr1584Cys) of which p.Cys1190Arg wild-type and variant peptide aligned with the zygosity at DNA level (Figure X). Based on spike-in with stable isotope-labeled peptides, the variant-to-wildtype ratios varied among VWD patients with distinct pathogenic variants, ranging from 0.5-2.5 in type 2M (p.Phe1293Leu) and type 1 patients (p.Tyr1584Cys), &amp;lt;1 in type 2A patients (p.Arg1374His), to &amp;gt;8 in type 2B patients (p.Arg1306Trp), suggesting distinct stoichiometry of VWF variants among these patients (Figure 2). Conclusion: Our study highlights the feasibility of VWF proteoform identification and quantification from plasma and suggest a potential role in unravelling their contribution to the biological variations and clinical diversity observed in VWD. 1 De Wee et al., Determinants of bleeding phenotype in adult patients with moderate or severe von Willebrand disease. Thromb Haemost. 2012 Oct;108(4):683-92.

Role of monocytes and dendritic cells in cardiac reverse remodelling after cardiac resynchronization therapy

Background and aimsMonocytes and dendritic cells (DC) are both key inflammatory cells, with recognized effects on cardiac repair. However, there are distinct subsets of monocytes with potential for beneficial or detrimental effects on heart failure (HF) pathogenesis. The connection between reverse cardiac remodelling, the potential anti-inflammatory effect of cardiac resynchronization therapy (CRT) and monocytes and DC homeostasis in HF is far from being understood. We hypothesized that monocytes and DC play an important role in cardiac reverse remodelling and CRT response. Therefore, we aimed to assess the potential role of baseline peripheral levels of blood monocytes and DC subsets and their phenotypic and functional activity for CRT response, in HF patients. As a secondary objective, we aimed to evaluate the impact of CRT on peripheral blood monocytes and DC subsets, by comparing baseline and post CRT circulating levels and phenotypic and functional activity.MethodsForty-one patients with advanced HF scheduled for CRT were included in this study. The quantification and phenotypic determination of classical (cMo), intermediate (iMo) and non-classical monocytes (ncMo), as well as of myeloid (mDC) and plasmacytoid DC (pDC) were performed by flow cytometry in a FACSCanto™II (BD) flow cytometer. The functional characterization of total monocytes and mDC was performed by flow cytometry in a FACSCalibur flow cytometer, after in vitro stimulation with lipopolysaccharide from Escherichia coli plus interferon (IFN)-γ, in the presence of Brefeldina A.Comparisons between the control and the patient group, and between responders and non-responders to CRT were performed.ResultsCompared to the control group, HF population presented a significantly lower frequency of pDC at baseline and a higher proportion of monocytes and mDC producing IL-6 and IL-1β, both before and 6-months after CRT (T6). There was a remarkable decrease of cMo and an increase of iMo after CRT, only in responders. The responder group also presented higher ncMo values at T6 compared to the non-responder group. Both responders and non-responders presented a decrease in the expression of CD86 in all monocyte and DC populations after CRT. Moreover, in non-responders, the increased frequency of IL-6-producing DC persisted after CRT.ConclusionOur study provides new knowledge about the possible contribution of pDC and monocytes subsets to cardiac reverse remodelling and response to CRT. Additionally, CRT is associated with a reduction on CD86 expression by monocytes and DC subsets and in their potential to produce pro-inflammatory cytokines, contributing, at least in part, for the well described anti-inflammatory effects of CRT in HF patients.

Molecular Mechanism Biomarkers Predict Diagnosis in Schizophrenia and Schizoaffective Psychosis, with Implications for Treatment.

Diagnostic uncertainty and relapse rates in schizophrenia and schizoaffective disorder are relatively high, indicating the potential involvement of other pathological mechanisms that could serve as diagnostic indicators to be targeted for adjunctive treatment. This study aimed to seek objective evidence of methylenetetrahydrofolate reductase MTHFR C677T genotype-related bio markers in blood and urine. Vitamin and mineral cofactors related to methylation and indolamine-catecholamine metabolism were investigated. Biomarker status for 67 symptomatically well-defined cases and 67 asymptomatic control participants was determined using receiver operating characteristics, Spearman's correlation, and logistic regression. The 5.2%-prevalent MTHFR 677 TT genotype demonstrated a 100% sensitive and specific case-predictive biomarkers of increased riboflavin (vitamin B2) excretion. This was accompanied by low plasma zinc and indicators of a shift from low methylation to high methylation state. The 48.5% prevalent MTHFR 677 CC genotype model demonstrated a low-methylation phenotype with 93% sensitivity and 92% specificity and a negative predictive value of 100%. This model related to lower vitamin cofactors, high histamine, and HPLC urine indicators of lower vitamin B2 and restricted indole-catecholamine metabolism. The 46.3%-prevalent CT genotype achieved high predictive strength for a mixed methylation phenotype. Determination of MTHFR C677T genotype dependent functional biomarker phenotypes can advance diagnostic certainty and inform therapeutic intervention.

Alpha-1 adrenergic signaling drives cardiac regeneration via extracellular matrix remodeling transcriptional program in zebrafish macrophages

The autonomic nervous system plays a pivotal role in cardiac repair. Here, we describe the mechanistic underpinning of adrenergic signaling in fibrotic and regenerative response of the heart to be dependent on immunomodulation. A pharmacological approach identified adrenergic receptor alpha-1 as a key regulator of macrophage phenotypic diversification following myocardial damage in zebrafish. Genetic manipulation and single-cell transcriptomics showed that the receptor signals activation of an "extracellular matrix remodeling" transcriptional program in a macrophage subset, which serves as a key regulator of matrix composition and turnover. Mechanistically, adrenergic receptor alpha-1-activated macrophages determine activation of collagen-12-expressing fibroblasts, a cellular determinant of cardiac regenerative niche, through midkine-mediated paracrine crosstalk, allowing lymphatic and blood vessel growth and cardiomyocyte proliferation at the lesion site. These findings identify the mechanism of adrenergic signaling in macrophage phenotypic and functional determination and highlight the potential of neural modulation for regulation of fibrosis and coordinationof myocardial regenerative response.

Differential Resistance of Borrelia burgdorferi Clones to Human Serum-Mediated Killing Does Not Correspond to Their Predicted Invasiveness.

Reservoir host associations have been observed among and within Borrelia genospecies, and host complement-mediated killing is a major determinant in these interactions. In North America, only a subset of Borrelia burgdorferi lineages cause the majority of disseminated infections in humans. We hypothesize that differential resistance to human complement-mediated killing may be a major phenotypic determinant of whether a lineage can establish systemic infection. As a corollary, we hypothesize that borreliacidal action may differ among human subjects. To test these hypotheses, we isolated primary B. burgdorferi clones from field-collected ticks and determined whether the killing effects of human serum differed among those clones in vitro and/or whether these effects were consistent among human sera. Clones associated with human invasiveness did not show higher survival in human serum compared to noninvasive clones. These results indicate that differential complement-mediated killing of B. burgdorferi lineages is not a determinant of invasiveness in humans. Only one significant difference in the survivorship of individual clones incubated in different human sera was detected, suggesting that complement-mediated killing of B. burgdorferi is usually similar among humans. Mechanisms other than differential human complement-mediated killing of B. burgdorferi lineages likely explain why only certain lineages cause the majority of disseminated human infections.

PB0642 Determinants of the Bleeding Phenotype in Persons with Non-Severe Hemophilia

PB0642 Determinants of the Bleeding Phenotype in Persons with Non-Severe Hemophilia

Up-Regulation of TCF21 Expression Reverses the Malignant Phenotype of Cancer-Associated Fibroblasts in Esophageal Squamous Cell Carcinoma

Cancer-associated fibroblasts (CAFs), as one major component of tumor microenvironment (TME), are closely associated with tumor initiation and progression. Our previous studies have discovered that CAFs induced the resistance of esophageal squamous cell carcinoma (ESCC) cells to a variety of chemotherapeutic drugs such as cisplatin and paclitaxel. Furthermore, CAFs attenuated ionizing irradiation (IR)-induced cancer cells death by regulating DNA damage response. CAFs themselves are highly resistant to death stimuli due to enhanced antioxidant potential and DNA repair capacity. How to inhibit the malignant phenotype of CAFs is critically important for the radical treatment of ESCC. By RNA-sequencing and DNA methylation analysis, the transcriptome and epigenome of CAFs and matched normal fibroblasts (NFs) have been integratively analyzed. By transfection of TCF21 cDNA plasmid, the expression of TCF21 in CAFs has been up-regulated. Using a cell counting kit and migration and invasion assay, the effect of TCF21 on the growth and migration and invasive ability of CAFs has been detected. Using immunofluorescence and flow cytometry (FCM) analysis and western blotting, the effect of TCF21 on the DNA damage repair and apoptotic death of CAFs following IR has been detected. TCF21 is one of the top ten down-regulated genes in CAFs compared with NFs due to promoter methylation. Up-regulation of TCF21 expression inhibited the growth rate and migration and invasive ability of CAFs. The expression of α-SMA, as an indicator of CAFs activation, was down-regulated in CAFs which were transfected with TCF21 cDNA. Furthermore, when TCF21 cDNA was transfected into CAFs, IR-induced DNA damage was increased while DNA repair was inhibited in CAFs, suggesting that TCF21 was involved in DNA damage response of CAFs following IR. FCM analysis showed that up-regulation of TCF21 expression promoted IR-induced apoptotic death of CAFs. TCF21 is a determinant of the malignant phenotype of CAFs in ESCC. Up-regulation of TCF21 expression is a promising approach of inhibiting the growth, migration and invasion, activation and radioresistance of CAFs in ESCC.

Pro-Apoptotic Activity of MCL-1 Inhibitor in Trametinib-Resistant Melanoma Cells Depends on Their Phenotypes and Is Modulated by Reversible Alterations Induced by Trametinib Withdrawal.

Although BRAFV600/MEK inhibitors improved the treatment of melanoma patients, resistance is acquired almost inevitably. Trametinib withdrawal/rechallenge and MCL-1 inhibition in trametinib-resistance models displaying distinct p-ERK1/2 levels were investigated. Trametinib withdrawal/rechallenge caused reversible changes in ERK1/2 activity impacting the balance between pro-survival and pro-apoptotic proteins. Reversible alterations were found in MCL-1 levels and MCL-1 inhibitors, BIM and NOXA. Taking advantage of melanoma cell dependency on MCL-1 for survival, we used S63845. While it was designed to inhibit MCL-1 activity, we showed that it also significantly reduced NOXA levels. S63845-induced apoptosis was detected as the enhancement of Annexin V-positivity, caspase-3/7 activation and histone H2AX phosphorylation. Percentages of Annexin V-positive cells were increased most efficiently in trametinib-resistant melanoma cells displaying the p-ERK1/2low/MCL-1low/BIMhigh/NOXAlow phenotype with EC50 values at concentrations as low as 0.1 μM. Higher ERK1/2 activity associated with increased MCL-1 level and reduced BIM level limited pro-apoptotic activity of S63845 further influenced by a NOXA level. Our study supports the notion that the efficiency of an agent designed to target a single protein can largely depend on the phenotype of cancer cells. Thus, it is important to define appropriate phenotype determinants to stratify the patients for the novel therapy.

Mortality Risk Factors in Pediatric Onco-Critical Care Patients and Machine Learning Derived Early Onco-Critical Care Phenotypes in a Retrospective Cohort.

To use supervised and unsupervised statistical methodology to determine risk factors associated with mortality in critically ill pediatric oncology patients to identify patient phenotypes of interest for future prospective study. This retrospective cohort study included nonsurgical pediatric critical care admissions from January 2017 to December 2018. We determined the prevalence of multiple organ failure (MOF), ICU mortality, and associated factors. Consensus k-means clustering analysis was performed using 35 bedside admission variables for early, onco-critical care phenotype development. Single critical care unit in a subspeciality pediatric hospital. None. There were 364 critical care admissions in 324 patients with underlying malignancy, hematopoietic cell transplant, or immunodeficiency reviewed. Prevalence of multiple organ failure, ICU mortality, determination of early onco-critical care phenotypes. ICU mortality was 5.2% and was increased in those with MOF (18.4% MOF, 1.7% single organ failure [SOF], 0.6% no organ failure; p ≤ 0.0001). Prevalence of MOF was 23.9%. Significantly increased ICU mortality risk was associated with day 1 MOF (hazards ratio [HR] 2.27; 95% CI, 1.10-6.82; p = 0.03), MOF during ICU admission (HR 4.16; 95% CI, 1.09-15.86; p = 0.037), and with invasive mechanical ventilation requirement (IMV; HR 5.12; 95% CI, 1.31-19.94; p = 0.018). Four phenotypes were derived (PedOnc1-4). PedOnc1 and 2 represented patient groups with low mortality and SOF. PedOnc3 was enriched in patients with sepsis and MOF with mortality associated with liver and renal dysfunction. PedOnc4 had the highest frequency of ICU mortality and MOF characterized by acute respiratory failure requiring invasive mechanical ventilation at admission with neurologic dysfunction and/or severe sepsis. Notably, most of the mortality in PedOnc4 was early (i.e., within 72 hr of ICU admission). Mortality was lower than previously reported in critically ill pediatric oncology patients and was associated with MOF and IMV. These findings were further validated and expanded by the four derived nonsynonymous computable phenotypes. Of particular interest for future prospective validation and correlative biological study was the PedOnc4 phenotype, which was composed of patients with hypoxic respiratory failure requiring IMV with sepsis and/or neurologic dysfunction at ICU admission.

Differentiated approach to glycemic management in type 2 diabetic patients during inpatient treatment of acute myocardial infarction: glycemic control and long-term prognosis

BACKGROUND: The risk of acute myocardial infarction (AMI) in patients with type 2 diabetes mellitus (T2DM) is 2 times higher than without DM. Previously an association between the time in the target range of glycemia during hospitalization of AMI in patients with T2DM (hTIR — «hospital time in range») and long-term prognosis was found. It is supposed that a differentiated approach to glycemic management will help to achieve a higher level of hTIR and will lead to an improved prognosis.AIM: To evaluate the effectiveness of differentiated approach to glycemic management in patients with T2DM during inpatient treatment of AMI and impact on long-term prognosis.MATERIALS AND METHODS: The study included patients with T2DM who were hospitalized with AMI with ST-segment elevation and underwent percutaneous coronary intervention. A total amount of patients was 161; 102 were in the main group, 59 in the control group. There was a differentiated approach to glycemia management based on phenotype determination in the main group. Insulin therapy was prescribed to patients with the 1st phenotype during the entire inpatient treatment. Patients with the second phenotype received insulin therapy followed by switching oral antidiabetic drugs (OAD). Patients of the 3rd phenotype were prescribed only OAD. In the control group, therapy of T2DM was prescribed according to the standard hospital procedures based on current clinical recommendations.RESULTS: In the main group, 22 patients (22%) were in the 1st phenotype, 54 (53%) –in the 2nd, 26 (25%) –in the 3rd. The average glycemia during hospitalization in the main group was lower than in the control group: 9.4±1.7 mmol/l vs 10.3±2.3 mmol/l (p=0.006). The hTIR in the main group was 58 [53; 71]% vs 46 [33; 63]% in the control group (p&lt;0.001). It was shown that when hTIR &gt; 55% is reached, the risk of cardiovascular death within a year after hospitalization is reduced by 80%.CONCLUSION: The differentiated approach to glycemic management in patients with T2DM leads to an improvement in glycemic control during inpatient treatment of AMI and is accompanied by a tendency to a lower incidence of cardiovascular death within a year after hospitalization. Achieving hTIR &gt; 55% is associated with a multifold reduction of the risk of cardiovascular death in a year. About 25% of patients with DM2 during inpatient treatment of AMI do not need the insulin therapy.