Negative Prognostic Indicator Research Articles

The Role of CLDN18.2 in Gastric Cancer Prognosis: A Systematic Review and Meta-Analysis

Background Gastric cancer (GC) is a major global cause of cancer mortality, with a median overall survival of just 12 months. CLDN18.2, a specific isoform of Claudin18 normally expressed in the gastric mucosa, has emerged as a potential therapeutic target and prognostic biomarker due to its exposure on the surface of tumor cells following malignant transformation. This exposure allows CLDN18.2's extracellular loops to bind monoclonal antibodies, presenting new opportunities for targeted therapy and improved prognostic assessment. Methods A comprehensive search of PubMed, EMBASE, Cochrane Library and Web of Science databases was conducted for studies that addressed the correlation of CLDN18.2 with: (1) Progression-free survival (PFS), and (2) Overall Survival (OS). Hazard ratio (HR) and odds ratio (OR) with 95% confidence intervals (CIs) were calculated using a fixed-effects model. Heterogeneity was examined with I2 statistics. P values of ≤ 0.05 were considered statistically significant. Statistical analyses were performed using RStudio, version 4.2.3. Results A total of 15 studies encompassing a total of 4,085 patients were included. There were 2,691 (65.8%) male, and 1,394 (34.2%) female patients. In the histologic GC analysis, there were 1,582 (38.7%) patients that had intestinal type and 1,280 (31.3%) with diffuse type. Patients with CLDN18.2 negative status exhibited a non-significant trend towards prolonged PFS (HR: 1.25; 95% CI: 0.98-1.61; p = 0.07; I2=18%) and a significant prolonged OS (HR: 1.20; 95% CI: 1.07-1.34; p < 0.01; I2=37%) when compared to CLDN18.2-positive patients. Conclusion Our findings establish CLDN18.2 as a robust negative prognostic indicator for overall survival in GC patients. While its impact on PFS was not statistically significant, the association with OS suggests CLDN18.2 may serve as a marker for complex biological processes underlying tumor advancement.

TGFB2 mRNA Levels Prognostically Interact with Interferon-Alpha Receptor Activation of IRF9 and IFI27, and an Immune Checkpoint LGALS9 to Impact Overall Survival in Pancreatic Ductal Adenocarcinoma

The treatment of pancreatic ductal adenocarcinoma (PDAC) is an unmet challenge, with the median overall survival rate remaining less than a year, even with the use of FOLFIRINOX-based therapies. This study analyzed archived macrophage-associated mRNA expression using datasets deposited in the UCSC Xena web platform to compare normal pancreatic tissue and PDAC tumor samples. The TGFB2 gene exhibited low mRNA expression levels in normal tissue, with less than one TPM. In contrast, in tumor tissue, TGFB2 expression levels exhibited a 7.9-fold increase in mRNA expression relative to normal tissue (p &lt; 0.0001). Additionally, components of the type-I interferon signaling pathway exhibited significant upregulation of mRNA levels in tumor tissue, including Interferon alpha/beta receptor 1 (IFNAR1; 3.4-fold increase, p &lt; 0.0001), Interferon regulatory factor 9 (IRF9; 4.2-fold increase, p &lt; 0.0001), Signal transducer and activator of transcription 1 (STAT1; 7.1-fold increase, p &lt; 0.0001), and Interferon Alpha Inducible Protein 27 (IFI27; 66.3-fold increase, p &lt; 0.0001). We also utilized TCGA datasets deposited in cBioportal and KMplotter to relate mRNA expression levels to overall survival outcomes. These increased levels of mRNA expression were found to be prognostically significant, whereby patients with high expression levels of either TGFB2, IRF9, or IFI27 showed median OS times ranging from 16 to 20 months (p &lt; 0.01 compared to 72 months for patients with low levels of expression for both TGFB2 and either IRF9 or IFI27). Examination of the KMplotter database determined the prognostic impact of TGFB2 mRNA expression levels by comparing patients expressing high versus low levels of TGFB2 (50th percentile cut-off) in low macrophage TME. In TME with low macrophage levels, patients with high levels of TGFB2 mRNA exhibited significantly shorter OS outcomes than patients with low TGFB2 mRNA levels (Median OS of 15.3 versus 72.7 months, p &lt; 0.0001). Furthermore, multivariate Cox regression models were applied to control for age at diagnosis. Nine genes exhibited significant increases in hazard ratios for TGFB2 mRNA expression, marker gene mRNA expression, and a significant interaction term between TGFB2 and marker gene expression (mRNA for markers: C1QA, CD74, HLA-DQB1, HLA-DRB1, HLA-F, IFI27, IRF9, LGALS9, MARCO). The results of our study suggest that a combination of pharmacological tools can be used in treating PDAC patients, targeting both TGFB2 and the components of the type-I interferon signaling pathway. The significant statistical interaction between TGFB2 and the nine marker genes suggests that TGFB2 is a negative prognostic indicator at low levels of the IFN-I activated genes and TAM marker expression, including the immune checkpoint LGALS9 (upregulated 16.5-fold in tumor tissue; p &lt; 0.0001).

The Prognostic Significance of CTSV Expression in Patients with Hepatocellular Carcinoma.

Cathepsin V (CTSV) is a cysteine protease peptidase, which is typically upregulated in cancer and is associated with various oncogenic processes, such as angiogenesis, proliferation, apoptosis, and invasion. The study explored the role of CTSV in hepatocellular carcinoma (HCC) and its potential as a potential biomarker. This study collected tumor and peritumoral archived specimens from 180HCC patients who underwent surgical resection at Zhongshan Hospital, Fudan University (Shanghai, China) between 2009 and 2010. We extracted data from the TCGA and GEO databases and conducted differential expression analysis, univariate Cox regression, and Kaplan-Meier analysis. Ultimately, we determined that CTSV may emerge as a potential biomarker. Then, immunohistochemical staining for CTSV was performed on tumors and adjacent tissues of HCC patients, and a Cox proportional hazards model was constructed to evaluate the prognostic significance of CTSV expression levels. Applied functional enrichment analysis to reveal the underlying molecular mechanisms. Utilized ssGSEA enrichment analysis and TIMER2.0 algorithm to explore the correlation between CTSV expression and immune cells in HCC. In vitro and in vivo experiments were conducted using human liver cancer cell lines to further validate the clinical application value of CTSV. In this study, we observed that CTSV expression was notably elevated in HCC (P < 0.001), and identified a significant association between elevated CTSV expression and reduced overall survival rates in patients. In vitro and in vivo experiments indicated that CTSV knockdown could significantly inhibit the proliferation, migration, and invasion of liver cancer cells, and it was found that the combination of CTSV knockdown with PD-1 inhibitors might enhance the therapeutic effect of PD-1 inhibitors in HCC. CTSV serves as a standalone negative prognostic indicator and possesses clinical significance in HCC.

Adaptation of a modified Elixhauser comorbidity index to isolate the impact of body mass index on survival outcomes in patients with cancer in a large integrated healthcare system.

124 Background: Variable impacts of body mass index (BMI) on overall survival (OS) and cancer-specific survival (CSS) have been described in patients with cancer, with existing data highlighting both detrimental effects and malignancy-specific benefits. Real-world data derived from integrated healthcare systems are underexplored in this context, and tight control of confounding using validated tools such as the Elixhauser Comorbidity Index (ECI) is rarely employed in pre-existing studies. Herein, we offer a highly refined description of the impact of BMI on cancer-specific outcomes in a large patient cohort using a targeted ECI model. Methods: 153,270 patient records were abstracted from an integrated healthcare system from 2010-2018, including descriptors of age, race/ethnicity, income, disease stage, and treatment regimens. Standardized derivation of BMI within one year prior to diagnosis was performed and categorized by World Health Organization definitions. A custom ECI was designed to exclude ICD codes for obesity to effectively isolate the impact of BMI on downstream analyses. Kaplan-Meier survival estimates of OS and CSS were constructed, and univariate and multivariate Cox proportional hazard (PH) modeling was performed to determine hazard ratios (HR). Results: When normalized to healthy BMI in multivariate modeling, we found a favorable dose-dependent relationship of overweight and obese BMI on OS and CSS independent of age, race/ethnicity, income, modified ECI score, cancer stage, or therapeutic regimen (Table). This advantage to OS and CSS was not as strongly reflected in severely obese patients, though still significant compared to healthy BMI counterparts. Being underweight was a negative prognostic indicator for OS and CSS in all patients compared to a healthy BMI. Conclusions: We demonstrate a beneficial impact of increased BMI on OS and CSS in a large cohort of patients treated in an integrated healthcare model independent of tightly controlled confounding. Specifically, we highlight the utility of isolating obesity using a targeted ECI model for comorbidity adjustment, which may provide a more robust measure of the influence of BMI on survival. Our findings raise questions regarding attention to adequate control of confounding in prior studies, as well as the potential benefit of removing variation in healthcare delivery in understanding these outcomes. Adjusted HR of OS and CSS stratified by BMI category (*95% Confidence Interval). BMI Category Adjusted OS Adjusted CSS Normal Weight(18.5 – 24.9) 1 (reference) 1 (reference) Underweight(&lt; 18.5) 1.35 (1.25 – 1.45*) 1.28 (1.15 – 1.41) Overweight(25 – 29.9) 0.83 (0.81 – 0.85) 0.84 (0.81 – 0.87) Obese(30 – 39.9) 0.81 (0.79 – 0.83) 0.81 (0.78 – 0.84) Severe Obesity(≥ 40) 0.86 (0.82 – 0.90) 0.82 (0.77 – 0.87) Unknown 1.33 (1.24 – 1.43) 1.19 (1.07 – 1.31)

Clinical presentation, outcome and prognostic factors in dogs with immune-mediated haemolytic anaemia: a retrospective single-centre study of 104 cases in Ireland (2002–2020)

BackgroundImmune-mediated haemolytic anaemia (IMHA) has a high mortality rate within the first weeks to months of diagnosis. Identifying dogs at increased risk of death may help guide decision-making for owners and veterinarians. Prior studies have identified several but inconsistent prognostic factors. The objectives of the study were to describe the clinical presentation and outcome of canine immune-mediated haemolytic anaemia in Ireland and to assess for independent factors associated with survival including long-term survival. Medical records from a single centre were reviewed between 2002 and 2020 to identify dogs with immune-mediated haemolytic anaemia using the American College of Veterinary Internal Medicine (ACVIM) consensus statement algorithm. Survival analysis was performed using univariable Cox proportional hazards regression models with Breslow method for ties to identify prognostic factors.ResultsOne hundred and four cases were included. The diagnosis of immune-mediated haemolytic anaemia was classified as definitive, supportive and suspicious in 42 (40%), 50 (48%), and 12 dogs (12%) respectively. Twenty-two dogs (21%) were diagnosed with associative IMHA and 82 dogs were diagnosed with non-associative IMHA (79%). 65% of the cases received more than one immunosuppressive medication during the course of treatment. The mortality rate at one and three months was 16% (95% confidence interval [CI] 9–26) and 31% (95% CI 21–43) respectively. Excluding dogs that died within three months, the median survival time was 2664 days. The relapse rate during the follow-up period was 7%. Survival did not improve over the course of the study period. Thrombocytopenia and hyperbilirubinaemia were identified as negative prognostic indicators (Hazard ratio 2.2 and 2.5, 95% CI 1.1–4.1 and 1.1–5.6, respectively).ConclusionsExcluding dogs that died within three months, the outcome was good in dogs with non-associative immune-mediated haemolytic anaemia in Ireland. The relapse rate was low regardless of the presence of associative causes. Thrombocytopenia and hyperbilirubinaemia were the only independent negative prognostic factors. The one-month and three-month mortality rates were similar compared to prior studies and survival did not improve over time during the study period: the mortality rate of canine immune-mediated haemolytic anaemia remains high in the acute phase.

The Spaghetti Sign: Nerve Root Redundancy in Lumbar Spinal Stenosis

Redundant nerve roots (RNRs) seen in conjunction with lumbar spinal stenosis (LSS) are well-described radiographic findings. Several studies suggest their presence may be a negative prognostic indicator of postoperative outcome. Our hypothesis was that severe RNR (informally known as the spaghetti sign [SS]) can serve as a reliable marker of LSS that would benefit from surgical decompression. We sought to evaluate a grading scale for RNR, characterize the association with stenosis, and investigate the clinical implications of RNR. We conducted a retrospective chart review of 72 patients who underwent lumbar spine surgery from 2016 to 2018 at 1 institution. Preoperative T2 magnetic resonance imaging scans were graded by 3 reviewers for severity of stenosis (0-4), severity of RNR (0-3), and rostral versus caudal RNR. SS was defined as RNR score ≥2 (clear-cut or marked nerve root irregularity). Preoperative and postoperative Oswestry Disability Index scores were analyzed by stenosis and RNR severity. Seventy-one (98%) patients had severe stenosis (score ≥3) and 25 (35%) had a SS. SS was 100% specific for high-grade stenosis. If patients had a SS, it was more likely rostral (P=0.02). Postoperative Oswestry Disability Index scores improved significantly, but there were no differences related to RNR score, presence of SS, or stenosis severity. The study demonstrated that there is a significant association between SS and severe LSS and that presence of RNR is not a negative prognostic indicator for postoperative outcomes.

Assessment of right ventricular function in healthy Great Danes and in Great Danes with dilated cardiomyopathy

IntroductionRight ventricular (RV) dysfunction is a significant negative prognostic indicator in human dilated cardiomyopathy (DCM). Many RV indices are weight-dependent, and there is a lack of reference values for the right heart in giant breed dogs (over 50 kg), including Great Danes (GDs). This study aimed to compare indices of RV function in echocardiographically normal GDs, those with preclinical DCM (PC-DCM), and those with DCM and congestive heart failure (DCM-CHF). AnimalsA total of 116 client-owned adult GDs: 74 normal, 31 with PC-DCM, and 11 with DCM-CHF. MethodsA retrospective, single-center cohort study assessed RV function using free-wall RV longitudinal strain (RVLS), strain rate, fractional area change (FAC), tricuspid annular plane systolic excursion (TAPSE), and pulsed-wave tissue Doppler imaging-derived systolic myocardial velocity of the lateral tricuspid annulus (TDI S’). Relationships between DCM status and RV function indices were analyzed. ResultsRV function, measured by TAPSE (P=0.001), FAC (P<0.001), and TDI S’ (P<0.001), decreased in dogs with PC-DCM and DCM-CHF compared to healthy dogs, with FAC being lower in DCM-CHF compared to PC-DCM (P=0.048). RVLS impairment was more significant in the DCM-CHF group than in the PC-DCM group (P=0.048). RVLS had the highest area under the curve (0.899) for differentiating between normal and DCM-CHF dogs. ConclusionAs DCM progresses, echocardiographic variables of RV function, including TAPSE, FAC, TDI S’, RVLS, and strain rate, worsen, indicating impaired RV systolic function in GDs affected by DCM.

CD38 regulates chronic lymphocytic leukemia proliferation via CD45 phosphatase activity

Chronic Lymphocytic Leukemia (CLL) growth is dependent on both B cell receptor (BCR) signaling and signals from microenvironmental T helper (Th) cells. We previously described a mechanism where Th cells enhance BCR signaling and proliferation through CD45 phosphatase activity regulation via Galectin-1 and CD43. The CLL negative prognostic indicator CD38 is linked to BCR signaling and proliferation, with its expression induced by Th cells. Here, we explore a link between CD38 and CD45 phosphatase activity regulation using patient-derived material in a Th-CLL cell co-culture model. Results demonstrate CD43 and Galectin-1 are co-expressed with CD38, defining proliferative CLL cells with augmented CD45 activity. CD38 enzymatic and receptor inhibition regulated CD43 and Galectin-1 expression, CD45 activityhi populations, and CLL proliferation, while leaving Th cells largely unaffected. Mechanistically, CD38- or LGALS1 (Galectin-1)-deficient malignant B cell lines further confirmed CD38-mediated regulation of CD45 activity and BCR signaling through CD43 expression and Galectin-1 surface binding, while Galectin-1 contributed to CD43/CD45 colocalization. Together, this highlights CD38 as an important regulator of CD45 activity via CD43 and Galectin-1, in turn acting as a positive modulator of CLL proliferation. Ultimately, the CD38/CD45 molecular hub could be an important therapeutic target in CLL.

Profiling of Tumor-Infiltrating Immune Cells and Their Impact on Survival in Glioblastoma Patients Undergoing Immunotherapy with Dendritic Cells.

Glioblastomas (GBM) are the most common primary malignant brain tumors, comprising 2% of all cancers in adults. Their location and cellular and molecular heterogeneity, along with their highly infiltrative nature, make their treatment challenging. Recently, our research group reported promising results from a prospective phase II clinical trial involving allogeneic vaccination with dendritic cells (DCs). To date, six out of the thirty-seven reported cases remain alive without tumor recurrence. In this study, we focused on the characterization of infiltrating immune cells observed at the time of surgical resection. An analytical model employing a neural network-based predictive algorithm was used to ascertain the potential prognostic implications of immunological variables on patients' overall survival. Counterintuitively, immune phenotyping of tumor-associated macrophages (TAMs) has revealed the extracellular marker PD-L1 to be a positive predictor of overall survival. In contrast, the elevated expression of CD86 within this cellular subset emerged as a negative prognostic indicator. Fundamentally, the neural network algorithm outlined here allows a prediction of the responsiveness of patients undergoing dendritic cell vaccination in terms of overall survival based on clinical parameters and the profile of infiltrated TAMs observed at the time of tumor excision.

Outcome in paraplegic dogs with or without pain perception due to thoracolumbar fibrocartilaginous embolic myelopathy or acute non-compressive nucleus pulposus extrusion.

Fibrocartilaginous embolic myelopathy (FCEM) and acute non-compressive nucleus pulposus extrusion (ANNPE) are common causes of acute spinal cord injury in dogs. Outcome among paraplegic deep pain positive (DPP) and deep pain negative (DPN) dogs with either condition and factors influencing recovery have not been clearly established. Dogs with thoracolumbar FCEM or ANNPE resulting in paraplegia presenting to university hospitals between 2012 and 2022 were retrospectively included. Diagnosis of FCEM or ANNPE was based on clinical and magnetic resonance imaging findings. Outcome was defined as successful (recovery of independent ambulation) or unsuccessful (non-ambulatory ≥3 months following diagnosis or at the time of death/euthanasia). Logistic regression analysis was performed to investigate associations between clinical or imaging variables and outcome. Thirty-one dogs were included. In total, 14 dogs were initially paraplegic DPP (8 FCEM, 6 ANNPE) and 17 dogs were paraplegic DPN (11 FCEM, 6 ANNPE). Outcome was available for 26 dogs (14 DPP, 12 DPN) with a median follow-up time of 182 days (range 0-2,311) including 2 dogs euthanized at the time of diagnosis; 1 of 12 DPN dogs (8.3%) regained independent ambulation, whereas 9 of 14 DPP dogs (64.3%) regained independent ambulation. DPN dogs had a significantly higher risk of not regaining independent ambulation compared with DPP dogs (OR: 47.40, 95% CI: 2.09-1073.99). No other variables were associated with outcome. While recovery of ambulation was possible, these results confirm that the absence of pain perception is a useful negative prognostic indicator in dogs with severe thoracolumbar FCEM or ANNPE.

Clinical characteristics, treatment patterns, and outcomes of hospitalized patients with acute heart failure in central Ethiopia: a retrospective observational study

BackgroundAcute heart failure is the rapid onset of new or worsening symptoms and signs of heart failure. Despite the increasing burden of heart failure in developing countries like Ethiopia, there is a paucity of comprehensive data regarding the clinical characteristics, treatment patterns, and outcomes of acute heart failure, especially in the selected study area. Therefore, this study aimed to assess the clinical characteristics, treatment patterns, and outcomes of hospitalized patients with acute heart failure at Yekatit 12 Hospital Medical College, Addis Ababa, Ethiopia.MethodsThis is a retrospective cross-sectional study of 303 acute heart failure patients who were admitted to the medical wards and intensive care unit of Yekatit 12 Hospital Medical College, Addis Ababa, central Ethiopia, from July 1, 2022, to July 1, 2023. A pretested data abstraction format was used for data extraction from electronic medical records, and SPSS version 26 was used for data analysis. Descriptive analysis was used to summarize sociodemographic data, clinical characteristics, treatment patterns, and outcomes of acute heart failure. Bivariate and multivariate logistic regression models were fitted to identify factors associated with in-hospital mortality. The odds ratio (OR) with the corresponding 95% confidence interval (CI) was calculated to show the strength of the association.ResultsOf the 303 patients, 51.5% were females, and the mean age was 56.7 years. The most frequent symptom and sign were dyspnea (98.7%) and peripheral edema (79%), respectively. The commonest underlying cause and precipitating factor of acute heart failure were cor pulmonale (22.8%) and pneumonia (35.3%), respectively. The commonest anti-remodeling medications prescribed on discharge were beta-blockers (47.9%), followed by mineralocorticoid receptor antagonists (42.8%) and angiotensin-converting enzyme inhibitors/angiotensin receptor blockers (38.6%), and the least prescribed were sodium-glucose cotransporter 2 inhibitors (8.3%). The in-hospital mortality rate was 8.6%, and the median length of hospital stay was 9 days. Based on the multivariate logistic regression analysis, the most important predictors of in-hospital mortality were systolic blood pressure (SBP) < 115 mmHg (adjusted odds ratio [AOR] = 6.28; 95% CI: 1.99, 19.78), chloride level < 96 mg/dL (AOR = 4.88; 95% CI: 1.30, 18.33), blood urea nitrogen (BUN) > 20 mg/dl (AOR = 5.48; 95% CI: 1.47, 20.49), and presence of dyslipidemia (AOR = 3.73, 95% CI: 1.15, 12.07).ConclusionsThis study has shown that systolic blood pressure (SBP) < 115 mmHg, blood urea nitrogen (BUN) > 20 mg/dL, chloride (Cl) level < 96 mg/dL, and the presence of dyslipidemia were statistically significant factors associated with in-hospital mortality among patients with acute heart failure. Hence, healthcare providers should stratify patients with acute heart failure upon admission based on their risk of in-hospital mortality and address those potential negative prognostic indicators accordingly.

Clinical Characteristics, Prognostic Factors and Outcomes of Paediatric Patients with Haemophagocytic Lymphohistiocytosis

Objective: Haemophagocytic lymphohistiocytosis (HLH) is a multi-system autoimmune disorder. The objective of this study was to find out the clinical characteristics and prognosis of paediatric patients with Haemophagocytic Lymphohistiocytosis. Study Design and setting: Cross sectional study conducted at Pak Emirates Military Hospital, Rawalpindi from July 2021 to June 2023. Methodology: Children with diagnosis of HLH were assessed by including patients who were aged = 13 years during hospitalization. All the patients who had not been diagnosed by using the HLH-2004 criteria were disqualified. Relevant findings were noted by evaluating records pertinent to physical examination, radiology and laboratory markers. Prognosis was assessed by determining the underlying clinical aetiology and whether patient-related factors modulated the overall life expectancy. Results: A total of 32 patient records were evaluated. Mean age at diagnosis was 44.3 ± 39.1 months (Range: 1-132 months) with majority being males [n=23 (71.9%)]. The common clinical characteristics included fever [n=29 (90.6%)], lymphadenopathy [n=27 (84.4%)], splenomegaly [n=23 (71.9%)] and hepatomegaly [n=23 (71.9%)]. Serum ferritin, bilirubin, ALT, AST, and LDH were also raised. All patients were followed for a mean period of 12 months and 18 (56.3%) children failed to survive. Negative prognostic indicators included severe anaemia (p=0.001), neutropenia (p=0.007), thrombocytopenia (p=0.033), and hyperferritinemia (p&lt;0.001). Elevation of liver enzymes (ALT: p&lt;0.001; AST: p=0.031), serum bilirubin (p=0.037), and LDH (p&lt;0.001) also indicated worse disease prognosis. Conclusion: HLH in childhood is a potentially life-threatening disease and carries a significant association with deranged liver function

Role of high-volume plasmapheresis in the management of paediatric acute liver failure.

Paediatric acute liver failure (PALF) is a life-threatening disease. Management aims to support hepatic regeneration or to bridge to liver transplantation. High-volume plasmapheresis (HVP) removes protein-bound substances, alleviates inflammation, and improves survival in adult acute liver failure. However, experience with HVP in PALF is limited. Aim of this study is to report on feasibility, safety, efficacy and outcomes of HVP in PALF. Retrospective observational study in children with PALF. HVP was performed upon identification of negative prognostic indicators, in toxic aetiology or multiorgan failure (MOF). Exchanged volume with fresh-frozen plasma corresponded to 1.5-2.0 times the patient's estimated plasma volume. One daily cycle was performed until the patient met criteria for discontinuation, that is, liver regeneration, liver transplantation, or death. Twenty-two children with PALF (body weight 2.5-106 kg) received 1-7 HVP cycles. No bleeding or procedure-related mortality occurred. Alkalosis, hypothermia and reduction in platelets were observed. Haemolysis led to HVP termination in one infant. Seven children (32%) survived with their native livers, 13 patients (59%) underwent liver transplantation. Two infants died due to MOF. Overall survival was 86%. International normalization ratio (INR), alanine aminotransaminases (ALT), bilirubin and inotropic support were reduced significantly (p < 0.05) after the first HVP-cycle (median): INR 2.85 versus 1.5; ALT 1280 versus 434 U/L; bilirubin 12.7 versus 6.7 mg/dL; norepinephrine dosage 0.083 versus 0.009 µg/kg/min. Median soluble-interleukin-2-receptor dropped significantly following HVP (n = 7): 2407 versus 950 U/mL (p < 0.02). HVP in PALF is feasible, safe, improves markers of liver failure and inflammation and is associated with lowering inotropic support. Prospective and controlled studies are required to confirm efficacy of HVP in PALF.

Factors associated with development of post-operative reflux in horses with large colon volvulus and association with complications and outcomes.

Post-operative reflux (POR) is rare following large colon volvulus (LCV) but does occur despite the absence of a small intestinal lesion. The prevalence, risk factors and association with survival of POR after LCV are currently unknown. To determine the prevalence of POR in horses with an LCV and its association with survival. A further objective was to assess factors which may predict POR. The hypothesis was that horses with POR following LCV surgery have a worse outcome for survival compared to those without POR. A retrospective cross-sectional study. Clinical data of client owned horses which underwent colic surgery at a single UK referral hospital between 2008 and 2021, where LCV was the primary finding, were retrieved from hospital records. Statistical analyses included chi-squared, t-tests and odds ratios (ORs). Horses with concurrent lesions, and those that did not survive past anaesthetic recovery, were excluded from analysis. POR was defined as ≥2 L of gastric reflux on at least one occasion. A total of 128 horses were included in the study, 23 of which had POR (18%). Overall survival to hospital discharge was 86%, 95% in the non-POR and 44% in the POR group. Horses with POR were less likely to survive to discharge than those without (OR = 26, 95% confidence interval [CI] [7.68-88.0], p < 0.001), and less likely to be alive 3 years after surgery (OR = 13.4, 95% CI [2.78-64.8], p < 0.001). Due to the retrospective study design, full data sets were not available for every case because clinical records were incomplete or, at that time, certain tests were not performed or clinical variables were not measured. POR in LCV cases is a negative prognostic indicator for survival.

Next-Generation Sequencing in Sporadic Medullary Thyroid Cancer Patients: Mutation Profile and Disease Aggressiveness.

Next-generation sequencing (NGS) analysis of sporadic medullary thyroid carcinoma (sMTC) has led to increased detection of somatic mutations, including RET M918T, which has been considered a negative prognostic indicator. This study aimed to determine the association between clinicopathologic behavior and somatic mutation identified on clinically motivated NGS. In this retrospective cohort study, patients with sMTC who underwent NGS to identify somatic mutations for treatment planning were identified. Clinicopathologic factors, time to distant metastatic disease (DMD), disease-specific survival (DSS), and overall survival (OS) were compared between somatic mutations. Somatic mutations were identified in 191 sMTC tumors, including RET M918T (53.4%), other RET codons (10.5%), RAS (18.3%), somatic RET indels (8.9%), and RET/RAS wild-type (WT) status (8.9%). The median age at diagnosis was 50 years (range, 11-83); 46.1% were female. When comparing patients with RET M918T, RET-Other, and RET WT (which included RAS and RET/RAS WT), there were no differences in sex, TNM category, systemic therapy use, time to DMD, DSS, or OS. On multivariate analysis, older age at diagnosis (HR 1.05, P < .001; HR 1.06, P< .001) and M1 stage at diagnosis (HR 3.17, P = .001; HR 2.98, P = .001) were associated with decreased DSS and OS, respectively, but mutation cohort was not. When comparing RET M918T to RET indels there was no significant difference in time to DMD, DSS, or OS between the groups. Somatic RET mutations do not portend compromised DSS or OS in a cohort of sMTC patients who underwent clinically motivated NGS.

Abstract 4278: Role of WNK1 in extracellular matrix modification in breast cancer

Abstract Introduction: Breast cancer (BCa) is the most common cancer and the second leading cause of cancer related deaths in women in the United States. Metastatic dissemination of cancer cells remains the leading cause of mortality in BCa. Therefore, determination of the mechanisms that promote metastasis is important for development of new therapeutic strategies. The extracellular matrix (ECM), which is the main non-cellular component of the tumor microenvironment (TME), plays an important role in BCa progression and metastasis. The cancer-associated fibroblasts (CAFs) are the principal cell types that together with BCa cells drive the modification of ECM during cancer development. Lysine deficient receptor protein kinase 1 (WNK1), a widely expressed serine threonine kinase that regulates blood pressure and electrolyte balance in the body, and is highly overexpressed in BCa microenvironment. The aim of the present study was to investigate the role of WNK1 in modification of ECM in BCa. Methods: The clinical relevance of WNK1 overexpression in BCa was determined by immunohistochemically staining human BCa sections, analyzing data from publicly available databases and creating orthotopic mouse models of BCa using 4T1 mouse BCa cells followed by treatment of the tumor bearing mice with WNK1 inhibitors, WNK-IN-11 and WNK463 (10 mg/kg , oral for 10 days). In vitro assays were conducted using 4T1 cells and mouse embryonic fibroblasts (MEFs) to determine the regulatory role of WNK1 on cancer cells and fibroblasts. Results and Conclusion: Our results indicated that WNK1 is strongly expressed in BCa cells and fibroblasts, the two most abundant cell types in the BCa microenvironment. Furthermore, WNK1 expression strongly correlated with expressions of matrix regulatory and cross-linking genes. Our results demonstrated that WNK1 inhibitors significantly reduced orthotopic 4T1 growth and metastasis. WNK1 inhibition also significantly inhibited collagen deposition in tumor tissues, which is a negative prognostic indicator in BCa. In vitro proliferation and migration assays using 4T1 cells and MEFs identified that WNK1 inhibitors (1-100nM) can significantly inhibit the proliferation and migration of 4T1 cells and MEFs. Results from co-culture studies with 4T1 cells and MEFs further indicated that WNK1 inhibition in MEFs could significantly reduce the invasive properties of 4T1 cells. Taken together, our results suggest that blocking the actions of WNK1 in BCa may be a promising therapeutic approach. Citation Format: Prabhat Suman, Sooraj Kakkat, Suhas S. Patil, Veronica Ramirez, Elba A. Turbat-Herrera, Seema Singh, Chandrani Sarkar, Debanjan Chakroborty. Role of WNK1 in extracellular matrix modification in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4278.

Recovery rates and long‐term olfactory dysfunction following COVID‐19 infection

AbstractObjectivesOlfactory dysfunction is one of the most recognized symptoms of COVID‐19, significantly impacting quality of life, particularly in cases where recovery is prolonged. This review aims to explore patterns of olfactory recovery post‐COVID‐19 infection, with particular focus on delayed recovery.Data SourcesPublished literature in the English language, including senior author's own work, online and social media platforms, and patients' anecdotal reports.MethodA comprehensive review of the literature was undertaken by the authors with guidance from the senior author with expertise in the field of olfaction.ResultsBased on self‐report, an estimated 95% of patients recover their olfactory function within 6 months post‐COVID‐19 infection. However, psychophysical testing detects higher rates of persistent olfactory dysfunction. Recovery has been found to continue for at least 2 years postinfection; negative prognostic indicators include severe olfactory loss in the acute phase, female sex, and older age. Variability in quantitative and qualitative disturbance in prolonged cases likely reflects both peripheral and central pathophysiological mechanisms. Limitations of many of the reviewed studies reflect lack of psychophysical testing and baseline olfactory assessment.ConclusionsPost‐COVID‐19 olfactory dysfunction remains a significant health and psychosocial burden. Emerging evidence is improving awareness and knowledge among clinicians to better support patients through their olfactory rehabilitation, with hope of recovery after several months or years. Further research is needed to better understand the underlying pathogenesis of delayed recovery, identify at risk individuals earlier in the disease course, and develop therapeutic targets.

Effects of Acipenserid herpesvirus 2 on the outcome of a Streptococcus iniae co-infection in white sturgeon (Acipenser transmontanus)

Acipenserid herpesvirus 2 (AciHV-2) is a large double-stranded DNA virus in the family Alloherpesviridae that causes catastrophic outbreaks in young naive white sturgeon (Acipenser transmontanus) populations, with mortalities of up to 80%. Survivors of these infections are suspected to remain latently infected. The gram-positive zoonotic bacterium Streptococcus iniae is another important sturgeon pathogen that causes severe myositis and up to 50% mortality during natural outbreaks. Throughout the last decade, co-infections of AciHV-2 and S. iniae have been reported in cultured white sturgeon in California resulting in severe presentations of piscine streptococcosis. This phenomenon of herpesvirus and streptococcus co-infection appears to span multiple taxa since in humans, it is recognized that a Human herpesvirus 3 infection (VZV) is a negative prognostic indicator for pediatric Invasive Group A Streptococcal infections (IGASI). While a decrease in humoral immunity caused by VZV has been hypothesized as a potentially important factor in IGASI cases, no natural animal model exists to study this process. Moreover, no studies have investigated these reported co-infections in white sturgeon. Therefore, the goal of this study was to investigate the effects of a recent AciHV-2 infection on the outcome of a subsequent S. iniae challenge in white sturgeon fingerlings. When fish were infected with 108 colony forming units (CFU) of S. iniae intramuscularly (IM), a statistically significant decrease in survival of 41% was detected in the co-infection group compared to the S. iniae group (p-value &amp;lt; 0.001). This difference was not observed when fish were infected with 106 CFU of S. iniae IM. At this lower infection dose, however, a statistically significant downregulation of tnfα was observed in the spleen of fish in the co-infection group compared to the S. iniae group (p-value = 0.0098). Analysis of serum from survivors revealed a statistically significant reduction in anti-S. iniae serum IgM and serum serotransferrin in fish from the co-infection group compared to the S. iniae group (p-value = 0.0134 and p-value = 0.0183, respectively). Further studies are indicated to determine what interactions lead to the decreased production of pathogen-specific IgM, serotransferrin, and TNFα in the host.

Predicting Time-to-Healing from a Digital Wound Image: A Hybrid Neural Network and Decision Tree Approach Improves Performance

Despite the societal burden of chronic wounds and despite advances in image processing, automated image-based prediction of wound prognosis is not yet in routine clinical practice. While specific tissue types are known to be positive or negative prognostic indicators, image-based wound healing prediction systems that have been demonstrated to date do not (1) use information about the proportions of tissue types within the wound and (2) predict time-to-healing (most predict categorical clinical labels). In this work, we analyzed a unique dataset of time-series images of healing wounds from a controlled study in dogs, as well as human wound images that are annotated for the tissue type composition. In the context of a hybrid-learning approach (neural network segmentation and decision tree regression) for the image-based prediction of time-to-healing, we tested whether explicitly incorporating tissue type-derived features into the model would improve the accuracy for time-to-healing prediction versus not including such features. We tested four deep convolutional encoder–decoder neural network models for wound image segmentation and identified, in the context of both original wound images and an augmented wound image-set, that a SegNet-type network trained on an augmented image set has best segmentation performance. Furthermore, using three different regression algorithms, we evaluated models for predicting wound time-to-healing using features extracted from the four best-performing segmentation models. We found that XGBoost regression using features that are (i) extracted from a SegNet-type network and (ii) reduced using principal components analysis performed the best for time-to-healing prediction. We demonstrated that a neural network model can classify the regions of a wound image as one of four tissue types, and demonstrated that adding features derived from the superpixel classifier improves the performance for healing-time prediction.

Clinical bleeding diathesis, laboratory haemostatic aberrations and survival in dogs infected with Angiostrongylus vasorum: 180 cases (2005-2019).

Bleeding diathesis is a complication in dogs infected with Angiostrongylus vasorum. This retrospective study investigated clinical and laboratory haemostatic differences in A. vasorum-positive dogs with and without signs of bleeding and impact of bleeding on survival. Demographics, type of clinical bleeding, haematocrit and a range of haemostatic tests, including thromboelastography and derived velocity curves were retrospectively registered from A. vasorum-positive dogs. All parameters were compared between dogs with and without signs of bleeding using univariable analyses. Binomial and multinomial regression models were applied to examine specific indicators in the bleeding dogs. P-values were false discovery rate adjusted, and adjusted P<0.05 was considered significant. One hundred and eighty dogs entered the study, including 65 dogs (36.1%) presenting with bleeding diathesis. Different types of cutaneous and mucosal bleeding were the most common clinical findings. Twenty dogs presented with neurological signs associated with intracranial and intra-spinal bleeding. One hundred and thirty-seven dogs had haematological and/or haemostatic laboratory analyses performed. Haematocrit, platelet count, thromboelastographic angle, maximum amplitude, global clot strength, maximum rate of thrombin generation and total thrombin generation were decreased, while prothrombin time was prolonged in bleeding dogs. Survival rate of bleeding dogs was lower at hospital discharge (76.9%) and 1 month after diagnosis (66.0%) than in dogs without signs of bleeding (94.8% and 90.1% at discharge and at 1 month, respectively). Several haemostatic aberrations were detected in A. vasorum-positive dogs with bleeding diathesis. Bleeding was identified as an important negative prognostic indicator in A. vasorum-positive dogs.