Indirect Marker Research Articles

Renal Afferent Nerve Activity and Association with Neurohumoral Dysregulation in a Model of Autosomal Recessive Polycystic Kidney Disease

Polycystic kidney disease (PKD) is a rare, genetic kidney disease characterized by cyst growth that leads to kidney failure. Renal nerves play an important role in the development and maintenance of hypertension and other renal diseases. Afferent (sensory) nerves convey information to the central nervous system in response to intrarenal stimuli. Our laboratory demonstrated that afferent renal nerve activity (ARNA) is elevated in 10-week-old PCK rats compared to non-cystic, age-matched Sprague-Dawley (SD) controls. Furthermore, ARNA positively correlated with cystic index (CI; a measure of cyst growth). To further elucidate the relationship between renal nerve activity, age, and cyst progression, we aimed to quantify both ARNA and renal sympathetic nerve activity (RSNA) in PCK and SD rats. We hypothesized that ARNA and RSNA would be greater in PCK than SD. Furthermore, arginine vasopressin (AVP) is elevated in PKD and increases cyst growth. There is evidence that activation of the afferent renal nerves can contribute to secretion of AVP. Thus, we propose that elevated ARNA increases AVP secretion, thereby increasing cyst growth. We also hypothesized that ARNA would positively correlate with age, CI, and copeptin (an indirect marker of AVP) in young PCK rats. To test our first hypothesis, we conducted direct renal nerve recordings in 10- to 12-week-old PCK (n=16;11M/5F) and 10-week-old SD (n=9;7M/2F) rats. For our second hypothesis, we conducted nerve recordings in 4- to 10-week-old PCK rats to evaluate the correlation of nerve activity with age, cystic index, and copeptin in animals ages 4- to 12-weeks-old (n=28;21M/7F). Multiunit nerve recording was performed under inactin anesthesia. As we previously published, a renal nerve bundle is isolated, placed on an iridium bipolar electrode, and encased in silicone. Resting RSNA was recorded following 10-minute stabilization period, then the nerve bundle was severed proximal to the electrode to isolate ARNA. ARNA recordings were normalized to maximal ARNA evoked by intrarenal 50 μM capsaicin. Comparisons between PCK and SD were analyzed by Student’s t-test (p<.05). Linear regression analyses were used to assess variable correlation (p<.05). Data presented as mean ± SEM. In contrast to our first hypothesis, RSNA did not differ between 10- to 12-week-old SD and PCK rats, whether measured via burst frequency (SD 6.92 ± 0.82; PCK *6.97 ± 0.34 Hz) or bursts/heart beat (SD 1.36±0.16; PCK 1.28 ± 0.06). ARNA was calculated as a percentage of maximal ARNA using either integrated ARNA (ARNAInt) or spikes/s (ARNASpikes). Baseline ARNAInt was greater in PCK rats (29.14±3.35%) than SD (13.46±2.08%, n=4), whereas PCK ARNASpikes trended (p=.06) higher than SD controls (SD 5.57±2.21; PCK 13.90±2.21%). In correlative analysis, RSNA trended (p=.05) to correlate with age in the PCK rat. However, there was no relationship between RSNA and (1) CI (p=0.29) or (2) copeptin (p=0.15). Similarly, there was no correlation detected between ARNA and age (ARNAInt: p=0.41; ARNASpikes: p=0.16) or serum copeptin (ARNAInt: p=0.96; ARNASpikes: p=0.20). ARNAInt trended (p=.07) to correlate with CI, but ARNASpikes did not correlate with CI (p=0.48). Overall, these data partially support our hypotheses. RSNA did not differ between SD and PCK rats, but ARNAInt was increased in PCK rats vs. SD. We failed to detect a correlation between either ARNA or RSNA and age, CI, or copeptin in PCK rats though, ARNAInt trended to correlate with CI. As ARNA does not appear to be correlated with serum copeptin, further studies are required to uncover the relationship between afferent renal nerves and cystogenesis in this model of PKD. PKD Foundation Research Award 1022534 (CB) NIDDK/PKD RRC Pilot and Feasibility Award U24DK126110 (CB) AHA 21PRE0000216068 (MG). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The Effect of Continuous Positive Airway Pressure (CPAP) Therapy on Serum Caspase-3 Level in Patients with Obstructive Sleep Apnea (OSA).

Intermittent hypoxemia has an important role in the physiopathogenesis of obstructive sleep apnea (OSA) complications. Increased apoptosis due to intermittent hypoxemia may be an important clinical entity in OSA. In this study, we aimed to evaluate caspase-3 enzyme level, which is an indirect marker of increased apoptosis in patients with OSA and to evaluate the effect of OSA treatment with continuous positive airway pressure on caspase-3 enzyme level. This study included 141 consecutive patients admitted to the sleep-disordered breathing laboratory within 6 months. Caspase-3 was measured in routine blood samples obtained on the morning of polysomnography (PSG) performed at night. The compliance of the patients to CPAP treatment was evaluated and caspase-3 levels were checked again after treatment. A total of 141 patients, 39 females (27,7%) and 102 males (72,3%) were included in the study. The mean age of the patients was 49 ± 12 years (min-17, max-77). According to PSG results, OSA was detected in 95.7% (135/141) of the cases. Mild OSA was 35 (24.8%), moderate OSA 39 (27.7%) and severe OSA 61 (43.3%) cases. Median caspase-3 enzyme levels were similar in men and women in the study group. There was no statistically significant difference in hemogram parameters and caspase-3 enzyme levels between the groups divided according to the presence and severity of OSA. It was determined that caspase-3 enzyme level did not change significantly after 3 months of CPAP treatment in OSA compared to pretreatment. Caspase-3 was found to have a negative correlation with both the percentage of daily use of CPAP therapy and the percentage of CPAP device use for more than 1 h per night. It was found that the control caspase-3 level decreased statistically significantly as the percentage of daily use of CPAP therapy increased (r = -0.397, p = 0.030). It was found that the control caspase-3 level decreased statistically significantly as the percentage of CPAP therapy use for more than 1 h per night increased (r = -0.411, p = 0.024). The results of this study did not reveal a relationship between the severity of OSA and caspase-3 levels. However, blood caspase-3 levels decreased as treatment compliance increased, suggesting that CPAP treatment may correct increased apoptosis in OSA. There is a need for more comprehensive studies on this issue.

Time-of-day effects on ex vivo muscle contractility following short-term satellite cell ablation.

Muscle isometric torque fluctuates according to time-of-day with such variation owed to the influence of circadian molecular clock genes. Satellite cells (SCs), the muscle stem cell population, also express molecular clock genes with several contractile-related genes oscillating in a diurnal pattern. Currently, limited evidence exists regarding the relationship between SCs and contractility, although long-term SC ablation alters muscle contractile function. Whether there are acute alterations in contractility following SC ablation and with respect to the time-of-day is unknown. We investigated whether short-term SC ablation affected contractile function at two times of day and whether any such alterations led to different extents of eccentric contraction-induced injury. Using an established mouse model to deplete SCs, we characterized muscle clock gene expression and ex vivo contractility at two times-of-day (morning: 0700 and afternoon: 1500). Morning-SC+ animals demonstrated ∼25%-30% reductions in tetanic/eccentric specific forces and, after eccentric injury, exhibited ∼30% less force-loss and ∼50% less dystrophinnegative fibers versus SC- counterparts; no differences were noted between Afternoon groups (Morning-SC+: -5.63 ± 0.61, Morning-SC-: -7.93 ± 0.61; N/cm2; P < 0.05) (Morning-SC+: 32 ± 2.1, Morning-SC-: 64 ± 10.2; dystrophinnegative fibers; P < 0.05). As Ca++ kinetics underpin force generation, we also evaluated caffeine-induced contracture force as an indirect marker of Ca++ availability and found similar force reductions in Morning-SC+ vs. SC- mice. We conclude that force production is reduced in the presence of SCs in the morning but not in the afternoon, suggesting that SCs may have a time-of-day influence over contractile function.NEW & NOTEWORTHY Muscle isometric torque fluctuates according to time-of-day with such variation owed to molecular clock regulation. Satellite cells (SCs) have recently demonstrated diurnal characteristics related to muscle physiology. In our work, force production was reduced in the presence versus absence of SCs in the morning but, not in the afternoon. Morning-SC+ animals, producing lower force, sustained lesser degrees of injury versus SC- counterparts. One potential mechanism underpinning lower forces produced appears to be lower calcium availability.

A Computed Tomography Study on the Prevalence of Lusorian Artery Among Hungarian Adults.

Introduction The aberrant right subclavian artery (ARSA), also called as lusorian artery (LA) is a developmental anomaly that exists in conjunction with a right non-recurrent laryngeal nerve (NRLN) in almost all cases. The average prevalence of such a vascular variation is estimated as 1%, although, studies have reported very different population means. Up to date, there is no available data on the frequency of this pattern in the Hungarian population. It can be treated as an indirect marker of a NRLN. Any preoperative information on the course of the inferior laryngeal nerves can help surgeons reduce the risk of an iatrogenic injury during thyroidectomies, especially in an environmentwhere access to intraoperative neuromonitoring is limited. Objectives The primary aims were to determine the prevalence of an ARSA, predict the existence of an NRLN in the Hungarian population, and provide demographic analysis. Methods A retrospective, computed tomography-based study was carried out. Demographic description and statistical analysis were provided where applicable. Detected anomalous vasculatures were visualized with 3D segmentation, and images were interpreted. Results The imaging database review identified three patients with ARSA out of 686 eligible recordings, resulting in a frequency of 0.437% in the study population. All three patients were female and had a retroesophageal LA. Two of them had a Kommerell's diverticulum. One patient had common carotid arteries with a single origin. Conclusions The frequency of an ARSA and a concomitant NRLN among Hungarians fits into the results of recent meta-analyses. Preoperative assessment of this anomaly may reduce vocal cord complication rates of thyroidectomies.

Functional analysis of postural spinal and pelvic parameters using static and dynamic spinometry

BackgroundSpinometry is a radiation-free method to three-dimensional spine imaging that provides additional information about the functional gait patterns related to the pelvis and lower extremities. This radiation-free technology uses the surface topography of the trunk to analyze surface asymmetry and identify bony landmarks, thereby aiding the assessment of spinal deformity and supporting long-term treatment regimes. Especially reliable dynamic spinometric data for spine and pelvis are necessary to evaluate the management of non-specific back pain. Research aimThis study aims to generate reliable dynamic spinometric data for spine and pelvis parameters that can serve as reference data for future studies and clinical practice. MethodsThis study assessed 366 subjects (185 females) under static and 360 subjects (181 females) under dynamic (walking on a treadmill at 3 km/h and 5 km/h) conditions. The DIERS Formetric 4Dmotion® system uses stripes of light to detect the surface topography of the spine and pelvis and identifies specific landmarks to analyze the spine during standing and walking. ResultsRelevant gender effects were calculated for lordotic angle (ηp2 = 0.22) and pelvic inclination (ηp2 = 0.26). Under static conditions, female subjects showed larger values for both parameters (lordotic angle: 41.6 ± 8.60°; pelvic inclination: 25.5 ± 7.49°). Regarding speed effects, three relevant changes were observed (sagittal imbalance: ηp2 = 0.74, kyphotic angle: ηp2 = 0.13, apical deviation: ηp2 = 0.11). The most considerable changes were observed between static condition and 3 km/h, especially for sagittal imbalance and lordotic angle. For these parameters, relevant effect sizes (d > 0.8) were calculated between static and 3 km/h for males and females. Concerning clinical vertebral parameters, only lordotic angle and pelvic inclination were correlated with each other (r = 0.722). ConclusionThis study generated a gender-specific reference database of asymptomatic individuals for static and dynamic spinometry. It demonstrated that the DIERS Formetric 4Dmotion® system could capture natural changes in static and dynamic situations and catalogue functional adaptations of spino-pelvic statics at different speeds. The lordotic angle is an indirect marker of pelvic inclination, allowing spinometry to identify individuals at risk even under dynamic conditions.

Obstrüktif Uyku Apnede (OUA) Ortalama Trombosit Hacmi (OTH) ve OUA'da Sürekli Pozitif Hava Yolu Basıncı (CPAP) Tedavisinin OTH Üzerine Etkisi

Aims: In this study, our objective was to evaluate mean platelet volume (MPV), an indirect marker of platelet activation, in patients with obstructive sleep apnea (OSA), and assess the effect of OSA treatment with continuous positive airway pressure (CPAP) on MPV&#x0D; Methods: In this study, records of consecutive patients who underwent polysomnographic evaluation for OSA symptoms in the Sleep Disorders Laboratory during a one-year period were reviewed retrospectively. Patients who had both complete blood count and MPV measurements were included in the study.&#x0D; Results: A total of 158 patients, including 51 females (32.3%) and 107 males (67.7%), were included in the study. The mean age of the patients was 51±13 (min-18, max-82) years. OSA was detected in 74.1% (117/158) of the patients. It was determined that as the severity of OSA increased, hemoglobin and hematocrit values increased significantly. There was no significant difference in platelet count according to the presence and severity of OSA. The MPV was significantly lower in severe OSA cases compared to those without OSA and mild OSA cases. A negative correlation was observed between MPV and the apnea-hypopnea index, desaturation index, and the amount of oxygen saturation below 90% during sleep. There was no significant difference in median erythrocyte and thrombocyte counts, hematocrit percentage and hemoglobin values before and after treatment in OSA patients who used CPAP therapy. However, a significant decrease in MPV was observed after OSA treatment compared to pre-treatment. (p=0.021).&#x0D; Conclusions: The results of the study do not support an increase in MPV and hence platelet activation in severe OSA patients compared with those without OSA. However, the results suggest that one month of CPAP treatment reduces MPV and thus platelet activation in severe OSA patients. Further controlled, prospective studies including treatment outcomes are needed on this subject.

Anti-Mullerian Hormone Assessment in Assisted Reproductive Technique Outcome and Natural Conception.

In recent years, the prevalence of infertility has increased, and appears to affect approximately one in six couples. Some of them must perform assisted reproductive techniques (ART) in order to achieve pregnancy. As a result, growing interest has arisen about predictive factors of pregnancy and live birth with and without ART. Anti-Mullerian hormone (AMH) is a glycoprotein discovered in the 1950s in male embryonic sexual differentiation. Later, in 1984, its role in folliculogenesis was reported: secreted by granulosa cells, this hormone is involved in the regulation of the recruitment of primordial follicles and in follicular growth. AMH assays were developed for women in 1990s, and the serum AMH level has rapidly become a crucial element in managing women's fertility. Based mainly on its ability to be a quantitative but indirect marker of ovarian reserve, the serum AMH assay is widely used in reproductive medicine and ART. This review summarizes current knowledge of the AMH assessment in the field of reproductive medicine. We focus on the role of AMH level to predict spontaneous pregnancy occurrence, ART outcomes, and fertility preservation outcomes.

Clinical predictive value of the initial neutrophils to lymphocytes and platelets ratio for prognosis of sepsis patients in the intensive care unit: a retrospective study.

Neutrophils to lymphocytes and platelets (N/LP) ratio has been confirmed as an indirect marker of inflammation. In this study, we aimed to further evaluate the prognostic significance of the N/LP ratio in sepsis patients admitted to the ICU. Sepsis patients from the Affiliated Hospital of Jiangsu University were retrospectively enrolled from January 2015 and July 2023. The primary outcomes were 30/60 days mortality. The secondary outcomes included the incidence of AKI, vasoactive drug, CRRT, invasive ventilation, length of ICU stay, length of hospital stay and ICU mortality. A total of 1,066 sepsis patients were enrolled with a median age of 75.0 (66.0, 85.0) years, and 62.5% of them being male. The 30 days and 60 days mortality rates were found to be 28.7 and 34.0%, respectively, while the incidence of AKI was 45.2%. Based on their N/LP ratios, we classified the sepsis patients into three groups: low, middle, and high, consisting of 266, 534, and 266 patients, respectively. According the Cox proportional hazard model, the middle and high N/LP groups were associated with a 1.990/3.106-fold increase in 30 days mortality risk and a 2.066/3.046-fold increase in 60 days mortality risk compared with the low N/LP group. Besides, multivariate logistic regression model suggested that the risk of AKI occurrence increased 2.460 fold in the high group compared to the low group. However, through subgroup analyses, we observed substantial variations in the association between N/LP ratios and 30/60 days mortality rates as well as the incidence of AKI among different populations. Notably, the N/LP ratio measured at ICU admission exhibited a higher AUC for predicting 30/60 days mortality (0.684/0.687). Additionally, we observed a good predictive power for the occurrence of AKI (AUC: 0.645) using the N/LP ratio measured at sepsis prognosis. Regarding the other secondary outcomes, the N/LP ratio was associated with disease severity in sepsis patients, including the need for vasoactive drugs, length of ICU stay, and ICU mortality. The N/LP ratio at ICU admission was found to have a significant independent association with 30/60 days mortality and the incidence of AKI in sepsis patients.

Significance of Chromogranin A and Synaptophysin in Gastroenteropancreatic Neuroendocrine Tumors

The two most commonly used immunohistochemical markers for neuroendocrine tumors are chromogranin A and synaptophysin. We had previously studied immunohistochemical staining on pancreatic neuroendocrine tumors: chromogranin A strongly positive tumors including gastrinomas, glucagonomas, pancreatic polypeptidomas were more malignant (&gt;50%) than chromogranin A weakly positive tumors of insulinomas (&lt;10%). With additional 40 cases of gastroenteric neuroendocrine tumors, formerly carcinoid tumors, we investigated chromogranin A and synaptophysin immunostaining: more aggressive neuroendocrine tumors of duodenum, small intestine and ascending colon were strongly positive for chromogranin compared to less aggressive neuroendocrine tumors of sigmoid colon and rectum. Immunohistochemical staining for chromogranin A represents a marker for the secretary granules with a possible marker of prognosis on all gastroenteropancreatic neuroendocrine tumors. Furthermore, serum CgA levels may be used as an indirect, independent diagnostic and prognostic marker for gastroenteropa-ncreatic neuroendocrine tumors in three folds: First to diagnose gastroenteropancreatic neuroendocrine tumors. Secondly, to assess the degree of malignancy by tissue and serum CgA levels and thirdly, evaluate increasing serum CgA levels as a prognostic indicator. Since there is no difference of immunostaining for synaptophysin between more aggressive neuroendocrine tumors and less aggressive tumors, immunostaining for synaptophysin is not a marker for aggressive tumors.

Tissue factor pathway-related biomarkers in liver cancer: activated factor VII–antithrombin complex and tissue factor mRNA levels are associated with mortality

BackgroundTissue factor (TF), the main initiator of the coagulation cascade, plays a role in cancer progression and prognosis. Activated factor VII–antithrombin complex (FVIIa-AT) is considered an indirect marker of TF exposure by reflecting TF-FVIIa interaction. ObjectivesTo assess the link between FVIIa-AT plasma levels, TF messenger RNA (mRNA) expression, and survival in cancer. MethodsTF pathway–related coagulation biomarkers were assessed in 136 patients with cancer (52 with hepatocellular carcinoma, 41 with cholangiocarcinoma, and 43 with colon cancer) undergoing surgical intervention with curative intent. TF mRNA expression analysis in neoplastic vs nonneoplastic liver tissues was evaluated in a subgroup of 91 patients with primary liver cancer. ResultsFVIIa-AT levels were higher in patients with cancer than in 136 sex- and age-matched cancer-free controls. In patients with cancer, high levels of FVIIa-AT and total TF pathway inhibitor were associated with an increased mortality risk after adjustment for confounders, but only FVIIa-AT remained a predictor of mortality by including both FVIIa-AT and total TF pathway inhibitor in Cox regression (hazard ratio, 2.80; 95% CI, 1.23-6.39; the highest vs the lowest quartile). This association remained significant even after adjustment for extracellular vesicle–associated TF-dependent procoagulant activity. In the subgroup of patients with primary liver cancer, patients with high TF mRNA levels had an increased mortality risk compared with that for those with low TF mRNA levels (hazard ratio, 1.92; 95% CI, 1.03-3.57), and there was a consistent correlation among high FVIIa-AT levels, high TF mRNA levels, and increased risk of mortality. ConclusionHigh FVIIa-AT levels may allow the identification of patients with cancer involving high TF expression and predict a higher mortality risk in liver cancer.

Fundoscopic changes in preeclampsia and eclampsia and their association with fetal outcome

Abstract Introduction: Hypertensive disorder is a major cause of maternal and perinatal mortality and morbidity, with preeclampsia and eclampsia being the most common. Preeclampsia is characterized by endothelial dysfunction and vasospasm of vessels, which can be observed by an ocular fundus examination. The fundus usually develops changes such as hypertensive retinopathy, papilledema, exudative retinal detachment, and vitreous and preretinal hemorrhages. Retinopathy is associated with placental insufficiency and intrauterine growth retardation. Therefore, this study has been undertaken to assess the prevalence of retinal changes in preeclampsia and their association with fetal parameters. Objective: To study the fundus changes in preeclampsia. To study the relationship between fundus changes and fetal outcome with respect to various parameters like birth weight, APGAR score, stillbirth, and perinatal death. To study the relationship between fundus changes with respect to proteinuria and serum uric acid. Study Design: Observational study. Methodology: One hundred patients presenting to the department of obstetrics and gynecology diagnosed with preeclampsia at a tertiary care center were included in the study. The patient recruited underwent an ocular examination using Snellen’s chart, a pupillary examination, a slit lamp examination, and a dilated fundus examination. The retinopathy was graded according to Keith and Wagner’s classification. The fetal parameters considered were birth weight, APGAR score, stillbirth, and neonatal death. Results: Patients ages ranged from 25 to 30 years old, with an average of 23.28 ± 3.37. Average systolic and diastolic blood pressures were 156.9 ± 17.961 and 104.88 ± 13.58, respectively. All patients had normal anterior segments. Visual acuity was6/6 in most of the patients. Fundus changes were noted in 48% of patients. Based on retinopathy classification, we found 66.7% patients with Grade 1 and 27.1% patients with Grade 2 hypertensive retinopathy. Only 6.3% of patients presented with Grade 3 hypertensive retinopathy. There was a significant association between retinopathy and proteinuria (P = 0.003), serum uric acid (P = 0.002), and the fetal birth weight (P = 0.001). Conclusion: Our findings suggest that progressive fundal changes in preeclampsia indicate worsening of the pathophysiological status of pregnancy-induced hypertension (PIH) and help in management. The degree of hypertensive retinopathy in women with preeclampsia is a valid and reliable prognostic factor that serves as an indirect marker of the severity of PIH and fetal outcome. Fundus examination is a simple, noninvasive, and cost-effective procedure that could be the initial finding in an asymptomatic patient who may require immediate management, which may help in saving the lives of both the mother and the baby. Fundus evaluation is essential for all patients with PIH.

Epigenomic Profiling Advises Therapeutic Potential of Leukotriene Receptor Inhibitors for a Subset of Triple-Negative Breast Tumors.

Triple-negative breast cancer (TNBC) is the most aggressive molecular subtype, with a poor survival rate compared to others subtypes. For a long time, chemotherapy was the only systemic treatment for TNBC, and the identification of actionable molecular targets might ultimately improve the prognosis for TNBC patients. We performed a genome-wide analysis of DNA methylation at CpG islands on a collection of one hundred ten breast carcinoma samples and six normal breast tissue samples using reduced representation bisulfite sequencing with the XmaI restriction enzyme (XmaI-RRBS) and identified a subset of TNBC samples with significant hypomethylation at the LTB4R/LTB4R2 genes' CpG islands, including CpG dinucleotides covered with cg12853742 and cg21886367 HumanMethylation 450K microarray probes. Abnormal DNA hypomethylation of this region in TNBC compared to normal samples was confirmed by bisulfite Sanger sequencing. Gene expression generally anticorrelates with promoter methylation, and thus, the promoter hypomethylation detected and confirmed in our study might be revealed as an indirect marker of high LTB4R/LTB4R2 expression using a simple methylation-sensitive PCR test. Analysis of RNA-seq expression and DNA methylation data from the TCGA dataset demonstrates that the expression of the LTB4R and LTB4R2 genes significantly negatively correlates with DNA methylation at both CpG sites cg12853742 (R = -0.4, p = 2.6 × 10-6; R = -0.21, p = 0.015) and cg21886367 (R = -0.45, p = 7.3 × 10-8; R = -0.24, p = 0.005), suggesting the upregulation of these genes in tumors with abnormal hypomethylation of their CpG island. Kaplan-Meier analysis using the TCGA-BRCA gene expression and clinical data revealed poorer overall survival for TNBC patients with an upregulated LTB4R. To this day, only the leukotriene inhibitor LY255283 has been tested on an MCF-7/DOX cell line, which is a luminal A breast cancer molecular subtype. Other studies compare the effects of Montelukast and Zafirlukast (inhibitors of the cysteinyl leukotriene receptor, which is different from LTB4R/LTB4R2) on the MDA-MB-231 (TNBC) cell line, with high methylation and low expression levels of LTB4R. In our study, we assess the therapeutic effects of various drugs (including leukotriene receptor inhibitors) with the DepMap gene effect and drug sensitivity data for TNBC cell lines with hypomethylated and upregulated LTB4R/LTB4R2 genes. LY255283, Minocycline, Silibinin, Piceatannol, Mitiglinide, 1-Azakenpaullone, Carbetocin, and Pim-1-inhibitor-2 can be considered as candidates for the additional treatment of TNBC patients with tumors demonstrating LTB4R/LTB4R2 hypomethylation/upregulation. Finally, our results suggest that the epigenetic status of leukotriene B4 receptors is a novel, potential, predictive, and prognostic biomarker for TNBC. These findings might improve individualized therapy for TNBC patients by introducing new therapeutic adjuncts as anticancer agents.

Input factors in the acquisition of evidentiality by Turkish heritage language children and adults in the United States

ABSTRACTAspectual and mood morphology are vulnerable domains in adult heritage speakers. This paper investigates the root of such vulnerability within the domain of Turkish evidentiality system by comparing 20 second-generation adult and 20 school-age child Turkish heritage speakers to 20 first-generation immigrants (main input providers for child heritage speakers) residing in the United States. A monolingual group from Turkey including 20 monolingual adults, 20 7-14-year-old and 20 3-6-year-old Turkish-speaking children were also tested. Results of a story-telling and a sentence selection task revealed weak mappings between the indirect evidential marker –mIş and its pragmatic use in story-telling as opposed to intact knowledge of the Turkish evidentiality system in the first-generation immigrant group. The school-age child and adult heritage speakers, on the other hand, were significantly the least accurate groups in both tasks, performing more variably than the 3-6-year-old monolingual children in Turkey. The individual analysis of the child heritage speaker data as well as a comparison of the child-parent dyads further showed that early heritage language experience and use rather than changes in the parental input are likely to be the source of the variability observed in child and eventually in adult Turkish heritage speakers in this domain.

Diffusion-weighted imaging in pediatric extracranial germ cell tumors.

Germ cell tumors (GCTs) comprise a rare and heterogeneous group of neoplasms presenting different clinical and histological characteristics, leading to a challenging scenario in clinical practice. Diffusion-weighted imaging (DWI) has been suggested as an indirect marker of tumor density and cellularity and could be used to monitor therapeutic response. However, its role in pediatric GCTs needs to be clarified. Here, we evaluated the features of DWI in pediatric extracranial GCTs in a reference Brazilian institution. We included 43 pediatric patients with primary GCTs treated between 2008 and 2022 in Hospital de Amor de Barretos. The patients' MRI images included T1-weighted without contrast, T2-weighted, DWI and apparent diffusion coefficient (ADC) maps. DWI was evaluated in the section that exhibited the greatest restricted diffusion in the largest hypersignal area of the image. The lowest ADC value was determined to define the region of interest (ROI). We used a small ROI, avoiding necrotic, adipose tissue, noisy or nonenhancing lesion voxels as recommended. ROI determination was established by visual inspection by two radiologists in accordance. We used two values of b (b = 50 mm2/s or b = 800) for ADC values. The highest mean ADC (mADC) value was observed in pure teratomas (1,403.50 ± 161.76 x10-3 mm2/s; mean ± SD) compared to other histologies (yolk sac, mixed teratoma, dysgerminoma and mixed GCT) of GCT (p<0.001). Furthermore, ROC analysis determined a cutoff mADC value of 1,179.00 x 10-3 mm2/s that differentiated pure teratomas from the other GCT histologies with a sensitivity of 95.8% and a specificity of 92.9% (AUC = 0.979; p<0.01). A significant increase in mADC was observed for malignant GCTs in treatment (1,197.00 ± 372.00 mm2/s; p<0.001) compared to that exhibited at the time of diagnosis (780.00 ± 168.00 mm2/s; mean ± SD. Our findings suggest that mADC assessment could be used as a tool to distinguish pure teratomas from malignant CGT histologies at diagnosis. Additionally, we demonstrated reasonable evidence that it could be used as a complementary tool to monitor treatment response in patients with malignant GCT.

1510. Evaluation of Quantitative (1,3)-β-D-glucan in Prediction of Pneumocystis jirovecii Pneumonia in Patients Living with HIV in New York

Abstract Background (1,3)-β-D-glucan (BDG) has been utilized as an indirect marker for Pneumocystis jirovecii pneumonia (PJP). The clinical utility of BDG, however, is limited because of uncertainty in interpreting quantitative values. A correlation was drawn between quantitative BDG and the diagnosis of PJP in hospitalized patients living with human immunodeficiency virus (HIV) at four affiliate campuses of the Montefiore Health System in the Bronx, NY. Methods All Pneumocystis direct fluorescent assays (DFA), polymerase chain reactions (PCR) either from bronchoalveolar lavage (BAL) fluid or sputum, and quantitative BDG assays either from serum or BAL fluid were retrieved from the Montefiore microbiology laboratory (date range: 8/1/18 - 8/31/22). Results from hospitalized patients with HIV whose CD4 count within 3 months were available, with either PJP DFA or PCR result within 7 days of serum BDG and lactate dehydrogenase (LDH) test were selected (n = 146). CD4 count was grouped into &amp;lt; 50 (n = 106), 50-100 (n = 15), 100-200 (n = 16), and &amp;gt;200 (n = 9). BDG, LDH, and CD4 were considered as input variables in a logistic regression model for true PJP status. The area under the curve (AUC) of the empirical receiver operating characteristic (ROC) curve with 95% confidence intervals (CI) was estimated based on trapezoidal area. Comparisons between ROC curve areas were performed via contrast matrix to take differences of the AUC of the empirical ROC curves and chi-square testing. Results Of the 146, 29 had PJP detected and 117 did not. When BDG only was considered, the AUC of the ROC curve was 0.78. When LDH quartiles (cutoffs: 232, 334, and 480 U/L) were considered with BDG, there was a statistically significant increase in AUC (0.84 vs. 0.78; p = 0.025). Adding CD4 group did not influence AUC. Figure 1 ROC curves for BDG only, BDG with CD4 count, BDG with LDH quartile, and BDG with CD4 group and LDH quartile combined Table 1 Area under the curve calculated for the ROC curves for BDG only, BDG with CD4 count, BDG with LDH quartile, and BDG with CD4 group and LDH quartile combined Conclusion The test characteristics (AUC) improved when LDH quartiles were used in conjunction with BDG, suggesting a better prediction of PJP diagnosis when BDG and LDH were considered together. This result supports further investigation of clinical markers of PJP given the limitations of establishing a microbiologic diagnosis. Further steps of the investigation will involve analyzing if other clinical factors including HIV viral load, oxygen saturation, and radiographical reading, influence predictability of PJP. Disclosures All Authors: No reported disclosures

Leaf explant based direct and indirect regeneration and SCoT marker assisted genetic fidelity analysis of endemic taxon Corynandra chelidonii var. pallae

Leaf explant based direct and indirect regeneration and SCoT marker assisted genetic fidelity analysis of endemic taxon Corynandra chelidonii var. pallae

Ticagrelor enhances the cardioprotective effects of ischemic preconditioning in stable patients undergoing percutaneous coronary intervention: the TAPER-S randomized study.

Ticagrelor improves clinical outcomes in patients with acute coronary syndromes compared with clopidogrel. Ticagrelor also inhibits cell uptake of adenosine and has been associated with cardioprotective effects in animal models. We sought to investigate the potential cardioprotective effects of ticagrelor, as compared with clopidogrel, in stable patients undergoing percutaneous coronary intervention (PCI). This was a Prospective Randomized Open Blinded End-points (PROBE) trial enrolling stable patients with coronary artery disease (CAD) requiring fractional flow reserve-guided PCI of intermediate epicardial coronary lesions. ST-segment elevation at intracoronary electrocardiogram (IC-ECG) during a two-step sequential coronary balloon inflations in the reference vessel during PCI was used as an indirect marker of cardioprotection induced by ischemic preconditioning (IPC). The primary endpoint of the study was the comparison of the delta (Δ) (difference) ST-segment elevation measured by IC-ECG during two-step sequential coronary balloon inflations. Fifty-three patients were randomized to either clopidogrel or ticagrelor. The study was stopped earlier because the primary endpoint was met at a pre-specified interim analysis. ΔST-segment elevation was significantly higher in ticagrelor as compared to clopidogrel arms (P<0.0001). Ticagrelor was associated with lower angina score during coronary balloon inflations. There was no difference in coronary microvascular resistance between groups. Adenosine serum concentrations were increased in patients treated with ticagrelor as compared to those treated with clopidogrel. Ticagrelor enhances the cardioprotective effects of IPC compared with clopidogrel in stable patients with CAD undergoing PCI. Further studies are warranted to fully elucidate the mechanisms through which ticagrelor may exert cardioprotective effects in humans. http://www.clinicaltrials.gov. Unique Identifier: NCT02701140.

Easix Score Correlates With Endothelial Dysfunction Biomarkers and Predicts Risk of Acute Graft-Versus-Host Disease After Allogeneic Transplantation

Plasma biomarkers of endothelial dysfunction have been postulated for the diagnosis and prognosis of acute graft-versus-host disease (aGVHD). However, their use is not validated in clinical practice yet. The endothelial activation and stress index (EASIX), a simple score based on routine laboratory parameters, is considered to be an indirect marker of endothelial damage. High value of EASIX was correlated with worse non-relapse mortality (NRM) and overall survival (OS) and a high risk of sinusoidal obstructive syndrome and transplant-associated thrombotic microangiopathy (TA-TMA). This study investigates the predictive value of plasma biomarkers and the EASIX score for the prediction of aGVHD. We assessed vascular cell adhesion molecule-1 (VCAM-1), tumor necrosis factor receptor 1 (TNFR1), and VWF:Ag plasma levels and the EASIX score before allogeneic hematopoietic stem cell transplantation (allo-HSCT) and on days 0, 3, 7, 14, and 21 in an experimental cohort (n = 33). EASIX was transformed to a base-2 logarithm to perform the analysis. For the most relevant biomarkers, we estimate the optimal cutoff values and the discriminatory ability to differentiate patients with high-risk of aGVHD. The conclusions obtained in the experimental cohort were validated in a large cohort of 321 patients at the same institution. Plasma biomarkers and EASIX showed similar post-transplantation dynamics consisting of a progressive increase. Multivariate analysis showed an association between high TNFR1 levels and Log-2 EASIX score on day 7 after transplantation with an increased likelihood of developing aGVHD (hazard ratio [HR] = 1, P = .002; HR = 2.31, P = .013, respectively). Patients with TNFR1 ≥1300 ng/mL (HR = 7.19, P = .006) and Log2-EASIX ≥3 (HR = 14.7, P <.001) at day 7 after transplantation were more likely to develop aGVHD with high predictive accuracy (C-index of 74% and 81%, respectively). In the validation cohort, patients with Log2-EASIX ≥3 on day 7 after transplantation presented a significantly higher incidence of grade II-IV aGVHD (HR = 1.94, P = .004) independent of GVHD prophylaxis (HR = 0.38, P = .004), conditioning regimen (HR = 0.59, P =.02) and type of donor (HR = 2.38, P = .014). Differential degree of endothelial damage can be measured using both EASIX score and plasma biomarkers in the early post-transplantation period. Patients at risk of developing aGVHD could be easily identified by a high EASIX score. Both indicators of endothelial activation represent a promising approach to predict aGVHD.

Improvement in Cardiac Morphology Demonstrated by Cardiac Magnetic Resonance Imaging and Echocardiography after Haploidentical Hematopoietic Cell Transplantation in Adults with Sickle Cell Disease

Cardiopulmonary complications account for approximately 40% of deaths in patients with sickle cell disease (SCD). Diffuse myocardial fibrosis, elevated tricuspid regurgitant jet velocity (TRV) and iron overload are all associated with early mortality. While HLA-matched sibling hematopoietic cell transplant (HCT) offers a potential cure, fewer than 20% of patients have a suitable donor. Haploidentical HCT allows for an increased donor pool and has recently demonstrated improved safety and efficacy. Our group has reported improved cardiac morphology via echocardiography at one year after HCT. Herein, we describe the first use of cardiac magnetic resonance imaging (CMR), the gold standard for measuring volume, mass, and ventricular function, to evaluate changes in cardiac morphology post-HCT in adults with SCD. We analyzed baseline and 1-year data from 12 adults with SCD who received nonmyeloablative haploidentical peripheral blood HCT at the NIH. Patients underwent non-contrast CMR at 3T, echocardiography, and laboratory studies. One year after HCT, patients showed marked improvement in cardiac chamber morphology by CMR, including left ventricular (LV) mass and volume (70.2 to 60.1 g/m2, p = 0.02; 114.5 to 90.6 mL/m2, p = 0.001). Furthermore, mean TRV normalized by 1 year, suggesting that HCT may offer a survival benefit. Fewer patients had pathologically prolonged native myocardial T1 times, an indirect marker of myocardial fibrosis at 1 year; these data showed a trend toward significance. In this small sample, CMR was very sensitive in detecting cardiac mass and volume changes after HCT and provided complementary information to echocardiography. Notably, post-HCT improvement in cardiac parameters can be attributed only in part to the resolution of anemia; further studies are required to determine the role of myocardial fibrosis reversal, improved blood flow, and survival impact after HCT for SCD.

Incidence and predictors of heart failure in a cohort of MINOCA patients

Abstract Background/Introduction A higher risk of death and major cardiovascular events, including Heart Failure (HF), after a Myocardial Infarction with Non-Obstructive Coronary Arteries (MINOCA) has been described. In particular, HF has a negative impact not only on survival but also on quality of life. In this context, strategies to identify subjects with a greater risk of developing HF are needed. Purpose. Assess incidence and predictors of HF developing subsequently a MINOCA. Methods 461 subjects with suspected MINOCA from 2016 to 2021 were assessed. Acute myocarditis, Takotsubo syndromes, cardiomyopathies and patients with incomplete data were excluded. The final cohort consisted of 188 patients with a confirmed MINOCA diagnosis. After a mean follow-up of 36±14.8 months, 20 patients (10.6%) had hospital readmission for HF. Demographic, clinical, laboratory, and instrumental data were collected and analyzed. Results None among demographic characteristics, cardiovascular risk factors or comorbidities showed correlation with HF occurrences. Paroxysmal or permanent atrial fibrillation was more frequent among patients who developed HF (p .005), such as ST-elevation at onset (p .001). Higher glycemia at admission correlated with HF (p .012). In this regard, hyperglycemia could act as a direct toxic agent or as an indirect marker of a greater stress response. Cardiac troponin I (cTnI) at peak, but not first cTnI measurement, was significantly higher in patients who developed HF (p&amp;lt;.001), confirming the prognostic value of troponin, even in this context. Concerning echocardiography, HF incidence was higher in patients with any wall motion abnormalities (p .037), lower ejection fraction (EF; p .046), especially if EF≤40% (p&amp;lt;.001), higher atrial maximum diameter (p .005), end-diastolic diameter (p. 001) or normalized end-diastolic volume (p&amp;lt;.001). HF occurred more often (p&amp;lt;.001) in patients with a transmural pattern of late gadolinium enhancement (LGE), which resulted as an independent risk factor for HF developing in multivariate analysis (HR 15.32, 95% CI 5.13-45.75, p&amp;lt;.001). 32 patients (17%) were affected by non-obstructive coronary arteries stenosis (i.e., &amp;lt;50%). This finding, but not re-infarction, was also associated with HF development (p .004). Only 2 out of the 68 patients dismissed with DAPT developed HF (p .010). In our study beta-blockers, RAS inhibitors and statins at dismission had not reduced the risk of HF. Nevertheless, the low sample size forbids inferring conclusions about the therapy. Conclusion In our cohort of patients with MINOCA, several clinical, laboratory and echocardiographic characteristics correlated with a higher frequency of HF occurrences. Moreover, a broader extent of cardiac damage, as testified by cTnI, wall motion abnormalities, reduced EF and transmural LGE, could predict HF development. Further studies to establish adequate follow-up programs and therapy to prevent HF progression in these patients are needed.Figure 1