Routine Protocol Research Articles

Abstract B022: Immediate post-treatment detection of minimal residual disease in HPV-Driven oropharyngeal cancer is associated with high risk of recurrence

Abstract Purpose: Detecting minimal residual disease (MRD) has emerged as a key strategy in oncology for identifying those at high risk of relapse. Circulating tumor tissue modified viral (TTMV)-HPV DNA has emerged as a promising biomarker in HPV-driven oropharyngeal cancer. Prior studies have largely focused on detection beyond three months post-treatment. This study evaluated the ability of TTMV-HPV DNA to detect MRD immediately after initial or salvage treatment. Patients and Methods: This IRB-approved, retrospective cohort study included 171 patients across eight U.S. centers who had pathologically confirmed HPV-driven oropharyngeal cancer. Patients received at least one TTMV-HPV DNA test (NavDx®, Naveris, Inc.) done between zero and three months of treatment between February 2020 and April 2022. Minimum four months of follow-up was required for patient inclusion. Test results were correlated with physician-reported exams, imaging studies, and biopsies. Results: Patients were primarily White (94%) and male (86%) and were a median age at diagnosis of 60 years (range: 31 - 81). Primary tumors were located in the tonsil or base of tongue (94%) and were mostly T1-T2 (77%), N1 (66%), and M0 (98%). Patients had a median follow-up of 17 months after the completion of initial definitive treatment (range: 4-96 months). In this cohort, 198 tests conducted in 171 patients were evaluable. A total of 14 patients had both biopsy-confirmed pathologic recurrences and relevant post-treatment testing data in relation to the recurrence, with a median disease-free interval of three months (range: 1-16). Within this cohort, TTMV-HPV DNA testing demonstrated a per-patient sensitivity of 85.7% (12/14, 95% CI: 67-100). The positive predictive value (PPV) of a single TTMV-HPV DNA test within the first three months following treatment was 86.7% (13/15, 95% CI: 69.5-100). Six of the positive tests (46.2%) preceded clinical recurrence detection by a median lead-time of 36 days (range: 21-97). Test positivity following initial definitive treatment was significantly associated with worse recurrence-free survival (log-rank test, p<0.0001; hazard ratio: 21). The two patients with false positive test results had their TTMV-HPV DNA testing performed just four days after completing CRT. Their test scores returned to negative on the subsequent test, and both patients have remained disease-free. Of the 157 patients who remained TTMV-HPV DNA negative throughout the duration of follow-up, only two (1.3%) had biopsy-confirmed, HPV-positive recurrence before or within three months of a negative test. The specificity and negative predictive value (NPV) per-test were each 98.9% (181/183, 95% CI: 97.4-100). Conclusion: TTMV-HPV DNA demonstrates strong potential as a prognostic biomarker for detecting MRD in the immediate post-treatment setting, with a high PPV and NPV for recurrence. These findings support the integration of TTMV-HPV DNA into routine surveillance protocols to enhance early detection of recurrence and guide timely interventions. Citation Format: Scott A Roof, James Jabalee, Eleni M Rettig, Rocco Ferrandino, Sida Chen, Marshall R Posner, Krzysztof J Misiukiewicz, Eric M Gender, Raymond L Chai, John Sims, Elaine Thrash, Scott J Stern, Noah S Kalman, Sreenija Yarlagadda, Adam Raben, Lydia Clements, Abie Mendelsohn, John M Kaczmar, Yadav Pandey, Mihir Bhayani, Catherine DV Fitz, Glenn J Hanna. Immediate post-treatment detection of minimal residual disease in HPV-Driven oropharyngeal cancer is associated with high risk of recurrence [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B022.

Recommendations for the use of natriuretic peptides for early diagnosis of heart disease in patients with diabetes: A consensus report by SPEDM, SPC, NEDM-SPMI and APMGF.

Diabetes is a significant risk factor for the development of heart disease (HD), with heart failure (HF) being one of the early manifestations of the disease in this population. The diagnostic process is challenging and contributes to a significant number of undiagnosed cases of HD among individuals with diabetes. This is largely due to the non-specific nature of symptoms and signs in the initial stages of disease, making early detection elusive. Timely identification and prevention of HD in patients with diabetes have the potential to significantly improve patient prognosis and alleviate the growing burden of this population on the national healthcare system. Natriuretic peptides (BNP and NT-proBNP) have been recognized as reliable, cost-effective biomarkers for detecting HD and can be further used as risk assessment biomarkers in asymptomatic patients. Despite being recommended in several European and American guidelines to rule out and rule in HF, the routine use of these biomarkers for the diagnosis of HDs in patients with diabetes has only recently been proposed by the American Diabetes Association (ADA) and has not yet been implemented in Portugal. Therefore, a multidisciplinary panel of experts from four medical societies, the Portuguese Society of Endocrinology, Diabetes and Metabolism, the Portuguese Journal of Cardiology, the Diabetes Study Group from the Portuguese Society of Internal Medicine and the Portuguese Association of Family Medicine were convened to discuss and establish consensus recommendations for natriuretic peptide screening in patients with diabetes and its integration into routine diabetes management protocols. This manuscript draws on the consensus recommendations from four Portuguese medical societies, offering clear guidance on natriuretic peptides use tailored to Portuguese clinical practice. Accordingly, this consensus advises the use of NT-proBNP analysis for all patients with diabetes aged 50 years and older, or under 50 if they have risk factors and/or comorbidities. Adjusted rule-out and rule-in values for age, sex and risk factors are provided. NT-proBNP levels above 125 pg/mL should prompt additional testing and cardiovascular investigation. Routine evaluation every two to three years for low-risk patients and annually for high-risk patients is proposed when NT-proBNP is below 125 pg/mL and in the absence of suspected heart disease.

IGLON5 Frequency in Idiopathic REM Sleep Behavior Disorder: A Multicenter Study.

Idiopathic/isolated REM sleep behavior disorder (iRBD) has been strongly linked to neurodegenerative synucleinopathies such as Parkinson disease, dementia with Lewy bodies, and multiple system atrophy. However, there have been increasing reports of RBD as a presenting feature of serious and treatable autoimmune syndromes, particularly IGLON5. This study's objective was to investigate the frequency of autoantibodies in a large cohort of participants with iRBD. Participants were enrolled in the North American Prodromal Synucleinopathy cohort with polysomnography-confirmed iRBD, free of parkinsonism and dementia. Plasma samples were systematically screened for the autoantibodies IGLON5, DPPX, LGI1, and CASPR2 using plasma IgG cell-based assay. Positive or equivocal results were confirmed by repeat testing, plus tissue-based indirect immunofluorescence assay for IGLON5. Of 339 samples analyzed, 3 participants (0.9%) had confirmed positive IGLON5 autoantibodies in the cell-based assay, which were confirmed by the tissue-based assay. An additional participant was positive for CASPR2 with low titer by cell-based assay only (of lower clinical certainty). These cases exhibited a variety of symptoms including dream enactment, cognitive decline, autonomic dysfunction, and motor symptoms. In 1 IGLON5 case and the CASPR2 case, evolution was suggestive of typical synucleinopathy, suggesting the possibility that findings were incidental. However, 2 participants with IGLON5 died before diagnosis was clinically suspected, with a final clinical picture highly suggestive of autoimmune disease. Our finding that nearly 1% of a large iRBD cohort may have a serious but potentially treatable autoantibody syndrome has important clinical implications. In particular, it raises the question of whether autoantibody testing for IGLON-5-IgG should be widely implemented for participants with iRBD, considering the difficulty in diagnosis of autoimmune diseases, their response to treatment, and the potential for rapid disease progression. However, any routine testing protocol will also have to consider costs and potential adverse effects of false-positive findings. NCT03623672.

Elucidating genetic variability between randomly bred domestic cats and Persian domestic cats from different geographical locations using microsatellite markers.

The domestic cat (Felis catus) is a newly evolved species in the family Felidae that has developed some great features among mammals. It is critical to conserve these species and prevent inbreeding from reducing their genetic diversity by understanding their genetic relationships and applying the information to breeding management. The diverse population was an excellent choice for studying genetic diversity and inbreeding phenomena. To conduct this research, 128 individuals from 8 populations, including Azerbaijan, Persian, Ahar, Uermia, Tehran, Karaj, Turkish and Shop cat (both genders), were randomly selected from different geographical regions. We selected eight STR markers with different chromosomal locations based on polymorphism and observed allele numbers in the next step. DNA extraction was performed using tail hair root, PCR and electrophoresis, and gel staining was performed according to routine laboratory protocol. For statistical analysis, CONVERT versions POPGENE, ARLEQUIN GenAlEx and R script analysis. Remarkably, our results showed that 23 alleles were identified in 128 samples. The highest number of alleles belonged to the FCa096 locus (eight alleles) in the Persian population, followed by FCa045 (seven alleles) in the Persian and Ahar populations. Another new finding is that the lowest number of alleles belonged to the 35 and FCa77 locus (two alleles). In addition, pairwise differentiation between and within populations was examined using the genetic distance index. Overall, the results showed that the degree of differentiation within the population is high in the Turkish population compared to other population groups and lower in the Azerbaijan population. In addition, principal component discriminant analysis-based analysis based on the ADAGENET package shows the distribution of samples by geographical location. The results show that genetic mixing between populations is high. On this basis, we conclude that randomly bred domestic cats have a higher level of diversity than Persian domestic cats. This is an interesting topic for future work.

Myocardial parametric mapping in patients with suspected acute myocarditis: a prognosis study

Abstract Background T1 mapping and T2 mapping have become essential components of the 2018 Lake Louise criteria and obtained excellent results in diagnosing myocarditis. However, their prognostic value in acute myocarditis has not yet been well recognized or established. Purpose Our study aims to investigate the prognostic value of T1 mapping, T2 mapping, and extracellular volume fraction (ECV) in a large cohort of patients with suspected acute myocarditis. Materials Patients meeting the recommended clinical criteria for suspected acute myocarditis were consecutively enrolled. The primary endpoint is the composite of cardiac death, heart failure hospitalization, heart transplantation, sustained ventricular arrhythmia, and recurrent myocarditis, defined as major adverse cardiovascular events (MACE). All patients underwent CMR at 3.0 T scanner using a standardized, routine imaging protocol, and the three short-axis view slices (basal, mid-ventricular, and apical) were divided into 16 segments according to the American Heart Association 17-segment model (apex excluded). We also averaged the native T1 values, ECV, and T2 values of all 16 segments to get global T1 and ECV fraction values, respectively. Statistically significant parameters from univariate Cox analyses were put into multivariate Cox analysis. Results A total of 235 patients (150 men; 32±13 years) were enrolled in this study. During a mean follow-up of 43.0±3.6 months, MACE occurred in 37 patients (15.7%) and was univariably associated with heart failure presentation, left ventricular ejection fraction, left ventricular end-systolic volume index, left ventricular end-diastolic volume index, native T1 and ECV. Patients with MACE showed higher global native T1, ECV, and T2 values (1348±59 vs 1266±51, p=<0.001; 40.0±8.7 vs 32.8±5.9, p<0.001; 63.4±12.1 vs. 55.5±9.1, p=0.011), and were more likely to have tissue changes in the interventricular septum and anterior wall (Figure 1). In a series of nesting multivariable Cox regression models, the addition of native T1 (per 10-ms increase: HR, 1.128; 95% CI: 1.043, 1.220; p = 0.003; Harrell’s C-index = 0.833) or ECV (per 5% increase: HR, 1.382; 95% CI: 1.060, 1.802; p = 0.017; Harrell’s C-index = 0.847) improved prognostication compared with the model based on clinical variables, left ventricular ejection fraction, and septal late gadolinium enhancement (Harrell’s C-index = 0.762) (Figure 2). T2 in multivariate Cox regression analysis didn’t show predictive value, but his inclusion increased the C-index of the model to 0.814. Conclusions Myocardial parametric mapping not only holds substantial diagnostic value but also plays a critical role in the prognostic assessment and risk prediction of myocarditis. Their utilization in these contexts enhances the accuracy and effectiveness of clinical evaluations and decision-making processes.

Adult mental healthcare professionals' experiences of family centred conversations with patients who are parents: a qualitative study.

Many parents with a mental illness report a desire for both recognition of their parental role and support for themselves and their children. However, parents are often fearful of negative judgements from professionals about their ability to be a parent, which inhibits raising concerns about their children with clinical teams. Consequently, an essential first step to supporting families is for professionals to proactively identify which patients are parents, although evidence indicates this is not consistently part of services. Professionals could play a pivotal role in guiding parents about how to talk to their children about their mental illness; this in turn can facilitate family functioning and enable children to access evidence-based interventions. This approach is crucial for mitigating the well-documented intergenerational risk of poorer outcomes and mental illness in children of affected parents. This study aimed to explore adult mental healthcare professionals' beliefs and experiences of talking with patients about: i) their role as parents ii) communication with their children about mental illness. Semi-structured qualitative interviews were conducted with 19 adult mental healthcare professionals with 4-30 years' experience of working with adult patients under the care of NHS adult mental healthcare services in England. Audio recordings were transcribed verbatim and analysed using an inductive coding approach following the principles of thematic analysis. All participants recognised their responsibility to identify patients' children through routine safeguarding protocols, but supporting patients around telling their children the diagnosis was less common. Many participants expressed concerns that raising the topic of children with patients could conflict with other parts of their professional role or would not be welcomed. Obstacles to these conversations were identified, across individual, environmental and organisational factors. Potential facilitators included specific staff training and resources for clinical teams and families around how to tell children about mental illness. Adult mental healthcare professionals would benefit from enhanced training on the importance and methods of guiding parents to communicate with their children about mental illness. This should include routine identification of which patients are parents and recognition of the impact of an adult's mental illness on the wider family system.

Phrenic Nerve Sonography Alterations in Patients with ALS: Insight with Clinical and Neurophysiological Findings.

Background: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder, and although the diagnosis is primarily based on clinical criteria, ENMG, as the "gold standard", does not always show detectable changes. Therefore, our study suggests that alterations in echogenicity and heterogeneity of the phrenic nerve (PN) may serve as potential additional diagnostic tools for ALS. Methods: Our study included 32 patients in the ALS group and 64 individuals in the control group. Each participant underwent an interview and completed questionnaires to collect clinical and demographic data, including age, gender, height, body mass index (BMI), hip and waist circumference, duration of illness, ALS-FRS-R score, comorbidities, and medication use. Ultrasound examinations of the PN were performed by two authors using a high-resolution "Philips EPIQ 7" ultrasound machine equipped with a linear 4-18 MHz transducer. The ALS group participants underwent PN sonography and conduction examinations, arterial blood gas (ABG) analysis, respiratory function tests (RFT), and electroneuromyography (ENMG). Results: The study demonstrated that the phrenic nerve is significantly smaller on both sides in patients with ALS compared to the control group (p < 0.01). Changes in the homogeneity and echogenicity of the PN were also observed on both sides. On the right side, 43.8% of the nerves showed heterogeneity, 40.6% were isoechoic, and 21.9% were hyperechoic. On the left side, 59.4% of the nerves exhibited heterogeneity, 34.4% were isoechoic, and 28.1% were hyperechoic. Moreover, sonography on both sides showed significant correlation with ALS-FRS-R, COMPASS-31, and ENMG results. Conclusions: Our study highlights the importance of phrenic nerve ultrasound as a promising supplementary diagnostic tool for ALS. The significant differences in phrenic nerve size, echogenicity, and homogeneity between patients with ALS and the control group demonstrate that ultrasound imaging can detect morphological changes in the phrenic nerve. Incorporating phrenic nerve ultrasound into routine diagnostic protocols could improve early detection, enhance disease monitoring, and offer a more comprehensive understanding of the neurodegenerative processes in ALS.

Multicenter study on Clostridioides difficile infections in Mexico: exploring the landscape.

This study aims to outline Clostridioides difficile infection (CDI) trends and outcomes in Mexican healthcare facilities during the COVID-19 pandemic. Observational study of case series. Sixteen public hospitals and private academic healthcare institutions across eight states in Mexico from January 2016 to December 2022. CDI patients. Demographic, clinical, and laboratory data of CDI patients were obtained from clinical records. Cases were classified as community or healthcare-associated infections, with incidence rates calculated as cases per 10,000 patient days. Risk factors for 30-day all-cause mortality were analyzed by multivariate logistic regression. We identified 2,356 CDI cases: 2,118 (90%) were healthcare-associated, and 232 (10%) were community-associated. Common comorbidities included hypertension, diabetes, and cancer. Previous high use of proton-pump inhibitors, steroids, and antibiotics was observed. Recurrent infection occurred in 112 (5%) patients, and 30-day mortality in 371 (16%). Risk factors associated with death were a high Charlson score, prior use of steroids, concomitant use of antibiotics, leukopenia, leukocytosis, elevated serum creatine, hypoalbuminemia, septic shock or abdominal sepsis, and SARS-CoV-2 coinfection. The healthcare-associated CDI incidence remained stable at 4.78 cases per 10,000 patient days during the pre-and pandemic periods. However, the incidence was higher in public hospitals. Our study underscores the need for routine epidemiology surveillance and standardized CDI classification protocols in Mexican institutions. Though CDI rates in our country align with those in some European countries, disparities between public and private healthcare sectors emphasize the importance of targeted interventions.

An Observational Study on Clinical Benefits of CytoSorb in Patients with Severe Sepsis

Background and Aim: Sepsis is a well-recognized healthcare issue worldwide. Despite research, the ability to positively influence outcome remains limited. Newer evidences have pointed that using adsorption of cytokines is beneficial during endotoxemia and sepsis. Therefore, this study was aimed to determine the clinical benefits of CytoSorb usage in patients with severe sepsis admitted in intensive care units (ICU). Study Design: Prospective, observational, comparative study. Material and Methods: Forty patients with sepsis admitted to ICU were included. The CytoSorb group included 20 patients who received CytoSorb therapy in addition to Standard-care (SOC) and 20 patients in SOC group who received SOC alone as per routine ICU protocols. Clinical and laboratory parameters were analyzed pre/post-treatment in both groups and compared. Results: CytoSorb and SOC groups were comparable at baseline. There was significant reduction in serum creatinine (2.59±2.0 vs. 2.89±1.2, mg/dl; P=0.042), serum lactate (3.26±1.1 vs. 3.27±0.9, mmol/lit; P&lt;0.001), serum procalcitonin (2.75±2.4 vs. 3.39±3.5, ng/ml; P&lt;0.001), serum CRP (90.2±57.4 vs. 175.6±100.9, mg/dl; P&lt;0.001) and serum IL6 (1769.7±4444.1 vs. 256.8±392.4, pg/ml; P=0.03) levels in CytoSorb compared to SOC. There was significant improvement in mean arterial pressure (MAP) (65.75±1.8 vs. 62.75±2.8, mmHg; P&lt;0.001) and reduction in norepinephrine dose (6.47±2.7 vs.10.65±3.6, mcg/min; P&lt;0.001) in CytoSorb group reflecting better hemodynamic stability. The post-treatment increase in SOFA score was lesser in CytoSorb group (11.85±2.9) than SOC group (12.3±2.3), but was not significant (P=0.135). Conclusion: CytoSorb therapy along with SOC was associated with significant improvements in hemodynamic stability, MAP and Norepinephrine requirements than SOC alone in severe sepsis.

Mycobacterium tuberculosis complex sample processing by mechanical lysis, an essential step for reliable whole genome sequencing

Mycobacterium tuberculosis complex (MTBC) whole genome sequencing (WGS) turnaround time and WGS success rates are highly influenced by DNA extraction protocols even from cultures. Efficient mycobacterial lysis is crucial for obtaining sufficient DNA from cultures to facilitate reliable genomic drug susceptibility prediction and accurate genotyping with WGS. We compared four DNA extraction protocols from BD BACTEC™ Mycobacterial Growth Indicator Tubes (MGIT) for WGS with a focus on the lysis step: protocol A) column-based protocol without mechanical lysis; protocol B) an adapted protocol including a bead beating step; protocol C) DNA extraction from primary received cultures using bead beating: and protocol D) DNA extraction from pre MGIT-positive (enriched) cultures. Protocol B increased DNA yield approximately 60-fold, and significantly improved the sequencing success rate. The increased yield also allowed DNA extraction from primary cultures with high success rates (protocol C). Additionally, by using pre-positive enriched MGIT cultures, we demonstrated that bead beating opens the possibility of reliable WGS up to five days before a MGIT tube would be flagged positive (protocol D). The most optimal bead beating-based DNA extraction was also evaluated for Nanopore sequencing. Shortening bead beating duration to 15 s resulted in longer read lengths (N50 from 1.4 kb to 2.6 kb) while still providing efficient lysis. Furthermore, AmpureXP bead beating-based DNA capture / purification proved to be as efficient as Qiagen column-based DNA extraction, further simplifying and shortening the DNA extraction protocol. Adding a mechanical lysis step to our routine MTBC DNA extraction protocol has allowed us to reduce the turnaround time while maintaining DNA quality sequencing success rates.

Quantitative brain T1 maps derived from T1-weighted MRI acquisitions: a proof-of-concept study.

Longitudinal T1 relaxation time is a key imaging biomarker. In addition, T1 values are modulated by the administration of T1 contrast agents used in patients with tumors and metastases. However, in clinical practice, dedicated T1 mapping sequences are often not included in brain MRI protocols. The aim of this study is to address the absence of dedicated T1 mapping sequences in imaging protocol by deriving T1 maps from standard T1-weighted sequences. A phantom, composed of 144 solutions of paramagnetic agents at different concentrations, was imaged with a three-dimensional (3D) T1-weighed turbo spin-echo (TSE) sequence designed for brain imaging. The relationship between the T1 values and the signal intensities was established using this phantom acquisition. T1 mapping derived from 3D T1-weighted TSE acquisitions in four healthy volunteers and one patient with brain metastases were established and compared to reference T1 mapping technique. The concentration of Gd-based contrast agents in brain metastases were assessed from the derived T1 maps. Based on the phantom acquisition, the relationship between T1 values and signal intensity (SI) was found equal to T1 = 0.35 × SI-1.11 (R2 = 0.97). TSE-derived T1 values measured in white matter and gray matter in healthy volunteers were equal to 0.997 ± 0.096 s and 1.358 ± 0.056 s (mean ± standard deviation), respectively. Mean Gd3+ concentration value in brain metastases was 94.7 ± 30.0 μM. The in vivo results support the relevance of the phantom-based approach: brain T1 maps can be derived from T1-weighted acquisitions. High-resolution brain T1 maps can be generated, and contrast agent concentration can be quantified and imaged in brain metastases using routine 3D T1-weighted TSE acquisitions. Quantitative T1 mapping adds significant value to MRI diagnostics. T1 measurement sequences are rarely included in routine protocols. T1 mapping and concentration of contrast agents can be derived from routine standard scans. The diagnostic value of MRI can be improved without additional scan time.

CT perfusion imaging in aneurysmal subarachnoid hemorrhage. State of the art.

CT perfusion (CTP) images can be easily and rapidly obtained on all modern CT scanners and have become part of the routine imaging protocol of patients with aneurysmal subarachnoid haemorrhage (aSAH). There is a growing body of evidence supporting the use of CTP imaging in these patients, however, there are significant differences in the software packages and methods of analysing CTP. In. addition, no quantitative threshold values for tissue at risk (TAR) have been validated in this patients' population. Here we discuss the contribution of the technique in the identification of patients at risk of aSAH-related delayed cerebral ischemia (DCI) and in the assessment of the response to endovascular rescue therapy (ERT). We also address the limitations and pitfalls of automated CTP postprocessing that are specific to aSAH patients as compared to acute ischemic stroke (AIS).

B-241 Next-generation monitoring: M-inSightclinical application in multiple myeloma

Abstract Background The improvement of multiple myeloma (MM) therapy effectiveness has increased the need for precise, sensitive, and dynamic monitoring of minimal residual disease (MRD). Current MRD assessment methods that are based on bone marrow aspirates, pose limitations due to the invasiveness of the procedure and potential heterogeneous sampling. Recognizing the need for non-invasive, frequent testing, a targeted mass spectrometry, M-InSight®, (MS)-based MRD blood-test was developed. This innovative assay identifies and tracks clonotypic peptides from the patient's monoclonal immunoglobulin, offering a blood-based alternative with equivalent sensitivity to existing bone marrow reliant techniques.Here we employed M-InSight® to sequence and select clonotypic peptides, enabling highly specific, ultra-sensitive quantification of the M-protein. This study examines the correlation between M-protein quantification using M-InSight® and conventional serum protein electrophoresis (SPEP) values routinely obtained in clinical settings. Given the established role of M-protein as a biomarker for MM diagnosis and longitudinal monitoring, ensuring concordance between emerging methodologies and existing standards is vital. Methods Serum samples were obtained from the clinical laboratory at the University of Miami and characterized using SPEP (capillary electrophoresis, Sebia), immunotyping (Sebia), and free light chain quantitation (The binding site). Patients with M-protein concentrations ≥0.2 g/dL by SPEP were included. Longitudinal samples were collected. Subsequently, all samples were analyzed using M-InSight®. The initial sample for each patient underwent de novo sequencing of the M-protein by mass spectrometry and was also used to calibrate the concentration with the known value from SPEP. An in-house bioinformatics algorithm aligned, sequenced, and selected patient-specific clonotypic peptides. These peptides were targeted for the M-inSight® follow up assay allowing the quantification of M-protein. Results A variety of M-protein isotypes were successfully sequenced (IgGK, IgGL, IgAK, IgAL, IgMK, IgML, free K, free L, IgDL, biclonal) at concentrations ranging from 0.2 to 2.9 g/dL. M-protein was successfully sequenced and clonotypic peptides were identified in 57 out of 59 patients using the initial sample. A total of 109 serum samples collected during maintenance or post treatment were analyzed, 50 of them were follow-up samples and were used to correlate M-protein values from both techniques. M-protein quantification by M-Insight showed high concordance with the one obtained by SPEP, with a correlation value of 0.88 (intercept set to 0, slope 0.91. Pearson = 0.886, p-value &amp;lt;0.001). Out of 57 patients whose M-protein was sequenced, 55 were positive by SPEP and were successfully quantified by M-InSight®. Furthermore, M-InSight® detected and quantified M-protein on samples of the 2 remaining patients that showed M-protein values below SPEP detection limits, highlighting the higher sensitivity of M-InSight®. Conclusions M-inSight® assay successfully identified clonotypic peptides in 57 out of 59 patients which were used to quantify M-protein concentration and showed a strong concordance with SPEP and affirming the utility of M-InSight® as a reliable method for routine monitoring of MM. The ability to seamlessly integrate M-InSight® into routine monitoring protocols offers clinicians a valuable tool for MRD monitoring, tracking disease progression and response to therapy with enhanced sensitivity and specificity.

Chitinase and proteinase K treatments enhance the DNA yield of microsporidium Ecytonucleospora hepatopenaei spores

Microsporidium Ecytonucleospora hepatopenaei (EHP) spores were purified from the hepatopancreas of Penaeus vannamei infected with EHP by percoll density gradient centrifugation and differential centrifugation. The EHP spores contain a thick chitin wall and might not rupture using the routine DNA extraction protocol. In this study, three enzymes were used, including chitinase, proteinase K, and DNase I. Chitinase or proteinase K digestions caused weakened fluorescence of chitin showing by a blurred edge of EHP spores stained with calcofluor white under a fluorescence microscope. Different combinations of these enzymes followed by DNA extraction with phenol–chloroform from EHP spores showed significant increases in the copy number of the EHP SSU gene per spore. The combination of the chitinase and proteinase K treatments resulted 4.46 ± 1.07 copies/spore detected, which is 31.6 ± 20.7 folds of no treatment groups, accounting to (55.7 ± 13.4)% of the total copies of the gene in the spore. The additional treatment with chitinase to the conventional extraction protocol with a proteinase K digestion step for feces and hepatopancreas samples of P. vannamei resulted in a significant difference in EHP copies in the DNA of (83.8 ± 64.1)% and (55.3 ± 88.0)% increases. The study proved that chitinase and proteinase K treatment enhance the DNA extraction from microsporidian spores resulting in high yield.

Evaluating The Diagnostic Value Of Red Cell Distribution Width (RDW) In Identifying Iron Deficiency Anemia: A Retrospective Analysis

Background: Iron Deficiency Anemia (IDA) is the most prevalent form of anemia globally, with a significant impact on public health, especially in low-resource settings like Bangladesh. Accurate and early diagnosis of IDA is essential for effective treatment. Red Cell Distribution Width (RDW) has emerged as a promising, accessible diagnostic tool for IDA, yet its utility remains underexplored in this context. Methods: This retrospective cross-sectional study, conducted between July 2022 and July 2024 at Addin Barrister Rafique-ul Huq Hospital in Bangladesh, included 106 patients diagnosed with microcytic hypochromic anemia. Key hematological parameters, including hemoglobin, RBC count, MCV, and RDW, were analyzed. Correlation analyses were performed to assess the relationships between RDW and other parameters such as hemoglobin, MCV, RBC count, and Erythrocyte Sedimentation Rate (ESR). Results: RDW was elevated (&gt;14.5%) in 92.45% of the participants, with a mean RDW of 17.59±3.33%. Significant negative correlations were observed between RDW and hemoglobin (r = -0.435, p &lt; 0.001) and between RDW and MCV (r = -0.664, p &lt; 0.001). RDW showed positive correlations with RBC count (r = 0.315, p = 0.001) and ESR (r = 0.225, p = 0.041). The study confirmed RDW’s strong diagnostic value in distinguishing IDA, particularly in resource-constrained settings. Conclusion: RDW, combined with other hematological parameters, is a sensitive and reliable marker for diagnosing IDA. Its incorporation into routine diagnostic protocols could significantly enhance IDA detection and management in low-resource settings like Bangladesh.

EVALUATION OF REMOTE CHECK IN CHILDREN WITH COCHLEAR IMPLANTS

ObjectiveThis study evaluates the feasibility of reducing in-person appointments by utilizing Remote Check (RC) in pediatric cochlear implant (CI) recipients and investigates the most suitable age range for implementing RC. MethodsFifty-eight pediatric CI recipients, aged 1 to 17 years, were evaluated using a web-based RC application by audiologists. Most (84.5%) used verbal communication. Inclusion criteria required stable map status, caregiver tech proficiency, and a compatible phone. ResultsRemote assessments were beneficial, with 89.7% of participants successfully completing RC. Aided threshold tests (ATT) were completed in 73.2% of participants, while speech-in-noise tests (Digit Triplet Test, DTT) were completed in 47.3%. Children as young as 4 successfully completed ATT, while a minimum age of 7 was required for DTT. A significant relationship between age and test success was demonstrated. Verbally communicative children had higher completion success rates. Parental education was significantly correlated with DTT success, with higher success rates in children whose parents had undergraduate degrees. With RC use, 69% of children did not need clinic visits, emphasizing the need for personalized and remote support for optimal cochlear implant performance. In-person appointments were primarily required due to concerns about the duration of device usage. ConclusionThe study confirms the effectiveness and feasibility of using RC in pediatric CI users, even as young as one year old with stable aided hearing. The findings advocate for the integration of RC into routine follow-up protocols, offering a more accessible option for enhanced patient care.

The Importance of Cited-1 and HIF-1α Immune Activity of Granulosa Cells in IVF Treatment

Aim: The aim of this study was to investigate the Cited-1 and HIF-1α immune activity in granulosa cells in follicular development in patients who underwent IVF for infertility. Materials and Methods: This study was conducted on 40 patients who were admitted to the assisted reproductive program with the complaint of infertility at the Gazi Yaşargil Training and Research Hospital Obstetrics and Gynecology Clinic IVF center between January 2022 and November 2022 and had primary or secondary infertility while starting the Ovum-Pick-Up (OPU) procedure. The fluid containing the granulosa cells was centrifuged at 3000 rpm for 10 min. The samples were fixed and processed for routine paraffine wax tissue embedding protocol. Sections were taken from paraffin blocks and immune stained with Cited-1 and HIF-1α. The preparations were examined under the microscope. Results: HIF-1α expression was positive in membrane of granulosa cells. The nuclei were apoptotic and pyknotic. Cited1 expression was positive in membrane of granulosa cells. the cells were pyknotic. Conclusion: The high level of HIF-1α immunopositivity and negative Cited1 immunoreactivity in the immunohistochemical staining after the granulosa cells around the oocytes collected from female patients admitted to the IVF clinic and diagnosed with infertility showed that granulosa cell viability may be important on oocyte quality.

Dietary Interventions in Irritable Bowel Syndrome: A Systematic Review of Clinical Outcomes, Microbiota Changes, and Inflammatory Markers.

This systematic review synthesizes findings from 12 studies to evaluate the effectiveness of dietary interventions in managing irritable bowel syndrome (IBS), with a focus on low-FODMAP (fermentable oligosaccharides, disaccharides, monosaccharides, and polyols) diets, probiotics, and prebiotics. The review rigorously follows the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines and includes studies selected through comprehensive database searches. In adults diagnosed with IBS, this review assesses how effective dietary interventions, specifically low-FODMAP diets, probiotics, and prebiotics, are compared to standard management or placebo in improving clinical outcomes, modifying gut microbiota composition, and reducing inflammatory markers. Our analysis reveals that low-FODMAP diets consistently alleviate IBS symptoms and improve quality of life. However, the effectiveness of probiotics and prebiotics varies, with outcomes dependent on specific strains and individual patient microbiota profiles. The studies demonstrate significant improvements in gastrointestinal symptoms and microbiota composition, highlighting the potential of dietary strategies to beneficially modify gut health. However, the research points to the necessity of personalizing dietary approaches based on individual responses and microbiota profiles to optimize treatment efficacy. The risk of bias was assessed using the Cochrane risk-of-bias 2 tool for randomized controlled trials (RCTs) and the AMSTAR 2 tool for systematic reviews, with varying degrees of bias across the studies. This review identifies gaps in the long-term efficacy of these interventions and calls for more extensive trials to explore their sustained impacts. Our findings suggest that dietary management should be integrated into routine IBS treatment protocols and emphasize the need for further research to establish standardized dietary recommendations tailored to patient-specific characteristics.

Role of diffusion-weighted imaging in the diagnosis of pituitary region tumors.

This study aimed to assess the role of Diffusion-Weighted Imaging (DWI) in routine pituitary Magnetic Resonance Imaging (MRI) protocols for distinguishing sellar and parasellar tumors, addressing the lack of clear guidelines in contemporary literature. A retrospective analysis of 242 pituitary MRI scans with DWI sequences was conducted in a single-center study using a 1.5T scanner and standard DWI sequence parameters. Measurements of both absolute and relative mean apparent diffusion coefficient (ADC) values, along with minimal ADC values within tumors, were performed. The adopted region of interest (ROI) based method used for these measurements was validated. Invasive pituitary adenomas exhibited significantly lower min ADC and min rADC than meningiomas, with optimal cut-off points of 0.64 (sensitivity 73%, specificity 82%) and 0.78 (sensitivity 73%, specificity 89%), respectively. Post-hemorrhagic pituitary adenomas demonstrated lower ADC values than adamantinomatous craniopharyngiomas, with an AUC of 0.893 for min rADC = 1.07, and Rathke's Cleft Cysts with mucous content, AUC 0.8 for min rADC = 1.01. Specific differentiation with high sensitivity and specificity based on diffusion parameters was observed for these tumor groups. Cystic pituitary non-functional adenomas obtained significantly lower ADC values compared to the adamantinomatous type of craniopharyngiomas and serous Rathke's Cleft Cysts (AUC up to 0.942). The study concludes that integrating DWI into routine pituitary MRI protocols enhances diagnostic accuracy in distinguishing sellar and parasellar tumors. The short scan time of one minute makes DWI a valuable and precise tool, supporting its recommendation as a standard component of pituitary MRI examinations.

Stress Echocardiography in the Follow-Up of Young Patients with Repaired Aortic Coarctation.

Background: Aortic coarctation (CoA) is a congenital heart disease affecting 5-8% of patients, with long-term complications persisting despite successful correction. Stress echocardiography (SE) is increasingly used for evaluating cardiac function under stress, yet its role in repaired CoA remains under-explored. Objective: This study aimed to assess the predictive value of SE and myocardial strain in repaired CoA patients with a history of hypertension without significant gradients or with borderline gradients at rest. Methods: Between June 2020 and March 2024, we enrolled 35 consecutive CoA patients with successful repairs and either a history of hypertension or borderline Doppler gradients. Baseline and peak exercise echocardiographic measurements, including left ventricular mass index (LVMi) and global longitudinal strain (LVGLS), were recorded. Patients were followed for up to 4 years. Results: At baseline, the positive SE group had higher systolic blood pressure (SBP) and diastolic blood pressure (DBP) compared to the negative SE group. The positive SE group also exhibited significantly higher basal and peak trans-isthmic gradients. Positive SE was found in 45.7% of patients, with 68.7% of these requiring re-intervention during follow-up. A peak trans-isthmic gradient > 61 mmHg during exercise predicted recoarctation with 100% sensitivity and 71% specificity (AUC = 0.836, p < 0.004). Conclusions: SE identifies at-risk patients post-CoA repair, aiding in early intervention. A peak trans-isthmic gradient > 61 mmHg during exercise is a strong predictor of recoarctation. These findings support incorporating SE into routine follow-up protocols for CoA patients, particularly those with a history of hypertension and borderline gradients, to improve long-term outcomes and quality of life.