Serum Collection Research Articles

P-2215. Intestinal microbiome and metabolome fluctuations in patients receiving intensive chemotherapy for AML: Results from a pilot, prospective observational study

Abstract Background Loss of a diverse intestinal microbiome, particularly through the depletion of anaerobic bacteria, is associated with poor outcomes in people undergoing treatment for leukemia. We present preliminary results from a pilot, prospective observational study characterizing longitudinal changes in the fecal microbiome and metabolome in patients undergoing intensive chemotherapy for newly diagnosed acute myeloid leukemia (AML). Methods We recruited 10 patients with newly diagnosed AML who were hospitalized for intensive chemotherapy. The subjects underwent daily serum and stool collections during the admission for induction chemotherapy and periodic serum and stool collections during subsequent admissions. Metabolome profiling was conducted by targeted GC- and LC-mass spectrometry of serum and stool specimens, and fecal microbiome composition was determined by Shotgun metagenomic sequencing. Clinical characteristics, including responses to chemotherapy and development of infections, were monitored. Results Metagenomic sequencing demonstrated marked variations in microbiome compositions between patients in our cohort. However, microbiome compositions and diversities remained stable within patients who maintained a higher prevalence of obligate anaerobes belonging to the Lachnospiraceae and Bacteroidaceae families. Moreover, despite stable microbiome compositions, we detected large fluctuations in fecal metabolite concentrations, particularly among secondary bile acids and conjugated and unconjugated bile acids, over the course of induction chemotherapy. Our results suggest that preservation of intestinal anaerobes enhances the stability of the microbiome over time, and the metabolic output of an individual’s microbiome is substantially impacted during cancer treatment. Conclusion Studying dynamics of the intestinal microbiome and metabolome during cancer therapy will help identify signals associated with microbiome diversity or disruption. We also plan to use broad targeted and untargeted platforms to identify metabolites in stool or serum that can be used as biomarkers to correlate with microbiome compositions and to identify patients who are at risk for adverse clinical outcomes associated with disrupted microbiomes. Disclosures All Authors: No reported disclosures

P-2134. Development and Validation of a Quantitative PCR Assay for the Detection of Mucorales in Human Serum Specimens

Abstract Background Mucormycosis are diseases with high morbidity and mortality caused by fungal species belonging to the order Mucorales. The intended use of a Mucorales qPCR assay is as an adjunct to microbiological, clinical, and radiologic means of diagnosing invasive mucormycosis (IM). Serum collection is a non-invasive alternative to BAL and biopsy. Our design process included sequences from all species within the Mucorales genera: Cunninghamella, Lichtheimia, Absidia, Apophysomyces, Mucor, Rhizomucor, Rhizopus and Saksenaea. The Mucorales qPCR assay is designed to detect and quantify Mucorales DNA extracted from human serum. Methods Primers and probes were designed to target the 18S regions of Mucorales species. Nucleic acid was extracted using the MagMAX™ Cell-Free DNA Isolation Kit and KingFisher™ Flex system. Amplification and detection were performed using TaqMan™ Fast Universal Master Mix and the Applied Biosystems™ QuantStudio 7 Pro. Quantification was performed using Rhizopus oryzae target-bearing linearized plasmid standards with results reported in copies/mL. Analytical validation was performed by spiking negative human serum with four different Mucorales species’ DNA: Rhizopus oryzae, Rhizomucor miehei, Lichtheimia corymbifera, and Cunninghamella bertholletiae. Results The limit of detection was 181 copies/mL, and the lower limit of quantification was determined to be 253 copies/mL as shown in Table 1. Linear regression of dilution data produced an R2=0.997, slope=1.029, and y-intercept=0.212 as show in Figure 1. The Mucorales target detection was 100% (60 of 60) of blinded positive samples with all observed log10 copies/mL results within ±0.5 log10 copies of expected values as shown in Table 2. All (20 of 20) blinded negative samples were negative for Mucorales and positive for Internal Control (IC). Intra-assay precision ranged from 5 – 22 %CV and inter-assay precision ranged from 12 – 17 %CV as Shown in Table 3. Conclusion The validated assay described here demonstrates excellent sensitivity, linearity, precision, and accuracy, providing a reliable means of detecting Mucorales DNA in human serum. This assay represents a non-invasive, rapid, and sensitive method to guide clinical decisions on patient populations at high risk of IM. Disclosures Jerod Davidson, B.A., Eurofins Viracor: Employee James Grantham, B.S., Eurofins Viracor: Employee Clayton Thomas, B.S., Eurofins Viracor: Employee Jamie Nutt, n/a, Eurofins Viracor: Employee Pam Morris, M.S., PMP, Eurofins Viracor: Employee Zachary Kockler, PhD, Eurofins Viracor: Employee Steve Kleiboeker, PhD, Eurofins: employee|Eurofins: Stocks/Bonds (Public Company)

P-1972. SARS-CoV-2 Vaccine–Induced IgG4 Response Negatively Correlates with Neutralizing Antibodies and Fc Effector Functions

Abstract Background IgG4, the least abundant human IgG subtype, increases after repetitive exposure to some antigens, and may induce immune tolerance. Studies have found that repeat mRNA SARS-CoV-2 vaccination leads to large proportional increases in spike (S)-specific IgG4. By contrast, increased IgG4 has not been observed following repeat vaccination with recombinant SARS-CoV-2 S (rS) protein (NVX-CoV2373, Novavax). Whether vaccine-induced anti-S IgG4 might impair SARS-CoV-2 immunity remains unknown.Figure:Serum anti-Spike IgG1 (A), IgG3 (B), and IgG4 (C) concentrations (ng/mL) measured by ELISA were correlated with neutralizing antibody titers (MN50) measured by infectious virus microneutralization assay. Methods Sera were collected from study participants in 2019nCoV-307 (NCT05463068) and 2019nCoV-301 (NCT04611802), including 2 groups from 2019nCoV-307 who received 3 homologous mRNA doses followed by 1 NVX-CoV2373 dose (mRNA-1273, Moderna, n=10; or BNT162b2, Pfizer, n=10), and a 3rd group from 2019nCoV-301 who received 4 homologous NVX-CoV2373 doses (n=18). Sera collected ≥6 months after the last dose in the first 2 groups and ∼4 weeks after the last dose in the 3rd group were assessed for neutralizing antibodies (nAb), IgG subclass profiles (anti-S total IgG, IgG1, IgG2, IgG3, and IgG4), and Fc effector activities (cell-based ADCP assay; surrogate ADCC by FcγRIIIa binding; and ADCD by C1q binding) after repeated NVX-CoV2373 or a single NVX-CoV2373 dose following repeated mRNA vaccine.Table:Summary of correlations between IgG1, IgG3, and IgG4 with neutralizing antibody titers (ancestral) and Fc effector functions in 2019nCoV-301 and 2019nCoV-307 studies. Results Increased nAb titers after NVX-CoV2373 were observed irrespective of the prior vaccine background, whereas increased IgG4 levels were only observed in recipients of mRNA vaccines. Spearman's rank correlation between nAb and Fc functions found significant positive correlations with anti-S IgG1 and IgG3 (Figure A, B), and significant negative correlation with anti-S IgG4 (Figure C) antibody responses (Table). Conclusion These are the first data to demonstrate the negative correlation of increased SARS-CoV-2 vaccine–induced anti-S IgG4 levels and nAb titers and Fc effector functions. The limitations of our data include small sample sizes, and potential differences attributable to vaccination and serum collection intervals. IgG4-mediated suboptimal nAb and reduced Fc effector function could reduce the vaccine effectiveness of the SARS-CoV-2 response, though more work is needed to understand its impact on the optimization of vaccine platform choice. Disclosures Raj Kalkeri, PhD, Novavax: Employee|Novavax: Stocks/Bonds (Public Company) Mingzhu Zhu, n/a, Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company) Shane Cloney-Clark, n/a, Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company) Anand Parekh, M.Sc., Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company) Drew Gorinson, B.S., Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company) Rongman Cai, PhD, Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company) Soham Mahato, PhD, Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company) Anthony M. Marchese, PhD, Novavax Inc: Employee|Novavax Inc: Stocks/Bonds (Public Company) Louis F. Fries, III, MD, Novavax, Inc.: contractor for Novavax|Novavax, Inc.: Stocks/Bonds (Public Company) Lisa M. Dunkle, MD, Novavax, Inc: Employee|Novavax, Inc: Ownership Interest|Novavax, Inc: Stocks/Bonds (Public Company) Amy W. Chung, PhD, Novavax: Grant/Research Support Joyce S. Plested, n/a, Novavax, Inc.: employee|Novavax, Inc.: Stocks/Bonds (Public Company)

Novel Method for Routine Ultrasound-Guided Serum Collection for Biomarker Analysis Around the Knee Joint

Background Biomarkers are essential tools in modern medicine, allowing stratification and monitoring of clinical care and treatment response. While systemic blood biomarkers, typically collected from the antecubital vein (AV), are widely used, their sensitivity for joint-specific pathologies such as osteoarthritis (OA) may be limited due to systemic dilution. At present, no serum biomarker reliably reflects the microenvironment of an affected joint in clinical practice. Although synovial fluid (SF) assessment can provide insights into localised pathology and are clinically used for diagnosis in crystal arthropathies and joint infections, their collection is invasive, painful, and carries risks, including infection, making repeated sampling impractical, limiting utility for monitoring treatment responses. This study introduces a novel ultrasound-guided venous sampling technique targeting the saphenous vein (SV) proximal to the knee joint as a less invasive alternative to SF aspiration, hypothesising that it may better reflect the joint-specific microenvironment. Methods A standardised gel model was used to train medically qualified researchers in ultrasound-guided venepuncture of vescles around 2-15 mm in diameter. Subsequently, 32 participants consented to blood sampling from the SV above the knee, with a proximally applied tourniquet to dilate vein diameter for easier venepuncture collection. Results The technique achieved serum collection in over 80% of consented individuals with minimal adverse effects (including n=2 minor bruising and n=1 transient nerve irritation). Key procedural insights included optimal site selection, appropriate pressure application, and effective tourniquet use. Discussion This method demonstrates feasibility, acceptability, and the potential for more localised sample collection, advancing biomarker research. Further validation, including paired SF comparisons, is required to confirm diagnostic utility and develop therapeutic strategies for joint-specific conditions.

Longitudinal analysis of SARS-CoV-2 IgG antibody durability in Puerto Rico

Understanding the dynamics of antibody responses following vaccination and SARS-CoV-2 infection is important for informing effective vaccination strategies and other public health interventions. This study investigates SARS-CoV-2 antibody dynamics in a Puerto Rican cohort, analyzing how IgG levels vary by vaccination status and previous infection. We assess waning immunity and the distribution of hybrid immunity with the aim to inform public health strategies and vaccination programs in Puerto Rico and similar settings. We conducted a prospective, longitudinal cohort study to identify SARS-CoV-2 infections and related outcomes in Ponce, Puerto Rico, from June 2020–August 2022. Participants provided self-collected nasal swabs every week and serum every six months for RT-PCR and IgG testing, respectively. IgG reactivity against nucleocapsid (N) antigens, which generally indicate previous infection, and spike (S1) and receptor-binding domain (RBD) antigens, which indicate history of either infection or vaccination, was assessed using the Luminex Corporation xMAP® SARS-CoV-2 Multi-Antigen IgG Assay. Prior infection was defined by positive RT-PCRs, categorized by the predominant circulating SARS-CoV-2 variant at the event time. Demographic information, medical history, and COVID-19 vaccination history were collected through standardized questionnaires. Of 882 participants included in our analysis, 34.0% experienced at least one SARS-CoV-2 infection, with most (78.7%) occurring during the Omicron wave (December 2021 onwards). SARS-CoV-2 antibody prevalence increased over time, reaching 98.4% by the final serum collection, 67.0% attributable to vaccination alone, 1.6% from infection alone, and 31.4% from both. Regardless of prior infection status, RBD and S1 IgG levels gradually declined following two vaccine doses. A third dose boosted these antibody levels and showed a slower decline over time. N-antibody levels peaked during the Omicron surge and waned over time. Vaccination in individuals with prior SARS-CoV-2 infection elicited the highest and most durable antibody responses. N or S1 seropositivity was associated with lower odds of a subsequent positive PCR test during the Omicron period, with N antibodies showing a stronger association. By elucidating the differential decay of RBD and S1 antibodies following vaccination and the complexities of N-antibody response following infection, this study in a Puerto Rican cohort strengthens the foundation for developing targeted interventions and public health strategies.

Replacing serum with dried blood microsampling for pharmacokinetics, viral neutralisation and immunogenicity bioanalysis supporting future paediatric development of RSM01, a candidate respiratory syncytial virus neutralising monoclonal antibody

BackgroundVirus neutralising antibodies in serum are considered key correlates of protection for vaccines and monoclonal antibodies against respiratory syncytial virus (RSV). RSM01 is a novel, highly-potent, half-life-extended and fully-human monoclonal antibody candidate targeting the RSV prefusion F protein. Currently in Phase 1 development, RSM01 is primarily being developed to potentially provide an effective and affordable RSV prevention strategy in low- and middle-income countries. To evaluate the ability of dried blood collection to generate data sets and conclusions comparable to serum collection, we compared pharmacokinetics (PK) of RSM01, immunogenicity, and virus neutralisation for dried capillary blood samples with serum samples.MethodsRSM01 PK, anti-drug antibodies (ADA), and RSV-neutralising antibodies from the Phase 1 trial were analyzed and compared between matched serum and dried blood samples. Deming regression analysis was performed using baseline-corrected values to evaluate correlation between measurements in liquid serum versus dried blood.ResultsThe analysis showed good correlation (R2 > 0.95) between individual RSM01 concentrations measured in both serum and capillary blood. Analysis of RSM01 PK parameters in capillary blood yielded equivalent conclusions as from serum. A strong correlation (R2 > 0.95) was observed between RSV neutralising activity measured in both serum and capillary blood. In addition, RSV neutralising activity was correlated with RSM01 concentrations in both serum and capillary blood data sets. For ADA, individual sample results had 96% agreement (290/302) and overall participant ADA status had 93% agreement (52/56).ConclusionsBoth RSM01 concentrations and RSV neutralising activity showed a strong correlation between the serum and blood measurements. ADA measurements also had an agreement of > 90% for individual samples and overall participant status. Our results demonstrate that dried blood is a suitable specimen type for collection and evaluation in the RSM01 clinical development program and shows promise as a useful approach to reduce patient burden in clinical trials, particularly for infants in low- and middle-income countries.Trial RegistrationClinicaltrials.gov NCT05118386 November 12, 2021.

The role of neurofilaments light chain in discriminating behavioral variant frontotemporal dementia from non‐neurodegenerative mimics

AbstractBackgroundThe measurement of serum and cerebrospinal fluid (CSF) neurofilaments light chain (NfLs) has been proven promising in differentiating the behavioral variant frontotemporal dementia (bvFTD) from non‐neurodegenerative mimics, including primary psychiatric disorders and non‐progressive cognitive/behavioral changes. However, studies on this topic are based on clinical diagnosis, which remains challenging and potentially confounded by the overlapping clinical phenotypes. We investigated the role of NfLs in this field by classifying patients based on the presence/absence of pathological longitudinal brain volume changes.MethodWe prospectively recruited subjects with early‐onset (< 65 yo) mild cognitive/behavioral impairment suggestive of potential bvFTD. At baseline they underwent neuropsychological assessment, magnetic resonance imaging (MRI) brain imaging, and CSF and serum collection for NfLs measurement. They were followed‐up clinically and repeated the MRI scan at 18 months. Single‐subject yearly percentage brain volume change (PBVC/y) was calculated using the FMRIB Software Library tool SIENA and compared to normative values. Subjects with a PBVC/y exceeding the 95th normative percentile were classified as Progressive, in line with a bvFTD diagnosis. Otherwise, they were indicated as Non‐Progressive, suggestive of a non‐neurodegenerative condition. Differences in NfLs levels between Progressive and Non‐Progressive individuals and their predictive value were evaluated through non‐parametric tests and univariate logistic regression models.Result30 patients were recruited: 19 were classified as Non‐Progressive and 11 as Progressive. Both CSF and serum NfLs values were significantly higher in patients with atrophy progression compared to the stable ones (2992,27 vs 558,53 pg/dl, p < 0.001; 55,91 vs 13,47 pg/dl, p < 0.001). Higher serum NfLs levels at baseline were predictors of atrophy progression (p = 0.041, OR = 1.301), whereas only a trend of significance was seen for CSF NfLs (p = 0.055, OR = 1.006). A cut‐off of 21.5 pg/dl for serum NfLs differentiated the two groups with 81.82% sensitivity and 94.74% specificity (AUC = 0.949); a cut‐off of 972 pg/dl for CSF NfLs yielded discriminative sensitivity of 81.82% and specificity of 100% (AUC = 0.928).ConclusionWe confirmed that NfLs measurement in biological fluids at baseline is a reliable biomarker to discriminate between bvFTD patients and non‐neurodegenerative mimics, objectively distinguished based on longitudinal atrophy progression.

A survey of Canadian neurologists’ perspectives and preferences for laboratory reporting of CSF oligoclonal banding

Cerebrospinal fluid (CSF) oligoclonal banding (OCB) analysis aids in the diagnosis of multiple sclerosis (MS). Despite its clinical importance, there is profound variation in processes, reporting, and interpretation of CSF OCB and associated tests/indices across Canadian laboratories. This is likely due to the lack of clear, evidence-based recommendations on CSF OCB analysis processes and reporting. Here, we assessed the CSF OCB reporting needs and preferences of Canadian neurologists as a first step in clinical stakeholder engagement to aid in the development of CSF OCB reporting recommendations. A 16-question survey was sent to neurologists across Canada in January 2022, and it closed in March 2022. The survey included questions regarding location and length of clinical practice; preferred maximum time limit for paired CSF and serum samples; reporting preferences for CSF-specific OCB, banding patterns, and associated tests/indices; as well as the clinical utility of CSF OCB and associated tests/indices. Twenty-two neurologists from nine provinces participated, with a median practice length of 13years. Most (64%) preferred a 24-hour limit for paired serum and CSF sample collection. The majority (73%) favored a cutoff of≥2 CSF-specific bands for positivity, aligning with the 2017 McDonald criteria. Opinions varied on reporting the number of bands and listing specific conditions in the interpretive comments. Some highlighted the need for further research on band count interpretation and its clinical implications. All respondents found CSF OCB results useful, with 64% valuing it more than other CSF tests for MS evaluation. Our survey reveals diverse preferences among Canadian neurologists for CSF OCB reporting. Stakeholder engagement and further research are crucial for standardized, improved MS diagnostic practices.

Application of Integrated Optical Density in Evaluating Insulin Expression in the Endocrine Pancreas During Chronic Ethanol Exposure and β-Carotene Supplementation: A Novel Approach Utilizing Artificial Intelligence.

β-carotene is an essential antioxidant, providing protection against type 2 diabetes mellitus, cardiovascular illnesses, obesity, and metabolic syndrome. This study investigates the impact of β-carotene on biochemical parameters and pancreatic insulin expression in mice exposed to ethanol. Thirty-six C57BL/6 mice (Mus musculus) were divided into six groups: 1. C (control), 2. LA (3% alcohol dose), 3. MA (7% alcohol dose), 4. B (0.52 mg/kg body weight/day β-carotene), 5. LA+B (3% alcohol dose + 0.52 mg/kg body weight/day β-carotene), and 6. MA+B (7% alcohol dose plus 0.52 mg/kg body weight/day β-carotene). After 28 days, the animals were euthanized for serum and pancreatic tissue collection. Biochemical analysis and pancreatic insulin expression were performed. One-way ANOVA was used. The B, LA+B, and MA+B groups improved insulin levels and decreased HOMA-β versus the C group, with the LA+B and MA+B groups also showing lower ADH and ALDH levels than their nonsupplemented counterparts (p < 0.05). The B, LA+B, and MA+B groups showed a greater β-cell mass area compared to the unsupplemented groups. Additionally, the LA+B and MA+B groups demonstrated significantly increased β-cell area and integrated optical density compared to the LA and MA groups, respectively (p < 0.001). In mice, β-cell loss led to increased glucose release due to decreased insulin levels. β-carotene appeared to mitigate ethanol's impact on these cells, resulting in reduced insulin degradation when integrated optical density was used. These findings suggest that antioxidant supplementation may be beneficial in treating ethanol-induced type 2 diabetes in animal models.

8665 Female Fertility In Patients With Classic And Non-Classic Congenital Adrenal Hyperplasia Under Modified-Release Hydrocortisone (MR-HC)

Abstract Disclosure: L. Tschaidse: None. H.F. Nowotny: None. M. Auer: None. C. Lottspeich: None. M. Bidlingmaier: None. J. Hawley: None. B.G. Keevil: None. N. Reisch: None. Background: Female patients with classic and non-classic (NC) CAH struggle with fertility, due to androgen excess, elevated progesterone and 17-hydroxyprogesterone (17OHP) levels, causing anovulation, impairment of endometrial development and menstrual irregularities. The typical approach to treating infertility often includes an increase in glucocorticoid (GC) dose. However, this poses a potential risk of side effects. Limited observational data on modified-release hydrocortisone (MR-HC), which better mimics the physiological diurnal rhythm of cortisol secretion, indicates improved fertility in female and male patients with classic CAH. Therefore, the aim of this study was to investigate fertility in women with classic and non-classic CAH before and after switching to MR-HC. Methods: 21 adult female patients with CAH (14 classic; 5 NC) with a median (range) age of 33.0 (30.0) years, premenopausal and without hormonal contraception, were enrolled in this prospective, observational, single-centre study. The data collection took place before (conventional GC preparation) and after dose-equivalent switching the medication to MR-HC. In addition to clinical parameters such as menstrual cycle and desire to conceive, the collection of morning serum and daily saliva profiles was conducted, measuring progesterone, 17OHP, androstenedione (A4), testosterone, 11-ketotestosterone (11KT) as well as 11-hydroxyandrostenedione (11OHA4). Results: Before study inclusion, 4/21 patients had an irregular cycle, 5/21 were amenorrhoeic and 10/21 patients had a desire to have children. There was no difference in median (range) hydrocortisone dose equivalent before and after switching to MR-HC (25.0 (50.0) vs. 25.0 (30.0), p=.441). Median (range) morning serum concentrations between 8-10 am were not different before and after switch to MR-HC for progesterone (5.9 (55.5) vs. 5.0 (58.8), p=.823), 17OHP (5.9 (362.1) vs. 5.7 (73.7), p=.280), A4 (2.9 (25.6) vs. 2.5 (6.9), p=.106), testosterone (0.8 (3.5) vs. 0.7 (1.5), p=.063), 11KT (0.4 (7.4) vs. 0.2 (2.6), p=.242) and 11OHA4 (0.8 (12.9) vs. 0.9 (10.4), p=.231). Concerning saliva day profiles, we found a significantly lower median (range) 17OHP level in early morning saliva (43.0 (526.9) vs. 19.4 (264.3), p=.049). 5/9 patients with menstrual irregularities reported menstrual regularisation within and 5/10 patients with a desire to have children became pregnant after switching to MR-HC. Conclusion: Our preliminary data show clinical improvement in fecundity and fertility under MR-HC in both women with classic and NC CAH without a dose increase of glucocorticoids. Under MR-HC replacement lower 17OHP concentrations in early morning saliva reflect better hormonal control over night contributing to ovulatory cycle regulation and improved fertility. Presentation: 6/1/2024

Can Peripheral Arterial Tonometry and Biomarkers Help Identify Women Who Will Have Progressively Worsening Hypertensive Disorders of Pregnancy?

This study aimed to (1) evaluate whether endothelial dysfunction, as measured by peripheral arterial tonometry (PAT) indices and biomarker (soluble fms-like tyrosine kinase-1 [sFLT], brain natriuretic peptide [BNP]) levels at 34 weeks gestation, can predict progression from nonsevere to severe hypertensive disorders of pregnancy (HDPs); and (2) develop a clinical risk model for prediction of progression from nonsevere to severe HDP. We prospectively enrolled patients with a singleton gestation carrying a nonsevere HDP diagnosis. Forty-five participants were enrolled for PAT evaluation and serum collection between 340/7 and 366/7 weeks. PAT indices (e.g., Augmentation Index normalized to a heart rate of 75 bpm [AI75]) and biomarker concentrations were assessed at enrollment. The primary outcome was progression from a nonsevere diagnosis in the late preterm period to a diagnosis of preeclampsia with severe features or superimposed preeclampsia. Statistical analyses included two-sample t-tests, Fisher's exact tests, and multivariate modeling. Thirteen subjects (30%) progressed to severe disease. No significant differences in mean PAT indices between the outcome groups were found. We found a significant difference in mean sFLT values between the two groups (p = 0.02, area under the curve [AUC] of 0.609), but not in mean BNP values. An endothelial dysfunction index (presence of fetal growth restriction, "high" AI75, and positive systolic blood pressure slope) discriminated between progression and nonprogression (p = 0.03, AUC of 0.707). sFLT level was a marker of progression from nonsevere to severe HDP. Further, a novel endothelial dysfunction index discriminated between progression and nonprogression to severe disease with good performance. · HDPs are important causes of morbidity and mortality.. · The sequelae of HDPs are not limited to pregnancy.. · Developing accurate tools to predict severe HDPs is of great clinical importance.. · Our index shows promising performance for predicting progression from nonsevere to severe HDPs..

The Effect of Hypnotherapy Methods on The Modification Scale of Menopause-Specific Quality of Life (Menqol) and Cortisol in Menopausal Women

A decrease in stressors will be followed by a decrease in cortisol, a basic principle used by researchers to apply hypnotherapy to menopausal women who experience complaints. Judging from treatment techniques, hypnotherapy is very far from pharmacological treatment. The role of the new instrument in measuring the quality of life of menopausal women uses a modified menopause quality of life scale. The general objective of this study is to prove the improvement of the scale of modification of menopausal quality of life with the influence of hypnotherapy through cortisol changes in menopausal women. The design of this study uses quasi-experimental, which is almost similar to the actual experimental method. The approach with the pretest-posttest control group was by comparing between two groups, namely the control group and the treatment group, and analyzed to see the effect of hypnotherapy on the quality of life of menopausal women through the MenQoL modification scale in accordance with the hypothesis that had been proposed previously. Both groups observed changes in the MenQoL modification scale through questionnaire collection and blood serum collection for cortisol examination (during the trance state cessation phase of the hypnotherapy group). After 4 weeks, both groups were again given hypnotherapy and healthy living education script (in the hypnotherapy group) and only given a healthy living education script (control group), then the cortisol test and the second MenQoL modification scale were repeated. The hypothesis is expected to have significant differences.

Clinical performance and application of novel serum collection “Ser-Col” device in the practice of laboratory diagnosis of infection diseases and several other immunochemical tests

Dried blood collection devices might be beneficial for diagnosing infectious diseases in areas far from the medical facilities and in lockdown situations. There are several reports on the efficacy of such applications for qualitative tests. Here we demonstrated the feasibility of a novel Ser-Col blood collection device as a standardized approach for qualitative and quantitative detection of infectious markers and several over immunochemical tests. In the current study, we included 395 adult participants, 191 men and 204 women, with a median age of 41years, as well as 75 children with a median age of 3years. Serological status was determined by testing serum samples for three groups of infection diseases: hepatitis A and C, SARS-CoV-2, and herpes family viruses, as well as for thyroid peroxidase (TPO), prolactin, vitamin B12, and folate. Blood collected on the Ser-Col device (Labonovum) was eluted using an automated system (SCAUT Ser-Col automation, Blok System Supply) and manually. Ser-Col results were compared with serum sampled via standard venipuncture considered as the reference. High correlation coefficients (r=0.95-0.99) were observed between serum samples collected with Ser-Col and via standard venipuncture for the following tests: anti-HCV, anti-SARS-CoV-2 IgG, anti-HSV-2 IgG, and anti-CMV IgM. Correlation coefficients between Ser-Col and standard venipuncture serum for anti-HSV-1 IgG, anti-CMV IgG, and anti-EBV tests were relatively low (r=0.73-0.77). Correlation coefficients for anti-TPO, prolactin, vitamin B12, and folate were also characterized with high values (r=0.97-0.99). High accuracy and quantitative correlation were demonstrated between Ser-Col and samples collected by standard venipuncture. Hence, the Ser-Col blood collection device should be considered as a promising alternative for blood collection, storage, and transportation in both adult and pediatric populations.

Evaluation of the clinical efficacy of Pediococcus acidilactici CCFM6432 in alleviating depression.

Aim: Accumulating evidence highlights the crucial role of the "gut-brain axis" and emphasizes the potential of dietary interventions to improve brain health through this pathway. This study assesses the effects of the probiotic Pediococcus acidilactici CCFM6432 on mood, sleep, and gastrointestinal function in patients with depressive disorder. Methods: This clinical trial is a randomized, placebo-controlled study (Registration: ChiCTR2300071025). It enrolled 39 adult patients diagnosed with depressive disorder, who were randomly assigned to either the placebo control group (n = 19) or the CCFM6432 intervention group (n = 20). The intervention period spanned four weeks. Assessments were conducted at both the beginning and end of the trial, including comprehensive questionnaire evaluations and the collection of serum and fecal samples. Results: In comparison to the placebo, treatment with CCFM6432 significantly decreased depression and anxiety scores, as well as ameliorated gastrointestinal dysfunction and poor sleep quality commonly associated with mood disorders. Microbiota analysis revealed an increase in species richness without notable changes in overall diversity, yet Pediococcus species was found to be more abundant post-treatment. Functional analysis indicated reduced activity in the NOD-like receptor signaling pathway, suggesting anti-inflammatory effects induced by the probiotic. Metabolomic profiling identified elevated levels of fecal lactic acid, which correlated with lower Hospital Anxiety and Depression Scale (HADS) scores, thereby linking probiotic metabolism to mood enhancement. Conclusion: These findings imply that CCFM6432 may improve brain function by modulating gut microbiota and their mediated immune homeostasis, underscoring its potential as an adjunctive treatment for mental disorders.

Characterization of the SARS-CoV-2 antibody landscape in Norway in the late summer of 2022: high seroprevalence in all age groups with patterns of primary Omicron infection in children and hybrid immunity in adults

BackgroundAccording to Norwegian registries, 91% of individuals ≥ 16 years had received ≥ 1 dose of COVID-19 vaccine by mid-July 2022, whereas less than 2% of children < 12 years were vaccinated. Confirmed COVID-19 was reported for 27% of the population, but relaxation of testing lead to substantial underreporting. We have characterized the humoral immunity to SARS-CoV-2 in Norway in the late summer of 2022 by estimating the seroprevalence and identifying antibody profiles based on reactivity to Wuhan or Omicron-like viruses in a nationwide cross-sectional collection of residual sera, and validated our findings using cohort sera.Methods1,914 anonymized convenience sera and 243 NorFlu-cohort sera previously collected from the Oslo-area with reported infection and vaccination status were analyzed for antibodies against spike, the receptor-binding domain (RBD) of the ancestral Wuhan strain and Omicron BA.2 RBD, and nucleocapsid (N). Samples were also tested for antibodies inhibiting RBD-ACE2 interaction. Neutralization assays were performed on subsets of residual sera against B.1, BA.2, XBB.1.5 and BQ.1.1.ResultsThe national seroprevalence estimate from vaccination and/or infection was 99.1% (95% CrI 97.0-100.0%) based on Wuhan (spike_W and RBD_W) and RBD_BA2 antibodies. Sera from children < 12 years had 2.2 times higher levels of antibodies against RBD_BA2 than RBD_W and their seroprevalence estimate showed a 14.4 percentage points increase when also including anti-RBD_BA2 antibodies compared to Wuhan-antibodies alone. 50.3% (95% CI 45.0-55.5%) of residual sera from children and 38.1% (95% CI 36.0-40.4%) of all residual sera were positive for anti-N-antibodies. By combining measurements of binding- and ACE2-RBD-interaction-inhibiting antibodies, reactivity profiles indicative of infection and vaccination history were identified and validated using cohort sera. Residual sera with a profile indicative of hybrid immunity were able to neutralize newer Omicron variants XBB.1.5 and BQ.1.1.ConclusionsBy late summer of 2022, most of the Norwegian population had antibodies to SARS-CoV-2, and almost all children had been infected. Antibody profiles indicated that children mostly had experienced a primary Omicron infection, while hybrid immunity was common among adults. The finding that sera displaying hybrid immunity could neutralize newer Omicron variants indicates that Wuhan-like priming of the immune response did not have a harmful imprinting effect and that infections induce cross-reacting antibodies against future variants.

Using Dried Blood Spots to Quantitatively Detect Anti-SARS-CoV-2 IgG Antibodies by ELISA: A Validation Study.

SARS-CoV-2 serological testing is useful to determine seroprevalence, epidemiological trends, and the extent of transmission. The collection and transport of serum samples can be logistically challenging, especially in remote underserved areas. Dried blood spots (DBSs) would allow easier sample collection and logistical handling compared with standard serum collection, particularly for extensive and repeated SARS-CoV-2 serosurveys. We evaluated the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the IgG ELISA (Wantai, Beijing, China) using DBSs against sera for the quantitative detection of SARS-CoV-2 IgG antibody. The IgG ELISA was used to test paired sera and DBSs obtained from individuals with recent virologically confirmed COVID-19 illness and banked paired sera and DBSs collected before the COVID-19 pandemic. We found that 100/100 (100%) seropositive samples were positive using DBSs, and 193/194 (99%) seronegative samples were negative using DBSs. Compared with sera, the DBS method had a 100% sensitivity, 99% specificity, 99% PPV, and 100% NPV. Use of DBSs for SARS-CoV-2 household or population serosurveys may be considered in situations with limitations in sample collection, shipment, and storage.

Modeling hidden hunger in toddlers to determine the most influential micronutrients in stunting

Background: Risk factors that directly influence the incidence of stunting are the level of macronutrient and micronutrient intake. Micronutrient deficiencies cause about 1.1 million of the 3.1 million annual child deaths. This condition leads to hidden hunger, a condition of insufficient intake of micronutrients (especially iron, zinc, and iodine deficiencies). This study aimed to analyze hidden hunger in stunted and non-stunting toddlers with a multivariate model. Methods: The method used was a case-control study. A total of 71 toddlers were taken as respondents for the stunting group and 71 toddlers for the non-stunting group. The study used data collection was 2×24 hour food recall, FFQ, and blood serum collection to check zinc deficiency, iodine, hemoglobin, and urine tests. For data analysis, multivariate logistic regression and then bivariate analysis were used. Using multivariate modeling, and consideration of OR changes, the final model was obtained, including the variables of iron intake, zinc intake, iodine intake, zinc deficiency, anemia status, and hidden hunger had a significant relationship with the incidence of stunting in toddlers. Results: Toddlers with stunting were much more likely to have inadequate iron (92.95%) and zinc intake (91.54%) compared to the non-stunting group (78.87% and 77.46% respectively). A significantly higher proportion of the stunting group (40.84%) had anemia compared to the non-stunting group (5.63%). Unlike iron, zinc, and anemia, there wasn't a statistically significant relationship between iodine deficiency (p = 0.459) or hidden hunger (p = 0.058) and stunting. The results of the multivariate analysis suggest that iron intake, anemia status, and zinc deficiency are all important risk factors for stunting in toddlers. The anemia status variable was the most dominant cause of stunting because it had the highest OR value of 41.733. Conclusion: The strongest predictor was anemia status, followed by iron intake and zinc deficiency

Effects silymarin and rosuvastatin on amyloid-carriers level in dyslipidemic Alzheimer’s patients: A double-blind placebo-controlled randomized clinical trial

The production/excretion rate of Amyloid-β (Aβ) is the basis of the plaque burden in alzheimer's disease (AD), which depends on both central and peripheral clearance. In this study, the effect of silymarin and rosuvastatin on serum markers and clinical outcomes in dyslipidemic AD patients was investigated. Participants (n=36) were randomized to silymarin (140 mg), placebo, and rosuvastatin 10 mg orally three times a day for 6 months. Serum collection and clinical outcome tests were performed at baseline and after completion of treatment. Lipid profile markers, oxidative stress markers, Aβ1-42/Aβ1-40 ratio, and Soluble Low-density lipoprotein receptor-Related Protein-1 (sLRP1)/Soluble Receptor for Advanced Glycation End Products (sRAGE) ratio were measured. There was a statistically significant increase in Δ-high density lipoprotein (ΔHDL) between silymarin and placebo (P<0.000) and also between rosuvastatin and placebo (p=0.044). The level of Δ-triglycerides (ΔTG) in the silymarin group has a significant decrease compared to both the placebo and the rosuvastatin group (p<0.000 and p=0.036, respectively). The Δ-superoxide dismutase (ΔSOD) level in the silymarin group compared to placebo and rosuvastatin had a significant increase (p<0.000 and p=0.008, respectively). The ΔAβ1-42/Aβ1-40 in the silymarin group compared to both the placebo and rosuvastatin groups had a significant increase (p<0.05). There was an inverse relationship between ΔTG and ΔAβ1-42/Aβ1-40 (p=-0.493 and p=0.004). ΔAβ1-42/Aβ1-40 has a direct statistical relationship with ΔSOD marker (p=0.388 and p=0.031). Also, there was a direct correlation between the level of ΔAβ1-42/Aβ1-40 and ΔsLRP1/sRAGE (p=0.491 and p=0.005). Our study showed the relationship between plasma lipids, especially ΔTG and ΔHDL, with ΔAβ1-42/Aβ1-40 in dyslipidemic AD patients, and modulation of these lipid factors can be used to monitor the response to treatments.

Infliximab serum concentrations and disease activity in perianal fistulizing Crohn's disease: a cross-sectional study.

Several studies associate the presence of higher serum concentrations of infliximab (IFX) with fistula healing in perianal Crohn's disease (CD). This study aimed to evaluate serum IFX concentrations in patients with perianal fistulizing CD (PFCD) in the presence or absence of general, clinical, and radiological activities. This was a cross-sectional study in patients with PFCD during maintenance treatment with IFX from two centers. Serum IFX concentrations were measured before their next infusion and anal fistulas were evaluated by clinical examination and magnetic resonance imaging (MRI), whenever possible, performed 90days before or after serum collection. According to clinical scores, radiological activity, and disease markers, patients were classified as in remission or active disease.Mean serum IFX concentrations were compared between the groups. Thirty-eight patients with PFCD were included. Demographic characteristics were similar in patients with remission or active disease. The overall mean serum IFX concentration of the entire sample (n = 38) was 5.21 ± 4.75 μg/mL (median 3.63; IQR 1.44-8.82). Serum IFX levels were 6.25 ± 5.34 μg/mL (median 3.62; IQR 1.95-11.03) in the 23 (60.5%) patients in remission and 3.63 ± 3.24 μg/mL (median 3.63; IQR 1.32-6.43; p = 0.226) in the 15 (39 .5%) who presented active disease. When evaluating general, clinical, and radiological activity of PFCD, and deep remission in isolation, no statistical difference between the groups was observed (p = 0.226, p = 0.418, p = 0.126, and p = 0.232, respectively). The 13 (34.2%) patients with an optimized dose of IFX had significantly higher serum concentrations than the remaining 25 (65.8%) with a standard dose: 8.33 ± 4.41 μg/mL (median 8.36; IQR 3.82-11.20) vs. 3.59 ± 4.13 μg/mL (median 1.97; IQR 1.18-3.85)-p = 0.002. Patients in remission and with an optimized IFX dose had significantly higher serum IFX concentrations than those with a standard dose (p = 0.006), whereas no significant difference was observed among those with active disease (p = 0.083). There were no differences in IFX serum concentrations in patients with clinical or radiological active PFCD as compared with those in remission. Patients with an optimized IFX dose had significantly higher serum concentrations than those with a standard dose. Patients in remission and with an optimized IFX dose had significantly higher serum concentrations than those with a standard dose.

Association of Serum Metabolites with Serum Indices and Preanalytical Factors of Biobanked Serum Samples.

Background: Serum indices (hemolysis, icterus, and lipemia; HIL) are known to impact clinical chemistry assay results. This study aimed to investigate the impact of HIL indices on serum metabolite profiles and the association of serum metabolite levels with pre-analytical factors of serum samples. Methods: A cohort of serum samples (n = 12,196) from the Korean Genome and Epidemiology Study (KoGES) was analyzed for HIL indices and the pre-analytical variables (SPRECs) which were generated in the process of serum collection. We further performed targeted metabolomics on a subset comprising hemolyzed (n = 60), icteric (n = 60), lipemic (n = 60) groups, and a common control group of non-HIL samples (n = 60) using the Absolute IDQ p180 kit. Results: We found 22 clinical chemistry analytes significantly associated with hemolysis, 25 with icterus, and 24 with lipemia (p < 0.0001). Serum metabolites (n = 27) were associated with all of hemolysis, icterus, and lipemia (p < 0.05). The PC ae C36 2 had exhibited a significant association with pre-analytical factors corresponding to the third (pre-centrifugation delay between processing) and sixth (post-centrifugation) elements of the SPREC. Conclusions: This study showed the association of the serum index and pre-analytical factors with serum metabolite profiles. In addition, the association of pre-analytical factors with serum metabolite concentrations would corroborate the utility of SPRECs for the quality control of biobanked serum samples.