Combined Screening Research Articles

Abstract 17407: A Small-molecule Screening Strategy to Promote Cardiomyocyte Proliferation That Protects the Heart From Ischemic Injury

Restoring the ability of adult cardiac myocytes (ACMs) to proliferate and promote heart regeneration have been proposed as a promising therapeutic strategy. We have demonstrated that selective depletion of H3K9me3 by lysine demethylase 4D (KDM4D) overexpression increases ACM cell cycle activity, but proliferation was limited. To improve this, we performed a small-molecule screening in combination with KDM4D on neonatal rat cardiomyocytes (NRVMs), using a library of kinase inhibitors targeting different signaling pathways. Our study revealed that compounds targeting the PI3K and JAK signaling pathways significantly potentiated the effects of KDM4D and promote NRVM proliferation in vitro. Specifically, a JAK inhibitor (JAKi2) in combination with Ad-KDM4D treatment resulted in approximately 100%, 34%, and 62% increase in EdU+, phospho-H3+, and total cell number compared to control, respectively. To validate these findings in vivo, we tested JAKi2 in an inducible KDM4D transgenic mouse model (iKDM4D). The animals underwent ischemia/reperfusion (I/R) to induce infarction, and JAKi2 was administrated orally for up to 8 weeks. We found both early and late effects. 48 hours after I/R, the infarct size in iKDM4D+JAKi2 hearts was approximately 4-fold smaller than that in vehicle-treated or JAKi2 alone groups, and the combination increased EdU+, Ki67+, and phospho-H3+ CM number separately by 3.2-fold, 2.9-fold, and 6.5-fold. At 2 weeks post-I/R, the scar size was smaller in iKDM4D+JAKi2 hearts compared to vehicle-treated mice (9.0% vs 1.8%, P <0.05), and the combination resulted in a 3-fold, 5.5-fold, and 8.5-fold increases in EdU+, Ki67+, and phospho-H3+ CMs, respectively. Moreover, the iKDM4D+JAKi2 hearts demonstrated progressive improvement in left ventricular ejection fraction (26.8%, 27.3%, and 25.5%) and fraction shorten (37.9%, 38.1%, and 35.0%) at 2 weeks, 4 weeks, and 8 weeks post-I/R. The increased estimated ACM number and mononucleated CMs at 8 weeks post-I/R support that CM division was increased in the iKDM4D+JAKi2 hearts. These findings suggest that a combination therapy involving JAKi2, along with KDM4D, promotes robust CM proliferation and protects the heart after ischemic injury.

Trisomy 21 screening with αlpha software and the Fetal Medicine Foundation algorithm

Background. Screening for trisomy 21 provides pregnant women with accurate risk information. Different algorithms are used to screen for trisomy 21 in South Africa (SA). The Fetal Medicine Foundation (FMF) provides software to screen for trisomy 21 in the first trimester by ultrasound or a combination of ultrasound and biochemistry (combined screening), and requires regular and stringent quality control. With αlpha software, first trimester combined screening and screening with biochemistry alone in the first or second trimester are possible. The αlpha screening requires quality control of biochemical tests, but not of ultrasound measurements. Ideally, a screening test should have a high detection and a low screen positive rate. Despite the availability of these screening programmes, only a minority of infants with trisomy 21 are detected prenatally, raising questions about the effectiveness of screening. Objectives. To determine the screen positive and detection rates of prenatal screening for trisomy 21 in the SA private healthcare system. Methods. Data from the three largest laboratories collected between 2010 and 2015 were linked with genetic tests to assess screen positive and detection rates. Biochemical screening alone with αlpha software (first or second trimester) and combined screening using either FMF or αlpha software were compared. Results. One-third of an estimated 675 000 pregnancies in private practice in the 6-year study period underwent screening. There were 687 cases of trisomy 21 in 225 021 pregnancies, with only 239 (35%) diagnosed prenatally. The screen positive rates were 11.8% for first trimester biochemistry, 7.6% for second trimester biochemistry, 7.3% for first trimester FMF software ultrasound alone, 3.7% for combined first trimester screening with FMF software, and 3.5% for combined first trimester screening with αlpha software. The detection rates for a 5% false positive rate were 63% for first trimester biochemistry, 69% for second trimester biochemistry, 95% for combined first trimester screening with FMF software and 80% for combined first trimester screening with αlpha software. Detection and confirmation rates were highest with FMF software. Conclusion. Screening with FMF software has a similar screen positive rate and better detection rate than screening with αlpha software. The low prenatal detection rate of trisomy 21 is mainly due to a low prevalence of screening. More research is needed in the SA setting to explore why screening and confirmatory testing after high-risk results are not performed in many pregnancies.

An explainable artificial intelligence framework for risk prediction of COPD in smokers

BackgroundSince the inconspicuous nature of early signs associated with Chronic Obstructive Pulmonary Disease (COPD), individuals often remain unidentified, leading to suboptimal opportunities for timely prevention and treatment. The purpose of this study was to create an explainable artificial intelligence framework combining data preprocessing methods, machine learning methods, and model interpretability methods to identify people at high risk of COPD in the smoking population and to provide a reasonable interpretation of model predictions.MethodsThe data comprised questionnaire information, physical examination data and results of pulmonary function tests before and after bronchodilatation. First, the factorial analysis for mixed data (FAMD), Boruta and NRSBoundary-SMOTE resampling methods were used to solve the missing data, high dimensionality and category imbalance problems. Then, seven classification models (CatBoost, NGBoost, XGBoost, LightGBM, random forest, SVM and logistic regression) were applied to model the risk level, and the best machine learning (ML) model’s decisions were explained using the Shapley additive explanations (SHAP) method and partial dependence plot (PDP).ResultsIn the smoking population, age and 14 other variables were significant factors for predicting COPD. The CatBoost, random forest, and logistic regression models performed reasonably well in unbalanced datasets. CatBoost with NRSBoundary-SMOTE had the best classification performance in balanced datasets when composite indicators (the AUC, F1-score, and G-mean) were used as model comparison criteria. Age, COPD Assessment Test (CAT) score, gross annual income, body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), anhelation, respiratory disease, central obesity, use of polluting fuel for household heating, region, use of polluting fuel for household cooking, and wheezing were important factors for predicting COPD in the smoking population.ConclusionThis study combined feature screening methods, unbalanced data processing methods, and advanced machine learning methods to enable early identification of COPD risk groups in the smoking population. COPD risk factors in the smoking population were identified using SHAP and PDP, with the goal of providing theoretical support for targeted screening strategies and smoking population self-management strategies.

Molecular Determinants of Sensitivity to Polatuzumab-Vedotin in Diffuse Large B-Cell Lymphoma

Purpose: Polatuzumab Vedotin (Pola-V) is an antibody-drug conjugate directed to the B cell surface antigen CD79B. When combined with conventional immunochemotherapy, Pola-V improves outcomes in DLBCL overall; however, there is noted heterogeneity in response to Pola-V, with germinal center b-cell (GCB) DLBCL showing no added benefit to the addition of Pola-V compared to standard immunochemotherapy. We aimed to identify molecular determinants of sensitivity or resistance to Pola-V. We hypothesized that these might lead us to innovative strategies to improve on-target tumor killing by Pola-V, or to find novel biomarkers that predict drug resistance. Methods: We employed combined drug-sensitization and CD79B-sorted CRISPR-Cas9 screening to identify molecular determinants of sensitivity to CD79B-directed, tumor killing by Pola-V in 9 cell lines representing different molecular subtypes of DLBCL. Results: Our results reveal the striking impact of epitope glycosylation, specifically a2,6 sialylation, on the binding of Pola-V to CD79B and thereby its ability to kill tumor cells. Specifically, we identify the exact glycosylated residues on CD79A and CD79B which create a glycan shield around the Pola-V binding site, precluding binding to its target. We show how genetic, pharmacological and enzymatic approaches that remove terminal sialic acid residues from these N-linked glycans lead to enhanced tumor killing by Pola-V. We hypothesize and test multiple methods of targeting this pathway in order to enhance Pola-V killing both in vitro and in vivo. Finally, we reveal a previously unappreciated role for the ubiquitin ligase KLHL6 in regulating CD79B protein abundance and surface expression of the B cell antigen receptor (BCR), including how this pathway is used by physiological germinal center B cells and how it is corrupted to enhance BCR expression in GCB DLBCL. Conclusions: These findings unravel the molecular basis of response heterogeneity to Pola-V and identify approaches that might be deployed therapeutically to enhance the efficacy of CD79B-specific tumor killing. In addition, we identify how KLHL6 determines expression of the BCR in both physiological and malignant germinal center B cells, and how KLHL6 mutation may modulate sensitivity of GCB DLBCL tumors to Pola-V.

Venetoclax Synergizes with IMGN632, a Novel CD123-Targeting Antibody Conjugated to a DNA Alkylating Payload, By Suppressing DNA Damage Response and Potentiating Apoptosis in Acute Myeloid Leukemia in Vitro Models

CD123, a component of interleukin-3 receptor, is overexpressed on the surface of acute myeloid leukemia (AML) blasts and leukemic stem cells compared with normal hematopoietic progenitors, which makes this antigen an attractive therapeutic target. IMGN632 is a novel antibody-drug-conjugate comprising a high affinity CD123-targeting antibody and DNA alkylating cytotoxic payload. In preclinical studies, IMGN632 demonstrated high potency and selective cytotoxicity against AML leukemic cells vs normal myeloid progenitors, suggesting reduced likelihood of myelosuppression (Kovtun, 2018). Importantly, we previously showed high synergy of IMGN632 combination with BCL-2 inhibitor venetoclax (VEN) and DNA hypomethylating azacytidine (AZA) in AML cell lines and xenograft models (ASH 2020, #617). IMGN632 demonstrated encouraging single agent activity and favorable tolerability in relapsed/refractory AML patients (ASH 2019 #734, ASCO 2020 TPS7563, NCT03386513) and is currently tested in combination with VEN and/ or AZA in relapsed and frontline CD123-positive AML patients (NCT04086264). In this study, we provide new insight into molecular mechanisms underlying the synergistic efficacy of these combinations using AML models in vitro. AML cells with FLT3-ITD mutations expressed higher levels of CD123, as previously reported (Han 2013). FLT3-ITD cell lines were particularly sensitive to IMGN632 with IC 50 values at low ng/ml range. Correspondingly, as evidenced by large scale drug combination screen using BLISS independence model, triple IMGN632/VEN/AZA treatment was synergistic at multiple dosage levels and ratios, suggesting a favorable potency of triple combination in FLT3-mutated subtypes of AML associated with poor prognosis. The efficacy of IMGN632/VEN/AZA combination was however decreased in cells lacking functional TP53. shRNA-mediated knockdown of TP53 in FLT3-ITD MOLM13 cells resulted in increased cell viability, reduced apoptosis and downregulation of cleaved caspase-3 and PARP. While both treated wild-type and TP53 KD cells showed robust overexpression of phosphorylated histone H2AX (p-H2AX), TP53 KD cells expressed higher levels of anti-apoptotic MCL-1 and lower levels of pro-apoptotic BAX, indicative of their intrinsic inability to fully execute apoptosis and resulting in suppressed response to triple combination. Importantly, in sensitive AML cells, IMGN632 induced profound DNA damage, G 1 (MOLM13 and MOLM14 cells) or G 2/M (MV4-11 cells) phase cell cycle arrest and decline in S-phase population, all above augmented by VEN/AZA co-treatment. Observed increase in p-H2AX was accompanied by apoptotic caspase-3 and PARP cleavage and was fully prevented by QVD-OPh caspase inhibitor, suggesting that double-stranded DNA damage and p-H2AX triggered by triple combination were secondary events associated with apoptosis but not the initial cause of cell cycle arrest. Thus, given that cytotoxic payload of IMGN632 is a DNA alkylator, we next tested its impact on mediators of the DNA damage response (DDR). IMGN632 alone induced phosphorylation of p53 at Ser15, a step required for p53 transcriptional activity, and triggered activating phosphorylation of checkpoint kinase Chk1 (Ser345), known to facilitate cell cycle arrest and DNA damage repair, thus preventing treated cells from progressing through S-phase but also increasing the potential of repairing IMGN632-induced DNA damage (Fig. 1A). Upon combined IMGN632+VEN treatment, p53 phosphorylation was sustained, but p-ChK1 decreased, suggesting that VEN may impair protective DDR triggered by IMGN632, resulting in greater cell death. To support this hypothesis, we showed that IMGN632 treatment prior to VEN produced significant synergistic activity (Fig. 1B). Together, these results suggest that VEN, apart from its canonical inhibitory effect on anti-apoptotic BCL-2, exerts previously unrecognized ability to suppress DDR program in AML and augments activity of DNA damaging IMGN632. Failure of cells to sustain DDR in the presence of VEN constitutes a key aspect of high efficacy of IMGN/VEN/AZA combination in AML.

A compact online proton spectrometer for diagnosis of picosecond intense-laser accelerated protons

A compact online proton spectrometer based on a CMOS plate detector was proposed for the diagnosis of picosecond intense-laser accelerated protons. Inside the CMOS plate detector, a composite structure that composed of a scintillator screen, a fiber optic plate (FOP), and a CMOS sensor was designed to detect the protons safely and efficiently. The scintillator screen converted the incident protons to visible light, which, in turn, was collected by using the FOP and detected by using the CMOS sensor. An additional aluminum housing was designed to decrease the strong electromagnetic pulse (EMP) impact on the electronic readout system from picosecond laser–plasma interaction. This online spectrometer is compact and cost-effective than the system based on the micro-channel plate or scintillator screen in combination with an electron multiplying CCD camera and has better resistance to radiation damage than the system based on a single CMOS sensor.

COVID-19 infection and hypertension disorders during pregnancy

The objective: to determine the impact of COVID-19, which occurred before or during pregnancy, on the frequency of development and features of the course of gestational hypertension and preeclampsia.Materials and methods. An observational retrospective study with parallel control one was conducted. The pregnancy course and its outcomes in 178 women during the period of the COVID-19 pandemic – from March 2020 to January 2022 – who were observed in the women’s consultation of a private medical center, were analyzed. It is important to note that the medical documentation was taken one after the other, without being specially selected according to any principle, that is, the analysis carried out had the character of continuous screening.Out of 178 pregnant women, 125 (70.2%) persons were sick with COVID-19, they were included in the main group; 53 (29.8%) women who were part of the comparison group did not get sick. The groups are comparable by age and body mass index. The age of the women was 19–51 years; in the main group – 30.5±4.9 years, in the comparison group – 30.9±5.6 years. Body mass index (kg/m2) at the beginning of pregnancy in women of the main group was 21.6±4.3; comparison group – 22.9±4.8. There were 104 (58.4%) primigravida, 74 (41.6%) – multigravida. 117 (65.7%) women were primipara, 61 (34.3%) – multipara.All pregnant women were performed with a standard examination in accordance with the Order No. 417 of the Ministry of Health of Ukraine dated 15.07.2011. Clinical and anamnestic risk factors for the development of preeclampsia were assessed in all women without exception, on the basis of which 3 women were assigned to the risk group. In addition, 158 (88.8%) pregnant women as part of the first combined screening had an additional examination to determine the risk of developing preeclampsia, as a result of which additional 23 women were included in the risk group. As a result, 26 (14.6%) pregnant women from 12 weeks of gestation were prescribed prophylaxis using acetylsalicylic acid.Results. In women who have had COVID-19, hypertensive disorders of pregnancy develop statistically significantly less often than in those who have not had the disease. After mildly symptomatic COVID-19 the risks of hypertensive disorders in pregnant women are statistically significantly reduced: gestational hypertension – RR=1.15, 95% CI: 1.0–1.3; preeclampsia – RR=1.1, 95% CI: 1.0–1.2, all hypertensive disorders – BP=1.3, 95% CI: 1.1–1.6.The groups did not differ among themselves in the prevalence of generally accepted risk factors. The frequency of hypertensive disorders depending on the vaccination was analyzed. The risk of gestational hypertension in vaccinees who did not suffer from COVID-19 is reduced by 1.4 times (RR=1.4, 95% CI: 1.0–1.9), the risk of all hypertensive disorders in pregnant women in this same group is reduced 2 times (RR=2.0, 95% CI: 1.2–3.5).The development of preeclampsia also probably depends on the vaccination status (χ2=4.0; p=0.04; φ=-0.17). In non-vaccinated persons it increases by 1.1 times (RR=1.1, 95%CI: 1.2–1.8). The risk of all hypertensive disorders is also higher in the group with a negative vaccine status (RR=1.3, 95%CI: 1.0–1.7) only at the level of significance p=0.06, that is, it appears as a certain tendency.Conclusions. Hypertensive disorders associated with pregnancy are less likely to develop in women who have experienced COVID-19 in the preconception period or during pregnancy. Vaccination against COVID-19 reduces the frequency of hypertensive disorders in pregnant women, regardless of the disease presence.Since immune mechanisms, along with placental-vascular mechanisms play a significant role in the pathogenesis of preeclampsia, let’s assume that the experienced coronavirus infection or vaccination against it disrupts certain links of immunity, which results in a decrease in the frequency of hypertensive disorders in pregnant women.

Impact of Endometrial Preparation on the Maternal and Fetal Cardiovascular Variables of the First Trimester Combined Screening Test

The modality of endometrial preparation for the transfer of frozen-thawed embryos may influence maternal and fetal adaptation to pregnancy and could thus impact the results of the first trimester combined screening test. We conducted a retrospective cross-sectional study on singleton pregnancies achieved by embryo transfer of a single frozen-thawed blastocyst, comparing two different endometrial preparation protocols: natural cycle (n = 174) and hormone replacement therapy (HRT) (n = 122). The primary outcome was the risk of preeclampsia at the first trimester combined screening test. Secondary endpoints included variable reflecting fetal cardiac function (nuchal translucency and fetal heart rate), maternal adaptation (median arterial blood pressure—MAP and uterine arteries pulsatility index—UtA-PI), and placentation (pregnancy associated plasma protein A and placental growth factor). The risk of early preeclampsia was comparable in the two groups (38% vs. a 28%, p = 0.12). However, women in the natural cycle group showed lower fetal heart rate (159 [155–164] vs. 164 [158–168], p = 0.002) and higher UtA-PI (0.96 [0.74–1.18] vs. 0.72 [0.58–0.90], p < 0.001). The frequency of a screening test at high risk for aneuploidies was similar. The modality of transfer of frozen-thawed embryos is associated with changes in the variables reflecting maternal and fetal cardiovascular function.

Combination of QSAR Modeling and Hybrid-Based Consensus Scoring to Identify Dual-Targeting Inhibitors of PLK1 and p38γ.

Polo-like kinase 1 (PLK1) and p38γ mitogen-activated protein kinase (p38γ) play important roles in cancer pathogenesis by controlling cell cycle progression and are therefore attractive cancer targets. The design of multitarget inhibitors may offer synergistic inhibition of distinct targets and reduce the risk of drug-drug interactions to improve the balance between therapeutic efficacy and safety. We combined deep-learning-based quantitative structure-activity relationship (QSAR) modeling and hybrid-based consensus scoring to screen for inhibitors with potential activity against the targeted proteins. Using this combination strategy, we identified a potent PLK1 inhibitor (compound 4) that inhibited PLK1 activity and liver cancer cell growth in the nanomolar range. Next, we deployed both our QSAR models for PLK1 and p38γ on the Enamine compound library to identify dual-targeting inhibitors against PLK1 and p38γ. Likewise, the identified hits were subsequently subjected to hybrid-based consensus scoring. Using this method, we identified a promising compound (compound 14) that could inhibit both PLK1 and p38γ activities. At nanomolar concentrations, compound 14 inhibited the growth of human hepatocellular carcinoma and hepatoblastoma cells in vitro. This study demonstrates the combined screening strategy to identify novel potential inhibitors for existing targets.

Zebrafish: A Smart Tool for Heart Disease Research.

The increasing prevalence of heart disease poses a significant threat to human survival and safety. However, the current treatments available for heart disease are quite limited. Therefore, it is of great importance to utilize suitable animal models that can accurately simulate the physiological characteristics of heart disease. This would help improve our understanding of this disease and aid in the development of new treatment methods and drugs. Zebrafish hearts not only exhibit similarities to mammalian hearts, but they also share approximately 70% of homologous genes with humans. Utilizing zebrafish as an alternative to costly and time-consuming mammalian models offers numerous advantages. Zebrafish models can be easily established and maintained, and compound screening and genetic methods allow for the creation of various economical and easily controlled zebrafish and zebrafish embryonic heart disease models in a short period of time. Consequently, zebrafish have become a powerful tool for exploring the pathological mechanisms of heart disease and identifying new effective genes. In this review, we summarize recent studies on different zebrafish models of heart disease. We also describe the techniques and protocols used to develop zebrafish models of myocardial infarction, heart failure, and congenital heart disease, including surgical procedures, forward and reverse genetics, as well as drug and combination screening. This review aims to promote the utilization of zebrafish models in investigating diverse pathological mechanisms of heart disease, enhancing our knowledge and comprehension of heart disease, and offering novel insights and objectives for exploring the prevention and treatment of heart disease. This article is protected by copyright. All rights reserved.

Flexible Anionic Groups-Activated Structure Dissymmetry for Strong Nonlinearity in Ln2 Ae3 MIV 3 S12 Family.

Structural dissymmetry and strong second-harmonic generation (SHG) responses are key conditions for nonlinear optical (NLO) crystals, and targeted combinatorial screening of suitable anionic groups has become extremely effective. Herein, optimal combination of flexible SnSn (n=5, 6) groups and highly electropositive cations (lanthanides (Ln3+ ) and alkaline earth (Ae2+ : Sr, Ca) metals) affords the successful synthesis of 12 NLO thiostannates including Ln2 Sr3 Sn3 S12 (Pmc21 ) and Ln2 Ca3 Sn3 S12 (P-62m); whereas 17 rigid GeS4 or SiS4 tetrahedra-constructed Ln2 Ae3 Ge3 S12 and Ln2 Ae3 Si3 S12 crystallize in the centrosymmetric (CS) Pnma. This unprecedented CS to noncentrosymmetric (NCS) structural transformation (Pnma to P-62m to Pmc21 ) in the Ln2 Ae3 MIV 3 S12 family indicates that chemical substitution of the tetrahedral GeS4 /SiS4 units with SnSn breaks the original symmetry to form the requisite NCS structures. Remarkably, strong polarization anisotropy and hyperpolarizability of the Sn(4+) S5 unit afford huge performance improvement from the nonphase-matching (NPM) SHG response (1.4×AgGaS2 and Δn=0.008) of La2 Ca3 Sn3 S12 to the strong phase-matching (PM) SHG effect (3.0×AgGaS2 and Δn=0.086) of La2 Sr3 Sn3 S12 . Therefore, Sn(4+) S5 is proven to be a promising "NLO-active unit." This study verifies that the coupling of flexible SnSn building blocks into structures opens a feasible path for designing targeted NCS crystals with strong nonlinearity and optical anisotropy.

Accelerated multi-property screening of Fe–Co–Ni alloy libraries by hyper-heuristic combinatorial flow synthesis and high-throughput spark plasma sintering

High-throughput (HT) chemical synthesis facilitates accelerated materials discovery products. However, HT methods are limited by the need for expensive robotic systems, complicated methodology, and low yield. Hence, we developed a hyper-heuristic combinatorial flow synthesis (HCFS) device capable of composition gradient generation and production of an adequate mass of Fe–Co–Ni alloy nanoparticles. A library of 91 Fe–Co–Ni powder compositions was synthesized using this technique. A high-throughput spark plasma sintering (HT-SPS) methodology, along with the die design, was developed for combinatorial screening of multiple properties. 56 compositions were down-selected and consolidated into compositionally graded bulk samples using HT-SPS and subsequent annealing. The crystallographic, magnetic, electrical, and magnetic properties of the bulk library were assessed. The saturation magnetization (Ms) varied from 83.3 emu/g to 225.2 emu/g, coercivity (Hc) from 17.5 Oe to 78.4 Oe, resistivity (ρ) from 17.2 μΩ·cm to 986.7 μΩ·cm, and Vickers hardness (HV) from 41.9 HV to 281.7 HV. Novel Fe–Co–Ni compositions, e.g., Fe36.5Co55.1Ni8.4 and Fe22.6Co73.4Ni4, with a promising multi-property set, were identified for the first time. This study demonstrated that promising new compositions exhibiting multi-property optimization can be successfully discovered by our hyper-heuristic combinatorial chemical synthesis methodology.

Newborn genetic screening is highly effective for high-risk infants: A single-centre study in China.

Newborn genetic screening (NBGS) is promising for early detection of genetic diseases in newborns. However, little is known about its clinical effectiveness in special groups like high-risk infants. To address this gap, we aimed to investigate the impact of NBGS on high-risk infants. We screened 10 334 healthy newborns from the general maternity unit and 886 high-risk infants from the neonatal ward using both traditional newborn screening (tNBS) and NBGS, and collected clinical data from electronic medical records. We found that high-risk infants had a higher proportion of eutocia (P < 0.01) and prematurity (P < 0.01). For high-risk infants vs healthy newborns screened by tNBS, the primary screening positive rate was 3.84% vs 1.31%, the false positive rate (FPR) was 3.62% vs 1.18% (P < 0.001), and the positive predictive value (PPV) was 5.88% vs 8.27%. For NBGS vs tNBS in high-risk infants, the primary screening positive rate was 0.54% vs 3.68%, the FPR was 0.22% vs 3.47%, and the PPV was 60.00% vs 5.88%. We found that combined newborn screening can effectively reduce the FPR caused by the high-risk symptoms and improve the PPV in high-risk infants, sufficient for more accurately showing the true status of the disease.

Implementation of hospital-based sickle cell newborn screening and follow-up programs in Haiti

AbstractOne in 120 children are born with sickle cell disease (SCD) in Haiti. However, health care challenges include isolated newborn screening (NBS) activities and lack of transcranial Doppler (TCD) ultrasound to assess stroke risk. The implementation activities of the Comparative Study of Children in Haiti and Miami with Sickle Cell Disease involved both NBS and TCD ultrasound implementations at 4 Haitian clinical sites. We hypothesized that hospital-based newborn SCD screening and follow-up programs would be feasible at Haiti. A traditional NBS laboratory method with dried blood samples was performed at 3 Port-au-Prince sites, and the traditional method plus point-of-care (POC) testing was used at the 2 northern sites. The rate of clinical follow-up for newborns with SCD as the outcome for the NBS intervention was compared with that of the NBS method. The NBS programs identified SCD in 0.77% of 8224 newborns over a 24-month period. In the rural hospital assigned to the combination screening, 56% of newborns identified with POC testing returned for follow-up, compared with 0% when POC was not available (P = .044). Newborns who tested positive for SCD and children aged <6 years with SCD at the clinical sites were eligible for study follow-up. Accrual was successful: 165 participants (mean age, 42 months; 53% males; 93% hemoglobin SS) were recruited and received oral penicillin. TCD ultrasound screening was hampered by poor internet connections and trained staff leaving Haiti, with only 1 active site conducting screening. Despite challenges, the implementation of NBS and sickle cell programs in Haiti is feasible. We are in the process of understanding how to mitigate implementation limitations.

Case Report: Three cases of suspected female genital schistosomiasis and precancerous lesions for cervical cancer in a highly endemic country—from clinical management to public health implications

Female genital schistosomiasis (FGS) is a chronic manifestation of schistosomiasis, usually caused by Schistosoma haematobium infection, which can be responsible for infertility, ectopic pregnancy, and abortion, and is associated with an increased prevalence of HIV infection. No screening programs are currently recommended for FGS. Colposcopy, the conventionally suggested diagnostic tool for FGS, is also considered a crucial screening tool for cervical cancer (CC). We performed an experimental screening via colposcopy for FGS at primary healthcare centers (PHCCs) in the Boeny region of Madagascar, allowing for the detection of patients with both FGS signs and HPV-related dysplasia (HPV-dy). All suspected FGS cases were treated with praziquantel on the day of colposcopy, and all images of suspected CC or HPV-dy were re-assessed by a gynecologist and, if needed, patients were then provided with additional colposcopy for histologic diagnosis and treatment. We describe three cases of FGS and HPV-related precancerous lesions detected during the project, discussing the state of art of the relationship between CC, FGS and HPV and the real-life challenges encountered in terms of both patient compliance and the diagnostic and treatment cascade. Despite the current diagnostic limitations, a screening for FGS via colposcopy may contribute to the early identification of CC or precancerous lesions. The addition of visual inspection with acetic acid (VIA) during colposcopy for FGS screening could improve its impact on CC screening. In addition, although there is limited evidence of the effectiveness of praziquantel in FGS, treatment should in any case be proposed for suspicious lesions, given its safety and ease of administration. The benefit of combined screening could be maximised by increasing the availability of good quality services and improve awareness of both diseases among women

In Vitro and In Silico Antiviral Activity of Di-Halogenated Compounds Derived from L-Tyrosine against Human Immunodeficiency Virus 1 (HIV-1).

HIV-1 infection is considered one of the major public health problems worldwide. Due to the limited access to antiretroviral therapy, the associated side effects, and the resistance that the virus can generate, it has become necessary to continue the development of new antiviral agents. The study aimed to identify potential antiviral agents for HIV-1 by evaluating the in vitro and in silico activity of 16 synthetic di-halogenated compounds derived from L-Tyrosine. The compounds were tested for cytotoxicity, which was determined using MTT, and a combined antiviral screening strategy (pre- and post-infection treatment) was performed against R5 and X4 strains of HIV-1. The most promising compounds were evaluated against a pseudotyped virus (HIV-GFP-VSV-G), and the effectiveness of these compounds was measured through GFP flow cytometry. Also, the antiviral effect of these compounds was evaluated in PBMCs using flow cytometry and ELISA for p24. The TODB-2M, TODC-2M, TODC-3M, and YDC-3M compounds showed low toxicity and significant inhibitory activity against HIV-1. In silico docking and molecular dynamics assays suggest that the compounds' antiviral activity may be due to interaction with reverse transcriptase, viral protease, or envelope gp120.

Extract2Chip-Bypassing Protein Purification in Drug Discovery Using Surface Plasmon Resonance.

Modern drug discovery relies on combinatorial screening campaigns to find drug molecules targeting specific disease-associated proteins. The success of such campaigns often relies on functional and structural information of the selected therapeutic target, only achievable once its purification is mastered. With the aim of bypassing the protein purification process to gain insights on the druggability, ligand binding, and/or characterization of protein-protein interactions, herein, we describe the Extract2Chip method. This approach builds on the immobilization of site-specific biotinylated proteins of interest, directly from cellular extracts, on avidin-coated sensor chips to allow for the characterization of molecular interactions via surface plasmon resonance (SPR). The developed method was initially validated using Cyclophilin D (CypD) and subsequently applied to other drug discovery projects in which the targets of interest were difficult to express, purify, and crystallize. Extract2Chip was successfully applied to the characterization of Yes-associated protein (YAP): Transcriptional enhancer factor TEF (TEAD1) protein-protein interaction inhibitors, in the validation of a ternary complex assembly composed of Dyskerin pseudouridine synthase 1 (DKC1) and RuvBL1/RuvBL2, and in the establishment of a fast-screening platform to select the most suitable NUAK family SNF1-like kinase 2 (NUAK2) surrogate for binding and structural studies. The described method paves the way for a potential revival of the many drug discovery campaigns that have failed to deliver due to the lack of suitable and sufficient protein supply.

Identification of Fetuses at Increased Risk of Trisomies in the First Trimester Using Axial Planes

Introduction: The measurement of nuchal translucency (NT) is crucial for assessing risk of aneuploidies in the first trimester. We investigate the ability of NT assessed by a transverse view of the fetal head to detect fetuses at increased risk of common aneuploidies at 11–13 weeks of gestation. Methods: We enrolled a nonconsecutive series of women who attended our outpatient clinic from January 2020 to April 2021 for aneuploidy screening by means of a first trimester combined test. All women were examined by operators certified by the Fetal Medicine Foundation. In each patient, NT measurements were obtained both from the median sagittal view and transverse view. We calculated the risk of aneuploidy using NT measurements obtained both with sagittal and axial scans, and then we compared the results. Results: A total of 1,023 women were enrolled. An excellent correlation was found between sagittal and transverse NT measurements. The sensitivity and specificity of the axial scan to identify fetuses that were deemed at risk of trisomy 21 using standard sagittal scans were 40/40 = 100.0% (95% confidence interval [CI]: 91.2–100.0) and 977/983 = 99.4% (95% CI: 98.7–99.7), respectively. The sensitivity and specificity of the axial scan to identify fetuses at risk of trisomy 13 or 18 were 16/16 = 100.0% (95% CI: 80.6–100.0) and 1,005/1,007 = 99.8% (95% CI: 99.3–99.9). Conclusions: When the sonogram, a part of combined test screening, is performed by an expert sonologist, axial views can reliably identify fetuses at increased risk of trisomies without an increase in false negative results.

RECOVER identifies synergistic drug combinations in vitro through sequential model optimization

For large libraries of small molecules, exhaustive combinatorial chemical screens become infeasible to perform when considering a range of disease models, assay conditions, and dose ranges. Deep learning models have achieved state-of-the-art results in silico for the prediction of synergy scores. However, databases of drug combinations are biased toward synergistic agents and results do not generalize out of distribution. During 5 rounds of experimentation, we employ sequential model optimization with a deep learning model to select drug combinations increasingly enriched for synergism and active against a cancer cell line-evaluating only ∼5% of the total search space. Moreover, we find that learned drug embeddings (using structural information) begin to reflect biological mechanisms. In silico benchmarking suggests search queries are ∼5-10× enriched for highly synergistic drug combinations by using sequential rounds of evaluation when compared with random selection or ∼3× when using a pretrained model.

OC13.01: Enhancing prenatal risk assessment: atypicality Index (AcFTS) as an adjunct to the combined first trimester screening in dichorionic twins*

OC13.01: Enhancing prenatal risk assessment: atypicality Index (AcFTS) as an adjunct to the combined first trimester screening in dichorionic twins*