Takes Two to Tangle: A Rare Case of Type IV Dual Left Anterior Descending Artery (LAD) and Dual Ostia of the LAD and Left Circumflex (LCx) Artery.

The dual left anterior descending (LAD) artery is a rare anatomic variant of the LAD artery that refers to the duplication of the LAD into a short and long LAD. These two vessels, differentiated based on their lengths, ultimately provide blood supply to the areas normally covered by the LAD. In this case report, we describe an unusual case of a type IV dual LAD system with an additional finding of a separate origin for the short LAD and left circumflex (LCx) artery. These two findings have not been reported together in the literature previously. During diagnostic procedures like coronary angiography or when interpreting cardiac imaging, awareness of these anomalies prevents confusion with pathological conditions such as coronary artery disease or stenosis. Additionally, it is crucial for cardiologists and surgeons to identify these aberrant vessels to avoid any wrongful interventions.

Influence of multi-stressor combinations of pCO2, temperature, and salinity on the toxicity of Heterosigma akashiwo (Raphidophyceae), a fish-killing flagellate.

Climate change and global warming have led to more frequent harmful algal blooms in the last decade. Among these blooms, Heterosigma akashiwo, a golden-brown phytoflagellate, is one of the 40 species with a high potential to form harmful blooms, leading to significant fish mortality. Climate change leads to rising atmospheric and ocean temperatures. These changes, along with altered rainfall patterns and meltwater input, can cause fluctuations in ocean salinity. Elevated atmospheric carbon dioxide (CO2) levels increase water acidity as oceans absorb CO2. This study investigated the effects of temperature, salinity, and CO2 levels on lipid production, hemolytic activity, and toxicity of H. akashiwo using the design of experiment approach, which can be used to investigate the effect of two or more factors on the same response simultaneously in a precise manner with fewer experiments and materials but in a larger region of the factor space. The lipid content was measured using a high-throughput Nile Red method, and the highest level of lipid content was detected at 25°C, a salinity of 30, and a CO2 concentration of 400 ppm. Hemolytic activity was assessed using rabbit blood erythrocytes in a 96-well plate, and the optimal conditions for achieving the highest hemolytic activity were determined at 15°C, a salinity of 10, and a CO2 concentration of 400 ppm. As the chemical structure of the toxin is not known, we used the toxicity against the cell line RTgill-W1 as the cell toxicity proxy. The maximum toxicity was identified at 15°C, a salinity of 10, and a CO2 level of 700 ppm.

Clinical effect of regional citrate anticoagulation continuous renal replacement therapy in three patients with hypercalcemic crisis.

In this study, we investigated the effectiveness of regional citrate anticoagulation continuous renal replacement therapy (RCA-CRRT) in reducing blood calcium levels in three patients with hypercalcemia crisis caused by different etiologies. The sodium citrate chelation of calcium ions was utilized as an anticoagulant for treating severely affected patients. By adjusting the citrate anticoagulant dose and monitoring treatment indicators, RCA-CRRT parameters were actively modified to alleviate the hypercalcemia crisis and provide time for surgery or specialized treatment. Two patients experienced rapid and effective reductions in blood calcium levels, allowing for further treatment, while the third patient exhibited a repeated increase in blood calcium, which eventually decreased after parathyroid adenoma resection, leading to clinical discharge. Our findings suggest that RCA-CRRT can help alleviate hypercalcemia crisis, stabilize the patient's internal environment, and provide valuable time for clinical treatment in cases of various medical conditions causing abnormal blood calcium elevations.

Portable Aptasensor Based on Parallel Rolling Circle Amplification for Tumor-Derived Exosomes Liquid Biopsy.

Here, a separation-free and label-free portable aptasensor is developed for rapid and sensitive analysis of tumor-derived exosomes (TEXs). It integrated a parallel rolling circle amplification (RCA) reaction, selective binding of metal ions or small molecules to nucleic acid-specific conformations, and a low-cost, highly sensitive handheld fluorometer. Lung cancer, for example, is targeted with two typical biomarkers (mucin 1 and programmed cell death ligand 1 (PD-L1)) on its exosomes. The affinity of aptamers to the targets modulated the amount of RCA products (T-Hg2+-T and cytosine (C)-rich single-stranded DNA), which in turn affected the fluorescence intensity of quantum dots (QDs) and methylene blue (MB). The results revealed that the limit of detection (LOD) of the handheld fluorometer for cell-derived exosomes can be as low as 30 particles mL-1. Moreover, its specificity, sensitivity, and area under the curve (AUC) are 93% (14/15), 92% (23/25), and 0.956, as determined by the analysis of 40 clinical samples. Retesting 16 of these samples with the handheld fluorometer yielded strong concordance between the fluorometer results and those acquired from clinical computed tomography (CT) and pathology.

Screening and functional verification of drought resistance-related genes in castor bean seeds.

Drought is one of the natural stresses that greatly impact plants. Castor bean (Ricinus communis L.) is an oil crop with high economic value. Drought is one of the factors limiting castor bean growth. The drought resistance mechanisms of castor bean have become a research focus. In this study, we used castor germinating embryos as experimental materials, and screened genes related to drought resistance through physiological measurements, proteomics and metabolomics joint analysis; castor drought-related genes were subjected to transient silencing expression analysis in castor leaves to validate their drought-resistant functions, and heterologous overexpression and backward complementary expression in Arabidopsis thaliana, and analysed the mechanism of the genes' response to the participation of Arabidopsis thaliana in drought-resistance.Three drought tolerance-related genes, RcECP 63, RcDDX 31 and RcA/HD1, were obtained by screening and analysis, and transient silencing of expression in castor leaves further verified that these three genes corresponded to drought stress, and heterologous overexpression and back-complementary expression of the three genes in Arabidopsis thaliana revealed that the function of these three genes in drought stress response.In this study, three drought tolerance related genes, RcECP 63, RcDDX 31 and RcA/HD1, were screened and analysed for gene function, which were found to be responsive to drought stress and to function in drought stress, laying the foundation for the study of drought tolerance mechanism in castor bean.

Novel Anatomical Variation of Left Coronary Artery Origin at the Sinotubular Junction Level With Coeval Hypoplastic Left Circumflex, Superdominant Right Coronary Artery, and Obstructive Coronary Artery Disease: A Case Report.

The isolated origin of the left coronary artery (LCA) ostium at the level of the sinotubular junction (STJ) has been described previously. Congenital absence of the left circumflex (LCx) coronary artery has also been documented with superdominant right coronary arterial circulation, either in the presence or absence of coronary artery obstruction. Earlier literature has linked the association of an absent LCx coronary artery with a superdominant right coronary artery (SRCA) but not with a hypoplastic LCx coronary artery (HLCx). The present case report details the case of a 37-year-old thin, athletic male with the risk factors of diabetes and hypertension who was admitted to the emergency unit of our hospital for losing consciousness while bicycling in the street. The current report establishes a combined association of LCA anomaly origin at STJ level along with HLCx and SRCA condition with the burden of mild to moderate coronary artery disease involving proximal left anterior descending artery, LCx, and mid right coronary artery in the literature for the first time. Further, the case report advocated that the presented case carries the risk of malignancy. Hence, with the advancement of modern imaging technologies, computed tomography angiography should be the first choice of imaging modality rather than coronary angiography to prevent fatal outcomes. Interventional cardiologists, cardiothoracic surgeons, and radiologists should have properly defined knowledge of coronary artery anatomy and associated pathology, as it is important for coronary cannulation or any coronary interventions.

Ultra-Sensitive Detection of the SARS-CoV-2 Nucleocapsid Protein via a Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a-Mediated Immunoassay.

Tracking trace protein analytes in precision diagnostics is an ongoing challenge. Here, we developed an ultrasensitive detection method for the detection of SARS-CoV-2 nucleocapsid (N) protein by combining enzyme-linked immunosorbent assay (ELISA) with the clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas) system. First, the SARS-CoV-2 N protein bound by the capture antibody adsorbed on the well plate was sequentially coupled with the primary antibody, biotinylated secondary antibody, and streptavidin (SA), followed by biotin primer binding to SA. Subsequently, rolling circle amplification was initiated to generate ssDNA strands, which were targeted by CRISPR/Cas12a to cleave the FAM-ssDNA-BHQ1 probe in trans to generate fluorescence signals. We observed a linear relationship between fluorescence intensity and the logarithm of N protein concentration ranging from 3 fg/mL to 3 × 107 fg/mL. The limit of detection (LOD) was 1 fg/mL, with approximately nine molecules in 1 µL of the sample. This detection sensitivity was 4 orders magnitude higher than that of commercially available ELISA kits (LOD: 5.7 × 104 fg/mL). This method was highly specific and sensitive and could accurately detect SARS-CoV-2 pseudovirus and clinical samples, providing a new approach for ultrasensitive immunoassay of protein biomarkers.

Parasite-derived microRNA let-7-5p detection for metacestodiasis based on rolling circular amplification-assisted CRISPR/Cas9.

Metacestodiasis is an infectious disease caused by the larval stage of cestode parasites. This disease poses a serious health hazard to wildlife, livestock, and humans, and it incurs substantial economic losses by impacting the safety of the livestock industry, the quality of meat production, and public health security. Unfortunately, there is currently no available molecular diagnostic method capable of distinguishing cysticercus- and Echinococcus-derived microRNAs (miRNAs) from other helminthes and hosts in the plasma of metacestode-infected animals. This study aims to develop a specific, sensitive, and cost-efficient molecular diagnostic method for cysticercosis and echinococcosis, particularly for early detection. The study developed a rolling circular amplification (RCA)-assisted CRISPR/Cas9 detection method based on parasite-derived miRNA let-7-5p. Using a series of dilutions of the let-7 standard, the limit of detection (LOD) of the qPCR, RCA, and RCA-assisted CRISPR/Cas9 methods was compared. The specificity of qPCR and CRISPR/Cas9 was evaluated using four artificially synthesized let-7 standards from different species. A total of 151 plasma samples were used to evaluate the diagnostic performance. Additionally, the study also assessed the correlation between plasma levels of let-7-5p, the number of Taenia pisiformis cysticerci, and the weight of Echinococcus multilocularis cysts. The results demonstrated that the RCA-assisted CRISPR/Cas9 assay could significantly distinguish let-7 from cestodes and other species, achieving a LOD of 10 aM; the diagnostic sensitivity and specificity for rabbit cysticercosis and mouse E. multilocularis were 100% and 97.67%, and 100% and 100%, respectively. Notably, let-7-5p gradually increased in the plasma of T. pisiformis-infected rabbits from 15 days post infection (dpi), peaked at 60 dpi, and persisted until 120 dpi. In E. multilocularis-infected mice, let-7-5p gradually increased from 15 dpi and persisted until 90 dpi. Furthermore, the expression of let-7-5p positively correlated with the number of cysticerci and cyst weight. These results indicated that the let-7-5p-based RCA-assisted CRISPR/Cas9 assay is a sensitive and specific detection method that can be used as a universal diagnostic method for metacestodiasis, particularly for early diagnosis (15 dpi).

Anomalous Right Coronary Artery With Interarterial Course: Risk Stratification and Surgical Decision-Making Using Coronary Computed Tomography Angiography-Derived Fractional Flow Reserve.

An anomalous right coronary artery (RCA) takeoff, a rare congenital condition often characterized by an interarterial RCA course between the pulmonary artery and the ascending aorta, can lead to symptoms of angina pectoris (chest pain) or even sudden cardiac death (SCD) due to compression of the RCA, although most patients remain asymptomatic. In this case report, we highlight the utility of computed tomography angiography (CTA)-derived fractional flow reserve (FFR), a minimally invasive technique used to assess the hemodynamic significance of coronary lesions, in the risk stratification and surgical decision-making process for a 46-year-old female patient presenting with exertional dyspnea and an anomalous RCA takeoff with an interarterial course. The information obtained from this imaging modality was instrumental in determining that surgical repair did not need to be performed urgently and could be scheduled as an elective case in the future.

Target proteins-regulated DNA nanomachine-electroactive substance complexes enable separation-free electrochemical detection of clinical exosome.

Simple and reliable profiling of tumor-derived exosomes (TDEs) holds significant promise for the early detection of cancer. Nonetheless, this remains challenging owing to the substantial heterogeneity and low concentration of TDEs. Herein, we devised an accurate and highly sensitive electrochemical sensing strategy for TDEs via simultaneously targeting exosomal mucin 1 (MUC1) and programmed cell death ligand 1 (PD-L1). This approach employs high-affinity aptamers as specific recognition elements, utilizes rolling circle amplification and DNA nanospheres as effective bridges and signal amplifiers, and leverages methylene blue (MB) and doxorubicin (DOX) as robust signal reporters. The crux of this separation- and label-free method is the specific response of MB and DOX to G-quadruplex structures and DNA nanospheres, respectively. Quantifying TDEs using this strategy enabled precise discrimination of lung cancer patients (n = 25) from healthy donors (n = 12), showing 100% specificity (12/12), 92% sensitivity (23/25), and an overall accuracy of 94.6% (35/37), with an area under the receiver operating characteristic curve (AUC) of 0.97. Furthermore, the assay results strongly correlated with findings from computerized tomography and pathological analyses. Our approach could facilitate the early diagnosis of lung cancer through TDEs-based liquid biopsy.

Machine learning model based on RCA-PDCA nursing methods and differentiating factors to predict hypotension during cesarean section surgery.

BACKGROUND: Intraoperative hypotension during cesarean section has become a serious complication for maternal and fetal healthy. It is commonly encountered by subarachnoid anesthesia. However, currently used control methods have varying degrees of side effects, such as drugs. The Root Cause Analysis (RCA) - Plan, Do, Check, Act (PDCA) is a new model of care that identifies the root causes of problems. The study aimed to demonstrate the usefulness of RCA-PDCA nursing methods in preventing intraoperative hypotension during cesarean section and to predict the occurrence of intraoperative hypotension through a machine learning model. METHODS: Patients who underwent cesarean section at Traditional Chinese Medicine of Southwest Medical University from January 2023 to December 2023 were retrospectively screened, and the data of their gestational times, age, height, weight, history of allergies, intraoperative vital signs, fetal condition, operative time, fluid out and in, adverse effects, use of vasopressor drugs, anxiety-depression-pain scores, and satisfaction were collected and analyzed. The statistically different features were screened and five machine learning models were used as predictive models to assess the usefulness of the RCA-PDCA model of care. RESULTS: (1) Compared with the general nursing model, the RCA-PDCA nursing model significantly reduces the incidence of intraoperative hypotension and postoperative complications in cesarean delivery, and the patient experience is comfortable and satisfactory. (2) Among the five machine learning models, the RF model has the best predictive performance, and the accuracy of the random forest model in preventing intraoperative hypotension is as high as 90%. CONCLUSION: Through computer machine learning model analysis, we prove the importance of the RCA-PDCA nursing method in the prevention of intraoperative hypotension during cesarean section, especially the Random Forest model which performed well and promoted the application of artificial intelligence computer learning methods in the field of medical analysis.

Enhancing the engineering characteristics of sustainable recycled aggregate concrete using fly ash, metakaolin and silica fume.

The present global environment is facing growing issues linked to production of concrete, mostly due to high consumption of concrete as the dominating construction material globally. In today's climate of environmental sustainability, recycled concrete production using RA (recycled aggregates) requires a more holistic approach. This study examined how recycled aggregate (RA), metakaolin, silica fume and fly ash affect mechanical characteristics (compressive strength, split tensile strength), and durability characteristics (acid resistance, absorption, sorptivity) of concrete. The objective of this investigation is to figure out how the potential of RA based concrete can be improved so that they can be used to their maximum extent. To achieve the desired outcome, the study involved testing three distinct groups of concrete samples, each containing different percentages (25%, 50%, and 75%) of recycled aggregate (RA) with a constant amount of silica fume, (25 kg/m3). The first group was used as the control mix, while the second group incorporated 10% fly ash, and the third group included 15% metakaolin. The findings of this research show that the RA concrete mechanical properties as well as durability can be significantly improved by incorporating 15% metakaolin and 10% fly ash. The investigation involves the examination of all ternary blends within two distinct acidic environments, specifically a 5% hydrochloric acid (HCl) solution and sulfuric acid (H2SO4). Both the ternary mixes (metakaolin and fly ash) with the combination of silica improve all characteristics. The quality assurance, cost analysis and the reduction of CO2 emissions are carried out, utilizing RA (recycled aggregates) as a substitute for NA (natural aggregate). In case of producing superior structural concrete, it is recommended based on results to limit the replacement percentage of recycled aggregate (RA) to a maximum of 50% in presence of 10% fly ash and 15% metakaolin, otherwise the RA replacement percentage should not be more than 25%.

Right Coronary Artery-to-Right Atrial Fistula Accompanied by Multiple Right Coronary Artery Aneurysms: A Case Report.

A coronary artery aneurysm (CAA) denotes a localized dilation of the coronary artery, while a coronary artery fistula signifies an aberrant connection between a coronary artery and a cardiac chamber or adjacent vessel. Here, we present a case study of a 68-year-old female with a previously diagnosed right coronary artery-to-right atrial fistula concomitant with multiple right coronary artery aneurysms. Initially asymptomatic, the patient subsequently manifested atrial fibrillation. Management involved augmenting the patient's home regimen with metoprolol tartrate, followed by successful cardioversion and restoration of sinus rhythm. Given the stability of the fistula and the absence of symptomatic exacerbation, no further interventional measures were undertaken. The patient was discharged with an adjusted metoprolol regimen and scheduled follow-up with her cardiologist. Subsequent imaging assessments unveiled progressive fistula expansion alongside the development of concurrent CAA, inciting deliberations concerning optimal treatment modalities.

Engineering of a DNAzyme-Based dimeric G-quadruplex rolling circle amplification for robust analysis of lead ion.

Detecting heavy metal pollution, particularly lead ion (Pb2⁺) contamination, is imperative for safeguarding public health. In this study, we introduced an innovative approach by integrating DNAzyme with rolling circle amplification (RCA) to propose an amplification sensing method termed DNAzyme-based dimeric-G-quadruplex (dimer-G4) RCA. This sensing approach allows for precise and high-fidelity Pb2⁺ detection. Strategically, in the presence of Pb2⁺, the DNAzyme undergoes substrate strand (S-DNA) cleavage, liberating its enzyme strand (E-DNA) to prime isothermal amplification. This initiates the RCA process, producing numerous dimer-G-Quadruplexes (dimer-G4) as the signal reporting transducers. Compared to conventional strategies using monomeric G-quadruplex (mono-G4) as the reporting transducers, these dimer-G4 structures exhibit significantly enhanced fluorescence when bound with Thioflavin T (ThT), offering superior target signaling ability for even detection of Pb2⁺ at low concentration. Conversely, in the absence of Pb2⁺, the DNAzyme structure remains intact so that no primers can be produced to cause the RCA initiation. This nucleic acid amplification-based Pb2⁺ detection method combing with the high specificity of DNAzymes for Pb2⁺ recognition ensures highly sensitive detection of Pb2+ with a detection limit of 0.058 nM, providing a robust tool for food safety analysis and environmental monitoring.

Detection of Cadmium(II) in Aquatic Products Using a Rolling-Circle Amplification-Coupled Ratio Fluorescent Probe Based on an Aptamer-Peptide Conjugate.

The existing aptamers for cadmium (Cd2+), the common toxic heavy metal contaminant in food, cannot meet the requirements for detecting Cd2+ in rapid detection methods. In previous work, we found that coupling aptamer-peptide conjugates (APCs) with peptides and aptamers can provide a less disruptive method with a significantly improved affinity. Moreover, we found that the spatial conformation of aptamers and peptides is crucial for obtaining proper affinity in APC. Therefore, we describe a simple design strategy to obtain a series of APCs with different affinities by designing peptide orientations (N-terminal, C-terminal). The best affinity was found for APC(C1-N) with a binding constant (Ka) of 2.23 × 106 M-1, indicating that the APC(C1-N) affinity was significantly increased by 829.17% over aptamer. Finally, a rolling-circle amplification (RCA)-coupled ratio fluorescence-based biosensor for Cd2+ detection was established with a detection limit of 0.0036 nM, which has great potential for practical aquatic product detection.

Percutaneous Coronary Intervention in a Patient Presenting With Inferior Myocardial Infarction and an Anomalous Left Main Artery Originating From the Right Coronary Sinus.

We present a case of a patient with inferior myocardial infarction (MI) and anomalous left main artery originating from the right coronary sinus. The left main artery and right coronary artery originated from the right coronary sinus but with separate ostia. The patient underwent revascularization of the right coronary artery with balloon angioplasty and a drug-eluting stent. Despite being rare, these anomalies can be life-threatening depending on the course of the artery, and when atherosclerotic disease is present, a revascularization strategy can be challenging. Knowing the existence of the left main artery anomaly is important to choose the right guide catheter to achieve successful cannulation and decrease the risk of complications, radiation exposure, and contrast usage.

Molecular characterization and expression pattern of Rubisco activase gene GhRCAß2 in upland cotton (Gossypium hirsutum L.).

BACKGROUND: Rubisco activase (RCA) is a pivotal enzyme that can catalyse the activation of Rubisco in carbon assimilation pathway. Many studies have shown that RCA may be a potential target for genetic manipulation aimed at enhancing photosynthetic efficiency and crop yield. OBJECTIVE: To understand the biological function of the GhRCAß2 gene in upland cotton, we cloned the coding sequence (CDS) of the GhRCAß2 gene and investigated its sequence features, evolutionary relationship, subcellular localization, promoter sequence and expression pattern. METHODS: The bioinformatics tools were used to analyze the sequence features of GhRCAß2 protein. Transient transformation of Arabidopsis mesophyll protoplasts was performed to determine the subcellular localization of the GhRCAß2 protein. The expression pattern of the GhRCAß2 gene was examined by analyzing transcriptome data and using the quantitative real-time PCR (qRT-PCR). RESULTS: The full-length CDS of GhRCAß2 was 1317 bp, and it encoded a protein with a chloroplast transit peptide. The GhRCAß2 had two conserved ATP-binding domains, and did not have the C-terminal extension (CTE) domain that was unique to the RCA α-isoform in plants. Evolutionarily, GhRCAß2 was clustered in Group A, and had a close evolutionary relationship with the soybean RCA. Western blot analysis demonstrated that GhRCAß2 was immunoreactive to the RCA antibody displaying a molecular weight similar to that of the RCA ß-isoform. The GhRCAß2 protein was found in chloroplast, aligning with its role as a vital enzyme in the process of photosynthesis. The GhRCAß2 gene had a leaf tissue-specific expression pattern, and the yellow-green leaf mutant exhibited a decreased expression of GhRCAß2 in comparison to the wild-type cotton plants. The GhRCAß2 promoter contained several cis-acting elements that respond to light, phytohormones and stress, suggesting that the expression of GhRCAß2 may be regulated by these factors. An additional examination of stress response indicated that GhRCAß2 expression was influenced by cold, heat, salt, and drought stress. Notably, diverse expression pattern was observed across different stress conditions. Additionally, low phosphorus and low potassium stress may result in a notable reduction in the expression of GhRCAß2 gene. CONCLUSION: Our findings will establish a basis for further understanding the function of the GhRCAß2 gene, as well as providing valuable genetic knowledge to improve cotton photosynthetic efficiency and yield under challenging environmental circumstances.

Enhanced selective discrimination of point-mutated viral RNA through false amplification regulatory direct insertion in rolling circle amplification.

Coronaviruses are single-stranded RNA viruses with high mutation rates. Although a diagnostic method for coronaviruses has been developed, variants appear rapidly. Low test accuracy owing to single-point mutations is one of the main factors in the failure to prevent the early spread of coronavirus infection. Although reverse transcription-quantitative polymerase chain reaction can detect coronavirus infection, it cannot exclude the possibility of false positives, and an additional multiplexing kit is needed to discriminate single nucleotide polymorphism (SNP) variants. Therefore, in this study, we introduced a new nucleic acid amplification method to determine whether an infected person has a SNP mutation using a lateral flow assay (LFA) as a point-of-care test. Unlike traditional DNA amplification methods, direct insertion into rolling circle amplification amplifies the target genes without false amplification. After SNP-selective nucleic acid amplification, nuclease enzymes are used to make double-stranded DNA fragments that the LFA can detect, where specific mismatched DNA is found and cleaved to show different signals when a SNP-type is present. Therefore, wild- and SNP-type variants can be selectively detected. In this study, the limit of detection was 400 aM for viral RNA, and we successfully identified a dominant SNP variant selectively. Clinical tests were also conducted.