The Evolution of Nucleic Acid-Based Diagnosis Methods from the (pre-)CRISPR to CRISPR era and the Associated Machine/Deep Learning Approaches in Relevant RNA Design.

Nucleic acid tests (NATs) are considered as gold standard in molecular diagnosis. To meet the demand for onsite, point-of-care, specific and sensitive, trace and genotype detection of pathogens and pathogenic variants, various types of NATs have been developed since the discovery of PCR. As alternatives to traditional NATs (e.g., PCR), isothermal nucleic acid amplification techniques (INAATs) such as LAMP, RPA, SDA, HDR, NASBA, and HCA were invented gradually. PCR and most of these techniques highly depend on efficient and optimal primer and probe design to deliver accurate and specific results. This chapter starts with a discussion of traditional NATs and INAATs in concert with the description of computational tools available to aid the process of primer/probe design for NATs and INAATs. Besides briefly covering nanoparticles-assisted NATs, a more comprehensive presentation is given on the role CRISPR-based technologies have played in molecular diagnosis. Here we provide examples of a few groundbreaking CRISPR assays that have been developed to counter epidemics and pandemics and outline CRISPR biology, highlighting the role of CRISPR guide RNA and its design in any successful CRISPR-based application. In this respect, we tabularize computational tools that are available to aid the design of guide RNAs in CRISPR-based applications. In the second part of our chapter, we discuss machine learning (ML)- and deep learning (DL)-based computational approaches that facilitate the design of efficient primer and probe for NATs/INAATs and guide RNAs for CRISPR-based applications. Given the role of microRNA (miRNAs) as potential future biomarkers of disease diagnosis, we have also discussed ML/DL-based computational approaches for miRNA-target predictions. Our chapter presents the evolution of nucleic acid-based diagnosis techniques from PCR and INAATs to more advanced CRISPR/Cas-based methodologies in concert with the evolution of deep learning (DL)- and machine learning (ml)-based computational tools in the most relevant application domains.

Long Chain Base Profiling with Multiple Reaction Monitoring Mass Spectrometry.

Sphingolipids (SLs) are essential lipids with important functions in membrane formation and cell signaling. The presence of a long chain base (LCB) structure is common to all SLs. De novo SL synthesis is initiated by the enzyme serine-palmitoyltransferase (SPT), which forms an LCB by the conjugation from serine and fatty acyl-CoAs. SPT can metabolize a variety of acyl-CoA substrates, which form diverse LCB structures within and across species. The LCB then undergoes further metabolic modifications resulting in an extraordinarily diverse spectrum of sphingolipids formed. SL analysis, using liquid chromatography-mass spectrometry (LC-MS)-based methods, poses challenges due to the diverse range of frequently isobaric species. This complexity complicates the identification of underlying LCB structures using standard lipidomics approaches. Here, we describe a simplified method to analyze the LCB profile in cells, tissue, and blood. The procedure involves chemical hydrolysis to remove the conjugated headgroups and N-acyl chains, allowing to specifically resolve the underlying LCB structures by LC-MS. This method can also be combined with an isotope labeling approach to determine in vivo SPT activity and total SL de novo synthesis over time.

Immunoglobulin D-Lambda Multiple Myeloma Initially Presenting in the Sphenoid Sinus, Orbital Apex, and Skull Base: A Systematic Review with a Case Report.

Objectives Multiple myeloma (MM) with initial manifestations in the sphenoid sinus, orbital apex, and skull base is exceedingly rare. A systematic review was conducted to investigate the epidemiology and advancements . Methods Relevant cases were identified by searching CNKI, WanFang Data, CQVIP databases, PubMed, Embase, and Web of Science. Additionally, we present a case of IgD-λ (immunoglobulin D-lambda) MM with initial symptoms of dizziness, unilateral pain, blindness, and ophthalmoplegia, leading to a 4-month overall survival. Strictly based on PRISMA standards, we included and summarized existing cases and reflected our case. Results Our systematic review includes 34 case reports, revealing 67.6% of patients initially presented with diplopia and 44.1% underwent endoscopic procedures, notably with only two cases of IgD-λ subtype. In our case, we performed an endoscopic wide trans-ethmoidal sphenoidotomy and biopsy of the skull base and orbital apex lesion. Postoperative pathology confirmed a highly active plasmacytoma, clinically diagnosed as IgD-λ MM with a TP53 deletion mutation and multiple extramedullary metastases. A range of diagnostic tools was employed, including hemoglobin, immunoglobulin, urinary protein analysis, positron emission tomography-computed tomography (CT), bone marrow cytology, and gene detection. Conclusion The subtle clinical manifestations of IgD-λ MM in the paranasal sinuses and skull base hinder early diagnosis. There is a paucity of literature describing MM initially presenting in these locations. CT/magnetic resonance scans are necessary to identify characteristic bone destruction. An endoscopic approach is popular for tissue biopsy. Bone marrow biopsy with a smear, serum or urine protein electrophoresis, and immunofixation electrophoresis are crucial upon the appearance of target organ damage.

Mesenteric Base Width: Abdominal Wall Diameter Correlates With Intestinal Rotational Anomalies in Infants.

INTRODUCTION: Symptomatic malrotation requires urgent Ladd procedure. Patients with incidental or indeterminate findings have historically been managed with observation or operatively. Fluoroscopic identification of the duodenojejunal junction (DJJ) and ileocecal junction (ICJ) can guide operative decision-making, but algorithms have not been validated. This study aimed to determine whether fluoroscopic mesenteric base width (MBW) standardized to abdominal wall diameter (AWD) correlates with intraoperative anatomy in infants. METHODS: We retrospectively reviewed patients between 2013 and 2023 who were <1 year with fluoroscopy identifying DJJ and ICJ. Infants with normal rotation evaluated for digestive concerns were included. Congenital conditions with intestinal nonrotation were excluded. Two radiologists independently measured MBW as a diagonal line from DJJ to ICJ and maximal transverse AWD from inferior ribs. A ratio was calculated and compared between groups. Wilcoxon rank-sum and Kruskal-Wallis tests with p < 0.05 were considered significant. Area under the receiver operating characteristic curve (AUROC) was used to identify optimal ratio cutoff. RESULTS: Fifty-eight patients, 22 normally rotated and 36 with intestinal rotational abnormality (IRA), met inclusion criteria. Preoperative radiographic concern for malrotation differed between groups (p < 0.0001). Median MBW:AWD was significantly lower in IRA than normal rotation based on imaging (0.31 vs 0.65, p < 0.0001). Optimal MBW:AWD of 0.55 had an AUROC of 0.9578. CONCLUSIONS: Radiographic measurement of MBW:AWD accurately predicted IRA from normal rotation with an optimal ratio cutoff of 0.55. Further validation will determine whether this ratio should play a role in management of incidental IRA or indeterminate findings on UGI. LEVEL OF EVIDENCE: III.

Epitope prime editing shields hematopoietic cells from CD123 immunotherapy for acute myeloid leukemia.

Acute myeloid leukemia (AML) is a malignant cancer characterized by abnormal differentiation of hematopoietic stem and progenitor cells (HSPCs). While chimeric antigen receptor T (CAR-T) cell immunotherapies target AML cells, they often induce severe on-target/off-tumor toxicity by attacking normal cells expressing the same antigen. Here, we used base editors (BEs) and a prime editor (PE) to modify the epitope of CD123 on HSPCs, protecting healthy cells from CAR-T-induced cytotoxicity while maintaining their normal function. Although BE effectively edits epitopes, complex bystander products are a concern. To enhance precision, we optimized prime editing, increasing the editing efficiency from 5.9% to 78.9% in HSPCs. Epitope-modified cells were resistant to CAR-T lysis while retaining normal differentiation and function. Furthermore, BE- or PE-edited HSPCs infused into humanized mice endowed myeloid lineages with selective resistance to CAR-T immunotherapy, demonstrating a proof-of-concept strategy for treating relapsed AML.

Quantifying vulnerability to plant invasion across global ecosystems.

The widely referenced "tens rule" in invasion ecology suggests that approximately 10% of established, non-native species will become invasive. However, the accuracy of this estimate has been questioned, as the original analysis focused on small groups of plant species in Great Britain and Australia. Using a novel database of 9501 established plants and 2924 invasive plants, we provide a comprehensive evaluation of the tens rule and the first empirical analysis of how invasion rates vary across spatial scales, islands/mainlands, and climate zones. We found that invasion rates (the percentage of established species with negative impacts) are highly variable across the globe. Well-sampled environments (those with at least 2000 total non-native species recorded) had invasion rates that ranged from 7.2% to 33.8%. Invasion rates were strongly scale-dependent, averaging 17% at the country scale and 25% at the continental scale. We found significantly higher invasion rates on islands when compared with mainlands, regardless of scale. Tropical ecosystems are often considered to be resistant to invasion; however, our results showed significantly higher invasion rates on both tropical islands and mainlands, suggesting unexpectedly high vulnerability of these species-rich ecosystems. We conclude that the tens rule is a poor general estimate of invasion rates for plants, as calculated invasion rates vary widely and are frequently much higher than 10%. Most locations would be better served by using invasion rates that vary based on the recipient environment. Our updated estimates of invasion rates should be highly relevant for invasive species management strategies, including weed risk assessments, which can be adjusted to identify more species as high-risk in areas where invasion rates are higher. Assuming that 10% of established species will become invasive is likely to substantially underestimate invasion rates in most geographies.

Sensitive Detection and Cell Imaging of Ca2+ Based on a "Turn-On" Schiff Base Fluorescent Probe.

Ca2+ ion as a second messenger in signaling pathway plays many vital roles in many biological phenomena. Thus, it is of significance for developing effective probes to detect Ca2+ ion specifically. Herein, a new Schiff base fluorescent probe FPH, fluorescein monoaldehyde (2-aminomethylpyridine) hydrazone, was designed and synthesized to identify Ca2+ in DMSO aqueous solution. The probe FPH revealed significant responses to Ca2+ with a fluorescence enhancement at 540 nm, exhibiting an evident fluorescence change from ultraweak luminescence to bright green. Otherwise, the FPH displayed a good linear range of 0.67 × 10-6 to 3.33 × 10-6 mol/L with a lower detection limit at 7.02 × 10-8 mol/L. The probe FPH were further successfully utilized to detect Ca2+ in living cells by an increased bright green fluorescence.

Artificial Intelligence in Temporal Bone Imaging: A Systematic Review.

OBJECTIVE: The human temporal bone comprises more than 30 identifiable anatomical components. With the demand for precise image interpretation in this complex region, the utilization of artificial intelligence (AI) applications is steadily increasing. This systematic review aims to highlight the current role of AI in temporal bone imaging. DATA SOURCES: A Systematic Review of English Publications searching MEDLINE (PubMed), COCHRANE Library, and EMBASE. REVIEW METHODS: The search algorithm employed consisted of key items such as 'artificial intelligence,' 'machine learning,' 'deep learning,' 'neural network,' 'temporal bone,' and 'vestibular schwannoma.' Additionally, manual retrieval was conducted to capture any studies potentially missed in our initial search. All abstracts and full texts were screened based on our inclusion and exclusion criteria. RESULTS: A total of 72 studies were included. 95.8% were retrospective and 88.9% were based on internal databases. Approximately two-thirds involved an AI-to-human comparison. Computed tomography (CT) was the imaging modality in 54.2% of the studies, with vestibular schwannoma (VS) being the most frequent study item (37.5%). Fifty-eight out of 72 articles employed neural networks, with 72.2% using various types of convolutional neural network models. Quality assessment of the included publications yielded a mean score of 13.6 ± 2.5 on a 20-point scale based on the CONSORT-AI extension. CONCLUSION: Current research data highlight AI's potential in enhancing diagnostic accuracy with faster results and decreased performance errors compared to those of clinicians, thus improving patient care. However, the shortcomings of the existing research, often marked by heterogeneity and variable quality, underscore the need for more standardized methodological approaches to ensure the consistency and reliability of future data. LEVEL OF EVIDENCE: NA Laryngoscope, 2024.

Current knowledge of antisense long non-coding RNA in the occurrence and prognosis of skull base tumors.

Antisense long non-coding RNA (AS-lncRNA) represents a novel class of RNA molecules. In recent years, it has been discovered that AS-lncRNAs play crucial roles in various biological processes, particularly in the onset and progression of tumors. Skull base tumors, originating from the base of the brain, exhibit specific expression patterns of AS-lncRNA which correlate significantly with clinical characteristics. This makes AS-lncRNA a promising candidate as a tumor marker. Functional studies have revealed that AS-lncRNAs can regulate gene expression by acting as miRNA sponges and interacting with RBPs. Consequently, they play pivotal roles in tumor cell cycle, apoptosis, angiogenesis, invasion, and metastasis processes. Further exploration into the mechanisms of AS-lncRNA in tumors holds substantial theoretical significance for deeper insights into the etiology, pathogenesis, and RNA dynamics of skull base tumors. Moreover, AS-lncRNA could serve as molecular markers or potential targets for early diagnosis. Their potential extends to efficacy assessment, prognosis prediction, and gene therapy, suggesting broad clinical applications. In summary, AS-lncRNA emerges as a promising molecular marker implicated in the onset and progression of skull base tumors.

Anatomical Features and Morphometric Characteristics of the Sphenoidal Sinus in MRI Studies.

Differential access to pathological sellar processes and adjacent regions is determined by the anatomic structures identified through diagnostic imaging. Both direct endonasal access (microscopic or endoscopic) and sublabial access utilize the sphenoid sinus (SS) as the primary surgical pathway. Critical factors include the pneumatization of the sinus, its intermediate septa, and the presence of a double wall, consisting of a connective tissue membrane along the dorsal wall of the SS. The present study aims to demonstrate the significance of the size and type of the SS based on MRI measurements. The type of SS, its pneumatization, and the proximity of adjacent brain structures are crucial for different surgical approaches to the SS and pituitary fossa. In neurosurgical practice, six main types of sinuses are recognized: sphenoid body type, lateral type, clival type, lesser wing type, anterior type, and combined type. Failure to consider these variations can lead to damage to the cavernous sinus, Meckel's cave, nerve structures in the middle cranial fossa, planum sphenoidale, suprasellar region, and vital brainstem structures located on the clivus. Randomly included MRI measurements were conducted on 112 patients from Pulmed University Hospital, Plovdiv, Bulgaria, categorized into two cohorts based on gender, with mean ages of 51 years for men and 47.8 years for women. The measurements, recorded in centimeters, were obtained using two imaging software programs, RadiAnt DICOM Viewer (Medixant, Poznan, Poland) and Weasis DICOM Viewer (Nicolas Roduit, https://github.com/nroduit/Weasis). No statistically significant differences were observed between the measurements produced by the two programs. Measurements of the SS were taken in two equal groups, using three different projections: axial, sagittal, and coronal. The results for height, width, and depth showed average sizes of 2.73-3.04 cm in axial projections, 1.70-2.64 cm in sagittal projections, and 2.86-3.03 cm in coronal projections. The minor differences between axial and coronal measurements of the same parameters (height and width) are statistically acceptable and attributed to the varying angles of the MRI scans. These measurements are crucial for planning surgical access to the sellar and parasellar regions, determining the necessary bony resection of the posterior wall of the SS, and preventing complications from excessive bony trepanation.

Synthesis, biological and computational evaluation of benzoxazole hybrid analogs as potential anti-Alzheimer's agents.

Aim: Current study aims exploration of bis-benzoxazole bearing bis-Schiff base scaffolds (1-16) as anti-Alzheimer's agents.Materials & methods: 2-aminophenol is used as starting materials which react with different reagents in different step to give us bis-benzoxazole bearing bis-Schiff base analogs. NMR and HREI-MS techniques were used for characterization. All derivatives demonstrated varied range of activities with IC50 values 1.10 ± 0.40-24.50 ± 0.90 µM against acetylcholinesterase (AChE) and 1.90 ± 0.70-28.60 ± 0.60 µM against butyrylcholinesterase (BuChE) in contrast to donepezil. In both cases, analog-3 was found most potent. Molecular docking explored modes of interactions between scaffolds and receptor sites of targeted enzymes.Conclusion: This study offering promising approach for optimization and development of potent inhibitors of cholinesterase enzymes.

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Operative versus non-operative treatment of ulnar styloid process base fractures: a systematic review and meta-analysis.

PURPOSE: Ulnar styloid process (USP) fractures are present in 40-65% of all distal radius fractures (DRFs). USP base fractures can be associated with distal radioulnar joint (DRUJ) instability and ulnar sided wrist pain and are treated by conservative management and surgical fixation, without consensus. This systematic review and meta-analysis compares operative to non-operative treatment of concomitant ulnar styloid base fractures in patients with distal radius fractures. METHODS: PubMed/Medline/Embase/CENTRAL databases were searched identifying RCTs and comparative observational studies. Effect estimates were extracted and pooled using random effect models to account for heterogeneity across studies. Results were presented as (standardized) mean differences (SMD or MD) or odds ratios (OR) and corresponding 95% confidence intervals (95%CI). RESULTS: Two RCTs (161 patients) and three observational studies (175 patients) were included. Tension band wiring was used for surgically treated USP fractures. Results were comparable across the different study designs and hence pooled across studies. Non-surgically treated patients had better wrist function at 6 months (SMD 0.57, 95%CI 0.30; 0.90, I2 = 0%). After 12 months there was no observed difference (MD 2.31, 95%CI -2.57; 7.19, I2 = 91%). Fewer patients had USP non-unions in the operative group (OR 0.08, 95%CI 0.04; 0.18, I2 = 0%). More patients suffered complications in the operative group (OR 14.3; 95%CI 1.08; 188, I2 = 89%). CONCLUSION: Routinely fixating USP base fractures as standard of care is not indicated. Surgery may be considered in selective cases (e.g. persistent DRUJ instability during ballottement test after fixation of the radius).

The BCL11A transcription factor stimulates the enzymatic activities of the OGG1 DNA glycosylase.

The BCL11A transcription factor has previously been shown to interact with and stimulate the enzymatic activities of the NTHL1 DNA glycosylase and Pol ß polymerase. Here we show that BCL11A and a smaller peptide encompassing amino acids 160 to 520 can interact with the 8-oxoguanine DNA glycosylase, OGG1, increase the binding of OGG1 to DNA that contains an 8-oxoguanine base and stimulate the glycosylase activity of OGG1. Following BCL11A knockdown, we observed an increase in oxidized purines in the genome using comet assays, while immunoassays reveal an increase in 8-oxoG bases. Structure-function analysis indicates that the stimulation of OGG1 by BCL11A requires the zinc fingers 1, 2 and 3 as well as the proline-rich region between the first and second zing finger, but a glutamate-rich region downstream of zinc finger 3 is dispensable. Ectopic expression of a small peptide that contains the three zinc fingers can rescue the increase in 8-oxoguanine caused by BCL11A knockdown. These findings, together with previous results showing that BCL11A stimulates the enzymatic activities of NTHL1 and the Pol ß polymerase, suggest that high expression of BCL11A is important to protect cancer cells against oxidative DNA damage.

ANXA5 and VEGFA Gene Variants in Women with Early Pregnancy Losses from North Macedonia.

Early pregnancy loss (EPL) is the most common pregnancy complication, found in approximately 15% of all clinically recognized pregnancy complications. Up to date, various maternal as well as fetal factors are reported as a cause of EPLs. However, in approximately 50% of EPL cases, the exact cause is not clearly identified and these cases are referred as idiopathic. The aim of our study was to examine the association of four distinct variants in the ANXA5 gene and two variants within the VEGFA gene in a cohort of women with EPLs from North Macedonia. This group was compared to a control group of women matched by ethnic background without pregnancy loss and at least one live birth. We also aimed to establish an effective and cost-efficient method for their detection based on multiplex single-base extension. Among 190 women experiencing EPLs, and 190 samples from women without a history of pregnancy loss (control group), our results demonstrated a statistically significant prevalence of heterozygotes for the M2/ANXA5 haplotype in women with EPLs, compared to the control group (p=0.0006). In the analyses comparing genotypic frequencies for the variants in the VEGFA gene, higher frequencies were generally observed among women experiencing EPLs, however without statistical significance. Our study aligns with multiple studies showing that M2 and M1 ANXA5 haplotypes are more prevalent in patients with pregnancy loss and presents an affordable genotyping technique for the specific ANXA5 and VEGFA variants.

l-Arginine and l-lysine improve the emulsifying and dissolution properties of pale, soft, exudative-like chicken myofibrillar proteins by modifying their conformations.

Herein, we investigated the effect and potential mechanisms of l-arginine (Arg) and l-lysine (Lys) on the emulsifying and dissolution properties of pale, soft, exudative (PSE)-like chicken myofibrillar proteins (MPs). The findings revealed that Arg/Lys effectively enhanced the emulsion activity and emulsion stability indexes of PSE-like MPs, resulting in smaller and more uniform PSE-like MP-soybean oil emulsions. Arg/Lys increased the solubility, absolute potential, hydrophobicity, fluorescence intensity, and ß-sheet content and decreased the turbidity, particle size, and ß-turn and random coil content of PSE-like MPs. Additionally, Arg/Lys did not significantly affect the Schiff base, carbonyl group, and total sulfhydryl contents, but caused a red shift of the band near 299 nm, indicating conformational rather than primary structural changes. Altogether, these findings indicate that Arg/Lys improves the emulsifying and dissolution performances of PSE-like MPs by adjusting conformation and contributes to a better understanding of how Arg/Lys enhances the physicochemical properties of PSE-like sausages.

Reconstruction of Anterior Skull Base After Cleaver Trauma: Combined Use of Fascia Lata and Middle Turbinate Flap in a Patient with Concha Bullosa.

This video article presents a 47-year-old male patient who presented to the emergency department after a cleaver bounced off the grinding wheel and lodged between his eyebrows. The patient complained of clear nasal drainage since the trauma. Computed tomography showed a comminuted displaced fracture starting from the right frontonasal recess and extending along the right ethmoid roof. The patient underwent surgery. The skull base defect was reconstructed with a free fascia lata graft in the first operation. In the second session, the reconstruction of the skull base was reinforced with a flap that was prepared from the middle turbinate with the concha bullosa. The patient was followed for eight months. The patient's symptoms resolved completely and there was no evidence of rhinorrhoea or any other complication at the control examination. No complications were seen on control magnetic resonance imaging.

Urinary chloride excretion in critical illness and acute kidney injury: a paediatric hypothesis-generating cohort study post cardiopulmonary bypass surgery.

Renal chloride metabolism is currently poorly understood but may serve as both a diagnostic and a treatment approach for acute kidney injury. We investigated whether plasma chloride, ammonia and glutamine as well as urinary chloride, ammonium and glutamine concentrations may serve as markers for acute kidney injury in paediatric patients. We conducted a prospective observational trial in a tertiary care paediatric intensive care unit. Ninety-one patients after cardiopulmonary bypass surgery were enrolled. Plasma glutamine, creatinine, (serum) albumin, urinary electrolytes and glutamine were collected pre-cardiopulmonary bypass surgery, at paediatric intensive care unit admission, and at 6, 12, 24, 48 and 72 h after paediatric intensive care unit admission. The urinary strong ion difference was calculated. The median urinary chloride excretion decreased from 51 mmol/L pre-cardiopulmonary bypass to 25 mmol/L at paediatric intensive care unit admission, and increased from 24 h onwards. Patients with acute kidney injury had lower urinary chloride excretion than those without. The median urinary strong ion difference was 59 mmol/L pre-cardiopulmonary bypass, rose to 131 mmol/L at 24 h and fell to 20 mmol/L at 72 h. The plasma chloride rose from 105 mmol/L pre-cardiopulmonary bypass to a maximum of 109 mmol/L at 24 h. At 24 h the plasma chloride concentration was associated with the presence of acute kidney injury. There was no association between plasma or urinary amino acids and chloride excretion or kidney injury. In conclusion, renal chloride excretion decreased in all patients, although this decrease was more pronounced in patients with acute kidney injury. Our findings may reflect a response of the kidneys to critical illness, and acute kidney injury may make these changes more pronounced. Targeting chloride metabolism may offer treatment approaches to acute kidney injury.

Skull Base Osteomyelitis Due to Staphylococcus epidermidis in the Absence of Indwelling Medical Devices Presenting With Bilateral Internal Carotid Artery Thrombosis.

Skull base osteomyelitis (SBO) is a severe and uncommon infection that typically affects the skull base and may arise from undiagnosed otogenic or sinonasal infection. This case describes a rare presentation of SBO, accompanied by thrombosis of the bilateral internal carotid artery with neurological deficits in a resource-limited environment, illustrating diagnostic and management dilemmas. A male patient aged 40 years with poorly controlled type 2 diabetes presented with sudden onset loss of consciousness and worsening right-sided weakness. MRI studies revealed SBO with cerebral involvement with thrombosis in major cerebral arteries and multiple brain infarcts. After receiving broad-spectrum antibiotics and supportive care shortly after admission, the patient developed septic shock and died two days after admission. The fast course of the disease in this case shows how severe SBO and its complications may be, calling for early diagnosis and intensive management of SBO, especially in diabetic patients. The fact that Staphylococcus epidermidis was established as a causative agent of disease in the absence of artificial heart valves or joints, it is becoming clear that there is a need to increase awareness of such rare pathogens, and probably new strategies for handling such infections should be developed. Additional research is required to elucidate the precise role of the pathogen and refine treatment approaches, especially for low-resource healthcare systems.

Investigation of the anticancer effect of newly synthesized palladium conjugate Schiff base metal complexes on non-small cell lung cancer cell line and mouse embryonic fibroblast cell line.

Lung cancer remains one of the leading causes of death worldwide. Due to the side effects of chemotherapeutic agents on normal cells and the development of resistance by cancer cells, there is an urgent need for alternative new pharmacological agents. Palladium (Pd)-conjugated Schiff base (SB) compounds represent an alternative approach with promising potential applications in cancer treatment. This study aims to identify novel therapeutic agents on A549 cells through the synthesis and characterization of Schiff base conjugated-Palladium complexes (Pd-L1 and Pd-L2). Additionally, it seeks to elucidate the mechanism of action of these compounds on both the A549 and NIH/3T3 cell lines. In the present study, two new Pd-L1 and Pd-L2 were synthesized for the first time and characterized mainly by single crystal X-ray diffraction and 1H, 13C, 31P NMR techniques. The cytotoxic effect of the compounds was evaluated by MTT assay on A549 and NIH/3T3 cell lines for 24 and 48 h. Cisplatin was used as a positive control group. Based on the cytotoxicity results, the complexes were evaluated for their anticancer activities against A549 cell lines for 48 h through reactive oxygen species (ROS), cell cycle, apoptotic, and necrotic cell analyses. The most potent cytotoxic effects were determined for Pd-L1 (IC50: 23.33 µM), Pd-L2 (IC50: 3.19 µM), and cisplatin (IC50: 33.27 µM) on A549 cells (p < 0.05). The compounds exhibited a significant cytotoxic effect at lower concentrations on A549 cells compared to NIH/3T3 cells (p < 0.05). All compounds showed a significant increase in ROS levels in A549 cells compared to the control group (p < 0.05). While necrosis and apoptosis was observed in A549 cells treated with cisplatin, induction of apoptosis was effective in cell death for A549 cells treated with Pd-L1 and Pd-L2 (p < 0.05). Additionally, it was observed that the compounds inhibited cell proliferation in the G0/G1 and G2/M cell cycle phases (p < 0.05). All compounds induced cell cycle arrest and cell death in A549 cells by increasing ROS levels. The results obtained in the present study could advance the utilization of the compounds as anticancer agents.