Bioanalytical HPLC method with fluorescence detector for determination of Entresto™ when co-administered with ibuprofen and fexofenadine: a pharmacokinetic study.

Entresto™ (LCZ696) has been approved globally for heart failure management. However, its lifelong use alongside over-the-counter (OTC) drugs like ibuprofen (IBU) and fexofenadine (FEX) necessitates an in-depth investigation of potential pharmacokinetic interactions, as they share the same metabolic and elimination pathways. This study aimed to develop a bioanalytical HPLC method with a fluorescence detector (FLD) to quantify LCZ696 analytes (valsartan, VAL; sacubitril, SAC; and sacubitril active metabolite, LBQ657) in rat plasma. Additionally, an in vivo study was performed to investigate the pharmacokinetic interactions of LCZ696 with IBU and FEX. Utilizing HPLC with a gradient-mode mobile phase of acetonitrile and 0.025 M phosphate buffer (pH 3), the study demonstrated a significant increase in the bioavailability of LCZ696 analytes (VAL and LBQ657) when co-administered with IBU (C max 0.23 ± 0.07 and 0.53 ± 0.21 µg mL-1, respectively) compared to the control (0.17 ± 0.03 and 0.33 ± 0.14 µg mL-1). A more significant increase in C max was noticed with FEX (0.38 ± 0.01 and 0.77 ± 0.18 µg mL-1, respectively). Moreover, a decrease in the clearance (Cl/F) of VAL and LBQ657 was observed (18.05 ± 1.94 and 12.42 ± 2.97 L h-1 kg, respectively) with a more pronounced effect in the case of FEX (30.87 ± 4.29 and 33.14 ± 9.57 L h-1 kg, respectively) compared to the control (49.99 ± 7.31 and 51.19 ± 9.12 L h-1 kg, respectively). In conclusion, our study underscores the importance of cautious administration and appropriate dose spacing of IBU and FEX in patients treated with LCZ696 to prevent elevated serum concentrations and potential toxicity. The novelty of this work lies in its dual contribution: developing a highly sensitive HPLC-FLD method and comprehensively elucidating significant pharmacokinetic interactions between LCZ696 and common OTC drugs.

Green and white MEKC for determination of different anti-diabetic binary mixtures and their triple-combo pill.

The work introduces green and white sustainable micellar electrokinetic chromatography (MEKC) procedure that could analyze therapeutically related drugs, empagliflozin (EMP), linagliptin (LIN) and metformin (MET) which are antidiabetic drugs with different mechanism of action, in their different pharmaceutical combinations. The method not only comply with the green analytical concepts, but also it is in line with sustainable analytical concepts as it is economic by applying the same operating conditions to analyze different pharmaceuticals in quality control (QC) labs which is a crucial step in QC labs and research centers to save time, effort, and money. Moreover, the method functionality regarding its scope with its achieved levels of accuracy, precision, low detection, and quantitation limits is tested using white assessment tool and compared with reported methods. The proposed MEKC coupled with a diode array detector (DAD) has been developed and validated for micro estimation of EMP and LIN in their low critical concentrations with MET in a ratio of (EMP: MET, 1:40) and (LIN: MET, 1:200). Separation was achieved within 6 min using fused silica capillary (40 cm × 50 µm id) using 20 mM Tris buffer (pH 10) in presence of 50 mM sodium dodecyl sulphate and 10% v/v methanol. The concentration ranges of the studied anti-diabetic drugs were 10-500, 10-100 and 2.5-100 µg. mL-1 for MET, EMP and LIN, respectively. The developed method is the first MEKC for concurrent determination of EMP, LIN and MET with high separation efficiency, low solvent consumption and regard as an easy green and white analytical tool. Moreover, Greenness and whiteness assessment were done via the most widely used Analytical Eco-Scale, the innovative AGREE tool and the RGB 12 algorithm.

Quantifying the impact of PFOA exposure on B-cell development and antibody production.

Per- and polyfluoroalkyl substances (PFAS) are synthetic chemicals; the vast majority are environmentally and biologically persistent, and some have demonstrated toxicity, including cancer, effects on metabolism, endocrine disruption, and immune dysfunction. Suppression of T-cell-dependent antibody responses (TDAR) has been observed in numerous studies of PFAS but mechanisms remain elusive. Evidence from our work suggests that B cells and how they use energy are impacted by PFAS exposure. We hypothesize that a well-studied and immunotoxic PFAS, perfluorooctanoic acid (PFOA), alters B-cell subclasses and markers of their metabolism. Adult male and female C57BL/6 mice were given PFOA (0 or 7.5 mg/kg) via gavage for 15 days, a duration and dose sufficient to suppress the TDAR. After dosing and immunization of subgroups, spleens were prepared to quantify B-cell subsets. Flow cytometric analysis revealed decreased numbers of plasmablasts, follicular, naïve, and overall B-cell subclasses in female PFOA-exposed groups. Male PFOA-exposed groups had a significant increase in follicular B cells and other subsets had decreases, including in the overall number of B cells. Twenty-four hours after naïve B-cell isolation and ex vivo activation, metabolic measurements revealed a 5-fold increase in metabolic markers in response to stimulation in PFOA-exposed groups compared with controls. These findings suggest that B-cell development and survival may be hindered by PFOA exposure, but that activation of the remaining B cells was not. Based on these findings, PFOA-mediated suppression of the primary IgM antibody response results changes to specific subsets of B cells.

Wernicke's Encephalopathy: A Precipitation of Glycogen Storage Disease.

A 16-year-old Saudi female who is a known case of glycogen storage disease type 1A (GSD1A), presented to the emergency department at King Faisal Specialist Hospital, Riyadh, Saudi Arabia on 15th January 2021 due to a complaint of persistent vomiting. Two weeks after admission, she began developing double vision and progressive leg weakness with intact sensation. She received the primary management to maintain good hydration and was admitted to the ICU for further workup. Over her hospital course, multiple investigations were conducted, the most significant of which was the MRI after sudden ocular deterioration. The result depicted findings classic for Wernicke's encephalopathy (WE) on MRI. The patient was then started on Thiamine supplementation and MRI performed three weeks later showed significant interval improvement of the parenchymal signal abnormality with complete resolution features of Wernicke's encephalopathy. This complex case emphasizes the need for early recognition and immediate treatment with IV thiamine in such a potential condition that can lead to permanent neurological deficits that present in a non-typical fashion.

The potential impact of insulin resistance and metabolic syndrome on migraine headache characteristics.

BACKGROUND & OBJECTIVES: Studying comorbidities with migraine aids in a better understanding of its pathophysiology and potential therapeutic targets. This case-control study aimed to study the impact of insulin resistance and metabolic syndrome on the characteristics of migraine headache attacks. METHODS: A case-control study was conducted on 30 migraine patients and 30 healthy controls. The following data were assessed in migraine patients: type of migraine, duration of attacks, Migraine Severity Scale (MIGSEV), and Headache Impact Test-6 (HIT-6). Both groups were assessed for waist circumference and underwent the following tests: fasting blood glucose, fasting insulin, high-density lipoprotein cholesterol level, and triglycerides, and homeostasis model assessment-insulin resistance (HOMA-IR) was applied. RESULTS: This study included age and sex-matched patients and controls. Migraine patients had significantly higher waist circumference, higher mean values of serum insulin, HOMA-IR and higher frequency of insulin resistance and metabolic syndrome than the control group (P-value = 0.005, 0.049, 0.01, 0.012, 0.024, respectively). Migraine patients with insulin resistance had significantly higher intensity and tolerability scores, MIGSEV total score, and HIT-6 total score compared to those without (P-value = 0.005, 0.005, 0.002, 0.018, respectively). There was a significantly positive correlation between the MIGSEV and HIT-6 scores and fasting insulin levels, and HOMA-IR value (P-value = 0.006, ≤ 0.001, 0.017, ≤ 0.001, respectively). CONCLUSION: Insulin resistance and metabolic syndrome are more common in migraine patients than in healthy controls. The severity and impact of migraine attacks are higher in patients with insulin resistance than in those without.

Empiric Anticoagulation Therapy in Hospitalized COVID-19 Patients: An Evaluation of Bleeding Risk Scores Performances in Predicting Bleeding Events.

Currently, there is no standardized consensus on anticoagulation (AC) among patients with coronavirus disease (COVID-19), which has an overwhelming bleeding risk. We aimed to compare the patterns of AC in COVID-19 patients and compare two validated risk scores in predicting bleeding events. A retrospective review of medical records was conducted for COVID-19 patients who received empiric anticoagulation therapy. The primary outcomes included bleeding events, survival, and mechanical ventilation needs. We applied the HAS-BLED and ORBIT bleeding risk scores to assess the predictive accuracy, using c-statistics and the receiver operating curve (ROC) method. Of the included patients (n = 921), with a mean age of 58.1 ± 13.2, 51.6% received therapeutic AC and 48.4% received a prophylactic AC dose. Significantly higher values of d-dimer and C-reactive protein (CRP) among the therapeutic AC users (p < 0.001) were noted with a significantly prolonged duration of hospital stay and mechanical ventilation (p < 0.001 and p = 0.011, respectively). The mean value of the HAS-BLED and ORBIT scores were 2.53 ± 0.93 and 2.26 ± 1.29, respectively. The difference between the two tested scores for major bleeding and clinically relevant non-major bleeding was significant (p = 0.026 and 0.036, respectively) with modest bleeding predictive performances. The therapeutic AC was associated with an increased risk of bleeding. HAS-BLED showed greater accuracy than ORBIT in bleeding risk predictability.

One-Pot Synthesis and Molecular Modeling Studies of New Bioactive Spiro-Oxindoles Based on Uracil Derivatives as SARS-CoV-2 Inhibitors Targeting RNA Polymerase and Spike Glycoprotein.

The first outbreak in Wuhan, China, in December 2019 was reported about severe acute coronaviral syndrome 2 (SARS-CoV-2). The global coronavirus disease 2019 (COVID-19) pandemic in 2020 resulted in an extremely high potential for dissemination. No drugs are validated in large-scale studies for significant effectiveness in the clinical treatment of COVID-19 patients, despite the worsening trends of COVID-19. This study aims to design a simple and efficient cyclo-condensation reaction of 6-aminouracil derivatives 2a-e and isatin derivatives 1a-c to synthesize spiro-oxindoles 3a-d, 4a-e, and 5a-e. All compounds were tested in vitro against the SARS-CoV-2. Four spiro[indoline-3,5'-pyrido[2,3-d:6,5-d']dipyrimidine derivatives 3a, 4b, 4d, and 4e showed high activities against the SARS-CoV-2 in plaque reduction assay and were subjected to further RNA-dependent-RNA-polymerase (RdRp) and spike glycoprotein inhibition assay investigations. The four compounds exhibited potent inhibitory activity ranging from 40.23 ± 0.09 to 44.90 ± 0.08 nM and 40.27 ± 0.17 to 44.83 ± 0.16 nM, respectively, when compared with chloroquine as a reference standard, which showed 45 ± 0.02 and 45 ± 0.06 nM against RdRp and spike glycoprotein, respectively. The computational study involving the docking studies of the binding mode inside two proteins ((RdRp) (PDB: 6m71), and (SGp) (PDB: 6VXX)) and geometrical optimization used to generate some molecular parameters were performed for the most active hybrids.

Stromal Derived Factor-1 Gene Polymorphism in Pediatric Immune Thrombocytopenia.

Immune thrombocytopenia (ITP) is a multifactorial disease in which both environmental and genetic factors have been implicated. The study aimed to investigate a possible association of single nucleotide polymorphisms (SNPs rs266085 and rs2839693) in the stromal derived factor-1 (SDF-1) gene and its association to ITP and effect on ITP severity and response to treatment. Genomic DNA was extracted from peripheral blood and polymorphism in SDF-1 gene rs266085 and rs2839693 was analyzed using PCR-restriction fragment length polymorphism technique in DNA extracted from 60 children with ITP together with 90 healthy controls. On analysis of SDF-1 rs266085 polymorphism, there was a high frequency of CC genotype in cases than controls and that difference was significant at codominant, overdominant, and dominant models (P<0.05). Furthermore, carriers of the CC genotype were more susceptible to severe ITP at onset, steroid dependency, and chronicity than carriers of other genotypes (P<0.05). Otherwise, no significant differences between ITP patients and controls as regard SDF-1 rs2839693 genotypes and alleles, and we did not find a relation between this polymorphism and ITP severity, steroid dependency, or duration. SDF-1 gene rs266085 SNP C allele is associated with susceptibility to develop ITP as well as increases the risk for severe ITP at onset, chronic ITP and steroid dependency.

Impact of nano-conjugate on Drosophila for early diagnosis of Alzheimer's disease.

Alzheimer's disease (AD) is an irreversible neurodegenerative disease. Recent identification of AD biomarkers has led to the diagnosis of AD before the onset of dementia. It has been shown that Drosophila melanogaster is a valuable model for studying human neurodegeneration, including AD. According to its properties, curcumin shows promising potential for the diagnosis of AD. In order to improve its use, new formulations, including nanotechnology-based delivery systems, have been applied. The current study aims to diagnose AD by detecting the accumulation of amyloid beta-peptide via carbon-dot-curcumin nanoparticle conjugation in Drosophila. The accumulation of amyloid beta-peptide has been detected via the conjugate using the fluorescence imaging technique. These results suggest that carbon-dot-curcumin nanoparticle conjugation could be used as a diagnostic tool for AD.

Voltammetric analysis of dantrolene and its active metabolite with indomethacin in rat plasma.

AIM: Differential pulse polarography was used for the concurrent analysis of the coadministered dantrolene (DAN) and indomethacin (IND) in plasma. MATERIALS & METHODS: DAN and IND, Hanging mercury drop electrode and Britton-Robinson buffer at pH 5 were used. In plasma, cathodic reduction of DAN nitro group and its active metabolite at -0.2 V was done. IND was analyzed after carbonyl group reduction at -1.1 V. RESULTS: Drugs determination in rat plasma with good recoveries and low limit of quantitation was done. Application to trace analysis of drugs in rat plasma was done with Cmax and Tmax determination. CONCLUSION: This technique shows high sensitivity, simplicity and low cost. The method is US FDA validated and it is applicable to human level.

High performance thin-layer and high performance liquid chromatography coupled with photodiode array and fluorescence detectors for analysis of valsartan and sacubitril in their supramolecular complex with quantitation of sacubitril-related substance in raw material and tablets.

Valsartan (VAL) and sacubitril (SAC) are combined in a supramolecular complex, LCZ696, which is a newly approved remedy for heart failure. SAC-related substance (biphenyl methyl pyrrolidinone [BMP]) which also appears as an intermediate during SAC synthesis is considered to be a suspected impurity for SAC and/or LCZ696 tablets. The study investigates the analysis of VAL and SAC in their supramolecular complex along with SAC-related substance, BMP, using high performance thin-layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) with two different detectors; fluorescence detector (FLD) and diode array detector (DAD). The work aimed at analyzing BMP at low levels in the presence of its parent drug, SAC. BMP was successfully analyzed at a level of 0.167, 1 and 3% of its parent drug, SAC upon using HPLC-FLD, HPLC-DAD and HPTLC, respectively. For HPLC-FLD, the detector was set at λex/λem (nm/nm): 0-4.5 min at 255/374; 4.5-6 min at 255/314, for achieving an adequate sensitivity of the method to monitor and quantify VAL and SAC in the presence of BMP. Low limits of detection (8.3, 3.3 and 1.7 ng mL-1) and limits of quantitation (25, 10 and 5 ng mL-1) values obtained for VAL, SAC and BMP, respectively, upon using FLD suggest that low level of baseline noise enables the detection and quantitation of low BMP concentration.

Photocatalytic properties of PbS/graphene oxide/polyaniline electrode for hydrogen generation.

In this work, roll-graphene oxide (Ro-GO), polyaniline (PANI) nano/microparticles, and PbS nanoparticles were prepared by modified Hammer, oxidative polymerization, and chemical bath deposition methods, respectively. These nano/microstructures were characterized, optimized, and designed to form PbS/Ro-GO/PANI nano/microcomposite. Also, the ratios of PbS and Ro-GO were optimized, and the optimized composition of the used composite was 0.4 g PANI, 0.125 g Ro-GO, and 0.075 g PbS. The band gap values for PANI, PbS, Ro-GO, and PbS/Ro-GO/PANI rocomposite were 3, 1.13, 2.86, (1.16, 2) eV, respectively. Two photoelectrode assemblies, Au/PbS/Ro-GO/PANI and PbS/Ro-GO/PANI/ITO/glass were used for the photoelectrochemical (PEC) hydrogen generation. In the first assembly 45 nm- Au layer was sputtered on the surface of a disk of PbS/Ro-GO/PANI composite. For the second assembly, a disk of PbS/Ro-GO/PANI composite was glued on ITO glass using Ag-THF paste. The lifetime efficiency values were 64.2 and 43.4% for the first and second electrode for 2 h, respectively. Finally, the incident photon-to-current conversion efficiency (IPCE) and photon-to-current efficiency (ABPE) were calculated under monochromatic illumination conditions. The optimum IPCE efficiency at 390 nm was 9.4% and 16.17%, whereas ABPE % efficiency was 1.01% and 1.75% for Au/PbS/Ro-GO/PANI and PbS/Ro-GO/PANI/ITO/glass, respectively.

First derivative emission spectrofluorimetric method for the determination of LCZ696, a newly approved FDA supramolecular complex of valsartan and sacubitril in tablets.

LCZ696 (sacubitril/valsartan, Entresto™) is a therapy lately approved by United States Food and Drug Administration (US FDA) as a heart failure therapy. It is claimed to decrease the mortality rate and hospitalization for patients with chronic heart failure. This study is considered as the first report to investigate the fluorimetric behavior of sacubitril in addition to pursuing all the different conditions that may affect its fluorescence. Various conditions were studied, for example studying the effects of organized media, solvents and pH, which may affect the fluorescence behavior of sacubitril. For the simultaneous determination of the newly approved supramolecular complex of valsartan (VAL) and sacubitril (SAC) in their tablets, a sensitive and simple first derivative spectrofluorimetric method was developed. The method involved the measurement of native fluorescence at 416 nm and 314 nm (λex 249 nm) for VAL and SAC, respectively. The first (D1) derivative technique was applied to the emission data to resolve a partial overlap that appeared in their emission spectra. The proposed method was successfully applied for the assay of the two drugs in their supramolecular complex LCZ696 with no interference from common pharmaceutical additives. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines were followed in order to validate the proposed method.

Carpal Tunnel Syndrome Grading Using High-Resolution Ultrasonography.

PURPOSE: Carpal tunnel syndrome (CTS) is a common entrapment neuropathy of the wrist. The diagnosis of CTS has been a concern for physicians for a long time. The aim of this study is to evaluate the use of the median nerve (MN) cross-sectional area (CSA) in the wrist compared with the CSA in the forearm to grade the severity of CTS in Egyptian patients. METHODS: The CSAs of the MN in the wrist and forearm were measured in 72 wrists that were diagnosed with CTS via nerve conduction studies and 80 healthy wrists. The CTS group was subdivided into three subgroups (mild, moderate, and severe CTS). The ratio of the CSA of the MN in the wrist to that in the forearm was used to calculate cutoff values for CTS grading. RESULTS: There were positive correlations between the CSAs of the MN in the wrist and MN conduction latency. At a wrist-forearm ratio of 1.7, the high-resolution ultrasonography showed 96.1% accuracy in the detection of CTS. CONCLUSIONS: High-resolution ultrasonography can be used in CTS grading.

Self-limiting caries therapy with proteolytic agents.

PURPOSE: To determine the extent to which artificial carious dentin can be removed by agents that do not seem to attack sound dentin such as pepsin, trypsin, collagenase and NaOCl, and to evaluate the effect of the enzyme pepsin and a new enzymatic solution SFC-V (pepsin in mild acidic buffer) as a self-limiting caries therapy in deep dentin carious lesions using our new model for artificial dentin caries. METHODS: Artificial dentin caries was used to investigate different proteolytic agents which have the potential to remove carious tissue. 408 slices of coronal dentin were subjected to a demineralization regime which produces dentin caries very similar to natural lesions: acetic acid (pH 5) or lactic acid (pH 4) were used (7 days). Subsequently, sodium hypochlorite, collagenase, trypsin and pepsin were dissolved each in a suitable buffer and the demineralized dentin was treated for 10 minutes or 24 hours with these solutions. To differentiate the influence of the acidic buffer in case of pepsin, a second experiment was performed. 192 slices were exposed to lactic acid for 1 week. Subsequently the demineralized dentin surfaces were treated with either the enzyme pepsin in its acidic buffer, the acidic buffer alone, and in addition a neutral buffer as a control. In addition a fourth group was added where a new enzyme-based solution SFC-V was used. This second experiment differentiated further the influence of "diffusion enhanced by agitation" versus "diffusion" alone. The application time of the solutions was 3 minutes with and without agitation using a stiff nylon brush. To obtain information on the morphology of the pre- and post-treatment dentin surfaces, high resolution FE-SEM was used. Descriptive statistics were used based on cross tabulation of the morphological criteria. RESULTS: Lactic acid produced demineralized dentin covered with a surface layer removable by proteolytic enzymes while acetic acid produced only demineralized dentin. The amount of tissue removed with the current proteolytic agents ranked as follows: trypsin < pepsin < collagenase < NaOCl. The neutral and the acidic buffers did not affect the surface precipitates while the enzyme pepsin and the solution SFC-V were effective in removing the degraded organic matrix.