Synergistic utility of NBD-Cl fluorogenic loading activity and salting-out assisted liquid-liquid extraction as sample pretreatment in rasagiline tracking in different matrices.

A typical drug used to treat Parkinson's disease is called rasagiline. It belongs to an assortment of drugs known as monoamine oxidase inhibitors, which function by raising dopamine levels in the brain. This work created a unique spectrofluorimetric method for the analytical assay of rasagiline for the first time. The approach utilized the synergistic utility of the fluorogenic properties of benzofurazan and salting-out assisted liquid-liquid extraction. By combining these techniques an ultrasensitive, and highly selective methodology for the assay of rasagiline was established. Measurements were made of the resultant yellow fluorescent product at 533 nm by applying an excitation wavelength of 475.3 nm. The calibration graph was examined to assess its linearity across a range of 30-600 ng/ml. Through estimation, the limit of detection was discovered to be 8.9 ng/ml, while the quantitation limit was estimated to be 27 ng/ml. All relevant parameters influencing the fulfillment of the developed method were thoroughly examined and tuned. Following the directives set by the (ICH) the suggested approach was confirmed and demonstrated its capability for the accurate determination of rasagiline in tablets, as well as for testing content uniformity. The incorporation of salting-out assisted liquid-liquid extraction technology enables effective tracking of rasagiline in plasma samples, providing a novel and innovative approach for its analysis in biological matrices.

Ultrasound-guided continuous erector spinae plane block vs continuous thoracic epidural analgesia for the management of acute and chronic postthoracotomy pain: a randomized, controlled,double-blind trial.

Introduction: Postthoracotomy pain (PTP) is a severe pain complicating thoracic surgeries and its good management decreases the risk of PTP syndrome (PTPS). Objectives: This randomized controlled study evaluated the efficacy of ultrasound-guided continuous erector spinae plane block (ESPB) with or without dexmedetomidine compared with thoracic epidural analgesia (TEA) in managing acute postoperative pain and the possible emergence of PTPS. Methods: Ninety patients with chest malignancies planned for thoracotomy were randomly allocated into 3 equal groups. Group 1: TEA (20 mL of levobupivacaine 0.25% bolus, then 0.1 mL/kg/h of levobupivacaine 0.1%), group 2: ESPB (20 mL of levobupivacaine only 0.1% bolus every 6 hours), and group 3: ESPB (20 mL of levobupivacaine 0.25% and 0.5 µg/kg of dexmedetomidine Hcl bolus every 6 hours). Results: Resting and dynamic visual analog scales were higher in group 2 compared with groups 1 and 3 at 6, 24, and 36 hours and at 8 and 12 weeks. Postthoracotomy pain syndrome incidence was higher in group 2 compared with groups 1 and 3 at 8 and 12 weeks, whereas it was indifferent between groups 1 and 3. The grading system for neuropathic pain score was higher in group 2 compared with groups 1 and 3 at 8 and 12 weeks, whereas it was indifferent between groups 1 and 3. Itching, pruritis, and urine retention were higher in group 1 than in ESPB groups. Conclusion: Ultrasound-guided ESPB with dexmedetomidine is as potent as TEA in relieving acute PTP and reducing the possible emergence of chronic PTPS. However, the 2 techniques were superior to ESPB without dexmedetomidine. Erector spinae plane block has fewer side effects compared with TEA.

Clinical Presentation and Management of Multisystem Inflammatory Syndrome in Children With COVID-19: A Systematic Review.

Multisystem inflammatory syndrome in children (MIS-C) is a relatively new syndrome associated with coronavirus disease 2019 (COVID-19) that is characterized by a severe clinical course compared to pediatric COVID-19. This review aimed to compile the available evidence on the clinical presentation and management of MIS-C in children with COVID-19. During this systematic review, a comprehensive search was performed in the following databases: PubMed, Embase, Medline, Google Scholar, Cochrane, and Scopus, using predetermined search terms, such as Medical Subject Headings (MeSH) and keywords to find relevant studies on the MIS-C. Relevant data were extracted, and the quality of the studies was evaluated using suitable methods. The collected findings were synthesized and discussed in the study. The World Health Organization's (WHO) definition of MIS-C was the most favored due to its precision and inclusiveness. MIS-C primarily affected children aged 6-12 years, with male predominance. MIS-C involves a range of systems, including gastrointestinal, cardiovascular, hematologic, mucocutaneous, and respiratory. Radiographic findings revealed cardiovascular abnormalities, solid visceral organ involvement, and bowel abnormalities, reflecting a systemic inflammatory process. Laboratory investigations unveiled elevated inflammatory markers, neutrophil activation, release of extracellular traps in vessels, elevated procalcitonin, hyponatremia, hypoalbuminemia, low hemoglobin, and thrombocytopenia. The inflammatory markers and autoantibody profiles are essential in differentiating MIS-C from COVID-19. The preferred treatment primarily involves immunomodulatory therapies like intravenous immunoglobulin (IVIG), glucocorticoids, and interleukin-6 or 1RA inhibitors or a combination of those. In severe cases, extracorporeal membrane oxygenation (ECMO) and mechanical ventilation are necessary, leading to reduced mortality and quick recovery. This review found that the average hospital stay was seven days, and most discharged children fully recovered within seven days. MIS-C is a life-threatening post-COVID-19 condition and involves multiple systems due to systemic inflammation, with elevated inflammation markers. Recognition of multisystem involvement is crucial, and prompt identification and multidisciplinary treatment are vital for optimal outcomes.

Structural and mechanistic insights into the CAND1-mediated SCF substrate receptor exchange.

Modular SCF (SKP1-CUL1-Fbox) ubiquitin E3 ligases orchestrate multiple cellular pathways in eukaryotes. Their variable SKP1-Fbox substrate receptor (SR) modules enable regulated substrate recruitment and subsequent proteasomal degradation. CAND proteins are essential for the efficient and timely exchange of SRs. To gain structural understanding of the underlying molecular mechanism, we reconstituted a human CAND1-driven exchange reaction of substrate-bound SCF alongside its co-E3 ligase DCNL1 and visualized it by cryo-EM. We describe high-resolution structural intermediates, including a ternary CAND1-SCF complex, as well as conformational and compositional intermediates representing SR- or CAND1-dissociation. We describe in molecular detail how CAND1-induced conformational changes in CUL1/RBX1 provide an optimized DCNL1-binding site and reveal an unexpected dual role for DCNL1 in CAND1-SCF dynamics. Moreover, a partially dissociated CAND1-SCF conformation accommodates cullin neddylation, leading to CAND1 displacement. Our structural findings, together with functional biochemical assays, help formulate a detailed model for CAND-SCF regulation.

Application of isoindole fluorophore formation for determination of linagliptin in the sole and co-formulated tablets: Application for plasma assay and content uniformity testing.

Linagliptin is a new medicament belonging to dipeptidyl peptidase-4 enzyme inhibitors group. The mentioned medication is used to cure type 2 diabetes and is taken orally as a monotherapy or in a co-formulation with metformin. or empagliflozin. Herein, a novel, straightforward, and cost-effective method for linagliptin assay was developed with a workable use of an isoindole derivative. The primary amine moiety present in linagliptin enables its condensation with o-phthalaldehyde to form a fluorescent product in the presence of a sulfhydryl group-containing compound (2-mercaptoethanol) 0.01 % V/V. The isoindole fluorophore yield was monitored at (λexcitation 337.8 nm, λemission 434.3 nm) and all experimental variables were meticulously checked and adjusted. Fluorescence intensity versus linagliptin concentration was plotted to construct the calibration graph, and excellent linearity was achieved at values between 50 and 2000 ng/mL. The validity of the method was verified through a rigorous examination of the ICH guidelines. The method application was successful for linagliptin in different dosage forms, content uniformity study, and monitoring in spiked plasma. The devised technique was demonstrated to be a promising, easy, and quick alternate method for linagliptin assayin clinical study and quality control.

The Role of Double Modality Ultrasonographic and Fluoroscopic Guided Superior Hypogastric Plexus Neurolysis in Treating Intractable Pelvic Cancer Pain: A Comparative Study.

BACKGROUND: Superior hypogastric neurolytic block is performed to block visceral pelvic pain. This could be performed through the anterior approach guided by CT or ultrasound and through a posterior approach, guided by fluoroscopy or CT. METHODS: Sixty adult patients with severe visceral pelvic pain (VAS>70 mm) were randomly divided into two groups. Group S: SHP block was done ultrasound guided using the anterior approach and confirmed by fluoroscopy. Group F: SHP block was done fluoroscopic guided using the posterior oblique approach. The VAS (visual analog scale), duration of the technique, time of X-ray exposure, patient satisfaction score, patient global impression of change (PGIC), quality of life score, and daily morphine consumption (mg/day) were measured pre-procedure and at the 1st, 4th, 8th, and 12th week after the procedure. In addition, any side effects of the procedure were recorded. RESULTS: There was a significant difference in VAS between the two groups (P<0.01) (better in group S). The quality of life score was improved from the pre-procedure in both groups (P<0.05), and morphine consumption was significantly lower in group S than in group F (P<0.05) at the 1st, 4th, and 8th week and not significant at the 12th week. The two groups show a statistically significant difference as regards the duration of the procedure and X-ray exposure (P<0.01). There was a statistically significant difference in the satisfactory score between the two groups at the 1st, 4th, 8th, and 12th week (P<0.01). As regards the PGIC score, there was no statistically significant difference between the two groups (P>0.05). In group S, no back pain was reported, while 11 patients of group F complained from post-procedure back pain (P<0.001). CONCLUSION: The anterior ultrasound guided SHPB aided by fluoroscopy is suggested to be more superior to the standard fluoroscopic guided technique in relieving pelvic cancer pain and decreasing morphine consumption.

Expression of Myoglobin in Normal and Cancer Brain Tissues: Correlation With Hypoxia Markers.

BACKGROUND: Myoglobin (MB) is increasingly recognized as a key player in cancer growth and metastasis. Low oxygen tensions, commonly associated with highly aggressive and recurrent cancers, have been shown to regulate its expression in several cancers such as lung, neck, prostate and breast cancer. However, it is not yet known whether it contributes to the growth and spread of brain cancers especially Glioblastoma multiforme (GBM). METHODS: Here we investigate the expression of MB, and its correlation with the hypoxia markers carbonic anhydrase IX (CAIX) and lactate dehydrogenase A (LDHA), in human tissue microarrays of multiple organ tumors, brain tumors, and GBM tumors, and their respective cancer-adjacent normal tissues. Correlation between MB protein expression and tumor grade was also assessed. RESULTS: We show that MB protein is expressed in a wide variety of cancers, benign tumors, cancer-adjacent normal tissues, hyperplastic tissue samples and normal brain tissue, and low oxygen tensions modulate MB protein expression in different brain cancers, including GBM. Enhanced nuclear LDHA immune-reactivity in GBM was also observed. Finally, we report for the first time a positive correlation between MB expression and brain tumor grade. CONCLUSION: Our data suggest that hypoxia regulate MB expression in different brain cancers (including GBM) and that its expression is associated with a more aggressive phenotype as indicated by the positive correlation with the brain tumor grade. Additionally, a role for nuclear LDHA in promoting aggressive tumor phenotype is also suggested based on enhanced nuclear expression which was observed only in GBM.

Cryo-EM reveals the transition of Arp2/3 complex from inactive to nucleation-competent state.

Arp2/3 complex, a crucial actin filament nucleator, undergoes structural rearrangements during activation by nucleation-promoting factors (NPFs). However, the conformational pathway leading to the nucleation-competent state is unclear due to lack of high-resolution structures of the activated state. Here we report a ~3.9 Å resolution cryo-EM structure of activated Schizosaccharomyces pombe Arp2/3 complex bound to the S. pombe NPF Dip1 and attached to the end of the nucleated actin filament. The structure reveals global and local conformational changes that allow the two actin-related proteins in Arp2/3 complex to mimic a filamentous actin dimer and template nucleation. Activation occurs through a clamp-twisting mechanism, in which Dip1 forces two core subunits in Arp2/3 complex to pivot around one another, shifting half of the complex into a new activated position. By showing how Dip1 stimulates activation, the structure reveals how NPFs can activate Arp2/3 complex in diverse cellular processes.

Immune Checkpoint Regulators: A New Era Toward Promising Cancer Therapy.

During the last century, our battle against cancer has been inaugurated upon three main approaches; surgery, radiation and chemotherapy. The latest findings on the effectiveness of immunotherapy in cancer management offer a ray of hope after decades of research and studies on the best treatment methods. Immunotherapy has proven effective in the surveillance and destruction of cancer- causing cells, demonstrating its ability to suppress cancer through controlling the wellestablished immune-editing process. Immuno-editing is a process that comprises three principal elements; elimination, equilibrium, and escape, and is paramount to the comprehension of checkpoint inhibition. Cancer cells employ various approaches to evade the elimination step leading to its immune- escape. The escape mechanism encompasses the up-regulation of negative co-signals that block successful activation of cancer-eradicating immune cells, developing cytokine background that favors the immunosuppressive tumor microenvironment (TME), or dropping the expression of tumor- specific proteins known as neo-antigens, therefore reducing the immunogenic activity against cancer cells. Today, checkpoint inhibitors are considered as a primary approach in our fight against cancer. Strategies targeting the inhibitory roles of checkpoint inhibitors have been shown effective against different cancer types and stages, and some already gained the FDA's approval. This review seeks to comprehensively cover the historical background as well as the most recent updates for the role of immune checkpoint regulators in the maintenance of immune homeostatic balance as well as keeping the tumorigenic cells in check.

Modified technique for thermal radiofrequency ablation of Thoracic dorsal root ganglia under combined fluoroscopy and CT guidance: a randomized clinical trial.

BACKGROUND: This study is comparing thermal radiofrequency ablation (TRFA) of the thoracic dorsal root ganglia (TDRG) guided by Xper CT and fluoroscopy with the standard fluoroscopy. METHODS: This randomized clinical trial included 78 patients suffering from chronic refractory pain due to chest malignancies randomly allocated into one of two groups according to guidance of TRFA of TDRG. In CT guided group (n = 40) TRFA was done under integrated Xper CT-scan and fluoroscopy guidance, while it was done under fluoroscopy guidance only in standard group (n = 38). The primary outcome was pain intensity measured by visual analog scale (VAS) score, functional improvement and consumption of analgesics. The secondary outcome measures were patient global impression of changes (PGIC) and adverse effects. RESULTS: VAS scores decreased in the two groups compared to baseline values (p < 0.001) and were lower in CT guided group up to 12 weeks. Pregabalin and oxycodone consumption was higher in the standard group at 1, 4 and 12 weeks (p < 0.001). Functional improvement showed near significant difference between the two groups (P = 0.06 at week 1, 0.07 at week 4 respectively) while the difference was statistically significant at week 12 (P = 0.04). PGIC showed near significant difference only at week 1 (P = 0.07) while the per-patient adverse events were lower in CT guided group (p = 0.027). CONCLUSIONS: Integrated modality guidance with Xper CT-scan and fluoroscopy together with suprapedicular inferior transforaminal approach may improve efficacy and safety of TRFA of TDRG for the treatment of intractable chest pain in cancer patients. TRIAL REGISTRATION: The study was retrospectively registered at clinicaltrials.gov on 04/22/2018 (Registration No.: NCT03533413).

The effect of leptin on maturation of the ovarian compartments of albino rats: an ultrastructural and histological study.

BACKGROUND: Leptin serves as a signal to the central nervous system with information on the critical amount of adipose tissue stores that is necessary for activation of the hypothalamic-pituitary-ovarian axis. OBJECTIVES: To document the histological and ultrastructural changes that occur in the ovarian follicles of immature albino rats treated with leptin when compared with controls. Furthermore, the endometrial histological and immunohistochemical, and vaginal cytological changes suggestive of ovulation were assessed. ANIMALS AND METHODS: The study was carried out on 50 immature female albino rats aged 22 days; 24 of them were injected with 5 microg leptin daily and 26 rats were taken as controls. Vaginal smears were taken daily, three animals were sacrificed every 2-4 days from each group, ovaries and uteri were dissected and specimens were prepared for electron microscopic, histological and/or immunohistochemical assessment. The research project was approved by The Histology Department Committee of Alexandria Medical School, which is licensed for animal care and use. RESULTS: Electron microscopic and histological examination confirmed the occurrence of maturational changes in various ovarian components from 26 days of age in leptin-treated rats, with ovulation occurring from the age of 30 days. The granulosa, theca and stroma cells showed signs of steroidogenesis, with increased mitosis within granulosa cells. The ooplasm showed an increased number of organelles, and annulate lamellae were demonstrated. The zona pellucida revealed microvilli, adhering junctions and gap junctions. Similarly, the endometrial histological and vaginal cytological maturational changes were detected in leptin-treated rats from 26 days of age. Furthermore, there was high expression of estrogen receptor-alpha in almost all columnar and stroma cells of the endometrium. However, the control rats ovulated around the normal age of maturation, i.e. 42 days. CONCLUSION: We documented ultrastructural, histological, immunohistochemical and cytological evidence that leptin accelerates the onset of puberty in female albino rats. The potential role of exogenous leptin, in cases of impaired reproductive function in humans, needs to be elucidated.