SARS-CoV-2 Mpro responds to oxidation by forming disulfide and NOS/SONOS bonds.

The main protease (Mpro) of SARS-CoV-2 is critical for viral function and a key drug target. Mpro is only active when reduced; turnover ceases upon oxidation but is restored by re-reduction. This suggests the system has evolved to survive periods in an oxidative environment, but the mechanism of this protection has not been confirmed. Here, we report a crystal structure of oxidized Mpro showing a disulfide bond between the active site cysteine, C145, and a distal cysteine, C117. Previous work proposed this disulfide provides the mechanism of protection from irreversible oxidation. Mpro forms an obligate homodimer, and the C117-C145 structure shows disruption of interactions bridging the dimer interface, implying a correlation between oxidation and dimerization. We confirm dimer stability is weakened in solution upon oxidation. Finally, we observe the protein's crystallization behavior is linked to its redox state. Oxidized Mpro spontaneously forms a distinct, more loosely packed lattice. Seeding with crystals of this lattice yields a structure with an oxidation pattern incorporating one cysteine-lysine-cysteine (SONOS) and two lysine-cysteine (NOS) bridges. These structures further our understanding of the oxidative regulation of Mpro and the crystallization conditions necessary to study this structurally.

Wound healing potential of Cystoseira/mesenchymal stem cells in immunosuppressed rats supported by overwhelming immuno-inflammatory crosstalk.

Wound healing, one of the most intricate and dynamic processes of the body, maintains skin integrity following trauma. One of the main issues that still exists is impaired wound healing, particularly for immunosuppressed patients. Recently, natural products from marine environments have been employed in wound-repairing activities. This work investigates the mesenchymal stem cells in the combined capacity of the bone marrow (BMMSC) for wound healing and Cystoseira sp. Algae extract in immunosuppressed rats. High-resolution liquid chromatography / MS investigation of Cystoseira extract revealed the prevalence of fatty acids that have wound-soothing potential. From constructed PPI network for wound healing and further analysis through molecular docking and molecular dynamics (MD) simulation experiments suggested that cystalgerone metabolite may be responsible for the wound healing-promoting effect of Cystoseira extract. According to the CD marker characterization of the BMMSC, 98.21% of them expressed CD90, and 97.1% expressed CD105. Sixteen d after immunity suppression (by 40 mg/kg hydrocortisone daily), an incision was made in the dorsal skin of the rat. The treatments were applied for 16 d and samples were taken from the tested groups on the 8th, 14th, and 16th days. The BMMSCs / Cystoseira group showed significantly improved wound closure, thickness, density of new layers, and skin elasticity than the control group (p < 0.001). The BMMSCs / Cystoseira combination significantly reduced the oxidative indicators, pro-inflammatory cytokines, and immune markers, according to the RT-PCR gene expression study. In order to delve deeper into the complex interconnections among wound healing-related biological targets and pinpoint key factors in this complex process, we engaged in network pharmacology and computational research. Subsequently, we conducted a comprehensive computational analysis, including reverse docking, free energy (ΔG) computation, and molecular dynamics simulations, on the molecular structures of the annotated compounds. The purpose of this investigation was to identify potential new targets for these chemicals as well as any potential interactions they may have with different signaling pathways related to the wound healing process. Our research indicates that the primary compounds of Cystoseira holds potential wound healing therapeutic activity. Although more safety testing and clinical studies are required, the combination has great potential for regenerative medicine and could be a revolutionary advance in the healing of the wounds of immunosuppressed patients.

Antiplasmodial potential of phytochemicals from Citrus aurantifolia peels: a comprehensive in vitro and in silico study.

Phytochemical investigation of Key lime (Citrus aurantifolia L., F. Rutaceae) peels afforded six metabolites, known as methyl isolimonate acetate (1), limonin (2), luteolin (3), 3`-hydroxygenkwanin (4), myricetin (5), and europetin (6). The structures of the isolated compounds were assigned by 1D NMR. In the case of limonin (2), further 1- and 2D NMR experiments were done to further confirm the structure of this most active metabolite. The antiplasmodial properties of the obtained compounds against the pathogenic NF54 strain of Plasmodium falciparum were assessed in vitro. According to antiplasmodial screening, only limonin (2), luteolin (3), and myricetin (5) were effective (IC50 values of 0.2, 3.4, and 5.9 µM, respectively). We explored the antiplasmodial potential of phytochemicals from C. aurantifolia peels using a stepwise in silico-based analysis. We first identified the unique proteins of P. falciparum that have no homolog in the human proteome, and then performed inverse docking, ΔGBinding calculation, and molecular dynamics simulation to predict the binding affinity and stability of the isolated compounds with these proteins. We found that limonin (2), luteolin (3), and myricetin (5) could interact with 20S a proteasome, choline kinase, and phosphocholine cytidylyltransferase, respectively, which are important enzymes for the survival and growth of the parasite. According to our findings, phytochemicals from C. aurantifolia peels can be considered as potential leads for the development of new safe and effective antiplasmodial agents.

Knowledge, Attitude, and Practice of Adult Diabetics Regarding Diabetic Foot Ulcers: A Cross-Sectional Study in Saudi Arabia.

Background Diabetic foot ulcers (DFUs) are one of the most common and fatal complications of diabetic patients with uncontrolled diabetes mellitus (DM) that may end with their feet amputation. These complications can be prevented through the adherence of diabetic patients to their diabetes management plan and by educating them about risk factors, complications of diabetic foot, and proper foot care. To develop effective health education programs to educate diabetic patients and caregivers regarding diabetic foot and its effective care, we should first identify gaps in patients' knowledge and perception of diabetic foot and evaluate their practice of foot care. Objectives This study aimed to evaluate knowledge and attitude toward DFUs and the practice of foot care among adult diabetic patients attending Al-Qunfudah Diabetes Center, Saudi Arabia, from October 2022 to March 2023. Methods A total of 403 adult diabetic patients were recruited in this cross-sectional study during their attendance at the diabetes center in Al-Qunfudah district, Saudi Arabia. The study researchers conducted a physical face-to-face interview with each diabetic patient using a validated questionnaire with closed-ended questions to collect their responses regarding their knowledge and attitude toward DFUs and their behavior toward foot care. The collected data were analyzed using IBM SPSS Statistics for Windows, version 23 (released 2015; IBM Corp., Armonk, New York, United States). Results Out of 403 diabetics, 50.4% (n = 203) had inadequate knowledge (knowledge score < 80%), 46.4% (n = 187) had negative attitudes (attitude score < 80%) toward DFUs, and the majority could not practice foot care well, with 77.7% (n = 313) giving a practice score less than 80%. Seventy patients (17.4%) self-reported a history of DFUs. Predictors of good knowledge about DFUs among diabetics include age between 30 and 59 years (odds ratio (OR) = 2.942, confidence interval (CI) 95% = 1.695-2.107, p < 0.001), marriage (OR = 3.101, CI 95% = 1.893-5.079, p < 0.001), working (OR = 5.325, CI 95% = 3.019-9.389, p < 0.001), diploma education (OR = 8.205, CI 95% = 3.332-20.203, p < 0.001), managing DM with oral drugs (OR = 2.219, CI 95% = 1.399-3.519, p < 0.001), and having no DFUs (OR = 2.712, CI 95% = 1.557-4.723, p < 0.001). Males were more likely to practice foot care well (OR = 1.925, CI 95% = 1.142-3.245, p = 0.013). Primary education (OR = 3.421, CI 95% = 1.655-7.073, p < 0.001) predicted appropriate foot care. Patients with DM for one to five years (OR = 1.995, CI 95% = 1.139-3.493, p = 0.016) and those on diet and metformin (OR = 2.133, CI 95% = 1.134-4.011, p = 0.019) were expected to have better foot care than other diabetics. Conclusion Diabetic patients in the Al-Qunfudah district of Saudi Arabia had inadequate knowledge and negative attitudes toward DFUs, and their foot care behaviors were inadequate. Furthermore, DFUs were self-reported in around 17% of diabetic patients. Specialized training programs are recommended to enhance knowledge regarding DFUs among diabetics and motivate and train them and caregivers on how to conduct proper foot care. These educational programs should target all diabetics, with an emphasis on those with DFUs, females, non-working patients, individuals who have had DM for a longer time, and illiterate diabetics. To understand the factors behind patients' negative attitudes toward diabetic foot, future qualitative research is required.

Knowledge and Perception of Radiation Risk From Computed Tomography Scans Among Patients Attending an Emergency Department.

To evaluate the level of knowledge about radiation dose and possible risks related to computed tomography (CT) scans among patients visiting emergency departments (EDs), a survey was conducted over a two-month period. A total of 357 adult patients (44% men and 56% women) presenting for diagnostic imaging in the ED answered a survey consisting of 15 questions. The survey included questions about the participants' demographics and knowledge of radiation. Most of the respondents (58.5%) reported that the physician did not explain the potential risk of radiation before the procedure. In addition, more than half of the respondents (58.1%) expressed feeling anxious about the potential risk of radiation. Most respondents (84.9%) stated that the potential radiation risk did not affect their decision to proceed with the procedure. Overall, the findings highlight a lack of information about radiation and its potential risks provided to patients prior to the diagnostic procedure. Increasing awareness and understanding of the risks associated with these imaging modalities should be considered essential in modern communities. Efforts should be made to ensure that patients undergoing diagnostic imaging are aware of the radiation risks they may encounter.

Learning curve of ultrasound-guided surgeon-administered transversus abdominis plane (UGSA-TAP) block on a porcine model.

Background: Surgeons commonly perform ultrasound-guided Transversus Abdominis Plane blocks to manage acute pain following abdominal surgeries. There is no consensus on whether surgeons should undergo basic hands-on training to perform TAP blocks or if video-based learning is sufficient. We theorized that simulation-based learning is superior to video-based learning. In the present study, we present the analysis of technical skills of UGSA-TAP block performance on a live porcine model by general surgery trainees after undergoing video or simulation-based learning. Methods: We performed a prospective, double-blinded, randomized study. Ten surgery residents and two surgery critical-care fellows (n = 12) without prior experience in performing the TAP block were recruited. The participants were randomized either into a video-based or simulation-based training group. After that, all participants performed a TAP block on a live anesthetized pig, which was recorded and scored by three blinded anesthesiologists. All participants completed a post-performance survey to assess their confidence in gaining competency in the UGSA-TAP block. Statistical analyses were performed to assess the differences between the two groups. P < 0.05 was considered statistically significant. Results: All simulation-based learning participants successfully performed a survey scan, identified the three muscular layers of the abdominal wall, and identified the transversus abdominis plane compared to 50 %, 50 %, and 33 % video-based learning group participants for the respective parameters (p < 0.05). While some performance metrics showed no statistically significant differences between the groups, substantial effect sizes (Cohen's ℎ up to 1.07) highlighted notable differences in participants' performance. Both groups exhibited confidence in core competencies, with varied rates of satisfactory skill execution. Performance assessed using a global rating scale revealed a higher passing rate for the simulation group (83 % vs. 33 %). Participant feedback via the Likert scale reflected confidence post-training. Inter-rater reliability (0.83-1) confirmed the robustness of study evaluations. Conclusion: The UGSA-TAP block curriculum should be introduced into the surgical residency programs with an emphasis on simulation-based learning to enhance the procedural skills of the trainees before transitioning to surgical patients.

Apium extract alleviates indomethacin-induced gastric ulcers in rats via modulating the VEGF and IK-κB/NF-κB p65 signaling pathway: insights from in silico and in vivo investigations.

BACKGROUND: Gastric ulcers represent a worldwide health problem, characterized by erosions that affect the mucous membrane of the stomach and may even reach the muscular layer, leading to serious complications. Numerous natural products have been assessed as anti-ulcerogenic agents, and have been considered as new approaches for treatment or prevention of gastric ulcers. The present research investigated the preventive benefits of Apium graveolens L. (Apiaceae), known as celery, seed extract towards indomethacin-induced ulceration of the stomach in rats. METHODS: Metabolomic profiling, employing liquid chromatography coupled to high-resolution electrospray ionization mass spectrometry (LC-HR-ESI-MS), was implemented with the aim of investigating the chemical profile of the seeds. Histopathological analysis of gastric tissues, as well as assessment of numerous inflammatory cytokines and oxidative stress indicators, confirmed the in vivo evaluation. RESULTS: The prior treatment with A. graveolens seed extract resulted in a substantial reduction in the ulcer index when compared to the indomethacin group, indicating an improvement in stomach mucosal injury. Moreover, the gastroprotective effect was demonstrated through examination of the oxidative stress biomarkers which was significantly attenuated upon pre-treatment with A. graveolens seed extract. Vascular endothelial growth factor (VEGF), a fundamental angiogenic factor that stimulates angiogenesis, was markedly inhibited by indomethacin. A. graveolens seed extract restored this diminished level of VEGF. The dramatic reductions in NF-κB protein levels indicate a considerable attenuation of the indomethacin-induced IKκB/NF-κB p65 signaling cascade. These activities were also correlated to the tentatively featured secondary metabolites including, phenolic acids, coumarins and flavonoids, previously evidenced to exert potent anti-inflammatory and antioxidant activities. According to our network pharmacology study, the identified metabolites annotated 379 unique genes, among which only 17 genes were related to gastric ulcer. The PTGS2, MMP2 and PTGS1 were the top annotated genes related to gastric ulcer. The top biological pathway was the VEGF signaling pathway. CONCLUSION: A. graveolens seed extract possesses significant anti-ulcer activity, similar to famotidine, against gastric lesions induced by indomethacin in rats. It is worth highlighting that the extract overcomes the negative effects of conventional chemical anti-secretory drugs because it does not lower stomach acidity.

3D atomic structure from a single X-ray free electron laser pulse.

X-ray Free Electron Lasers (XFEL) are cutting-edge pulsed x-ray sources, whose extraordinary pulse parameters promise to unlock unique applications. Several new methods have been developed at XFELs; however, no methods are known, which allow ab initio atomic level structure determination using only a single XFEL pulse. Here, we present experimental results, demonstrating the determination of the 3D atomic structure from data obtained during a single 25 fs XFEL pulse. Parallel measurement of hundreds of Bragg reflections was done by collecting Kossel line patterns of GaAs and GaP. To the best of our knowledge with these measurements, we reached the ultimate temporal limit of the x-ray structure solution possible today. These measurements open the way for obtaining crystalline structures during non-repeatable fast processes, such as structural transformations. For example, the atomic structure of matter at extremely non-ambient conditions or transient structures formed in irreversible physical, chemical, or biological processes may be captured in a single shot measurement during the transformation. It would also facilitate time resolved pump-probe structural studies making them significantly shorter than traditional serial crystallography.

Extracellular vesicles as tools and targets in therapy for diseases.

Extracellular vesicles (EVs) are nano-sized, membranous structures secreted into the extracellular space. They exhibit diverse sizes, contents, and surface markers and are ubiquitously released from cells under normal and pathological conditions. Human serum is a rich source of these EVs, though their isolation from serum proteins and non-EV lipid particles poses challenges. These vesicles transport various cellular components such as proteins, mRNAs, miRNAs, DNA, and lipids across distances, influencing numerous physiological and pathological events, including those within the tumor microenvironment (TME). Their pivotal roles in cellular communication make EVs promising candidates for therapeutic agents, drug delivery systems, and disease biomarkers. Especially in cancer diagnostics, EV detection can pave the way for early identification and offers potential as diagnostic biomarkers. Moreover, various EV subtypes are emerging as targeted drug delivery tools, highlighting their potential clinical significance. The need for non-invasive biomarkers to monitor biological processes for diagnostic and therapeutic purposes remains unfulfilled. Tapping into the unique composition of EVs could unlock advanced diagnostic and therapeutic avenues in the future. In this review, we discuss in detail the roles of EVs across various conditions, including cancers (encompassing head and neck, lung, gastric, breast, and hepatocellular carcinoma), neurodegenerative disorders, diabetes, viral infections, autoimmune and renal diseases, emphasizing the potential advancements in molecular diagnostics and drug delivery.

Acceptability of self-administered antigen test for COVID-19 in the Philippines.

OBJECTIVES: In response to the Omicron surge in early 2022, the HTA Philippines evaluated the acceptability of Filipinos in using self-administered antigen tests (SAAgTs) as part of COVID-19 HTAs in the Philippines. METHODS: Scoping review from literature databases was initially conducted to identify preset codes in the use of SAAgT. Preset codes were used to establish the questions for focus group discussions (FGDs). Semi-structured questionnaires were created through Delphi technique. FGDs with four stakeholder groups (i.e., nine healthcare workers [HCWs], seven representatives of at-risk groups, six economic frontliners, and seven representatives of micro-small-medium-sized enterprises) were conducted. RESULTS: Discomfort in being a target of stigma and being prescribed an "illness identity" when suspected or confirmed COVID-19-positive, along with lack of confidence to perform self-test, caused hesitancy in self-testing among participants. The need for subsidies for test kits from the government or employers was emphasized to increase its accessibility. Having a designated access point and reporting system for SAAgT was highlighted to avoid nepotism (padrino system attributed to debt of gratitude), inequitable distribution, and lapses in reporting. A participatory approach to education was perceived as crucial to reduce any misconceptions associated with the use of SAAgT. CONCLUSIONS: All FGD groups expressed favorable reviews on the implementation of SAAgT because it can potentially reduce the burden of health facility-administered tests. These findings were considered by the HTA Council in the recommendation of SAAgT as part of the overarching national strategies for the diagnosis and screening of COVID-19.

Protein supplement intake by non-athlete gym attendees in Jazan region: misconceptions and gender differences.

BACKGROUND: Protein supplements (PSs) have gained widespread popularity among non-athlete gym attendees, who often perceive them as essential tools for muscle growth and recovery. However, misinformation surrounding PSs may lead to inappropriate use and negative health consequences. This study aimed to assess whether non-athlete gym attendees using PSs have greater misconceptions than non-users while also examining the prevalence of PS consumption and gender differences. METHODS: A cross-sectional study of 387 participants in the Jazan region was conducted. Customers of 10 fitness centers were screened with a questionnaire comprising questions to measure PS misconceptions. Participants were divided into PS users and non-users. Data were analyzed using descriptive statistics and the Chi-square test to assess the associations between variables. An independent t-test was used to compare the PS Misconception Index Score between the two groups. RESULTS: A total of 82.4% of non-athlete gym attendees consumed at least one PS. Our findings revealed a significant association between gender and PS utilization (P<0.001), with a higher proportion of females (90.8%) consuming PS than males (77.6%). PS users had a significantly lower PS Misconception Index Score than non-users (26.8 vs. 28.3; P=0.006), indicating that PS users had a higher number of misconceptions. The internet (41.37%) and coaches (gym instructors/trainers) (34.48%) were the most common sources of information about PSs, with muscle gain being the primary reason for consumption (82.75%). CONCLUSIONS: PS consumption is highly prevalent among non-athlete gym attendees in the Jazan region, with many individuals having misconceptions about their benefits and potential risks. Targeted educational interventions are needed to promote evidence-based knowledge about PSs for gym attendees, as well as for coaches, given that they were among the primary sources of information on supplements.

Intralesional cidofovir vs. bevacizumab for recurrent respiratory papillomatosis: a systematic review and indirect meta-analysis.

BACKGROUND: Specific HPV types cause recurrent respiratory papillomatosis (R.R.P.). When administered intralesionally, cidofovir, an antiviral agent, has shown favorable outcomes in reducing papilloma. Bevacizumab, an angiogenesis inhibitor, has demonstrated improved R.R.P. However, both treatments lack FDA approval for R.R.P. Our study aims to evaluate the efficacy and safety of intralesional Cidofovir and Bevacizumab for R.R.P. and compare the two interventions. METHODS: We searched five electronic databases to find relevant studies. After the screening, data were extracted from the included studies. Pooled ratios with 95% confidence intervals (CIs) were used for categorical outcomes, and mean difference (MD) was used for continuous outcomes. Statistical heterogeneity was evaluated using the chi-squared test for I2 statistics. The Cochrane Risk of Bias assessment tool was used to assess the methodological quality of randomized controlled trials (RCTs), while the National Institutes of Health's tool was used for observational studies. Analysis was done by Review Manager software. RESULTS: In our comprehensive meta-analysis of 35 articles involving 836 patients, cidofovir demonstrated an overall remission ratio of (0.90 [95% CI: 0.83, 0.98], p = 0.01), while bevacizumab (0.92 [95% CI: 0.79, 1.07]), p = 0.3). The complete remission ratio for cidofovir was (0.66 [95% CI: 0.57, 0.75], p > 0.0001), while bevacizumab was (0.29 [95% CI: 0.12, 0.71], p = 0.07). In partial remission, Bevacizumab showed a higher ratio than Cidofovir 0.74 [0.55, 0.99] vs. 0.40 [0.30, 0.54]. Bevacizumab had a pooled ratio of 0.07 [95% CI: 0.02, 0.30] in terms of no remission, indicating better outcomes compared to Cidofovir with a ratio of 0.28 [95% CI: 0.16, 0.51]. Additionally, Cidofovir showed a favorable decrease in the Derkay Severity Score (DSS) with a mean difference (MD) of 1.98 [95% CI: 1.44, 2.52]. CONCLUSION: Cidofovir had a higher impact on complete remission compared to Bevacizumab. Both showed partial remission, with Bevacizumab having a higher ratio. Moreover, Cidofovir showed a significant decrease in DSS. Bevacizumab had lower rates of no remission and recurrence and fewer adverse events compared to Cidofovir. However, the difference between the two treatments was not significant, except for partial remission.

Large animal models to study effectiveness of therapy devices in the treatment of heart failure with preserved ejection fraction (HFpEF).

Our understanding of the complex pathophysiology of Heart failure with preserved ejection fraction (HFpEF) is limited by the lack of a robust in vivo model. Existing in-vivo models attempt to reproduce the four main phenotypes of HFpEF; ageing, obesity, diabetes mellitus and hypertension. To date, there is no in vivo model that represents all the haemodynamic characteristics of HFpEF, and only a few have proven to be reliable for the preclinical evaluation of potentially new therapeutic targets. HFpEF accounts for 50% of all the heart failure cases and its incidence is on the rise, posing a huge economic burden on the health system. Patients with HFpEF have limited therapeutic options available. The inadequate effectiveness of current pharmaceutical therapeutics for HFpEF has prompted the development of device-based treatments that target the hemodynamic changes to reduce the symptoms of HFpEF. However, despite the potential of device-based solutions to treat HFpEF, most of these therapies are still in the developmental stage and a relevant HFpEF in vivo model will surely expedite their development process. This review article outlines the major limitations of the current large in-vivo models in use while discussing how these designs have helped in the development of therapy devices for the treatment of HFpEF.

Efficacy of platelet-rich plasma (PRP) in benign vocal fold lesions: a systematic review and meta-analysis.

OBJECTIVES: To evaluate the efficacy of platelet-rich plasma (PRP) in benign vocal fold lesions. METHODS: MEDLINE, Cochrane Central, Web of Science, and Scopus databases were searched in April 2023 for relevant clinical trials. Inclusion criteria were clinical trials evaluating the efficacy of PRP in benign vocal fold lesions. We conducted a comparative double-arm analysis using the pooled mean difference (MD) and 95% confidence interval (CI). Outcomes of interest included the vocal handicap index (VHI), the Jitter and Shimmer percentages, and the noise-to-harmonic ratio (NHR). RESULTS: Six studies matched the inclusion criteria. The pooled analysis shows that PRP was associated with significantly lower VHI scores compared with the control (MD = - 5.06, p < 0.01). Regarding the Jitter percentage, the PRP group was not superior to the control group at 2 and 4 weeks. However, the results revealed that PRP significantly reduced the Jitter percentage at 3 months (MD = - 0.61, p = 0.0008). The overall analysis favored the PRP arm significantly (p < 0.001). As for the Shimmer percentage, the combined effect estimate favored the PRP group (MD = - 1.22, p = 0.002). Subgroup analysis according to the time did not reveal any significant differences between studies at 2 weeks, 4 weeks, and 3 months. The analysis of the NHR outcome revealed a significant difference between both groups (MD = -1.09, p = 0.01). However, at 4 weeks, the treatment group had a significantly lower NHR % compared to the control group (MD = - 0.61, p = 0.02). There was no significant difference at 3 months (MD = - 2.14, p = 0.14). CONCLUSIONS: Platelet-rich plasma is effective in reducing VHI scores, Jitter and Shimmer percentages, and NHR values. This effect is more evident after follow-up, especially 3 months.

New cytotoxic dammarane type saponins from Ziziphus spina-christi.

Cancer is the world's second-leading cause of death. Drug development efforts frequently focus on medicinal plants since they are a valuable source of anticancer medications. A phytochemical investigation of the edible Ziziphus spina-christi (F. Rhamnaceae) leaf extract afforded two new dammarane type saponins identified as christinin E and F (1, 2), along with the known compound christinin A (3). Different cancer cell lines, such as lung cancer (A549), glioblastoma (U87), breast cancer (MDA-MB-231), and colorectal carcinoma (CT-26) cell lines, were used to investigate the extracted compounds' cytotoxic properties. Our findings showed significant effects on all the tested cell lines at varying concentrations (1, 5, 10, and 20 µg/mL). The three compounds exhibited potent activity at low concentrations (< 10 µg/mL), as evidenced by their low IC50 values. To further investigate the complex relationships between these identified cancer-relevant biological targets and to identify critical targets in the pathogenesis of the disease, we turned to network pharmacology and in silico-based investigations. Following this, in silico-based analysis (e.g., inverse docking, ΔG calculation, and molecular dynamics simulation) was performed on the structures of the isolated compounds to identify additional potential targets for these compounds and their likely interactions with various signalling pathways relevant to this disease. Based on our findings, Z. spina-christi's compounds showed promise as potential anti-cancer therapeutic leads in the future.

3D-printed sheet jet for stable megahertz liquid sample delivery at X-ray free-electron lasers.

X-ray free-electron lasers (XFELs) can probe chemical and biological reactions as they unfold with unprecedented spatial and temporal resolution. A principal challenge in this pursuit involves the delivery of samples to the X-ray interaction point in such a way that produces data of the highest possible quality and with maximal efficiency. This is hampered by intrinsic constraints posed by the light source and operation within a beamline environment. For liquid samples, the solution typically involves some form of high-speed liquid jet, capable of keeping up with the rate of X-ray pulses. However, conventional jets are not ideal because of radiation-induced explosions of the jet, as well as their cylindrical geometry combined with the X-ray pointing instability of many beamlines which causes the interaction volume to differ for every pulse. This complicates data analysis and contributes to measurement errors. An alternative geometry is a liquid sheet jet which, with its constant thickness over large areas, eliminates the problems related to X-ray pointing. Since liquid sheets can be made very thin, the radiation-induced explosion is reduced, boosting their stability. These are especially attractive for experiments which benefit from small interaction volumes such as fluctuation X-ray scattering and several types of spectroscopy. Although their use has increased for soft X-ray applications in recent years, there has not yet been wide-scale adoption at XFELs. Here, gas-accelerated liquid sheet jet sample injection is demonstrated at the European XFEL SPB/SFX nano focus beamline. Its performance relative to a conventional liquid jet is evaluated and superior performance across several key factors has been found. This includes a thickness profile ranging from hundreds of nanometres to 60 nm, a fourfold increase in background stability and favorable radiation-induced explosion dynamics at high repetition rates up to 1.13 MHz. Its minute thickness also suggests that ultrafast single-particle solution scattering is a possibility.

Nutritional Composition and Anti-Type 2 Diabetes Mellitus Potential of Femur Bone Extracts from Bovine, Chicken, Sheep, and Goat: Phytochemical and In Vivo Studies.

Nutritional deficits in one's diet have been established as the key risk factor for T2DM in recent years. Nutritional therapy has been demonstrated to be useful in treating T2DM. The current study was carried out to assess the nutritional composition of bovine (12 months), chicken (4 months), sheep (13 months), and goat (9 months) femur bone extracts, as well as their potential therapeutic effects on T2DM regression in a Wistar albino rat model (500 mg/kg b.wt.). The proximate composition of the different extracts, their fatty acid composition, their amino acids, and their mineral contents were identified. In vivo data indicated considerably improved T2DM rats, as seen by lower serum levels of TL, TG, TC, ALT, AST, ALP, bilirubin, creatinine, urea, IL-6, TNF-α, sICAM-1, sVCAM-1, and MDA. Low levels of HDL-C, GSH, and total proteins were restored during this study. Histological investigations of liver and pancreatic tissue revealed that the distribution of collagen fibers was nearly normal. The bovine extract, on the other hand, was the most active, followed by the sheep, goat, and finally chicken extract. This research could result in the creation of a simple, noninvasive, low-cost, and reliable method for T2DM control, paving the way for potential early therapeutic applications in T2DM control.

Egyptian mandarin peel oil's anti-scabies potential via downregulation-of-inflammatory/immune-cross-talk: GC-MS and PPI network studies.

The current study investigated the scabicidal potential of Egyptian mandarin peel oil (Citrus reticulata Blanco, F. Rutaceae) against sarcoptic mange-in-rabbits. Analysis of the oil's GC-MS identified a total of 20 compounds, accounting for 98.91% of all compounds found. Mandarin peel oil topical application improved all signs of infection, causing a scabicidal effect three days later, whereas in vitro application caused complete mite mortality one day later. In comparison to ivermectin, histopathological analysis showed that the epidermis' inflammatory-infiltration/hyperkeratosis-had disappeared. In addition to TIMP-1, the results of the mRNA gene expression analysis showed upregulation of I-CAM-1-and-KGF and downregulation of ILs-1, 6, 10, VEGF, MMP-9, and MCP-1. The scabies network was constructed and subjected to a comprehensive bioinformatic evaluation. TNF-, IL-1B, and IL-6, the top three hub protein-coding genes, have been identified as key therapeutic targets for scabies. From molecular docking data, compounds 15 and 16 acquired sufficient affinity towards the three screened proteins, particularly both possessing higher affinity towards the IL-6 receptor. Interestingly, it achieved a higher binding energy score than the ligand of the docked protein rather than displaying proper binding interactions like those of the ligand. Meanwhile, geraniol (15) showed the highest affinity towards the GST protein, suggesting its contribution to the acaricidal effect of the extract. The subsequent, MD simulations revealed that geraniol can achieve stable binding inside the binding site of both GST and IL-6. Our findings collectively revealed the scabicidal ability of mandarin peel extract for the first time, paving the way for an efficient, economical, and environmentally friendly herbal alternative for treating rabbits with Sarcoptes mange.

Heterologous expression, purification and structural features of native Dictyostelium discoideum dye-decolorizing peroxidase bound to a natively incorporated heme.

The Dictyostelium discoideum dye-decolorizing peroxidase (DdDyP) is a newly discovered peroxidase, which belongs to a unique class of heme peroxidase family that lacks homology to the known members of plant peroxidase superfamily. DdDyP catalyzes the H2O2-dependent oxidation of a wide-spectrum of substrates ranging from polycyclic dyes to lignin biomass, holding promise for potential industrial and biotechnological applications. To study the molecular mechanism of DdDyP, highly pure and functional protein with a natively incorporated heme is required, however, obtaining a functional DyP-type peroxidase with a natively bound heme is challenging and often requires addition of expensive biosynthesis precursors. Alternatively, a heme in vitro reconstitution approach followed by a chromatographic purification step to remove the excess heme is often used. Here, we show that expressing the DdDyP peroxidase in ×2 YT enriched medium at low temperature (20°C), without adding heme supplement or biosynthetic precursors, allows for a correct native incorporation of heme into the apo-protein, giving rise to a stable protein with a strong Soret peak at 402 nm. Further, we crystallized and determined the native structure of DdDyP at a resolution of 1.95 Å, which verifies the correct heme binding and its geometry. The structural analysis also reveals a binding of two water molecules at the distal site of heme plane bridging the catalytic residues (Arg239 and Asp149) of the GXXDG motif to the heme-Fe(III) via hydrogen bonds. Our results provide new insights into the geometry of native DdDyP active site and its implication on DyP catalysis.

Mix-and-extrude: high-viscosity sample injection towards time-resolved protein crystallography.

Time-resolved crystallography enables the visualization of protein molecular motion during a reaction. Although light is often used to initiate reactions in time-resolved crystallography, only a small number of proteins can be activated by light. However, many biological reactions can be triggered by the interaction between proteins and ligands. The sample delivery method presented here uses a mix-and-extrude approach based on 3D-printed microchannels in conjunction with a micronozzle. The diffusive mixing enables the study of the dynamics of samples in viscous media. The device design allows mixing of the ligands and protein crystals in 2 to 20 s. The device characterization using a model system (fluorescence quenching of iq-mEmerald proteins by copper ions) demonstrated that ligand and protein crystals, each within lipidic cubic phase, can be mixed efficiently. The potential of this approach for time-resolved membrane protein crystallography to support the development of new drugs is discussed.