From Bulk to Single Molecules: Surface-Enhanced Raman Scattering of Cytochrome C Using Plasmonic DNA Origami Nanoantennas.

Cytochrome C, an evolutionarily conserved protein, plays pivotal roles in cellular respiration and apoptosis. Understanding its molecular intricacies is essential for both academic inquiry and potential biomedical applications. This study introduces an advanced single-molecule surface-enhanced Raman scattering (SM-SERS) system based on DNA origami nanoantennas (DONAs), optimized to provide unparalleled insights into protein structure and interactions. Our system effectively detects shifts in the Amide III band, thereby elucidating protein dynamics and conformational changes. Additionally, the system permits concurrent observations of oxidation processes and Amide bands, offering an integrated view of protein structural and chemical modifications. Notably, our approach diverges from traditional SM-SERS techniques by de-emphasizing resonance conditions for SERS excitation, aiming to mitigate challenges like peak oversaturation. Our findings underscore the capability of our DONAs to illuminate single-molecule behaviors, even within aggregate systems, providing clarity on molecular interactions and behaviors.

Clinical Significance of miRNA-145 and -126 in Chronic Obstructive Pulmonary Disease with Pulmonary Embolism.

BACKGROUND: Pulmonary embolism (PE) a consequence of hypercoagulability status associated with chronic obstructive pulmonary disease (COPD) and worsens its course. Recently, microRNAs (miRNAs) have been linked to PE in COPD settings. We aimed to measure expression levels of miRNAs145 and 126 in COPD patients with and without PE. METHODS: Herein, miRNA (145 and 126) expression levels were measured in 250 COPD patients with PE by quan-titative real-time PCR, and their data were compared with 300 COPD patients without PE. RESULTS: Our results showed that miRNA-145 expression was downregulated in COPD patients with PE compared to those without PE. The reverse was observed in miRNA-126 expression that was higher in COPD patients with PE than in those without PE. miRNA-145 correlated positively with FEV1/FVC and correlated negatively with D-dimer in all patients regardless of the presence of PE. In addition, miRNA-126 positively correlated with D-dimer and negatively correlated with FEV1/FVC in all studied COPD patients. CONCLUSIONS: Lower levels of miRNA-145 and higher levels of miRNA-126 associated with worse diagnosis PE in patients with COPD. Extensive studies are mandated to bring a better understanding of the role of these miRNAs in the mechanism of thrombosis in COPD patients.

Does local injection of long acting corticosteroid improve postoperative outcome of hypospadias repair? A randomized controlled trial.

PURPOSE: To assess the safety and efficacy of local corticosteroid injection during hypospadias repair. METHODS: Between May 2021 and March 2023 children less than 10 years who were admitted for hypospadias repair were divided by random allocation into two groups. We injected local corticosteroid 2 ml proximal to coronal sulcus in group A while in group B we didn't. All types of hypospadias were included in the study. We excluded patients older than 10 years and those with pre-existing complicated hypospadias (multiple fistulae and multiple surgeries), or bleeding diatheses. Pre-operative, intraoperative and postoperative variables were compared between the two groups. RESULTS: A total of 120 patients (60 in each group) were enrolled in the study. The mean ages and preoperative variables were not significantly different. The site of hypospadias and the type of surgery were comparable in both groups. (Table) There were no significant differences between both groups regarding average blood loss and operative time in each type of surgical repair. There was a significant higher incidence of intraoperative and postoperative penile oedema in group B (P-value < 0.001) while the incidence of skin discolouration was higher in group A. Postoperative complications, described as Clavian classification, were significantly higher in group B. The incidences of superficial skin infection, meatal stenosis, urethral fistula, and recurrence with the need for redo repair were significantly higher in group B (P-value: 0.002, 0.018, 0.032, and 0.001, respectively). CONCLUSION: Local corticosteroid injection during hypospadias repair minimize the penile oedema and decrease the incidence of postoperative functional and cosmetic complications.

Watching a Single Enzyme at Work Using Single-Molecule Surface-Enhanced Raman Scattering and DNA Origami-Based Plasmonic Antennas.

The detection of a single-enzyme catalytic reaction by surfaced-enhanced Raman scattering (SERS) is presented by utilizing DNA origami-based plasmonic antennas. A single horseradish peroxidase (HRP) was accommodated on a DNA origami nanofork plasmonic antenna (DONA) containing gold nanoparticles, enabling the tracing of single-molecule SERS signals during the peroxide reduction reaction. This allows monitoring of the structure of a single enzymatic catalytic center and products under suitable liquid conditions. Herein, we demonstrate the chemical changes of HRP and the appearance of tetramethylbenzidine (TMB), which works as a hydrogen donor before and after the catalytic reaction. The results show that the iron in HRP adopts Fe4+ and low spin states with the introduction of H2O2, indicating compound-I formation. Density functional theory (DFT) calculations were performed for later catalytic steps to rationalize the experimental Raman/SERS spectra. The presented data provide several possibilities for tracking single biomolecules in situ during a chemical reaction and further developing plasmon-enhanced biocatalysis.

Efficacy of neuromuscular electrical stimulation and interrupted serial casting in children with spastic diplegia.

Objectives: This research was aimed at comparing the effects of neuromuscular electrical stimulation (NMES) combined with interrupted serial casting (SC) versus SC alone on various aspects of lower limb function in children with diplegic cerebral palsy. SC is a clinical technique used to increase passive range of motion (ROM), decrease hypertonicity, and improve walking in children with cerebral palsy (CP). Methods: This randomized comparative trial involved 33 children with diplegic CP, who were randomly assigned to group A or group B at recruitment. Group A received SC along with a customized physical therapy program, whereas group B received the same interventions as group A along with NMES applied through cast windows during casting. Evaluations were based on ROM, the Modified Tardieu Scale, handheld dynamometer measurements, and the Observational Gait Scale. Assessments were conducted before and after 8 weeks of intervention. Results: Both groups exhibited significant improvements in dorsiflexion ROM, popliteal angle, gastrocnemius dynamic spasticity, and hamstring dynamic spasticity after the intervention (P = 0.0001 for all). However, significant differences (P < 0.05) in dorsiflexor strength, knee extensor strength, and observational gait scale score were observed between groups after the intervention, favoring group B. Conclusions: The use of NMES during SC may help overcome the substantial decrease in strength resulting from casting, thus achieving less reduction of tone, improving ROM without significantly decreasing strength, and attaining greater improvements in gait function.

Lab-on-a-DNA origami: nanoengineered single-molecule platforms.

DNA origami nanostructures are self-assembled into almost arbitrary two- and three-dimensional shapes from a long, single-stranded viral scaffold strand and a set of short artificial oligonucleotides. Each DNA strand can be functionalized individually using well-established DNA chemistry, representing addressable sites that allow for the nanometre precise placement of various chemical entities such as proteins, molecular chromophores, nanoparticles, or simply DNA motifs. By means of microscopic and spectroscopic techniques, these entities can be visualized or detected, and either their mutual interaction or their interaction with external stimuli such as radiation can be studied. This gives rise to the Lab-on-a-DNA origami approach, which is introduced in this Feature Article, and the state-of-the-art is summarized with a focus on light-harvesting nanoantennas and DNA platforms for single-molecule analysis either by optical spectroscopy or atomic force microscopy (AFM). Light-harvesting antennas can be generated by the precise arrangement of chromophores to channel and direct excitation energy. At the same time, plasmonic nanoparticles represent a complementary approach to focus light on the nanoscale. Plasmonic nanoantennas also allow for the observation of single molecules either by Raman scattering or fluorescence spectroscopy and DNA origami platforms provide unique opportunities to arrange nanoparticles and molecules to be studied. Finally, the analysis of single DNA motifs by AFM allows for an investigation of radiation-induced processes in DNA with unprecedented detail and accuracy.

Single-Molecule Surface-Enhanced Raman Scattering Measurements Enabled by Plasmonic DNA Origami Nanoantennas.

Surface-enhanced Raman scattering (SERS) has the capability to detect single molecules for which high field enhancement is required. Single-molecule (SM) SERS is able to provide molecule-specific spectroscopic information about individual molecules and therefore yields more detailed chemical information than other SM detection techniques. At the same time, there is the potential to unravel information from SM measurements that remain hidden in Raman measurements of bulk material. This protocol outlines the SM SERS measurements using a DNA origami nanoantenna (DONA) in combination with atomic force microscopy (AFM) and Raman spectroscopy. A DNA origami fork structure and two gold nanoparticles are combined to form the DONAs, with a 1.2-2.0 nm gap between them. This allows an up to 1011-fold SERS signal enhancement, enabling measurements of single molecules. The protocol further demonstrates the placement of a single analyte molecule in a SERS hot spot, the process of AFM imaging, and the subsequent overlaying of Raman imaging to measure an analyte in a single DONA.

Efficient Purification of Cowpea Chlorotic Mottle Virus by a Novel Peptide Aptamer.

The cowpea chlorotic mottle virus (CCMV) is a plant virus explored as a nanotechnological platform. The robust self-assembly mechanism of its capsid protein allows for drug encapsulation and targeted delivery. Additionally, the capsid nanoparticle can be used as a programmable platform to display different molecular moieties. In view of future applications, efficient production and purification of plant viruses are key steps. In established protocols, the need for ultracentrifugation is a significant limitation due to cost, difficult scalability, and safety issues. In addition, the purity of the final virus isolate often remains unclear. Here, an advanced protocol for the purification of the CCMV from infected plant tissue was developed, focusing on efficiency, economy, and final purity. The protocol involves precipitation with PEG 8000, followed by affinity extraction using a novel peptide aptamer. The efficiency of the protocol was validated using size exclusion chromatography, MALDI-TOF mass spectrometry, reversed-phase HPLC, and sandwich immunoassay. Furthermore, it was demonstrated that the final eluate of the affinity column is of exceptional purity (98.4%) determined by HPLC and detection at 220 nm. The scale-up of our proposed method seems to be straightforward, which opens the way to the large-scale production of such nanomaterials. This highly improved protocol may facilitate the use and implementation of plant viruses as nanotechnological platforms for in vitro and in vivo applications.

The feasibility and safety of one-shot dilatation compared to conventional sequential dilatation in tubeless percutaneous nephrolithotomy: a prospective randomized controlled study.

To study the feasibility and safety of One-Shot Dilatation (OSD), versus serial sequential dilatation in tubeless Percutaneous Nephrolithotomy (PCNL). One Hundred and Fifty patients were randomised into two groups; Group A (One-Shot Dilatation), Group B (Serial Dilatation). Twenty-one patients were excluded from the study. Detailed history was taken and full physical examination was performed. Pre-operative routine laboratory investigations were done. Also, non-contrast Computed Tomography of the Urinary Tract (CTUT) and plain urinary tract x-ray were done. Intra-operative assessments of dilatation, total operative, total fluoroscopy and fluoroscopy during dilatation durations were recorded, as well as estimated blood loss. Post-operatively haemoglobin, creatinine levels and CTUT were performed for all patients. Complications, as urinary leakage time, analgesic requirements and hospitalization time were measured. There were statistically significant differences in the intraoperative durations, where Group A had shorter dilatation time, fluoroscopy time during dilatation and total operative time. Group B had a higher complications rate than Group A; 37.9%, 11.3%, respectively. Also, Group B showed haemoglobin drop by 0.44 mg/dl higher than Group A. More doses of analgesia were required for Group B. Hospitalization time and rate of urinary leakage were both in favour of Group A. For patients undergoing Tubeless PCNL, we have concluded that one-shot dilatation seems to be a safer and more feasible technique than Serial dilatation.

Molecular states and spin crossover of hemin studied by DNA origami enabled single-molecule surface-enhanced Raman scattering.

The study of biologically relevant molecules and their interaction with external stimuli on a single molecular scale is of high importance due to the availability of distributed rather than averaged information. Surface enhanced Raman scattering (SERS) provides direct chemical information, but is rather challenging on the single molecule (SM) level, where it is often assumed to require a direct contact of analyte molecules with the metal surface. Here, we detect and investigate the molecular states of single hemin by SM-SERS. A DNA aptamer based G-quadruplex mediated recognition of hemin directs its placement in the SERS hot-spot of a DNA Origami Nanofork Antenna (DONA). The configuration of the DONA structure allows the molecule to be trapped at the plasmonic hot-spot preferentially in no-contact configuration with the metal surface. Owing to high field enhancement at the plasmonic hot spot, the detection of a single folded G-quadruplex becomes possible. For the first time, we present a systematic study by SM-SERS where most hemin molecule adopt a high spin and oxidation state (III) that showed state crossover to low spin upon strong-field-ligand binding. The present study therefore, provides a platform for studying biologically relevant molecules and their properties at SM sensitivity along with demonstrating a conceptual advancement towards successful monitoring of single molecular chemical interaction using DNA aptamers.

The Value of Ultrasonography Using Thyroid Imaging Reporting and Data Systems (TIRADS) in the Diagnosis of Thyroid Cancer Among the Population of Ha'il, Saudi Arabia.

BACKGROUND: Thyroid cancer is one of the common malignant conditions of the head and neck region, and it is considered as one of the most common cancers among endocrine tumors. Ultrasonography is widely used in order to assess thyroid nodules, Therefore, the aim of our study is to determine the accuracy of ultrasonography and fine needle aspiration biopsy (FNAB)-guided by ultrasonography in the diagnosis of thyroid malignancy among the population in the Ha'il region of Saudi Arabia. METHODS: A retrospective evaluation of 137 patients was undertaken at King Salman Specialist Hospital, Ha'il, Kingdom of Saudi Arabia. Patients who presented with thyroid swellings underwent ultrasonography and FNAB. RESULT: Our study results have concluded that the sensitivity of the ultrasonography in the confirmation of a malignant thyroid lesion is 59.4% and its specificity was found to be 74.3%. CONCLUSION: The study showed that ultrasonography using Thyroid Imaging Reporting and Data Systems (TIRADS) is a significant step in evaluating a thyroid nodule. Also, it is highly sensitive and specific, cost-effective, and convenient for the patient.

The superior photocatalytic performance and DFT insights of S-scheme CuO@TiO2 heterojunction composites for simultaneous degradation of organics.

The necessity to resolve the issue of rapid charge carrier recombination for boosting photocatalytic performance is a vigorous and challenging research field. To address this, the construction of a binary system of step-scheme (S-scheme) CuO@TiO2 heterostructure composite has been demonstrated through a facile solid-state route. The remarkably enhanced photocatalytic performance of CuO@TiO2, compared with single TiO2, which can consequence in the more efficient separation of photoinduced charge carriers, reduced the band gap of TiO2, improved the electrical transport performance, and improved the lifetimes, thus donating it with the much more powerful oxidation and reduction capability. A photocatalytic mechanism was proposed to explain the boosted photocatalytic performance of CuO@TiO2 on a complete analysis of physicochemical, DFT calculations, and electrochemical properties. In addition, this work focused on the investigation of the stability and recyclability of CuO@TiO2 in terms of efficiency and its physical origin using XRD, BET, and XPS. It is found that the removal efficiency diminishes 4.5% upon five recycling runs. The current study not only promoted our knowledge of the binary system of S-scheme CuO@TiO2 heterojunction composite photocatalyst but also shed new light on the design of heterostructure photocatalysts with high-performance and high stability.

The electronic structure of the metal-organic interface of isolated ligand coated gold nanoparticles.

Light induced electron transfer reactions of molecules on the surface of noble metal nanoparticles (NPs) depend significantly on the electronic properties of the metal-organic interface. Hybridized metal-molecule states and dipoles at the interface alter the work function and facilitate or hinder electron transfer between the NPs and ligand. X-ray photoelectron spectroscopy (XPS) measurements of isolated AuNPs coated with thiolated ligands in a vacuum have been performed as a function of photon energy, and the depth dependent information of the metal-organic interface has been obtained. The role of surface dipoles in the XPS measurements of isolated ligand coated NPs is discussed and the binding energy of the Au 4f states is shifted by around 0.8 eV in the outer atomic layers of 4-nitrothiophenol coated AuNPs, facilitating electron transport towards the molecules. Moreover, the influence of the interface dipole depends significantly on the adsorbed ligand molecules. The present study paves the way towards the engineering of the electronic properties of the nanoparticle surface, which is of utmost importance for the application of plasmonic nanoparticles in the fields of heterogeneous catalysis and solar energy conversion.

Increased percentage of apoptotic and CTLA-4 (CD152) expressing cells in CD4+/CD8+ cells in COVID-19 patients.

Coronavirus infectious disease 2019 (COVID-19) confirmed cases are characterized by T lymphopenia. Total apoptotic and cytotoxic T-lymphocyte antigen-4 (CTLA-4) expressing cells among CD4+/CD8+ cells were analyzed in 24 COVID-19 patients (16 out-patients and 8 in-patients) and 18 healthy volunteers using flow cytometry to detect their possible role in T lymphopenia. Hospitalized patients did not show significant difference compared to non-hospitalized patients. While the percentage and absolute count of CD4+/CD8+ cells were significantly reduced in COVID-19 cases compared to healthy control (P < .05), the proportion of apoptotic and CTLA-4 expressing CD4+/CD8+ cells were significantly up-regulated in COVID-19 patients (P < .05). In addition, apoptotic and CTLA-4+/CD4+ cells were directly related to dyspnea duration, chest CT score, ferritin, and C-reactive protein and inversely correlated with platelet count in COVID-19 patients. While apoptotic and CTLA-4+/CD8+ cells were directly related to lymphocyte count in COVID-19 patients. The apoptotic and CTLA-4+ cells were directly related to each other in CD4+/CD8+ cells (P < .05). White blood cells (WBCs) (×103/L), eosinophils (ratio and count), lymphocyte ratio, neutrophil ratio, neutrophil/lymphocyte ratio, neutrophil/CD4 ratio, neutrophil/CD8 ratio, CD4+ cells ratio, and CTLA-4+ cells percentage), and CD8+ cells (ratio, count, total apoptotic cell, and CD152 + cells) were all found to be significantly altered in association with COVID-19. Total lymphopenia and depletion of CD4+/CD8+ cells are characterizing COVID-19 patients. Increased apoptosis and CTLA-4 expression in CD4+/CD8+ cells in COVID-19 and their correlations with reduced cell count and severity indicators as CRP and ferritin can be used for diagnosis and follow up of the clinical severity. Our current study proposes promising future diagnostic and therapeutic targets.

Care for commercial living donors: the experience of an NGO's outreach in Egypt.

Follow-up care for living organ donors is inadequate in countries with advanced transplantation systems based on altruistic donation. In cases where financial incentives drive an organ donation, care for the live 'donor' is largely absent. Care must be provided not only to living altruistic organ donors but especially to victims of organ trafficking who are often not suitable candidates for a donation and subject to poor surgical practices and conditions. Such follow-up is essential not only as a basic right, but also as an important reconciliatory step to regain public trust in transplants where they have been characterized by commercialism. Yet, the question, who will provide care for the commercial living donor? persists. In the absence of public or private commitments to this care, the Coalition for Organ-Failure Solutions (COFS) conducts outreach programmes that include identifying victims of organ trafficking, assessing their consequences and arranging support services. This paper presents studies on consequences for commercial living donors (CLDs), describes the case of organ trafficking and COFS' care provision in Egypt and discusses why follow-up care for CLDs is not an appropriate ingredient for advancing regulation proposals but should be considered an essential component of the movement to end organ trafficking.

A Versatile DNA Origami-Based Plasmonic Nanoantenna for Label-Free Single-Molecule Surface-Enhanced Raman Spectroscopy.

DNA origami technology allows for the precise nanoscale assembly of chemical entities that give rise to sophisticated functional materials. We have created a versatile DNA origami nanofork antenna (DONA) by assembling Au or Ag nanoparticle dimers with different gap sizes down to 1.17 nm, enabling signal enhancements in surface-enhanced Raman scattering (SERS) of up to 1011. This allows for single-molecule SERS measurements, which can even be performed with larger gap sizes to accommodate differently sized molecules, at various excitation wavelengths. A general scheme is presented to place single analyte molecules into the SERS hot spots using the DNA origami structure exploiting covalent and noncovalent coupling schemes. By using Au and Ag dimers, single-molecule SERS measurements of three dyes and cytochrome c and horseradish peroxidase proteins are demonstrated even under nonresonant excitation conditions, thus providing long photostability during time-series measurement and enabling optical monitoring of single molecules.

Assessing the effects of a novel biostimulant to enhance leafminer resistance and plant growth on common bean.

The leafminer Liriomyza trifolii is one of the major insects that affect Phaseolus vulgaris production worldwide. Novel and safe biobased stimulator compound (BSTC) with micronutrient-amino acid chelated compounds was developed from natural compounds and was used for foliar spray of P. vulgaris. Treated plants showed significantly increased in quality and productivity as well as significant reduction in leafminer infestation by close the tunnel end resulting in larvae suffocation and death. BSTC contains chemical composition that has important function in inducing immunity and resistance against insects, enhance plant growth and production. Also, HPLC showed that the assembled BSTC is rich in nucleobases than yeast extract (> 56 fold). Aminochelation zinc enhanced the rate of absorption of nutrient compounds and could participate in safe biofortification strategy. The expression of plant defense related genes under BSTC treatment revealed strong correlations between the transcription rates of defense related genes. Based on binding energies and interacting residues of six vital insect proteins, the best-docked complexes was obtained with disodium 5'-inosinate, delphinidin 3-glucoside and hyperoside. Obtained findings indicate that the foliar application of BSTC can enhance plant growth and productivity, uptake of important elements, expression of defense related genes and inhibit insect essential genes.

Anticancer activity of milk fat rich in conjugated linoleic acid against Ehrlich ascites carcinoma cells in female Swiss albino mice.

BACKGROUND AND AIM: The major conjugated linoleic acid (CLA) isomers have anticancer effect, especially breast cancer cells, inhibits cell growth and induces cell death. Also, CLA has several health benefits in vivo, including antiatherogenesis, antiobesity, and modulation of immune function. The present study aimed to assess the safety and anticancer effects of milk fat CLA against in vivo Ehrlich ascites carcinoma (EAC) in female Swiss albino mice. This was based on acute toxicity study, detection of the tumor growth, life span of EAC bearing hosts, and simultaneous alterations in the hematological, biochemical, and histopathological profiles. MATERIALS AND METHODS: One hundred and fifty adult female mice were equally divided into five groups. Groups (1-2) were normal controls, and Groups (3-5) were tumor transplanted mice (TTM) inoculated intraperitoneally with EAC cells (2×106/0.2 mL). Group (3) was (TTM positive control). Group (4) TTM fed orally on balanced diet supplemented with milk fat CLA (40 mg CLA/kg body weight). Group (5) TTM fed orally on balanced diet supplemented with the same level of CLA 28 days before tumor cells inoculation. Blood samples and specimens from liver and kidney were collected from each group. The effect of milk fat CLA on the growth of tumor, life span of TTM, and simultaneous alterations in the hematological, biochemical, and histopathological profiles were examined. RESULTS: For CLA treated TTM, significant decrease in tumor weight, ascetic volume, viable Ehrlich cells accompanied with increase in life span were observed. Hematological and biochemical profiles reverted to more or less normal levels and histopathology showed minimal effects. CONCLUSION: The present study proved the safety and anticancer efficiency of milk fat CLA and provides a scientific basis for its medicinal use as anticancer attributable to the additive or synergistic effects of its isomers.

Surface Etching of 3D Printed Poly(lactic acid) with NaOH: A Systematic Approach.

The article describes a systematic investigation of the effects of an aqueous NaOH treatment of 3D printed poly(lactic acid) (PLA) scaffolds for surface activation. The PLA surface undergoes several morphology changes and after an initial surface roughening, the surface becomes smoother again before the material dissolves. Erosion rates and surface morphologies can be controlled by the treatment. At the same time, the bulk mechanical properties of the treated materials remain unaltered. This indicates that NaOH treatment of 3D printed PLA scaffolds is a simple, yet viable strategy for surface activation without compromising the mechanical stability of PLA scaffolds.