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Instruct-ERIC, "the European Research Infrastructure Consortium for Structural biology research," is a pan-European distributed research infrastructure making high-end technologies and methods in structural biology available to users. Here, we describe the current state-of-the-art of integrated structural biology and discuss potential future scientific developments as an impulse for the scientific community, many of which are located in Europe and are associated with Instruct. We reflect on where to focus scientific and technological initiatives within the distributed Instruct research infrastructure. This review does not intend to make recommendations on funding requirements or initiatives directly, neither at the national nor the European level. However, it addresses future challenges and opportunities for the field, and foresees the need for a stronger coordination within the European and international research field of integrated structural biology to be able to respond timely to thematic topics that are often prioritized by calls for funding addressing societal needs., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

SSR128129E (SSR) is a unique small-molecule inhibitor of fibroblast growth factor receptors (FGFRs). SSR is a high-affinity allosteric binder that selectively blocks one of the two major FGFR-mediated pathways. The mechanisms of SSR activity were studied previously in much detail, allowing the identification of its binding site, located in the hydrophobic groove of the receptor D3 domain. The binding site overlaps with the position of an N-terminal helix, an element exclusive for the FGF8b growth factor, which could potentially convert SSR from an allosteric inhibitor into an orthosteric blocker for the particular FGFR/FGF8b system. In this regard, we report here on the structural and functional investigation of FGF8b/FGFR3c system and the effects imposed on it by SSR. We show that SSR is equally or more potent in inhibiting FGF8b-induced FGFR signaling compared to FGF2-induced activation. On the other hand, when studied in the context of separate extracellular domains of FGFR3c in solution with NMR spectroscopy, SSR is unable to displace the N-terminal helix of FGF8b from its binding site on FGFR3c and behaves as a weak orthosteric inhibitor. The substantial inconsistency between the results obtained with cell culture and for the individual water-soluble subdomains of the FGFR proteins points to the important role played by the cell membrane., (© 2024 The Author(s). FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

The stem-loop 2 motif (s2m) in SARS-CoV-2 (SCoV-2) is located in the 3'-UTR. Although s2m has been reported to display characteristics of a mobile genomic element that might lead to an evolutionary advantage, its function has remained unknown. The secondary structure of the original SCoV-2 RNA sequence (Wuhan-Hu-1) was determined by NMR in late 2020, delineating the base-pairing pattern and revealing substantial differences in secondary structure compared to SARS-CoV-1 (SCoV-1). The existence of a single G29742-A29756 mismatch in the upper stem of s2m leads to its destabilization and impedes a complete NMR analysis. With Delta, a variant of concern has evolved with one mutation compared to the original sequence that replaces G29742 by U29742. We show here that this mutation results in a more defined structure at ambient temperature accompanied by a rise in melting temperature. Consequently, we were able to identify >90% of the relevant NMR resonances using a combination of selective RNA labeling and filtered 2D NOESY as well as 4D NMR experiments. We present a comprehensive NMR analysis of the secondary structure, (sub)nanosecond dynamics, and ribose conformation of s2m Delta based on heteronuclear 13 C NOE and T 1 measurements and ribose carbon chemical shift-derived canonical coordinates. We further show that the G29742U mutation in Delta has no influence on the druggability of s2m compared to the Wuhan-Hu-1 sequence. With the assignment at hand, we identify the flexible regions of s2m as the primary site for small molecule binding., (© 2024 Matzel et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)

Introduction: Treating deep vein thrombosis (DVT) using a once-daily dose of enoxaparin offers greater convenience and the possibility of home-based care for certain patients, as opposed to a continuous infusion of unfractionated heparin (UFH). The study aimed to determine the most cost-effective thromboprophylaxis between low-molecular-weight heparin (LMWH) and UFH for hospitalized patients., Materials and Methods: After obtaining clearance from the institutional ethical committee, the study was conducted in the Department of General Surgery, Sri Devaraj Urs Medical College, over a period of six months. Informed consent was obtained from all 46 patients included in this study. The participants were divided into two groups: group A received LMWH and group B received UFH., Results: The mean age in group A was 59.8 + 10.6 years and in group B was 54.9 + 12.3 years. There was no significant difference in the girth of the lower limb between the groups during the follow-up period (p > 0.05). In group A, there was a highly significant reduction in lower limb girth from day one to day five (p < 0.0001), day five to day 10 (p < 0.0001), and day one to day 10 (p < 0.0001). In group B, there was no significant reduction from day one to day five (p = 0.06), but there was a significant reduction from day five to day 10 (p = 0.001) and day one to day 10 (p = 0.001)., Conclusion: Treatment with LMWH as an anticoagulant significantly reduced the lower extremity girth and thrombus thickness in cases of DVT when compared to UFH., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Ullalkar et al.)

Introduction Necrotizing fasciitis (NF) is a grave and life-threatening infection of the soft tissues. It is defined by the gradual necrosis of the fascia and subcutaneous tissue, which spreads along the fascial planes. Cellulitis, a prevalent skin infection, has led to suggestions that procalcitonin could serve as a diagnostic tool to distinguish it from other inflammatory skin conditions that resemble cellulitis. The study aims to assess the procalcitonin (PCT) levels in individuals with NF and cellulitis and determine its effectiveness in early differentiation between these two conditions. Methods After obtaining clearance from the institutional ethical committee, the study was conducted in the Department of General Surgery, Sri Devaraj Urs Medical College, over six months. Informed consent was obtained from all 30 patients included in this study. The study compared PCT levels in patients diagnosed with NF and cellulitis. Statistical analysis was performed using SPSS version 22 software (IBM Corp., Armonk, NY, USA). Results The mean age of subjects was 53.23 ± 8.78 years. Among patients, 21 (70%) were diagnosed with cellulitis and 9 (30%) were diagnosed with NF. The mean PCT levels were 0.34 ± 0.32 and 4.89 ± 1.98 among the cellulitis and NF groups, respectively. There was a significant difference (p<0.05). PCT had a sensitivity of 100% and a specificity of 100%, in differentiating cellulitis and necrotizing fasciitis. Conclusion PCT levels were notably elevated in cases of NF compared to cellulitis. Despite the study's limited sample size, it represents the first report highlighting the value of PCT as an early diagnostic tool for identifying necrotizing fasciitis., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Vaibhavi et al.)

The main protease M pro , nsp5, of SARS-CoV-2 (SCoV2) is one of its most attractive drug targets. Here, we report primary screening data using nuclear magnetic resonance spectroscopy (NMR) of four different libraries and detailed follow-up synthesis on the promising uracil-containing fragment Z604 derived from these libraries. Z604 shows time-dependent binding. Its inhibitory effect is sensitive to reducing conditions. Starting with Z604, we synthesized and characterized 13 compounds designed by fragment growth strategies. Each compound was characterized by NMR and/or activity assays to investigate their interaction with M pro . These investigations resulted in the four-armed compound 35b that binds directly to M pro . 35b could be cocrystallized with M pro revealing its noncovalent binding mode, which fills all four active site subpockets. Herein, we describe the NMR-derived fragment-to-hit pipeline and its application for the development of promising starting points for inhibitors of the main protease of SCoV2.

The EU-OPENSCREEN (EU-OS) European Research Infrastructure Consortium (ERIC) is a multinational, not-for-profit initiative that integrates high-capacity screening platforms and chemistry groups across Europe to facilitate research in chemical biology and early drug discovery. Over the years, the EU-OS has assembled a high-throughput screening compound collection, the European Chemical Biology Library (ECBL), that contains approximately 100 000 commercially available small molecules and a growing number of thousands of academic compounds crowdsourced through our network of European and non-European chemists. As an extension of the ECBL, here we describe the computational design, quality control and use case screenings of the European Fragment Screening Library (EFSL) composed of 1056 mini and small chemical fragments selected from a substructure analysis of the ECBL. Access to the EFSL is open to researchers from both academia and industry. Using EFSL, eight fragment screening campaigns using different structural and biophysical methods have successfully identified fragment hits in the last two years. As one of the highlighted projects for antibiotics, we describe the screening by Bio-Layer Interferometry (BLI) of the EFSL, the identification of a 35 μM fragment hit targeting the beta-ketoacyl-ACP synthase 2 (FabF), its binding confirmation to the protein by X-ray crystallography (PDB 8PJ0), its subsequent rapid exploration of its surrounding chemical space through hit-picking of ECBL compounds that contain the fragment hit as a core substructure, and the final binding confirmation of two follow-up hits by X-ray crystallography (PDB 8R0I and 8R1V)., Competing Interests: X. J. and J. M. are currently employees of the company Chemotargets, of which J. M. is co-founder and co-owner., (This journal is © The Royal Society of Chemistry.)

The ephrin type-A 2 receptor tyrosine kinase (EPHA2) is involved in the development and progression of various cancer types, including colorectal cancer (CRC). There is also evidence that EPHA2 plays a key role in the development of resistance to the endothelial growth factor receptor (EGFR) monoclonal antibody Cetuximab used clinically in CRC. Despite the promising pharmacological potential of EPHA2, only a handful of specific inhibitors are currently available. In this concept paper, general strategies for EPHA2 inhibition with molecules of low molecular weight (small molecules) are described. Furthermore, available examples of inhibiting EPHA2 in CRC using small molecules are summarized, highlighting the potential of this approach., (© 2023 The Authors. ChemMedChem published by Wiley-VCH GmbH.)

Development of new antiviral medication against the beta-coronavirus SARS-CoV-2 (SCoV2) is actively being pursued. Both NMR spectroscopy and crystallography as structural screening technologies have been utilised to screen the viral proteome for binding to fragment libraries. Here, we report on NMR screening of elements of the viral RNA genome with two different ligand libraries using 1 H-NMR-screening experiments and 1 H and 19 F NMR-screening experiments for fluorinated compounds. We screened against the 5'-terminal 119 nucleotides located in the 5'-untranslated region of the RNA genome of SCoV2 and further dissected the four stem-loops into its constituent RNA elements to test specificity of binding of ligands to shorter and longer viral RNA stretches. The first library (DRTL-F library) is enriched in ligands binding to RNA motifs, while the second library (DSI-poised library) represents a fragment library originally designed for protein screening. Conducting screens with two different libraries allows us to compare different NMR screening methodologies, describe NMR screening workflows, validate the two different fragment libraries, and derive initial leads for further downstream medicinal chemistry optimisation., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Phenoloxidase (PO) is a significant biomolecule involved in humoral defence mechanism of invertebrates. Spontaneous melanization of insect haemolymph is the major hinderance for studying PO activity, as haemolymph was collected devoid of phenylthiourea. In the study, no visible melanization was observed in crude serum from the grub of Oryctes rhinoceros up to 30 min of incubation amongst crude haemolymph, diluted haemolymph, crude serum and diluted serum that were subjected to visual observation for spontaneous melanization reaction. Accordingly, crude serum was taken for evaluating PO activity. At the same time, as PO substrates tend to auto-oxidize and provide false optical density value, tris-buffered saline devoid of any substrates were used as blank for PO assays. The ideal wavelength at which maximum PO activity occurred for each substrate, namely, tyrosine, tyramine, dopamine, L-dopa, DL-dopa, catechol, protocatechuic acid and pyrogallol was determined as 407, 410, 429, 465, 403, 466, 428 and 400 nm, respectively. Additionally, time course of oxidation for each phenolic substrate by the serum PO were examined and DL-dopa was identified as the specific substrate for serum PO in the grub of O. rhinoceros . Furthermore, maximum PO activity was observed at 5 min of incubation for 10 mM of DL-dopa that was considered as optimum concentration. The ideal pH and temperature for serum PO activity was observed as 7.5 and 20°C, respectively. These results suggested that standardizing a suitable substrate is an essential prerequisite to evaluate the real PO activity of serum which might significantly fluctuate in each insect model.

Insects have evolved with effective strategies to utilize cellulose as an energy source by possessing cellulolytic enzymes which can be used as an optimal resource in the bioenergy sector. The study was aimed at evaluating the cellulolytic enzyme in the larval gut of the banana pseudostem weevil, Odoiporus longicollis Olivier (Coleoptera: Curculionidae). Primarily, cellulase activity was localized along the gut, in which the midgut showed the highest activity (2858 U/mg). The thermo-tolerance of cellulase activity was found to be up to 80°C (highest at 60°C), and the enzyme was stable at a pH between 5 and 6. Various concentrations of divalent cations (CaCl 2 , MgCl 2 , and CuCl 2 ) have differential enhancing and inhibitory effects on cellulase activity. The cellulase (OlCel) was purified using anion exchange chromatography. The molecular weight of the cellulase was determined to be 47 kDa. The physicochemical parameters of the purified enzyme were similar to that of enzyme activity of whole gut extract. Mass spectrometry results identified sequence similarities of purified cellulase to the glycosyl hydrolase family 5 (GHF5) family. The gut microbial cellulase activity as exogenous source showed no competence compared with the endogenous activity., (© 2023 Wiley Periodicals LLC.)

Introduction: The purpose of this study was to determine whether balance and harmony of the face, when seen as a whole, alter the smile perception as compared to a lone standing smile in female subjects of two age groups and to identify parameters of dental composition and dentofacial harmony associated with the smile perception of dental specialists (DS) and laypeople (LP)., Settings and Design: A cross-sectional study., Materials and Methods: Twenty-nine DS and 29 LP scored the smile attractiveness of 84 full-face images (FFIs) and corresponding smile images (SIs) of young adults (YA: 20-29 years) and old adults (OA: 50-59 years), using the Visual Analogue Scale. Smile analysis software (Planmeca Romexis®Smile-Design) was used to measure 10 parameters of dental composition and dentofacial harmony in FFI., Statistical Analysis Used: Independent t -test and Pearson's correlation coefficient (r) were used to analyze the data., Results: A significant difference was not seen between the smile attractiveness scores of FFI and SI in both raters and age groups. Significant differences were observed between DS and LP in the YA group. The OA group was rated significantly higher than the YA group. The correlation between smile attractiveness scores and each of the nine dentofacial parameters was not significant ( P < 0.05)., Conclusions: Framing of images did not significantly alter smile perception. FFI received higher scores than SI, and raters were more critical of the YA group. Professional training of raters and an interplay of smile and facial attractiveness of the female subjects contributed to the smile perception of raters. No parameters of dental composition or dentofacial harmony significantly associated with smile attractiveness were identified., Competing Interests: There are no conflicts of interest., (Copyright: © 2023 Journal of Pharmacy and Bioallied Sciences.)

The splicing isoform b of human fibroblast growth factor 8 (FGF8b) is an important regulator of brain embryonic development. Here, we report the almost complete NMR chemical shift assignment of the backbone and aliphatic side chains of FGF8b. Obtained chemical shifts are in good agreement with the previously reported X-ray data, excluding the N-terminal gN helix, which apparently forms only in complex with the receptor. The reported data provide an NMR starting point for the investigation of FGF8b interaction with its receptors and with potential drugs or inhibitors., (© 2023. The Author(s).)

Background Fine-needle aspiration cytology (FNAC) is one of the reliable methods in diagnosing breast cancers. Morphometric studies are done in benign and malignant neoplasms of various organs by using software, which measures cellular, cytoplasmic, and nuclear parameters. Nuclear parameters define the behavior of the neoplasm. This study aims to evaluate nuclear morphometry parameters in aspirated smears of breast lesions and determine the association between cytological findings with nuclear morphometry parameters. Methodology It's a retrospective cytology study from July 2020 to June 2022 conducted at a tertiary health care center in Kolar, Karnataka, India. The FNAC smears of breast mass were analyzed cytologically and were subjected to nuclear morphometry study. Nuclear parameters such as nuclear area, nuclear perimeter, nuclear Feret diameter, minimum Feret, and shape factor were captured in Zen software (Zeiss, Oberkochen, Germany) and ImageJ software (National Institutes of Health, Bethesda, MD, USA; Laboratory for Optical and Computational Instrumentation [LOCI], University of Wisconsin-Madison, Madison, WI, USA). The association between nuclear morphometric findings and cytological findings was noted. A descriptive statistical analysis was done. Results Sixty cases of mass in the breast were considered for the study of which 37 cases were benign and 23 were malignant. Nuclear morphometry parameters such as nuclear area, nuclear perimeter, nuclear Feret diameter, minimum Feret, and shape factor for benign breast lesions were 25.16 ± 3.2 µm 2 , 21.58 ± 1.89 µm, 6.5 ± 0.94 µm, 4.87 ± 0.50 µm, and 0.92 ± 0.02, respectively, and for malignant breast cases were 46.57 ± 12.24 µm 2 , 27.53 ± 3.26 µm, 10.08 ± 1.18 µm, 6.49 ± 0.88 µm, and 0.93 ± 0.01, respectively. The association of all nuclear parameters between benign and malignant lesions was statistically significant ( P  = 0.001). Conclusions Nuclear morphometric study in breast lesions is a concept that supplements FNAC findings in differentiating benign from malignant lesions., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Girdhar et al.)

Over the past years, molybdenum disulfide (MoS 2 ) has been the most extensively studied two-dimensional (2D) semiconductormaterial. With unique electrical and optical properties, 2DMoS 2 is considered to be a promising candidate for future nanoscale electronic and optoelectronic devices. However, charge trapping leads to a persistent photoconductance (PPC), hindering its use for optoelectronic applications. To overcome these drawbacks and improve the optoelectronic performance, organic semiconductors (OSCs) are selected to passivate surface defects, tune the optical characteristics, and modify the doping polarity of 2D MoS 2 . Here, we demonstrate a fast photoresponse in multilayer (ML) MoS 2 by addressing a heterojunction interface with vanadylphthalocyanine (VOPc) molecules. The MoS 2 /VOPc van der Waals interaction that has been established encourages the PPC effect in MoS 2 by rapidly segregating photo-generated holes, which move away from the traps of MoS 2 toward the VOPc molecules. The MoS 2 /VOPc phototransistor exhibits a fast photo response of less than 15 ms for decay and rise, which is enhanced by 3ordersof magnitude in comparison to that of a pristine MoS 2 -based phototransistor (seconds to tens of seconds). This work offers a means to realize high-performance transition metal dichalcogenide (TMD)-based photodetection with a fast response speed.

The ephrin type-A receptor 2 (EPHA2) kinase belongs to the largest family of receptor tyrosine kinases. There are several indications of an involvement of EPHA2 in the development of infectious diseases and cancer. Despite pharmacological potential, EPHA2 is an under-examined target protein. In this study, we synthesized a series of derivatives of the inhibitor NVP-BHG712 and triazine-based compounds. These compounds were evaluated to determine their potential as kinase inhibitors of EPHA2, including elucidation of their binding mode (X-ray crystallography), affinity (microscale thermophoresis), and selectivity (Kinobeads assay). Eight inhibitors showed affinities in the low-nanomolar regime (K D <10 nM). Testing in up to seven colon cancer cell lines that express EPHA2 reveals that several derivatives feature promising effects for the control of human colon carcinoma. Thus, we have developed a set of powerful tool compounds for fundamental new research on the interplay of EPH receptors in a cellular context., (© 2023 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)

Cardiac vascular diseases, especially acute myocardial infarction (AMI), are one of the leading causes of death worldwide. Therefore cardio-specific biomarkers such as cardiac troponin I (cTnI) play an essential role in the field of diagnostics. In order to enable rapid and accurate measurement of cTnI with the potential of online measurements, a chemiluminescence-based immunosensor is presented as a proof of concept. A flow cell was designed and combined with a sensitive CMOS camera allowing sensitive optical readout. In addition, a microfluidic setup was established, which achieved selective and quasi-online cTnI determination within ten minutes. The sensor was tested with recombinant cTnI in phosphate buffer and demonstrated cTnI measurements in the concentration range of 2-25 µg/L. With the optimized system, a limit of detection (LoD) of 0.6 µg/L (23 pmol/L) was achieved. Furthermore, the selectivity of the immunosensor was investigated with other recombinant proteins, such as cTnT, and cTnC, at a level of 16 µg/L. No cross-reactivity could be observed. Measurements with diluted blood plasma and serum resulted in an LoD of 60 µg/L (2.4 nmol/L) and 70 µg/L (2.9 nmol/L), respectively.

Background Colorectal cancer is a leading cause of cancer-related deaths worldwide, and epithelial-mesenchymal transition (EMT) plays an important role in cancer metastasis. In EMT, there is downregulation of E-cadherin, an intracellular adhesion molecule, as well as mutations in beta-catenin genes. On immunohistochemistry (IHC), the expression of CD44 portrays stem cell differentiation, which, in turn, is strongly associated with EMT. Thus, newer targeted therapies can be advised based on the expression of EMT and stem cell differentiation. Aims and objectives To determine the IHC expression of E-cadherin, beta-catenin, and CD44 in colorectal adenocarcinoma and find the association of the IHC expression of E-cadherin, beta-catenin, and CD44 with the histopathological grade, stage, lymph node metastasis, and lymphovascular invasion of colorectal adenocarcinoma. Materials and methods Fifty histologically proven cases of colorectal adenocarcinoma from 2016 to 2021 were included in this study, and clinicopathological data including age, gender, grading, TNM (tumour, node, and metastasis) staging, and lymph node metastasis were collected and hematoxylin and eosin slides were reviewed. IHC staining for E-cadherin, beta-catenin, and CD44 was done for all cases using the peroxidase and anti-peroxidase method, and the results were analysed. Results Peak incidence occurred in the 61-70 years age group (36%), and the most common site of the tumour was the rectal area (48%). The majority of the cases were in TNM stage II (37.3%), and a low expression of E-cadherin was found to be associated with higher T stage (p = 0.03), TNM staging (p = 0.04), as well as the presence of lymph node metastasis (p = 0.006). High beta-catenin expression was observed to have a significant correlation with a higher T stage (p = 0.006) and TNM staging (p = 0.005), while high CD44 expression was found to be associated with lymph node metastasis (p = 0.01). Altered expression of EMT-related proteins (E-cadherin and beta-catenin) showed a significant correlation with higher T stage (p = 0.03), TNM staging (p = 0.016), and lymph node metastasis (0.04). Conclusions EMT and cancer stem cell IHC markers are biomarkers for aggressive tumour growth and lymph node metastasis. Hence, EMT markers (E-cadherin and beta-catenin) and cancer stem cell markers (CD44) can be used as prognostic markers., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Tunuguntla et al.)

Replication of the coronavirus genome starts with the formation of viral RNA-containing double-membrane vesicles (DMV) following viral entry into the host cell. The multi-domain nonstructural protein 3 (nsp3) is the largest protein encoded by the known coronavirus genome and serves as a central component of the viral replication and transcription machinery. Previous studies demonstrated that the highly-conserved C-terminal region of nsp3 is essential for subcellular membrane rearrangement, yet the underlying mechanisms remain elusive. Here we report the crystal structure of the CoV-Y domain, the most C-terminal domain of the SARS-CoV-2 nsp3, at 2.4 Å-resolution. CoV-Y adopts a previously uncharacterized V-shaped fold featuring three distinct subdomains. Sequence alignment and structure prediction suggest that this fold is likely shared by the CoV-Y domains from closely related nsp3 homologs. NMR-based fragment screening combined with molecular docking identifies surface cavities in CoV-Y for interaction with potential ligands and other nsps. These studies provide the first structural view on a complete nsp3 CoV-Y domain, and the molecular framework for understanding the architecture, assembly and function of the nsp3 C-terminal domains in coronavirus replication. Our work illuminates nsp3 as a potential target for therapeutic interventions to aid in the on-going battle against the COVID-19 pandemic and diseases caused by other coronaviruses., (© 2023. The Author(s).)

Background and objectives Breast cancer is the most frequent cancer among women, globally. Postoperative pain after mastectomy not only causes slow recovery and prolonged hospital stay but can also increase the risk of chronic pain. For patients undergoing breast surgery, effective perioperative pain management is required. Various approaches have been introduced to overcome this, such as opioids, non-opioid analgesics, and regional blocks. The erector spinae plane block is a new regional anesthesia technique used in breast surgery to provide adequate intraoperative and postoperative analgesia. Opioid-free anesthesia is a multimodal analgesia technique that does not use opioids and thus prevents opioid tolerance after surgery. This study aims to investigate whether administering an opioid-free analgesic mixture lowers the pain score and the need for analgesics during and after surgery. Material and methods In this randomized prospective comparative clinical study, 66 patients of the American Society of Anesthesiologists (ASA) psychological status (PS) class 1 and 2, aged 18 to 80, were included. Group M received erector spinae plane block + general anesthesia + opioid-free analgesic mixture (1 mcg/cc dexmedetomidine + 1 mg/cc ketamine + 100 mg/cc magnesium sulfate prepared in a 20 ml syringe). Group N received erector spinae plane block + general anesthesia + 20ml of normal saline infusion. The primary outcome was to assess pain scores in the perioperative period. The secondary outcomes were to compare the time for the first rescue analgesia requirement perioperatively, intraoperative hemodynamic profile, and postoperative patient satisfaction. A p<0.05 was considered to be statistically significant. Results All patients were females undergoing modified radical mastectomy or breast conservative surgery + axillary sampling + latissimus dorsi flap reconstruction. The visual analog scale (VAS) scores were less than or equal to 3 in zero, first, and second hours postoperatively in both groups. The pain was moderate i.e., less than 4 in almost all time intervals in both groups. Group M had a better intraoperative hemodynamic profile, including mean arterial pressure and heart rate when compared to group N. In group M, the time of request for rescue analgesia was 726.67±390.99 minutes, while it was 468±278.79 minutes in group N. The total analgesic requirement was less in group M than in group N, but this was not statistically significant. Conclusion Multimodal analgesia with erector spinae plane block and opioid-free analgesic mixture provides effective perioperative analgesia and a better intraoperative hemodynamic profile in patients undergoing breast cancer surgery under general anesthesia., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, B et al.)

Insect life on earth is greatly diversified despite being exposed to several infectious agents due to their diverse habitats and ecological niche. One of the major factors responsible for their successful establishment is having a powerful innate immune system. The most common and effective method used by insects in recognizing pathogen and non-self-substances is the melanization process among others. The key enzyme involved in melanin biosynthesis is the copper containing humoral defense enzyme, phenoloxidase (PO). This review focused on understanding about PO and that had been in research for nearly a century. The review elaborates about evolutionary significance of PO in arthropods, its relationship with mammalian tyrosinases, various substrates, activators and inhibitors involved in the activation of phenoloxidase cascade, as it requires an integrated system of activation that vary among insect species. The enzyme also plays a vital role in insect immunity by involving in several other immune functions like sclerotization, wound healing, opsonization, encapsulation and nodule formation. Further, gene knock down or knock out of PO genes and inhibition of PO-melanization cascade by several mechanisms can also be considered as promising future alternative to control serious pests by making them highly susceptible to any targeted attack., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)