Tissue Disruption Research Articles

Optimization of a high-throughput shotgun immunoproteomics pipeline for antigen identification

Identification of proteins which initiate and/or perpetuate adaptive immune responses has potential to greatly impact pre-clinical and clinical work across numerous fields. To date, however, the methodologies available to identify antigens responsible for driving adaptive immune responses have been plagued by numerous issues which have drastically limited their widespread adoption. Therefore, in this study, we sought to optimize a shotgun immunoproteomics approach to alleviate these persistent issues and create a high-throughput, quantitative methodology for antigen identification. Three individual components of a previously published approach, namely the protein extraction, antigen elution, and LC-MS/MS analysis steps, were optimized in a systematic manner. These studies determined that preparation of protein extracts using a one-step tissue disruption method in immunoprecipitation (IP) buffer, eluting antigens from affinity chromatography columns with 1% trifluoroacetic acid (TFA), and TMT-labeling & multiplexing equal volumes of eluted samples for LC-MS/MS analysis, resulted in quantitative longitudinal antigen identification, with reduced variability between replicates and increased total number of antigens identified. This optimized pipeline provides a multiplexed, highly reproducible, and fully quantitative approach to antigen identification which is broadly applicable to determine the role of antigenic proteins in inciting (i.e., primary antigens) and perpetuating (i.e., secondary antigens) a wide range of diseases. SignificanceUsing a systematic, hypothesis-driven approach, we identified potential improvements for three individual steps of a previously published approach for antigen-identification. Optimization of each step created a methodology which resolved many of the persistent issues associated with previous antigen identification approaches. The optimized high-throughput shotgun immunoproteomics approach described herein identifies more than five times as many unique antigens as the previously published method, greatly reduces protocol cost and mass spectrometry time per experiment, minimizes both inter- and intra-experimental variability, and ensures each experiment is fully quantitative. Ultimately, this optimized antigen identification approach has the potential to facilitate novel antigen identification studies, allowing evaluation of the adaptive immune response in a longitudinal manner and encourage innovations in a wide array of fields.

Perineurial Window is Critical for Experimental Reverse End-to-Side Nerve Transfer.

The depth of connective tissue window in the side of a recipient nerve in reverse end-to-side transfers (RETS) remains controversial. To test whether the depth of connective tissue disruption influences the efficiency of donor axonal regeneration in the context of RETS. Sprague-Dawley rats (n = 24) were assigned to 1 of the 3 groups for obturator nerve to motor femoral nerve RETS: group 1, without epineurium opening; group 2, with epineurium only opening; and group 3, with epineurium and perineurium opening. Triple retrograde labeling was used to assess the number of motor neurons that had regenerated into the recipient motor femoral branch. Thy1-GFP rats (n = 8) were also used to visualize the regeneration pathways in the nerve transfer networks at 2- and 8-week time point using light sheet fluorescence microscopy. The number of retrogradely labeled motor neurons that had regenerated distally toward the target muscle was significantly higher in group 3 than that in groups 1 and 2. Immunohistochemistry validated the degree of connective tissue disruption among the 3 groups, and optical tissue clearing methods demonstrated donor axons traveling outside the fascicles in groups 1 and 2 but mostly within the fascicles in group 3. Creating a perineurial window in the side of recipient nerves provides the best chances of robust donor axonal regeneration across the RETS repair site. This finding aids nerve surgeons by confirming that a deep window should be undertaken when doing a RETS procedure.

Critical Evaluation of Embedding Media for Histological Studies of Early Stages of Chick Embryo Development.

A histological examination is an important tool in embryology, developmental biology, and correlated areas. Despite the amount of information available about tissue embedding and different media, there is a lack of information regarding best practices for embryonic tissues. Embryonic tissues are considered fragile structures, usually small in size, and frequently challenging to position correctly in media for the subsequent histological steps. Here, we discuss the embedding media and procedures that provided us with appropriate preservation of tissue and easier orientation of embryos at early development. Fertilized Gallus gallus eggs were incubated for 72 h, collected, fixed, processed, and embedded with paraplast, polyethylene glycol (PEG), or historesin. These resins were compared by the precision of tissue orientation, the preview of the embryos in the blocks, microtomy, contrast in staining, preservation, average time, and cost. Paraplast and PEG did not allow correct embryo orientation, even with agar-gelatin pre-embedded samples. Additionally, structural maintenance was hindered and did not allow detailed morphological assessment, presenting tissue shrinkage and disruption. Historesin provided precise tissue orientation and excellent preservation of structures. Assessing the performance of the embedding media contributes significantly to future developmental research, optimizing the processing of embryo specimens and improving results.

490 Beyond Placement of Pedicle Screws - New Applications for Robotics in Spine Surgery: A Multi-Surgeon, Single-Institution Experience

INTRODUCTION: Interest in robotic-assisted spine surgery has grown as surgeon comfort and technology have evolved to maximize the benefits of time-saving and precision. However, the Food and Drug Administration (FDA) has currently only approved robotics to assist in determining the ideal trajectory for pedicle screw placement after extensive research supporting its efficacy and efficiency. To be considered a durable and effective option, robotics need to expand beyond the indication of just placing pedicle screws. METHODS: We will explore accessing Kambin’s Triangle in percutaneous transforaminal interbody fusion (percLIF), iliac fixation in metastatic cancer, and sacroiliac (SI) fusions. Each of these topics will be covered in-depth with associated background information and subsequent discussion. RESULTS: Without the need for laminectomies or facetectomies, minimal tissue disruption was attained by using RA instrumentation finely tuned to each patient’s unique spinal landscape for percLIF. In the cases of iliac fixation, the use of S2AI screws was precluded by the extensive damage to the sacrum. Therefore, a modified iliac screw entry point was utilized with an entry point slightly more medial than the traditional method, with a pre-planned trajectory of the screws using robot assistance. For SI joint fusion, we review our group at Duke University's published case series displaying the “off-label” use of the robot, which is the first reported case series of its kind in the literature. CONCLUSIONS: We show that with a proper understanding of its limitations, robots can help surgeons perform difficult surgeries in a safe manner.

Intrascrotal Incision: An Alternative Technique for the Management of Inguinoscrotal Pathologies, Experience from 76 Cases

Abstract Introduction The aim of this clinical study was to present our experience gained by using the intrascrotal incision through the mid raphe for the management of a variety of inguinoscrotal pathologies. Materials and Methods A total of 76 male patients, between 2 and 16 years, underwent surgical treatment through a mid raphe intrascrotal incision, for a wide range of inguinoscrotal diseases, including torsion of the spermatic cord, torsion of the testicular appendages, non-communicating hydrocele, communicating hydrocele, ectopic testis, retractile testicles, palpable undescended testis, testicular trauma and testicular prosthesis placement. Results All the patients had an uneventful postoperative recovery, with none mentioned complication, and none of them required convention to the traditional inguinal method. The follow-up examination ranged from 6 months to 3 years, with no surgical complication highlighted. Conclusions We recommend that the intrascrotal incision through the mid raphe may be considered as an alternative technique to inguinoscrotal pathologies, instead of other approaches. The ability to treat a variety of pathologies regarding both two hemi-scrotums and the inguinal region at the same time, the provision of adequate surgical site, while succeeding much less dissection and disruption of tissue, the excellent cosmetic result, the greater comfort for the ‘day-case’ child, the ability to use the scrotal septum in order to fix the testis in the scrotum and the avoidance of an extra incision are the main advantages of the intrascrotal incision.

Paraconduit herniation - Invited editorial

Minimally invasive esophageal surgery is becoming increasingly common as the advantages of minimizing tissue disruption and bleeding are recognized. The embracement of new technology inevitably results in a change in the frequency and type of complications seen. In this edition, De Silva et al.1 highlights the increased incidence of para-conduit hiatal herniation after minimally invasive esophagectomy. This complication is well recognized in the UK, with centers reporting a higher rate of para-conduit herniation after performing minimally invasive techniques. The authors of this paper have reported the frequency of this problem across two high volume centers in Australia. The theory is that after minimally invasive surgery, there are less abdominal adhesions, which would normally prevent the free movement of intrabdominal viscera through the widened mediastinum. In contrast, during open abdominal upper gastrointestinal surgery, tissue trauma leads to the formation of adhesions in the upper abdomen, fixing structures within the abdominal cavity. It is clear that adhesion formation is important, but there are a few other factors that should be considered. For example, does the herniation occur immediately after surgery when adhesions have not yet formed? Or is it a result of pulling the conduit through into the chest? Our experience is that hernias can occur early and are sometimes in the presence of previous upper abdominal surgery.

QX-type infectious bronchitis virus infection in roosters can seriously injure the reproductive system and cause sex hormone secretion disorder

ABSTRACT Since its discovery, QX-type avian infectious bronchitis virus (IBV) has rapidly spread worldwide and become the most prevalent dominant genotype in Asia and Europe. Currently, although the pathogenicity of QX-type IBV in the reproductive system of hens is widely and deeply understood, its pathogenicity in the reproductive system of roosters remains largely unknown. In this study, 30-week-old specific pathogen-free (SPF) roosters were used to investigate the pathogenicity of QX-type IBV in the reproductive system after infection. The results showed that QX-type IBV infection caused abnormal testicular morphology, moderate atrophy and obvious dilatation of seminiferous tubules, and produced intense inflammation and obvious pathological injuries in the ductus deferens of infected chickens. Immunohistochemistry results showed that QX-type IBV can replicate in spermatogenic cells at various stages and in the mucous layer of the ductus deferens. Further studies showed that QX-type IBV infection affects plasma levels of testosterone, luteinizing hormone, and follicle-stimulating hormone as well as causes changes in transcription levels of their receptors in the testis. Furthermore, the transcription levels of StAR, P450scc, 3βHSD and 17βHSD4 also changed during testosterone synthesis after QX-type IBV infection, indicating that the virus can directly affect steroidogenesis. Finally, we found that QX-type IBV infection leads to extensive germ cell apoptosis in the testis. Collectively, our results suggest that QX-type IBV replicates in the testis and ductus deferens, causing severe tissue damage and disruption of reproductive hormone secretion. These adverse events eventually lead to mass germ cell apoptosis in the testis, affecting the reproductive function of roosters.

Development of an in vitro Functional Assay to Evaluate the Occlusive Properties of Moisturizers on Dry Skin

Introduction: Dry skin is a hallmark of impaired skin barrier function. Moisturizers are a mainstay of treatment to help the skin retain moisture, and there is a high consumer demand for effective products. However, the development and optimization of new formulations are hampered due to lack of reliable efficacy measures using in vitro models. Methods: In this study, a microscopy-based barrier functional assay was developed using an in vitro skin model of chemically induced barrier damage to evaluate the occlusive activity of moisturizers. Results: The assay was validated by demonstrating the different effects on barrier function between humectant (glycerol) and occlusive (petrolatum). Significant changes in barrier function were observed upon tissue disruption, which was ameliorated by commercial moisturizing products. Conclusion: This newly developed experimental method may be helpful to develop new and improved occlusive moisturizers for the treatment of dry skin conditions.

Arthroscopically Assisted Anterior Bone Block for Shoulder Instability

Background: Shoulder instability is a common complaint, with treatment depending on the severity of the bony defect. Advancements in arthroscopic techniques have allowed for a less invasive surgery with decreased postoperative pain, improved graft placement, and better visualization. Indications: Large bony defects of the glenoid require bone block augmentation to reduce recurrent instability. In this presentation, arthroscopically assisted anterior bone block was performed with a distal tibial allograft to address recurrent anterior shoulder instability with a large glenoid defect. Technique Description: The patient is placed in a lateral decubitus position. Standard arthroscopic portals are made. The bone block is fashioned from a distal tibial allograft. Using a guide, drill holes are placed in the bone block. The guide is inserted from the posterior portal and placed parallel to the joint surface. Using drill sleeves, superior and inferior screw holes are drilled and a metal cannula is left in place to allow for shuttling of suture. Two anchors are placed on the glenoid to allow for capsule labral junction to be brought up to the graft once the graft is in place. The bone block is delivered through a dilated anterior portal and reduced. The sutures are tensioned. A button is placed on each set of looped sutures, a Nice knot is placed into each, and a tensioning device is tensioned. Once compression is achieved, knots are tied to perform final fixation over the buttons. A suture passing device is used to grab inferior capsule and labrum to the priorly placed anchor on the inferior glenoid and tied. Results: Arthroscopically assisted anterior bone block for shoulder instability has been demonstrated to have similar clinical and radiographic outcomes when compared with open treatment. However, arthroscopically assisted Latarjet has been shown to have decreased soft tissue disruption and possible improvements in graft placement. Discussion/Conclusion: Arthroscopically assisted anterior bone block is a technically demanding surgery but has been shown to have similar outcomes when compared with open treatment. In this video, we demonstrate an arthroscopically assisted anterior bone block for recurrent anterior shoulder instability. Patient Consent Disclosure Statement: The author(s) attests that consent has been obtained from any patient(s) appearing in this publication. If the individual may be identifiable, the author(s) has included a statement of release or other written form of approval from the patient(s) with this submission for publication.

Whole-brain diffusion tensor imaging predicts 6-month functional outcome in acute intracerebral haemorrhage

IntroductionSmall vessel disease (SVD) causes most spontaneous intracerebral haemorrhage (ICH) and is associated with widespread microstructural brain tissue disruption, which can be quantified via diffusion tensor imaging (DTI) metrics: mean diffusivity (MD) and fractional anisotropy (FA). Little is known about the impact of whole-brain microstructural alterations after SVD-related ICH. We aimed to investigate: (1) association between whole-brain DTI metrics and functional outcome after ICH; and (2) predictive ability of these metrics compared to the pre-existing ICH score.MethodsSixty-eight patients (38.2% lobar) were retrospectively included. We assessed whole-brain DTI metrics (obtained within 5 days after ICH) in cortical and deep grey matter and white matter. We used univariable logistic regression to assess the associations between DTI and clinical-radiological variables and poor outcome (modified Rankin Scale > 2). We determined the optimal predictive variables (via LASSO estimation) in: model 1 (DTI variables only), model 2 (DTI plus non-DTI variables), model 3 (DTI plus ICH score). Optimism-adjusted C-statistics were calculated for each model and compared (likelihood ratio test) against the ICH score.ResultsDeep grey matter MD (OR 1.04 [95% CI 1.01–1.07], p = 0.010) and white matter MD (OR 1.11 [95% CI 1.01–1.23], p = 0.044) were associated (univariate analysis) with poor outcome. Discrimination values for model 1 (0.67 [95% CI 0.52–0.83]), model 2 (0.71 [95% CI 0.57–0.85) and model 3 (0.66 [95% CI 0.52–0.82]) were all significantly higher than the ICH score (0.62 [95% CI 0.49–0.75]).ConclusionOur exploratory study suggests that whole-brain microstructural disruption measured by DTI is associated with poor 6-month functional outcome after SVD-related ICH. Whole-brain DTI metrics performed better at predicting recovery than the existing ICH score.

Lateral lumbar interbody fusion (LLIF) reduces total lifetime cost compared with posterior lumbar interbody fusion (PLIF) for single-level lumbar spinal fusion surgery: a cost-utility analysis in Thailand

BackgroundLumbar interbody fusion techniques treat degenerative lumbar diseases effectively. Minimally invasive lateral lumbar interbody fusion (LLIF) decreases soft tissue disruption and accelerates recovery better than standard open posterior lumbar interbody fusion (PLIF). However, the material cost of LLIF is high, especially in Thailand. The cost-effectiveness of LLIF and PLIF in developing countries is unclear. This study compared the cost-utility and clinical outcomes of LLIF and PLIF in Thailand.MethodsData from patients with lumbar spondylosis who underwent single-level LLIF and PLIF between 2014 and 2020 were retrospectively reviewed. Preoperative and 1-year follow-up EuroQol-5D-5L and healthcare costs were collected. A cost-utility analysis with a lifetime time horizon was performed using a societal perspective. Outcomes are reported as the incremental cost-effectiveness ratio (ICER) and quality-adjusted life-year (QALY) gained. A Thai willingness-to-pay threshold of 5003 US dollars (USD) per QALY gained was used.ResultsThe 136 enrolled patients had a mean age of 62.26 ± 11.66 years. Fifty-nine patients underwent LLIF, while 77 underwent PLIF. The PLIF group experienced greater estimated blood loss (458.96 vs 167.03 ml; P < 0.001), but the LLIF group had a longer operative time (222.80 vs 194.62 min; P = 0.007). One year postoperatively, the groups’ Oswestry Disability Index and EuroQol-Visual Analog Scale scores were improved without statistical significance. The PLIF group had a significantly better utility score than the LLIF group (0.89 vs 0.84; P = 0.023). LLIF’s total lifetime cost was less than that of PLIF (30,124 and 33,003 USD). Relative to PLIF, LLIF was not cost-effective according to the Thai willingness-to-pay threshold, with an ICER of 19,359 USD per QALY gained.ConclusionsLLIF demonstrated lower total lifetime cost from a societal perspective. Regard to our data, at the 1-year follow-up, the improvement in patient quality of life was less with LLIF than with PLIF. Additionally, economic evaluation modeling based on the context of Thailand showed that LLIF was not cost-effective compared with PLIF. A strategy that facilitates the selection of patients for LLIF is required to optimize patient benefits.

Fatty acid-binding protein genes in gilthead seabream: molecular cloning and nutritional regulation under low water temperatures.

The molecular characteristics and tissue disruption of 10 fatty acid-binding protein (fabp) genes in gilthead seabream (Sparus aurata) were investigated, and their expression levels were found in the fish fed diets with different vegetable oil (VO) sources, which may explore the potential function of fabp genes in S. aurata. For this purpose, the open reading frames of fabp genes involved in the transport and ß-oxidation of fatty acids (FA) were molecularly cloned and characterized. S. aurata was then exposed to a two-staged feeding trial (the grow-out period following a wash-out period) at low water temperatures. In the grow-out period, the fish were fed diets containing 50% and 100% ratios of various VOs for 60 days, and in the wash-out period, the fish were fed a diet containing 100% fish oil (FO) for 30 days. It has been determined that (a) S. aurata and vertebrate fabp/FABP genes are orthologues; (b) spatio-temporal differences in tissue-specific patterns of fabp genes differ importantly; for instance, the difference between the highest and lowest values reaches 13 × 105 -fold in the fabp10a; and (c) VO-based diets upregulated fabp transcript levels in the liver and muscle with some exceptions, such as liver fabp11a and muscle fabp7a. Gene expressions of only the hepatic fabp7b and fabp10a genes were diminished at the end of the wash-out period. In this study, the authors provide further evidence that dietary FAs affect fabp mRNA expressions in S. aurata. This might be useful in the nutritional control of fabp genes to maintain lipid homeostasis in marine fish fed VO-based diets at low water temperatures.

P914 An adult zebrafish model for adherent-invasive Escherichia coli indicates protection from infection by treatment with probiotic E. coli Nissle

Abstract Background Adherent-invasive Escherichia coli (AIEC) is an opportunistic pathogen associated with major inflammatory bowel disease (IBD), Crohn’s disease (CD) and ulcerative colitis (UC). AIEC is a human commensal but during unfavorable gastric conditions it exhibits pathogenic properties and induces inflammation in the gut, sometimes progressing to inflammation-induced colon cancer. The exact mechanisms for AIEC pathogenesis are still being discovered. As a human opportunistic pathogen, AIEC has few animal models. In recent years, zebrafish have emerged as a useful model for studies of various human pathogens. Here, a zebrafish model to study AIEC infection was developed. Methods Colonization of AIEC from overnight culture was determined with different infectious doses (103, 104, 105 and 106 CFU/ml of infection water) in zebrafish, incubating fish with AIEC in water for 6 h and then in fresh water for 18 h at 28o C. The invasive property of AIEC in zebrafish gut was determine by gentamycin protection assay. Persistent colonization of AIEC in zebrafish gut was also assessed for 10 days. Induction of pro-inflammatory markers such as IL-1β, TNFα, IFNγ and S100A-10b (akin to human calprotectin) in zebrafish intestinal cells in response to AIEC infection was determined by qRT PCR and ELISA. A probiotic strain, E. coli Nissle 1917 (EcN) was used as a therapeutic and prophylactic against AIEC infection. Propionic acid was used to augment the AIEC infection and the efficacy of ECN was tested against the hyper-infectious condition of AIEC in zebrafish. Results Bath inoculation with AIEC resulted in colonization, invasion and tissue disruption in the zebrafish intestine. The AIEC infection induced neutrophil recruitment and showed significant pro-inflammatory responses in zebrafish gut. S100A-10b protein of zebrafish, equivalent of a human IBD marker, calprotectin, was significantly increased in the zebrafish gut during AIEC infection. The probiotic E. coli Nissle 1917 (EcN) reduced AIEC colonization, tissue damage and pro-inflammatory responses in zebrafish in both prophylactic and therapeutic ways. Furthermore, EcN diminished the propionic acid-augmented hyper infection of AIEC in the zebrafish model. Conclusion In this study a new zebrafish model was developed to study the infection and inflammation induced by AIEC. Probiotic strain, EcN was used successfully against AIEC to reduce the inflammation in AIEC-zebrafish model.

Inflammatory Treatment Used to Mimic Osteoarthritis and Patients' Synovial Fluid Have Divergent Molecular Impact on Chondrocytes In Vitro.

Osteoarthritis (OA) is a chronic disease characterized by joint tissue disruption and inflammation with a paucity of therapeutic options. Chondrocyte in vitro models are commonly used as the first step in evaluating new approaches and rely on the stimulation of an OA-like phenotype with inflammation often the method of choice. Inflammatory priming is frequently based on cytokines used at concentrations very far from the reality in the patients' synovial fluid (SF). The aim of this work was to compare the transcriptional response of chondrocytes to different inflammatory conditions: the high levels of IL1β that are used for standardized inflammation protocols, OA-SF, IL1β, IL6 and IFNγ at SF-like concentrations both individually and simultaneously to mimic a simplified "in vitro" SF. Both high IL1β and OA-SF strongly influenced chondrocytes, while SF-like concentrations of cytokines gave weak (IL1β alone or in combination) or no (IL6 and IFNγ alone) outcomes. Chondrocytes under the two most powerful polarizing conditions had a clearly distinct fingerprint, with only a shared albeit molecularly divergent effect on ECM stability, with IL1β mainly acting on ECM degrading enzymes and OA-SF accounting for a higher turnover in favor of fibrous collagens. Moreover, OA-SF did not induce the inflammatory response observed with IL1β. In conclusion, although partially similar in the endpoint phenotype, this work intends to encourage reflection on the robustness of inflammation-based in vitro OA models for molecular studies on chondrocytes.

Isolation of Mitochondria from Mouse Tissues for Functional Analysis.

Methods for isolating mitochondria from different rodent tissues have been established for decades. Although the general principles for crude mitochondrial preparations are largely shared across tissues - tissue disruption followed by differential centrifugation - critical differences exist for isolation from different tissues to optimize mitochondrial yield and function. This protocol offers a unified resource for preparations of isolated mitochondria from mouse liver, kidney, heart, brain, skeletal muscle, and brown and white adipose tissue suitable for functional analysis.

Probiotics and postbiotics play a role in maintaining dermal health.

Probiotics and postbiotics have emerged as an alternative to traditional antibiotics for the treatment of persistent skin infections. The use of probiotics and postbiotics has been shown to have a positive impact on the maintenance of skin health by promoting the growth of beneficial bacteria and inhibiting the growth of harmful bacteria. Probiotics work by adhering to the skin and mucosal membranes and competing with pathogenic organisms for nutrients, which prevent the growth of harmful bacteria. In addition, probiotics and postbiotics produce antimicrobial substances that help eliminate pathogenic bacteria, resulting in improved skin health. The skin is the largest organ in the body and serves as a protective barrier against external pathogens. When harmful bacteria colonize the skin, it can lead to tissue damage and disruption, which can cause chronic inflammatory, non-healing skin disorders such as dermatitis, psoriasis, and acne. Traditional treatments for persistent skin infections rely heavily on antibiotics, which can have several adverse effects on the body, including the development of antibiotic resistance. Moreover, pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus, which are often implicated in chronic skin infections, can form biofilms that are highly resistant to antibiotics and host immune responses. In recent years, research has shown that probiotics and postbiotics can play a significant role in the maintenance of dermal health. Probiotics and postbiotics can stimulate the immune system, enhance the production of skin barrier components, and modulate skin inflammation, all of which are critical for the maintenance of healthy skin. In this review, we have compiled the current literature on the therapeutic potential of probiotics and postbiotics for the treatment of persistent skin infections and their impact on maintaining dermal health.

The injectable local anaesthetics in the prevention of intraoperative pain in paediatric dentistry – practical remarks on agent choice and methods. Possible complications of local anaesthetic administration in paedodontics

In paedodontics, as in general dentistry, causal treatment plays a fundamental role in the management of pain present before the appointment, but the therapeutic procedures performed are often associated with the occurrence of acute pain. It is currently being accepted, that effective dental treatment of children is possible after obtaining the appropriate level of local anaesthesia. Local anaesthesia is one of the most delicate and difficult procedures in paediatric dentistry. Fear of anaesthesia and the accompanying pain is clearly marked in paediatric patients; therefore, the right attitude of a dentist plays a crucial role, while psychological strategies and pharmacological approach may be a valuable help. In the case of intraoperative pain prevention in paedodontics, injectable aminoamide derivatives-mainly lidocaine, articaine and mepivacaine in Polish conditions – are of fundamental importance. The success of the procedure is the result of the precision of the injection and administration of the solution, dependent on the practitioner, the reaction of the tissues into which the drug is deposited, their anatomical structure and the patient's stress level. The article discusses the clinically significant features distinguishing paediatric patient population and the impact of this specificity on the choice of methods of effective dental local anaesthesia. The agents used have been characterized, taking into account the dosage, the latency period and the resulting anaesthesia time for the dental pulp and soft tissues. Pulp anaesthesia allows for painless intervention within the tooth tissues, while soft tissue anaesthesia is necessary to undertake a therapeutic action requiring soft tissue disruption in dental surgery. Soft tissue anaesthesia, especially in the inferior alveolar nerve block, with long-acting local anaesthetics, is associated with a higher risk of self-injury in children with loss of sensation and proprioception in mucosa of the cheeks, lips and tongue. Local and general complications of dental anaesthesia in children were presented, along with their prevention and management.

Molecular mechanisms of neuronal death in brain injury after subarachnoid hemorrhage.

Subarachnoid haemorrhage (SAH) is a common cerebrovascular disease with high disability and mortality rates worldwide. The pathophysiological mechanisms involved in an aneurysm rupture in SAH are complex and can be divided into early brain injury and delayed brain injury. The initial mechanical insult results in brain tissue and vascular disruption with hemorrhages and neuronal necrosis. Following this, the secondary injury results in diffused cerebral damage in the peri-core area. However, the molecular mechanisms of neuronal death following an aneurysmal SAH are complex and currently unclear. Furthermore, multiple cell death pathways are stimulated during the pathogenesis of brain damage. Notably, particular attention should be devoted to necrosis, apoptosis, autophagy, necroptosis, pyroptosis and ferroptosis. Thus, this review discussed the mechanism of neuronal death and its influence on brain injury after SAH.

Anti-tumour efficacy of Musa paradisiaca flower extracts on DMBA induced mammary carcinogenesis in female wistar rats

Several reports have shown the beneficial advantages of Musa paradisiaca in wound healing activities and other ailments. Previously, our in vitro studies validated the anti-cancer activities of Musa flower extracts and confirmed the potential. This thread has led to our current investigation to explore the anticancer potential in vivo . This study focused on the efficacy of banana florets on DMBA induced breast cancer in female wistar rats. In our current study, we investigated that tumour induction has an effect in disrupting the estrous cycle in mice which may result by hormonal fluctuation levels. Antioxidant and hormonal analysis in vivo revealed the imbalance in estrogen and progesterone levels in untreated group compared to the treated ones. Floral crude extract treatment in vivo has shrunken the tumour volume in flower extract treatment group as well as in standard drug compared to the control. Histopathological staining confirms the disruption of epithelial tissues in tumour induced groups compared to the treated ones. Moreover, Musa floral treatment has shown to revert the damaged tissue morphology in the treated groups compared to the saline treated one. Tumour induction in mice has an effect in showing the disruption of estrous cycles. In vitro studies in MCF-7 and MDA-MB-231 breast cancer cell lines has shown the potent anti-tumorigenic activity using Musa floral extracts. Taken together, our finding confirmed that banana flower extracts showcase anti-carcinogenic activity against breast cancer both in vitro and in vivo . • Musa paradisiaca crude ethanolic flower extracts have a potential in reducing the tumour growth. • DMBA induced cancer has a deleterious effect on estrogen cycles in female wistar rats. • Disruption of epithelial morphology was observed in tumour induced wistar rats. • Treatment with crude Musa flower extracts on cancer induced rats shows the reduced level of damage and oxidative stress compared to the no treatment group.

Comparing the effect of several pretreatment steps, selected to steer (bio)chemical reactions, on the volatile profile of leek (Allium ampeloprasum var. porrum)

Volatile compounds in foods can witness the occurrence of (bio)chemical reactions, comprising both enzymatic and non-enzymatic reactions, which can be influenced by processing. This study investigated the effect of different pretreatments aimed at either minimizing, realized by a heat treatment, or inducing, realized by a tissue disruptive treatment, enzymatic reactivities, on the volatile profile of leek. The volatile profiles obtained were then linked to possible (bio)chemical reactions that could have occurred during the treatments. The study showed that different pretreatments led to markedly different volatile profiles, characterized by different (abundances of) volatile compounds. Partial and extensive tissue disruption was achieved by pulsed electric fields at low electric field strength and mixing, respectively. After these tissue disruptive treatments, the volatile compounds could mainly be related to the occurrence of several enzyme-substrate interactions, including conversions of alk(en)yl cysteine sulfoxides by alliinase and of unsaturated fatty acids by lipoxygenase and hydroperoxide lyase. Thermally-induced reactions were also observed to impact the resultant volatile profile. Present study revealed that targeted (pre)treatment allows to steer (bio)chemical reactions towards specific volatile compounds in leek products.