Background and objective: The osteoporotic vertebral compression fracture (OVCF) has an incidence of 7.8/1000 person-years at 55–65 years. At 75 years or older, the incidence increases to 19.6/1000 person-years in females and 5.2–9.3/1000 person-years in males. To solve this problem, percutaneous vertebroplasty (PVP) was developed in recent years and has been widely used in clinical practice to treat OVCF. Are the clinical effects of unilateral percutaneous vertebroplasty (UPVP) and bilateral percutaneous vertebroplasty (BPVP) the same? The purpose of this study was to compare biomechanical differences between UPVP and BPVP using finite element analysis.Materials and methods: The heterogeneous assignment finite element (FE) model of T11-L1 was constructed and validated. A compression fracture of the vertebral body was performed at T12. UPVP and BPVP were simulated by the difference in the distribution of bone cement in T12. Stress distributions and maximum von Mises stresses of vertebrae and intervertebral discs were compared. The rate of change of maximum displacement between UPVP and BPVP was evaluated.Results: There were no obvious high-stress concentration regions on the anterior and middle columns of the T12 vertebral body in BPVP. Compared with UPVP, the maximum stress on T11 in BPVP was lower under left/right lateral bending, and the maximum stress on L1 was lower under all loading conditions. For the T12-L1 intervertebral disc, the maximum stress of BPVP was less than that of UPVP. The maximum displacement of T12 after BPVP was less than that after UPVP under the six loading conditions.Conclusion: BPVP could balance the stress of the vertebral body, reduce the maximum stress of the intervertebral disc, and offer advantages in terms of stability compared with UPVP. In summary, BPVP could reduce the incidence of postoperative complications and provide promising clinical effects for patients. Osteoporosis (OP) is a systemic metabolic disease characterized by reduced bone mass and easily fractured bones, which causes approximately 8.9 million fractures worldwide each year, with an average of one every 3 s . The ostTo solve this problem, percutaneous vertebroplasty (PVP), a minimally invasive surgical technique, was developed in recent years. This technique can quickly stabilize the fracture, relieve pain, and reinforce the anterior column through the percutaneous injection of bone cement into the fractured vertebral body . MeanwhiCurrently, PVP has been widely used in clinical practice to treat OVCF  and can A three-dimensional FE model of T11-L1 was first reconstructed using Mimics 21.0  based on computed tomography (CT) scans with 0.2-mm intervals of a normal male adult without any lumbar disease . This stρ represents density, HU represents the threshold, and E represents elastic modulus.Three-dimensional solid elements with an isotropic character were used to model the vertebral body, endplate, and intervertebral disc . The eleThe materials were divided into ten categories and assigned according to the grayscale of the CT images .The element types of endplates were eight-node hexahedral elements (C3D8). Each intervertebral disc consisted of a nucleus pulposus surrounded by an annulus ground substance . The annTo get accurate results, the mesh grid of this patient-specific model was verified. In this study, the three-dimensional finite element model of T11-L1 was used for the mesh convergence test. The element size of T11-L1 was varied to yield four different mesh resolutions by keeping the very refined mesh as the reference for comparisons . The peaSpinal ligaments  were included in the model. These ligaments were modeled with two-node 1D spring elements applied only to the tension force in seven groups  Tan et . All matThe osteoporotic condition was modeled by decreasing the elastic moduli of each category of vertebral bodies by a set amount  Polikei. AccordiThe simulation of UPVP and BPVP was performed in Materialise Magics 21.0 . The cement cylinder was vertically implanted into one side of the fractured vertebra to simulate UPVP . AnotherThe lower surface of the L1 vertebral body was fixed in all directions throughout the simulation process. The upper surface of the T11 vertebral body was implemented with a 500 N axial compression load to simulate the weight of the human upper body segment , a pure The current FE model was validated in combined loading of both pure moment and follower load, and the predicted results of range of motion (ROM) was compared with the numerical data obtained under the identical condition to validate the FE model. The present model results were in good agreement with the results obtained from the literature , and theCompared with the fractured model, the maximum stress on T12 in UPVP increased under left lateral bending and left axial rotation . HoweverAs shown in The stress distributions at the T11-T12 disc and the T12-L1 disc did not change significantly in the fractured model, UPVP, and BPVP under six conditions , 10.For the T11-T12 intervertebral disc, the maximum stresses of a fractured model, UPVP, and BPVP had little difference under six loading conditions . HoweverUnder flexion, extension, right lateral bending, and right axial rotation, the maximum displacement of T12 in UPVP decreased by 47.81%, 48.55%, 0.82%, and 17.90%, respectively, compared with the fractured model. Under left lateral bending and left axial rotation, the maximum displacement of T12 in UPVP increased by 5.60% and 0.27%, respectively, compared with the fractured model. However, the maximum displacement of T12 in BPVP decreased under all loading conditions and was lower than UPVP, which decreased by 58.88%, 49.24%, 16.31%, 20.42%, 10.27%, and 20.13% compared with the fractured model .Complications of PVP include recompression of the PVP-operated vertebrae, new fractures at the neighborhood vertebrae, and degenerative disc disease (DDD) . All of First, for the recompression of the PVP-operated vertebrae, some studies have shown that the distribution of bone cement and the stiffness of the vertebral body, which affect stress balance on the latter, are important influencing factors . n this According to the three-column theory by Denis, recompression of the vertebrae could be affected by the stability of the spine . The pooIn conclusion, compared with UPVP, BPVP, with the symmetrical distribution of bone cement and superior stability, could reduce the incidence of the recompression of PVP-operated vertebrae and relieve pain caused by recompression.Second, for the new fractures at the neighboring vertebrae, some studies have shown that load transfer, which could be along the longitudinal axis of the spine to the adjacent vertebral body, is one of the important causal factors affecting the fracture risk of the untreated adjacent vertebrae . Kim et Third, for DDD, To sum up, this study found that compared with UPVP, BPVP with the symmetrical distribution of bone cement has certain advantages in restoring the stability of the spine and reducing the incidence of postoperative complications such as recompression of the PVP-operated vertebrae, new fractures at neighboring vertebrae, and DDD by better restoring the biomechanics of the spine. Therefore, BPVP can improve the quality of life of patients and bring satisfying long-term clinical effects.in vivo loadings. All of these might have an impact on stress and displacement. Third, for simulating bone cement with a vertical cylinder, the similar method to simulate bone cement had been reported by in vitro biomechanical experiments and clinical study to evaluate the findings from this study also would be conducted in the future.The finite element model has several limitations in this study. First, some simplifications were necessary for creating the model, such as the characteristics of intervertebral discs, ligaments and paraspinal muscles and he assumptions of linear, isotropic, and homogeneous material properties for the finite element model. However, these simplifications might have an impact on stress and displacement. via the finite element method to determine the relationship with complications. The results suggested that BPVP could balance the stress of the vertebral body, reduce the maximum stress of the intervertebral disc, and has advantages in stability compared with UPVP. Therefore, BPVP could reduce the incidence of postoperative complications, which could provide guidance for clinical application and promising clinical effects for patients.In this study, the biomechanical differences between BPVP and UPVP were compared


Background: Within the UK, a non-medical prescriber is a non-medical healthcare professional who has undertaken post-registration training to gain prescribing rights. Lack of post-qualification NMP training has previously been identified as a barrier to the development of oncology non-medical prescribing practice. Aim: To explore the experiences and opinions of multi-professional non-medical oncology prescribers on post-qualification training. Method: Nine out of 30 oncology non-medical prescribers  from a single cancer centre in Wales, were selected from a study site NMP database using randomisation sampling within Microsoft® Excel. Participants were interviewed using a validated and piloted semi-structured interview design on the topic of post-qualification training for non-medical prescribers. Participants were invited via organisational email. Interviews were audio-recorded and transcribed verbatim. Anonymised data were thematically analysed aided by NVivo® software. Results: Main themes identified: experience related to training, competency, support and training methods. Competency assessment methods discussed were the annual non-medical prescriber appraisal, peer review and a line manager’s overarching appraisal. Support requirements identified included greater consultant input to help non-medical prescribers identify training and peer support opportunities. Organisational support was requested regarding regular study leave and governance around clinical judgement and errors. The need for regular structured in-house training related to non-medical prescriber’s level of experience was identified. Conclusion: Development of organisation-led governance strategies and in-house training programmes will support training equity for all non-medical prescribers within the organisation. Non-medical prescribing competence incorporating a variety of assessment methods and tools, such as OSCEs, peer review and ‘portfolios of evidence’, will support maintenance of non-medical competence.Implementation of an organisation-wide oncology non-medical prescribing training programme, based on level of experience, will guide non-medical prescribers and their consultant mentors with support and aid transition from early career to advanced practice.Establishing consultant roles for each non-medical prescribing profession will provide leadership and development of non-medical prescribing governance strategies, to support training equity for all non-medical prescribers.Within the United Kingdom, a non-medical prescriber (NMP) is a non-medical healthcare professional, who has undertaken post-registration university training to gain prescribing rights .NMPs are either an ‘independent’ or ‘supplementary’ prescriber. An independent prescriber (IP) is defined as ‘a practitioner, who is responsible and accountable for the assessment and diagnosis of patients’ conditions and can make prescribing decisions to manage the clinical condition of the patient’. Professionals who can qualify as IPs in the UK are midwives, nurses, optometrists, paramedics, pharmacists, physiotherapists, podiatrists, and therapeutic radiographers . A supplThe main objectives of implementing non-medical prescribing were to improve access to medicines, utilise eligible professionals’ clinical skills and to reduce the workloads of medical colleagues so they could focus on more complicated patient cases . NMPs haWithin oncology, all treatments given to treat cancer are known as systemic anti-cancer therapy (SACT) . The NMPIn 2018, the study site had 30 non-medical prescribers: eight pharmacists, 18 nurses (including two nurse supplementary prescribers (SPs)) and four therapeutic radiographers. Nurse SPs were excluded from this study as their practise varied due to clinical restrictions within CMPs which may affect their training requirements.At the study site, the majority of nurse and pharmacist NMPs practise within medical consultant oncologist teams reviewing outpatients prior to their next cycle of SACT. NMPs prescribe SACT and supportive medicines, assess haematological tests and request scans and blood transfusions if appropriate. Three nurse NMPs review patients at ward level. Therapeutic radiographer NMPs review patients within radiotherapy review clinics, prescribing supportive medicines and SACT in combination with radiotherapy .NMPs have varying levels of experience at the study site. For this study, NMPs were classified as an ‘advanced practice’ NMP if they have practised within oncology as an NMP for five years or more; if the NMP has practised within oncology for less than five years, they were classified as an ‘early years’ NMP.Currently, there is no structured programme for NMP training at the study site, although ad-hoc NMP training sessions on various topics are arranged through quarterly NMP meetings, known as the ‘NMP meeting forum’ if needed and mostly arranged by NMP staff themselves. There is no automatic provision of study leave for NMPs, although some NMPs will undertake certain CPD training dependant on their day-to-day role, e.g., Clinical Nurse Specialists (CNS) have an annual CPD training day. All NHS employees at the study site have an annual appraisal with their departmental line manager. All practising NMPs also have an additional annual NMP appraisal with their consultant oncologist mentor as a governance requirement of the organisation. Previous studies undertaken at the study site by Harding and colleagues have explored the opinions and beliefs of all oncology prescribers and their senior managers. They concluded that skills and training of oncology NMPs across the study site organisation needed to be strengthened for NMP practice to develop further.To explore the experiences and opinions of multi-professional non-medical oncology prescribers on post-qualification training.Ethical approval for the study was obtained from Aston University Ethics Committee , on approval date 30/09/2016 (minor amendment in April 2021 adding therapeutic radiographers due to practice change) . OrganisA literature search (September 2021) via EMBASE and MEDLINE identified 28 relevant NMP studies published within the last 15 years  and focused on non-medical prescribing by pharmacists, nurses and radiographers and narrowed further using education and oncology. MeSH terms were used. Studies were included if they included the study of UK NMP practice and published in English, whilst conference abstracts and publications not published in English were excluded. Although many were predominantly focused on nurse prescribing, only three studies were identified related to oncology NMP practice; none was associated with oncology NMP training –11.The research study was set within one oncology centre in Wales, which employs 670 staff from a range of professions across all departments. Each year, 5000 new referrals are received and around 50,000 outpatients are treated. .n = 8), nurse group (n = 16) and radiographer group (n = 4)].Three SSIs from each of the three professional groups were included as this was considered sufficient to explore both the depth and width of the study site, and on analysis findings were considered to be reliable and valid. Further interviews were considered but believed unlikely to add anything of importance. SPs were excluded from the study. Each of the pre-selected nine participants were emailed by the research team using the organisation’s email directory for recruitment and contained both the participant information sheet and a consent form. Provisions were made to recruit more participants using the randomisation software if needed. Participants were asked to sign the written consent form and submit it before the start of the SSI. The study was designed and conducted in accordance with COREQ principles [Between January and June 2018, nine participants were randomly selected to participate in a 60-minute one-to-one face-to-face semi-structured interview (SSI). A randomisation calculation within Microsoft® Excel software version 15.0) was used to choose potential participants within each professional group. The interview schedule was designed for a 60-minute interview using McNamara guidelines [.0 was usroup n =  and radiFace-to-face SSIs were undertaken using reflexivity (by SH) at the study site. The SSIs were audio recorded and included pre-piloted topic questions which were validated by the R&D department at the study site and the pilot interviews were included in the main study data. Free discussion was encouraged around open questions on the topic of NMP training and aimed to explore topics identified within published literature  then anonymised and analysed using a thematic approach  , aided bAll participants were IPs practising within oncology at the study site at the time of the interviews. Participants were three nurse NMPs, three pharmacist NMPs and three radiographer NMPs.Analysis using a thematic approach of the data identified four main themes: experience relating to training, competency, support and training methods. Sub-themes were also identified , scope of practice and peer review. Exemplar quotes are provided in Box Nurse-1, Pharmacist-2 and Radiographer-1 described varying opinions on the use of annual revalidation appraisals as a method of ensuring competency governance at the study site. Nurse-1 discussed undertaking a combined appraisal method which involved NMPs proactively collecting a competency ‘portfolio of evidence’ which was reviewed by their nursing line manager and signed off by the consultant oncologist.An OSCE is a direct observation evaluation tool used to assess clinical staff . Nurse-1Nurse-1 suggested that each NMP’s scope is not clearly defined at the same level of detail across all oncology specialities, which may be due to variable expectations of consultant mentors.Pharmacist-1 and Pharmacist-3 believed that peer review could be used to assess competency. Nurse-1 recommended developing an oncology ‘independent prescribing advanced practice framework’ which could be used to peer review NMPs by reviewing their portfolios of evidence.Nurse-2 described how they audited themselves as a method of assessing competency, which could have been undertaken due to lack of peer review available at the study site. Collecting and recording NMP prescribing data is not current practice at the study site.All participants agreed that training support from all stakeholders is fundamental to the NMP role. Exemplar quotes are provided in Box Most participants stated that they did receive training support from their consultant oncologist, although they also described a need for more leadership and support with their NMP development. Pharmacist-3 explained that they used their own initiative to direct their own learning and training needs.Pharmacist-1 and Pharmacist-2 described a lack of support for attending NMP training from their line manager, due to their line manager not being an NMP themselves, but there was support from the NMP lead. Radiographer-1, Radiographer-2, Radiographer-3 and Nurse-2 described feeling supported by their line managers regarding NMP training. This may be due to their line manager also practising as an NMP. The consultant non-medical professional role was not discussed by pharmacists and nurses due to this role not being implemented within their departments at the study site. Radiographer-3 did describe their consultant radiographer currently providing NMP leadership within the radiotherapy department at the study site.All participants described how future in-house NMP training sessions are planned within the study site NMP meeting forum on an ‘ad hoc’ basis. Pharmacist-3 suggested that these meetings could be improved upon, especially regarding their frequency and efficacy. Nurse-2 suggested utilising the NMP meeting forum to facilitate peer support, i.e., more experienced NMPs sharing experiences with early career NMPs.Nurse-1 and Nurse-2 commented on organisational support. Nurse-1 described the need for support when NMPs make an error, making decisions and for regulation protection regarding their clinical judgement. Nurse-2 believed that regular study leave should be provided by the organisation for NMPs’ training mirroring the current medical prescribers’ training at the study site.Regarding professional support internal to the organisation, Radiographer-1 and Radiographer-2 described having internal radiographer department support sessions for case discussion and in-house NMP competencies led by the lead radiographer NMP (Consultant Radiographer) and believed these to be beneficial. Pharmacist-1, Pharmacist-2, Pharmacist-3 and Nurse-1, Nurse-2 and Nurse-3 did not describe any internal professional training support across the study site, although Nurse-2 described attending useful regular CPD sessions within their CNS role.Regarding professional support external to the organisation, Pharmacist-1 described utilising British Oncology Pharmacy Association (BOPA) events to keep up to date and network with external colleagues. Radiographer-1 described a radiographer national online forum for NMPs. Both Pharmacist-1 and Radiographer-1 found these beneficial to their practice.Main training methods described by participants are detailed below with exemplar quotes in Box All participants commented on various aspects of ‘learning from others’. Nurse-1 discussed how their practice benefitted by communicating with other nurses fulfilling the same role, both within and outside of their organisation. Learning from other NMP professional groups was described as beneficial by Nurse-3.Nurse-2 explained how they had not completed any formal training after qualifying as an NMP. Nurse-3 believed that in-house training should be available on a regular basis and Radiographer-3 was in favour of regular in-house training, but topics should be beneficial to all. Pharmacist-3 described how NMPs should be allocated time to attend established in-house junior medical training within the organisation. Participants made suggestions for the development of a structured in-house organisation-wide NMP training programme.Seven NMP participants described using their own initiative to undertake relevant training to develop their own NMP role. Nurse-2 had experienced a lack of guidance when extending their scope of practice and believed that more guidance should be available from the organisation.NMP appraisal was the main method of competency assessment described at the study site and its structure was believed to vary between different NMP professional roles or between NMP individuals. One participant described a line manager NMP appraisal method of assessing competency which could be made available to all NMPs. Another participant believed that an NMP’s scope of practice should be clearly defined; this does not currently occur, although it is requested within the organisation’s NMP guideline. Peer review of a portfolio of evidence was not currently utilised for NMP competency assessment at the study site, but participants believed it would be beneficial to implement.Most study participants felt supported by their consultant mentor but believed their consultant mentors should have more responsibility for sharing their views on an individual’s NMP training needs. The level of line manager NMP support experienced by participants was believed to be affected by line managers’ understanding of the NMP role. Participants believed organisational support for NMP training should be viewed as high importance if NMP practice is seen as important to the organisation’s strategy. Departmental support sessions were in place for radiographer NMPs and led by a radiographer lead NMP (Consultant Radiographer). However, nurses and pharmacists within the organisation did not describe any designated departmental professional NMP leads.Various learning methods for NMP training were described by each professional NMP group. Nurse NMPs train and discuss practice with their nursing colleagues within the same role, radiographers learn within departmental peer review sessions, and pharmacist NMPs learn from other pharmacist NMP peers outside of the organisation. No formal training after qualifying as an NMP was in place and developing an in-house rolling training programme incorporating training for both early years NMP practice and advanced level NMPs was recommended to aid oncology NMP development at the study site.Prior to this study, oncology NMP training was under-explored within published literature, especially the inclusion of equal representation of pharmacist, nurse and radiographer NMPs. Semi-structured interviews allowed the researcher to explore participants’ opinions in depth.Trustworthiness in qualitative studies involves ensuring credibility, transferability, dependability and confirmability , 20. TheAlthough participants were all employed and practised at one study site, experiences and opinions on NMP competency and support may vary within other organisations. Therefore, study findings may or may not be transferable to other organisations or other medical specialties. Triangulation was not used within the study methodology but could have established a higher level of trustworthiness of the study data.Organisation-wide adoption of the combined NMP and line manager appraisal method model, which involves a review of each individuals’ portfolio of evidence (previously signed off by the consultant mentor) within an annual line manager appraisal, is in line with BOPA recommendations on competency assessment . At the Consultant support is seen as beneficial to NMPs, the study by Latter and colleagues also described how the benefit of medical support can build NMP confidence . VaryingA need for a post-qualifying training programme related to competency was highlighted by study participants and the development of formal programmes of CPD have been recommended as a priority for all employers in two nursing studies exploring nurse prescriber CPD , 30. A UThemes identified could be further explored in more detail within focus groups, including representation from NMP stakeholders such as current NMP staff, medical staff, and hospital managers both across the organisation and within other organisations offering oncology services. Survey methods could also be used to obtain more generalisable data across a wider participant population and include greater participant numbers. Future development of an underpinning research framework would explore reasons for the lack of training implementation in secondary care settings.The combined NMP and line manager appraisal method model suggested should be adopted across the organisation incorporating the portfolio of evidence (which is in line with BOPA recommendations) .Establishing consultant posts for all NMP professional groups within the organisation may provide equitable and strengthened support to all professional NMP groups. These lead professionals could deliver a clinical educator role for oncology NMPs within their departments, but also form an NMP leadership panel. The leadership panel would lead on implementation of a NMP training programme for all professionals, as well as development of their own professional group. The panel could hold a central NMP training budget and manage training allocations and lead on the development of clear organisational NMP governance strategies. The panel would provide support and mentorship to address current NMP training inequalities identified by participants.Development of a core multidisciplinary NMP programme with separate training on certain topics for certain professional groups is recommended. Future training topics suggested in this study should be considered and incorporated into a training framework algorithm for oncology NMPs within the study organisation with consideration of the NMPs’ level of experience.This study explored experiences and opinions of oncology NMPs of current post-qualification NMP training across three professional groups. Lack of a multi-disciplinary standardised approach to NMP post-qualifying training needs to be addressed by the organisation. An organisation-led NMP training programme should be developed which incorporates NMP levels of experience and professional needs. The programme should be supported by a sound organisation-wide NMP training strategy incorporating factors such as support networks and competency assessment, in order to aid development and bolster resilience of future oncology NMP practice.


Polygonum minus (P. minus) aqueous extract on chronic ultra-mild stress (CUMS)-induced depressive mice model. Chronic ultra-mild stress can disturb the neurotransmitters levels and plasticity of the hippocampus. Balb/c male mice were used in this study, which consisted of six groups (n = 14). Treatment was given for eight weeks, and chronic ultra-mild stress was applied for six weeks. Commercially available P. minus extract (BioKesum®) was used in this study. The behavior and neurochemical parameters were investigated through behavioral Tests and ELISA assays. P. minus administration significantly (p < 0.05) restored CUMS-induced behavior abnormalities, decreased the immobility time, and increased the sucrose preference and increased the spatial memory. P. minus treatment also showed the decreased level of serum corticosterone and increased the level of hippocampal neurotransmitters (Serotonin and Norepinephrine) significantly (p < 0.05). The brain-derived neurotrophic factor (BDNF) level also increased significantly in both the prefrontal cortex and hippocampus (p < 0.05). P. minus treatment exhibited significant (p < 0.05) reduction of Monoamine Oxidase-A (MAO-A) in the hippocampus. These findings indicate that P. minus aqueous extract exhibits antidepressant effects, including decreased immobility time, increased spatial memory, reduced corticosterone, increased BDNF level, and reduced MAO-A enzyme level with increasing the monoamines (serotonin and norepinephrine) in the hippocampus.Depression is the most common behavior disorder that leads to many disabilities. The main aim of this study was to evaluate the effects of a  Depression is a common illness around the world. According to a WHO report, around 350 million people of all ages are influenced by this disorder. It is a heterogeneous syndrome and can lead to poorer quality of life. In the worst case, depression can also lead to suicide ,2. SeverStress is a common reason for depression. Chronic stress causes major depressive disorders by altering neurotrophic and neuroinflammatory factors in an organism . The chrPolygonum minus Huds, the synonym for Persicaria minor comes from the Polygonaceae family and is commonly known as Kesum or Laksa leaf in Malaysia. It is used as a flavoring in many dishes and is also consumed as ulam  for health care [P. minus leaf extract contained flavonoids such as quercitrin (quercetin 3-rhamnoside) and quercetin 3. glucuronide has been suggested to have antioxidant, anti-inflammatory, antimicrobial, and cognitive-enhancing properties. An aqueous extract P. minus showed significant enhancing effects on memory and cognitive functions with a 100 mg/kg dose in mic [P. minus extract has shown significant improvements in attention and memory, mood, and quality of life among middle-aged women after six weeks of administration. BioKesum is a Malaysian brand of P. minus leaf extract which consist of standardized flavonoids including quercitrin and quercetin 3. glucuronide. The latest research of six months of BioKesum® administration potentially improved visual memory, and negative mood among older patients but its mechanism was unknown. BioKesum® was developed and registered under the National Pharmaceutical Control Bureau (NPCB) with the registration number of MAL14015033T [lth care . The P. e in mic  e. Recen4015033T . The antPolygonum minus aqueous extract on stress-induced depressive, and anxiety behavior in mice through measurement of various neurochemicals.The main aim of this study was to evaluate the effects of Polygonum minus on chronic ultra-mild stress-induced mice models through various behavioral and experimental assays. Behavioral tests consisted of the sucrose preference test (SPT), Barnes Maze assay, open field test (OFT), and forced swimming test (FST). ELISA assays were performed for the measurement of the brain-derived neurotrophic factor (BDNF), serotonin (5HT), norepinephrine (NE), monoamine oxidase A  were used in this study because the latest research about the cognitive effect of this extract was on male mice [P. minus 50 mg,100 mg, and 200 mg with CUMS). Treatment was continued for eight weeks and chronic ultra-mild stress was applied for six weeks. The chronic ultra-mild stress protocol included some interventions like restraining stress, food, and water deprivation, wet bedding, cage tilting, tail pinching, foreign object incorporation, forced swimming, crowded housing, and continuous lighting. Male Balb/c mice , serotonin, norepinephrine, monoamine oxidase A, serum corticosterone, and brain-derived neurotrophic factor (BDNF) Elisa kits were purchased from . 70% Ethanol, Sucrose, and Phosphate buffer .BioKesumAdult male balb/c mice (22–26 g) were purchased from the Animal Research and Service Centre, Kelantan Campus, Universiti Sains Malaysia. All experimental procedures were carried out after getting the approval from USM Institutional Animal Care and Use Committee (USM IACUC) of Universiti Sains Malaysia (1075) & USM/IACUC/2021/ (127) (1129).The animals were housed under standard environmental conditions (24 ± 1 °C) in the Animal Transit Room at the School of Pharmaceutical Sciences, Universiti Sains, Malaysia. They were maintained on a 12-h light/dark cycle. The animals were randomly separated into six experimental groups. A chronic ultra-mild stress method was adopted to induce stress in the animals. The normal control group was without any chronic ultra- mild stress (CUMS), and the remaining groups were influenced by chronic ultra-mild stress (CUMS). Chronic ultra-mild stress remained for six weeks. Treatment remained for eight weeks means until the end of behavioral tests and sampling. n = 14). Group 2:  Chronic ultra- mild stress (CUMS) treated with 6 weeks of CUMS and distilled water for 8 weeks (n = 14). Group 3: CUMS + Amitriptyline 20 mg, a Positive Control (n = 14). Group 4: CUMS + P. minus treated group with 50 mg/kg (n = 14). Group 5: CUMS + P. minus treated group with 100 mg/kg (n = 14). Group 6: CUMS + P. minus treated group with 200 mg/kg (n = 14). Treatment was given through the oral route by using Oral gavage for mice. A complete design of the study is expressed in Group 1: Control group (Without CUMS) treated with distilled water only  and tap water for a 24 h period before 2 days from the experimental trial. Mice were food and water-deprived for 14 h before the start of the experiment. During the test, sucrose preference was evaluated for 6 h by using two bottles of 1% sucrose and tap water and the position of the bottles was switched after 3 h. The sucrose preference was calculated by the ratio of the consumed sucrose solution to the total amount of liquid consumed .n = 14/group) [The open-field test is a classic and well-validated test for detecting the anxiety and locomotion activity of animals. Many researchers explained that an anxious object shows less movement in the central zone as compared to normal, and they spend more time in the periphery and not in the central zone. Thigmotaxis theory and rearing can also provide an idea as an exploratory measure for anxiety ,25. An a4/group) .® (0.8.15) a video tracking software was used to track the path length.The Barnes Maze procedure was first developed by Carol Barnes. It is a behavioral test based on dry-land. It was developed to study spatial memory in rodents . The BarDuring this period, the subject was placed in the center area on the maze surface. A chamber was placed on mice for 10 s after it was removed, and the subject was allowed to explore the maze for 30 s, then was gently guided towards the escape hole. If the subject did not enter the escape hole, it was placed inside the black box. The hole was then covered and the mice remained there for 3 min. After this, the subject was returned to its original cage and the surface of the maze was cleaned with 70% ethanol.In this period, the subject could explore the maze for 8 min and then be gently guided to the escape hole. If the subject did not enter the escape hole, it was placed inside. The hole was then covered and the subject remained there for 1 min. The subject was returned to its home cage and the platform was cleaned with 70% ethanol. All movements of mice were recorded by the camera every day.In the probe trial, the subject was placed in the center zone of the maze surrounded by a chamber for 10 s. The chamber was removed and the subject was fully allowed to explore the area for sharp 90 s after the subject was returned and the surface was cleaned with 70% alcohol .Mice were introduced into a glass cylinder  filled with water up to a height of 40 cm from the base and forced to swim for 6-min. The swim session was recorded by a video recorder. After each swimming session, the mice were removed from the cylinder, dried with towels, and placed in a temporary resting cage for 20 min, and then mice were returned to their home cage. Water in the cylinder was renewed after every 6 subjects. The time of swimming and immobility was recorded using a stopwatch. The duration of each behavior was calculated .The blood samples were collected by cardiac puncture and were kept at room temperature for 30 min, then centrifuged at 3000 rpm for 20 min for serum collection, and stored at −80 °C for the corticosterone assays .w/v with phosphate buffer saline (PBS- Sigma 7.2–7.6). The supernatant was collected for further assays after centrifuging it at 12,000 rpm [The mice were sacrificed, and the brains were quickly removed on ice. The prefrontal cortex and hippocampus were isolated and stored at −80 °C. Further at the time of assays, the prefrontal cortex and hippocampus were weighed and homogenized by 1:4 ,000 rpm .Serum corticosterone level was measured by using an enzyme-linked immunosorbent assay (ELISA) kit  by following the manufacturer’s protocol at 450 nm absorbance in a microplate reader.The brain-derived neurotrophic factor BDNF level was measured from both hippocampus and prefrontal cortex separately by using an enzyme-linked immunosorbent assay kit (ELISA) kit  by following the manufacturer’s protocol at 450 nm absorbance in a microplate reader.The concentrations of serotonin, norepinephrine, and monoamine oxidase A (MAO-A) were measured from the hippocampus only by Elisa Kits  at 450 nm by using a microplate reader and all protocol was followed absolutely as provided by the manufacturer.p < 0.05 were considered as a level of significance in all tests. The means were calculated with standard error means (SEM).Statistical analysis was performed using SPSS version 26 statistical software . Most data were analyzed by one-way ANOVA followed by post hoc comparisons between all groups by using Tukey HSD, except for the Barnes Maze assay. For the Barnes Maze assay, data of three parameters  were analyzed by repeated measure ANOVA. Differences in mean values with P. Minus Aqueous Extract on Sucrose Preference Test.Effects of p < 0.05) decrease in sucrose preference in comparison to the control group, which was improved in all treatment groups significantly compared to CUMS group. CUMS+ Amitriptyline 20 mg showed larger, and CUMS + P. minus 50 mg showed minimum sucrose preference among all treated groups.As depicted in p < 0.05 level of significance as compared to the control, which was only reversed by CUMS + Amitriptyline 20 mg group by p < 0.05 significance. The remaining CUMS+ P. minus treated groups showed a non-significant increase in travelled distance as compared to CUMS group. For the percentage of time spent and covered distance in the central zone, results have been shown in p < 0.05) small residence time and shorter distance in the central zone in comparison to the control group. Only CUMS + Amitriptyline 20 mg treatment improved timings and travelled distance in the central zone significantly (p < 0.05), whereas the CUMS + P. minus 50 mg,100 mg and 200 mg groups showed non-significance improvement in both timings and distance as compared to CUMS group. Results for rearing behavior are presented in Open field test results have been expressed in four parts: total distance travelled by mice A, percenP. minus 100 mg, CUMS+ P. minus 200 mg) except CUMS + P. minus 50 mg showed a significant less travelled distance as compare to CUMS. On the last day of the acquisition trial (Day 5), only CUMS + Amitriptyline 20 mg and CUMS+ P. minus 200 mg showed a significant decrease in the travelled distance in comparison to CUMS group. Part B of P. minus treatment groups CUMS + P. minus 100 mg and CUMS + P. minus 200 mg showed significant less latency in comparison to CUMS group. The same pattern had been observed in CUMS+ P. minus 200 mg for the last 2 days (Day 4 and Day 5), whereas CUMS + P. minus 100 mg expressed this result only on the last day (Day 5). p < 0.05, whereas Day 5 was also significant. In the case of comparison between the treatment group and CUMS group, there was no significant difference in these groups from Day 1 to Day 3, at Day 4 CUMS + Amitriptyline showed a significantly small number of errors by p < 0.05, and on Day 5 this group also showed significance variance with decreased number of errors. Among P. minus treated groups, CUMS + P. minus 100 mg showed a significantly small number of errors on Day 5 with p < 0.05, and CUMS + P. minus 200 mg group also showed a significance in the decreased number of errors on Day 5. The results of the probe trial in form of percentage time spent by subjects to achieve their task in fixed provided time have been shown in the D part of p < 0.05) time to reach their destination in comparison of Control group. CUMS + Amitriptyline 20 mg, CUMS + P. minus 100 mg, and CUMS + P. minus 200 mg groups have been showing a significantly shorter time to escape from a hole than the CUMS group.The results of the Barnes maze are expressed in P. minus 50 mg group were significantly slower than other groups, whereas CUMS + Amitriptyline, CUMS + P. minus 200 mg and CUMS + P. minus 100 mg showed significantly better power of learning and efficient memory characteristics than CUMS group.As a whole, the results of Barnes maze analysis showed that all groups learnt about the task day by day, but CUMS group and CUMS + P. minus 200 mg reduced immobility by p < 0.05 level of significance, whereas CUMS + P. minus 100 mg and 50 mg treated groups also reduced immobility significantly. The results of swimming behavior during the forced swimming test are presented in (B) part of P. minus 200 mg and P. minus 100 mg. The significance level of improved swimming behavior in CUMS + P. minus 50 mg treated group was p < 0.05. The results demonstrate the anti-depressant effect of Amitriptyline and P. minus treated groups with CUMS induced depression model of mice.As sketched in P. minus treated groups, whereas CUMS + P. minus 50 mg treated group showed a minimum decrease as compared to other groups. The reduction pattern in CUMS+ P. minus 100 mg and CUMS + P. minus 200 mg was quite similar.As described in P. minus 200 mg and100 mg treated groups significantly, but CUMS + P. minus 50 mg treated group did not show any significant variations in hippocampal BDNF level. In P. minus treated groupsP. minus 200 mg than in other treatment groups.MAO-A level was measured from the hippocampus of the brain. P. minus 100 mg and P. minus, 200 mg groups showed a significant increase in serotonin (5-HT) level, whereas CUMS + P. minus 50 mg did not show a significant difference in serotonin level in comparison to CUMS group. These results indicate that administration with higher doses of P. minus can enhance the level of serotonin in the hippocampus.In this section serotonin results measured from the hippocampus are discussed, as per P. minus 100 and 200 mg treated groups showed a significant increase in norepinephrine level in comparison to CUMS group. While CUMS group decreases the norepinephrine level significantly as compared to the control group. In addition, CUMS + P. minus 50 mg has not shown any significant difference in comparison to CUMS groupNorepinephrine level was also observed in the hippocampus part of the brain, According to ® extract is registered under the National Pharmaceutical Control Bureau (NPCB) (Reg# MAL14015033T). BioKesum® contains bioactive compounds including quercetin-3-glucuronide (not less than 0.45%), and quercitrin (not less than 0.15%), These are the most abundant compounds in the total phenolic extract of P. minus aqueous extract of leaf part [BioKesumeaf part . P. minus aqueous extract (BioKesum®) has a significant role in improvement on CUMS induced cognition deficit and depression with maximum doses . A previously done work on the effects of P. minus extract on scopolamine-induced cognition deficit supports the results (P. minus aqueous extract is due to the presence of quercitrin and quercetin 3 glucuronide (Q3G) like flavonoids. Q3G is an abundant compound in this extract, which is a derivative of quercetin [P. minus extract can enhance mood and BDNF levels in a human trial [P. minus extract administration significantly enhanced the BDNF level in CUMS induced depressive mice. Our primary finding of this research work showed that administration with  results  of this uercetin ,31. Recean trial , throughP. minus aqueous extract. Despite all these findings and studies, there was a gap in research regarding its antidepressant effects because no proper mechanisms were found in all previously done work regarding its beneficial activities. So, the current study demonstrates some potential regarding anti-depressant and cognitive enhancing properties of P. minus has the potential to reverse the CUMS induced behavioral disturbances significantly by using the method of sucrose preference test, Barnes maze assay and forced swimming test with higher doses of P. minus 100 mg and P. minus 200 mg mostly. Our findings regarding antidepressant effect are strongly supported by previously done work on individual flavonoids which are present in P. minus extract, Quercitrin (quercetin 3-rhamnoside) alleviated depression-like behaviors with a maximum dose (10 mg/kg) among depressive mice in a previous study [Hypericum perforatum (St. John’s wort) through forced swimming test in mice, it can reduce the immobility time significantly which is similar to our result  only  induced depressive mice group  in P. minus aqueous extract and other studies showed that 0.6 mg/kg dose of this constituent had significant antidepressant potential which was near to concentration used in current research even P. minus 200 mg/kg (maximum dose group) had more concentration of Q3G as compare to that extract which was used in previous studies [P. minus aqueous extract) was not sufficient for significant anti-depressant potential. This research showed that us study . Whereasr result  of immobficantly , The sammg) only . These ron model . If we dce group . It is r studies , whereas studies ,36. BaseP. minus aqueous extract (BioKesum), it is strongly recommended to develop its planned and prescribed use for improving the memory and depression signs based on its beneficial mechanisms of action, observed in current research work.In the case of P. minus aqueous extract has an anti-depressant-like effect on behavior, it shows an improved spatial memory, reduced immobility, decreased corticosterone level, elevated serotonin, norepinephrine and BDNF level along with a significant reduction of MAO-A level compared to CUMS induced depressive mice model.It is concluded that


Background: People living with mental health disorders are at increased risk for developing obesity due to poor diet, physical inactivity, and antipsychotic medications. In the United States, the obesity rate is 36% in the general population and more than 50% for people living with mental health disorders. Although mental health clinicians concentrate on managing psychiatric disorders, they seldom recognize the gradual increase in body mass index of their patients. The result is a disconnection between the clinical management of psychiatric disorders and the medical management of obesity. Purpose: This study assessed the effectiveness of an evidence-based education program for improving the obesity management practices of mental health clinicians caring for residents at a state psychiatric hospital. Methods: This was a quasi-experimental study design with a pretest and posttest evaluation. Convenience sampling was used to recruit mental health professionals, or clinicians, at a large psychiatric hospital in the Southern region of the United States. Data was collected with the Advising and Treating Overweight and Obese Patient questionnaire (17 items). Data analysis included descriptive and inferential statistics. The findings were reported in accordance with the TREND and GREET guidelines. Results: The education program was completed by 50 MHCs. The pretest indicated that 76% of MHCs were not involved in helping obese residents manage their weight, but the posttest indicated 90% were involved. There was a significant increase in MHC knowledge about obesity management and reported actions 90-days after the program. MHCs were unable to arrange follow-up visits for residents, a task not directly within their control. Conclusions: Mental health clinicians reported increased knowledge and improved clinical practice after an education program. Because the outcomes were reported at 90-days after the program, further research needs to evaluate the longitudinal impact of this type of program, where the reported behaviors are correlated to process and clinical outcome measures for obesity. As the second leading cause of preventable death in the United States , obesityApproximately 1 in 25 people is diagnosed with a serious mental disorder , includiThe weight gain of residents in psychiatric hospitals has been discussed for decades  as a conThe risks associated with obesity at psychiatric hospitals may result from five causes addressed in other settings. First, residents enter the hospital with higher rates of chronic conditions than the general population . Second,The Promoting Action on Research Implementation in Health Care Services , or PARiThe current evidence and best practices about obesity management for residents of psychiatric hospitals suggest the topic has not been widely explored , as the This pilot study used a quasi-experimental design  with an The study was conducted at the beginning of 2017 at a large state psychiatric hospital, with approximately 200 beds, located in Southern region of the United States. The hospital is responsible for people living with mental disorders who cannot be cared for in an ambulatory setting because of the severity of their condition. Most residents had been hospitalized for at least six months, and the average length of stay was nine months. The resident population was almost equally men and women, aged between 18 and 65 years, and 55% White, 40% African American, and 5% Hispanic.A convenience sampling strategy was implemented using email invitations to reach a sample goal of 50 clinicians. The inclusion criteria were licensed mental health clinicians who had prescriptive authority and were credentialed to provide primary care and/or mental health services at the hospital. Clinician profile data was not collected because of restrictions for protecting participant confidentiality.All participants completed a pretest questionnaire, the one-hour educational program, and a posttest questionnaire in a conference room at the psychiatric hospital. The pretest questionnaire was completed before the educational program. Then, all the participants completed the educational program during the same session. Finally, the posttest questionnaire was completed 90-days after the educational program. Both pretest and posttest questionnaires were returned in sealed unmarked envelopes to a secure box. The participants were instructed to return a blank questionnaire in the envelope at any point during the study if they decided to stop participating. All participants gave written informed consent. No financial incentives were provided for participation.The continuing education program was developed from a scoping review  to identr = 0.81), including for efficacy expectation (r = 1.00), outcome expectation (r = 0.88), and barriers (r = 0.95). Two additional items were added after the ATOOP questionnaire to assess previous obesity management education and training. The complete 19-item instrument was validated, face and content [The Advising and Treating Overweight and Obese Patient (ATOOP) questionnaire , a valid content ,70, by a content ,72, an at and McNemar tests [The data were entered into an Excel spreadsheet to organize and clean. Analyses were performed in Statistical Analysis System  software . Descripar tests  were comar tests ,76. An αThe project was approved by the psychiatric hospital and the institutional review board at the participating university. Before data collection, all participants were provided with written information about the study. As required by the hospital, no personally identifiable information, including demographic data, was collected during the study. Individual data were aggregated for group reporting. The de-identified data was securely stored on a password-protected computer hard drive with restricted access. There were no breaches in confidentiality and no reported adverse events.A total of 50 mental health clinicians responded to the questionnaire before (pretest) and 90-days (posttest) after the educational program. This accounted for 100% of the total responses. There were no incomplete or blank questionnaire returned and no deviations from the study protocol. Although the pretest questionnaire indicated that most clinicians were not involved in helping overweight and obese residents manage their weight, the posttest questionnaire showed that most clinicians became involved in obesity management after the educational program, which was a statistically significant finding. Overall, 15 of the 16 items were significantly improved after the educational program see . AdditioThe first questionnaire item provided an overall assessment of the clinician’s perspective about assisting overweight and obese patients with losing weight. Before the educational program, 76% of participants most frequently indicated they had not seriously thought about assisting their overweight and obese patients with losing weight. After the educational program, 42% of the participants indicated they had been thinking about assisting their overweight an obese patient with losing weight within the next six months. Importantly, 48% of the participants had developed formal plans or actively began assisting their patients with losing weight. After the educational program, the participants were more confident in their ability to ask patients if they were concerned about their weight, advise patients about weight management, assess patient willingness to lose weight, assist patients with losing weight, and arrange for follow-up visits for patients; these findings were statistically significant. Although there was a statistically significant improvement in participant confidence in arranging follow-up visits for patients, half the participants were only slightly confident, and very few were confident (14%) or highly confident (2%). Similarly, after the educational program, the participants were statistically more likely to ask their patients if they were concerned about their weight, advise patients about weight management, assess patient willingness to lose weight, and assist patients with losing weight. In terms of the ability of participants to arrange for follow-up visits for patients, most were unlikely (64%) or not sure (30%) after the educational program. When considering the obesity management of different patients, participants were less likely to assist obese patients with comorbid conditions, such as diabetes, before the educational program. Half the participants did not assist obese patients with losing weight before the educational program, whereas almost all participants managed patients with comorbidities as often (30%) or more often (64%) than patients without comorbidities after the educational program. Twice as many participants responded more often (34% versus 64%) after the educational program. These findings were statistically significant. Participants were less familiar with patient compliance problems associated with weight gain related to psychotropic medications before the educational program. Before the program, almost all participants considered weight gain to be a minor (20%) to moderate (64%) barrier to medication compliance. After the educational program, all participants considered weight gain to be a moderate (48%) to major (52%) barrier to medication compliance; these findings were statistically significant. The recognition of this major barrier nearly tripled from 16% to 52% after the educational program. Before the educational program, all participants reported that they had no formal education about weight management for overweight and obese patients with mental disorders (item 18). After the educational program, all participants indicated they had received formal education. In terms of the adequacy of the educational program based on the learning objectives, all participants indicated that the program was adequate or very adequate for assisting them with helping their patients with weight management (item 19).The purpose of this study was to increase clinicians’ awareness, change perceptions, and improve clinical practice by including weight management as part of the daily clinical care for psychiatric hospital residents. The association between mental disorders and obesity in the context of a psychiatric hospital is complicated by intensive pharmacological management, sedentary lifestyle, and poor eating habits, including the limited availability of healthy foods . These on = 500) in the United States reported that most nutrition professionals (78%) received high-quality weight management training during their degree program, followed by nursing (53%), exercise (50%), pharmacy (47%), and mental health (32%) professionals [n = 281) reported additional training in obesity management, including pharmacy (77%), nutrition (76%), nursing (59%), mental health (55%), and exercise (50%) professionals who earned continuing education credits.An important finding from this study was that all participants reported no previous education or training for the clinical management of obesity in the context of mental health. In this regard, Brown and Flint  noted thssionals . More thBefore the educational program, the clinicians did not recognize obesity as an important part of daily practice, they did not notice resident weight gain, and they lacked the prerequisite knowledge to use basic medical weight reduction strategies to manage obese residents. Similarly, mental health clinicians surveyed in different countries reported limited awareness about patient obesity and reported insufficient knowledge to manage the physical health of people with severe mental illness ,81,82. MAlthough evidence-based programs with multiple strategies  can addrThe participants also encountered barriers when ordering external nutritional services and scheduling follow-up visits for specialized medical management. In this regard, psychiatric hospitals can assist clinicians by implementing clinical practice guidelines using a multidisciplinary approach to medical management. This approach would limit the environmental factors in psychiatric services  that canAfter retrospective assessment of the measurement instrument and applying the new “person first language” guidelines ,72, somen = 50) was small, but it was the target based on an a priori power analysis. Although response-shift bias is possible for the study design [This project has five limitations and three major strengths. First, the sample size (y design , observey design . Third, y design  can causy design . Howevery design ,97. Foury design . Finallyy design .Despite these limitations, this study addresses an important gap in the literature specific to obesity management at psychiatric hospitals. Furthermore, the educational program was developed with expert review from the evidence-based literature, best practices, and national guidelines to help clinicians identify and effectively manage obesity. The evaluation was completed using a validated instrument. Moreover, the evaluation was conducted 90 days after the educational program to reduce recall bias and to measure planned and actual changes to clinical practice. Because the education program was tailored for a psychiatric hospital, the content may be transferable to similar institutions in other areas of the United States.There is a considerable gap in the obesity literature about the medical management of obesity by clinicians in the context of psychiatric hospitals. Although evidence-based practices are reported in the literature to address gaps in obesity management in other environments, none were identified for public psychiatric hospitals. The results suggested that the educational program improved the current level of knowledge, skills, and attitudes of mental health clinicians employed at a public psychiatric hospital regarding the clinical obesity management of people living with mental health disorders. Additional research is necessary to correlate the reported behavior changes with process measures and clinical outcomes over time. This pilot study demonstrates that an inexpensive but evidence-based educational program can shift obesity management knowledge and the associated behaviors of clinicians practicing at a public psychiatric hospital.


Earthquakes are a natural hazard affecting millions of people globally every year. Researchers are working on understanding the mechanisms of earthquakes and how we can predict them from various angles, such as experimental work, theoretical modeling, and machine learning. We invited Marie Violay (EPFL Lausanne), Annemarie Baltay (USGS), Bertrand Rouet-Leduc (Kyoto University) and David Kammer (ETH Zürich) to discuss how such a multi-disciplinary approach can advance our understanding of Earthquakes. Can you give a brief overview of what your scientific work looks like and from what angle you approach Earthquakes?Bertrand: My research on earthquakes is focused on the topics of earthquake nucleation and the interaction between slip modes - the way tectonic stress is released. A variety of slip modes exist, with dynamic earthquakes and creep at both ends of a spectrum that encompasses slow slip events of varied duration and scale. Many questions remain on the interplay between the members of this spectrum, including what may determine how and why a slow slip event may degenerate into an earthquake.My research approaches these questions of earthquake nucleation and the interplay between slip modes from two angles: at multiple scales and using data science. I develop machine learning-based methods to detect seismic and geodetic signals from the scale of laboratory experiments, to the scale of subduction zones.Marie: My research aims to understand the physics of fluid-induced earthquakes. Anthropogenic fluid injections during hydraulic fracturing, reservoir impoundment, the injection of waste water or CO2 storage can induce small stress perturbations in the underground and lead to fault reactivation and enhanced seismic activity. Moreover, long-lasting regular natural earthquake sequences are often associated with elevated pore fluid pressures at seismogenic depths. The mechanisms that govern the nucleation, propagation and recurrence of fluid-induced earthquakes are poorly constrained, and our ability to assess the seismic hazard that is associated with natural and induced events remains limited. At EPFL, we aim to improve our knowledge of fluid-induced earthquake mechanisms through multi-scale experimental approaches.We apply cm-scale friction experiments to study the effect of fluid pressure on earthquake nucleation and propagation under crustal deformation conditions during the entire earthquake cycle. dm-scale dynamic rupture experiments are in turn applied on experimental faults to investigate the influence of fluid pressure on the nucleation and propagation of ruptures. Our analysis of post-mortem experimental faults is carried out with state-of-the-art microstructural techniques. We finally aim to calibrate the theoretical friction law with friction experiments and faulted rock microstructural observations.David: In my research, we aim to establish a fundamental understanding of tectonic fault ruptures as they occur during natural earthquakes. We develop theoretical and numerical models that describe the full cycle of an earthquake, including nucleation, propagation and arrest of the fault rupture, and help us to understand the mechanisms that govern earthquakes.We pursue our objectives along multiple research axes. First, we develop numerical methods that allow us to include more complexity into earthquake fault rupture models in order to build more realistic earthquake scenarios. Second, we calibrate our models with observations from friction experiments, as described by Marie, and use them to support the analysis of observations from large-scale laboratory earthquake experiments by giving access to quantities that are not easily measured in the experiments. Finally, we use our simulation results to develop fracture-mechanics-based theoretical models of laboratory earthquakes, which we then apply to upscale the knowledge gained from large-scale experiments to the field scale and natural earthquakes.Annemarie: I am an observational earthquake scientist at the U.S. Geological Survey, using seismograms recorded at various distances from the earthquakes to probe what we know about both the earthquake source as well as how seismic waves propagate through Earth. I am interested in how both earthquakes and Earth control ground motions which are measured at distance, and how these reveal the earthquake source and path. I am particularly interested in earthquake stress drop, which is the amount of tectonic stress released during an earthquake rupture, and which can be estimated from the radiated seismic waves.I further work on ground-motion models (GMMs) and their physical components and uncertainty. Reducing the latter, will ultimately lead to more precise and accurate seismic hazard maps. Currently, I am working towards physical explanations for variability in the source, site, and path components in ground motions. Ultimately, we will develop models for predicting those effects from geophysical observables, such as stress drop (for source), site velocity profiles and attenuation (for site), and whole-path attenuation (for path).What are the most impactful recent advances in your communities and how do they add to the bigger picture in Earthquake science?Bertrand: Recent physical models of the earthquake cycle and laboratory studies suggest earthquakes may nucleate during a preparatory aseismic phase of variable duration from minutes to years4. An aseismic phase is characterized through surface displacement, but the absence of notable earthquakes.6 to week-long slow slip events8. The systematic observation of deformation events on faults is getting closer and may soon give definite answers on the interaction between slip modes and on earthquake nucleation.Thanks to increasing deployments of seismic and GPS stations, as well as the development of Interferometric Synthetic Aperture Radar (InSAR), the observation of such aseismic deformation is becoming common, from continuous aseismic slipMarie: Aseismic slip plays an important role for us as well - recent laboratory and natural observations suggest it to be one of the triggering mechanisms of fluid-induced earthquakes. Whereas other trigger mechanisms do exist as well, aseismic slip has an important role insofar that it can induce seismicity in regions beyond the fluid pressurized zone and hence potentially increase the seismic hazard area. Thus, it is critical for us to not only understand the mechanisms that cause fault slip, but also the conditions that lead to (a)seismic slip.David: Our community is continuously pushing the theoretical and numerical approaches to create more realistic models for the full earthquake cycle. One important contribution in the large sense is the community code verification exercises9, in which various numerical codes are compared and benchmarked. This is a very important contribution to continue supporting rigor and reproducibility in our field, and I believe this will have long-lasting impact.11 is the development of theoretical models for elongated earthquake ruptures. Others include theoretical models for the propagation speed of frictional ruptures13, fluid-driven fault rupture15, and earthquake scaling17.Other recent advances that I am personally very excited about are efforts to use numerical simulations to make theoretical models, which are often very simple, a degree more realistic, but in a fundamental way. A very nice example22.Finally, there are exciting efforts to enhance numerical simulations with more complexity, such as realistic fault geometry, multi-physical fault phenomena, and fault heterogeneityAnnemarie: In earthquake seismology, we are starting to explore new ways to utilize the vast amounts of available data more efficiently. Novel machine learning (ML) techniques help us to improve our earthquake catalogs, in particular to understand seismic sequences for smaller and much more frequent events. ML is further applied to mine the ambient seismic wavefield to discover tectonic tremor which helps to track plate motions or map the Earth’s interior. This includes more effectively regressing instrumental records of moderate and large earthquakes which are spatially variable, to develop so-called non-ergodic ground-motion models, with increasing sophistication and customization; and even interpreting felt earthquake reports from citizen responders to get a better idea of how people experience shaking, a topic that we are currently working on now.What are the most pressing research questions your respective communities are working on at the moment?Bertrand: Systematically observing deformation events on faults may well be key to understanding the interaction between modes of slip and earthquake nucleation, and might provide observables that may allow discriminating between a harmless slow slip event and an aseismic precursor to a major earthquake.23 to earthquakes24, is among the most pressing research topics in the quest towards a better understanding of the spectrum of slip modes, the interaction between slip modes, and earthquake nucleation.However, current geodetic methods cannot always resolve small  day- to week-long events of slip, and doing so involves manual processing and analysis that cannot scale to the systematic and global observation of deformation events. Progress towards automatic detection of tectonic events, with recent successes from automatic detection of aseismic slipMarie: One major research task is to determine what controls the onset of dynamic instability, i.e. the competition between frictional aseimic preslip and fluid diffusion fronts. We further try to get a better handle on both what’s controlling the maximum magnitude of fluid-induced events, but also whether the maximum magnitude scales with a number of parameters . A final question is whether heterogeneity enters into the scaling.David: Physically speaking, there are many questions related to the earthquake cycle and the processes governing it. For instance, what is the exact nucleation process of an earthquake or how do natural fault ruptures arrest? Many of these questions are directly related to a need for a better understanding of fault friction properties  and multi-physical phenomena  under natural conditions, and for more information about fault heterogeneity and its effect on earthquake mechanics.From a theoretical perspective, there is an important question on reconciling observations from small-scale rock experiments, with large-scale laboratory earthquake experiments, and field observations. Can we build models that consolidate our knowledge from the lab with observations from the field?Are there specific research questions you would like to see addressed by another community?Bertrand: As progress towards automation of tectonic deformation is becoming a pressing issue to keep progressing towards a better understanding of earthquakes, the involvement of the data science and machine learning (ML) communities could make all the difference. Similar to how developments of ML for the life sciences have become ubiquitous, developments of ML specifically for the earth sciences will hopefully become another important area of applied ML research.Marie: As an experimentalist we always try to make our measurements as precise and fast as possible, as close to the fault, and on as many points as possible. Digital image correlation allows fast and precise measurements of displacement for experiments performed without confining pressure. The development of distributed fiber optic measurement has just started to produce excellent results in pressure and temperature, and we intend to deepen our collaboration with this community.David: As modelers we are always relying on experimental data for calibration and validation of our models . For this reason, more precise experimental observations of the local constitutive friction law at realistic conditions (e.g. high rupture speed and high contact pressure) would be very helpful. This is, of course, technically very challenging, but I would like to push for more direct collaboration between experimental and theoretical researchers, as this could lead to important progress in our fundamental understanding of earthquake mechanics.Annemarie: As an observational earthquake seismologist, I think we need to strengthen our link in two directions -- earthquake simulations, both dynamic and kinematic, and laboratory experiments. In both of those cases, inputs such as stress, slip, dimension or material properties can be set and controlled, parameters which we have difficulty resolving in detail or with reliability observationally. We need to continue to validate the simulations, to ensure that they are capturing the correct physics and earth properties, and on the lab side, push the scale of experiments to bridge the link to in-situ earthquakes. Of course, the collaboration between all the disciplines is essential to ensure results and interpretations are brought together.How would you like to see the link between earthquake policy and hazard mitigation strategies strengthened in regards to your research area?Bertrand: In the not so distant future, tectonic deformation may be continuously monitored using data science and ML models on both seismic and geodetic data, notably yielding improved mappings of fault locking and slip budget, with the potential to inform and improve models of seismic hazard.Marie: The reliability of natural hazard estimates needs to rely heavily on the definition of a faulting model, which needs to be underpinned by realistic physical constraints such as fault geometry, friction and rupture laws.David: I agree that data-driven and ML approaches have the potential to support the process of determining the seismic hazard. As nicely pointed-out by Marie, the models should be constrained by physical considerations. In addition to those already mentioned, I would also include constraints based on fault rupture processes, such as energy balance, rupture mode, and propagation/arrest conditions.Annemarie: As we continually refine and update our models of seismicity rates and occurrence, we have more detailed, specific, accurate models for seismic shaking, which also results in models that are more precise and less variable. Spatial and temporal dependence on finer scales could be incorporated into hazard and forecast products; in the case of USGS products such as Operational Aftershock Forecasting, we could give communities a more accurate and precise picture of what to expect after a large earthquake, which could quell anxiety and bring better preparedness.This interview was conducted by Sebastian Müller.


Drosophila melanogaster, also known as the fruit fly, is a widely used organism model, especially for genetic studies or as a model for pathologies. The Drosophila genome is well known and conserved within humans, thus allowing biologists to obtain numerous mutants and transgenic flies from the fruit flies. Gene function studies at the cellular and molecular levels are often performed using extracts of larval tissues. Due to their small size, it is difficult to dissect substantial amounts of these tissues for performing genomic or proteomic experiments. This paper develops a simple method to purify larval tissues en masse. This protocol preserves tissue integrity in the same way as manual dissection; the protocol is achievable by individual researchers and allows the purification of different samples simultaneously.Drosophila melanogaster imaginal discs are larval internal structures that become the external organs of the adult. They have been used to study numerous developmental processes for more than fifty years. Dissecting these imaginal discs for collection is challenging, as the size of third-instar larvae organs is typically less than 1 mm. Certain experimental applications of the organs require many cells, which requires researchers to spend several hours dissecting them. This paper proposes an alternative to dissection in the form of a mass enrichment protocol. The protocol enables the recovery of many wing imaginal discs by grinding large quantities of third-instar larvae and separating the organs using filtration and a density gradient. The wing imaginal discs collected with this protocol in less than three hours are as well preserved as those collected by dissection. The dissociation and filtration of the extract allow the isolation of a large amount of wing imaginal disc cells. Drosophila melanogaster for developmental studies. The structures are accessible, and most genetic tools engineered for this species are designed to be used in these organs, including drivers to use the UAS-GAL4 system [Drosophila melanogaster [Drosophila wing imaginal disc has proven to be a great model for genetic or cell biology approaches, its use is more difficult in biochemistry or molecular biology experiments that need large amounts of starting material. For instance, a chromatin immunoprecipitation (ChIP) requires 400 wing imaginal discs per sample [In Holometabola, imaginal discs are larval internal epithelial structures that are precursors of external organs in the adult, such as wings or legs. These structures are widely used in the model organism 4 system  or mitot4 system  enablingnogaster , making nogaster  and humanogaster ,7). Suchnogaster  and the nogaster . Howeverr sample , and thi®  to isolate wing imaginal discs according to their shape, size, or Green Fluorescent Protein (GFP) pattern. However, this device is expensive and not commonly available. Therefore, we wanted to develop a mass enrichment protocol routinely usable in any laboratory.Since imaginal discs such as wing ones are precursors of non-essential organs, they are instrumental in studying cell death as it is possible to induce cell death during the development in these structures without affecting the survival of the specimen . Our teaWe propose here an imaginal disc mass enrichment protocol without manual dissection that allows the recovery of a large number of discs in an optimized time range, doable entirely by one experimenter, using density gradients and sedimentation; see the overall protocol in vg-GAL4 strain is a generous gift from Joel Silber . The UAS-mCD8-GFP was obtained from the Bloomington Drosophila Stock Center (BL-32185).Flies were raised at 25 °C on a standard medium. The w/v) on top of a 15:20:25% gradient. After centrifugation, the 15:20% interface containing the enriched wing imaginal discs was collected and rehydrated in Ringer 1X  and filtered through a series of sieves. The resulting material was loaded in 10% Ficoll solution , pH 7.6. Discs obtained by mass enrichment or dissection were fixed with 3.7% formaldehyde in 1X PBS for 20 min at room temperature and washed three times for 10 min in PBST . Drosophila Dcp-1  at 4 °C. The following day, after 3 washes in PBST, wing discs were incubated for two hours with Alexa-568-coupled anti-rabbit secondary antibody  diluted to 1:400 in PBST. Finally, wing discs were mounted in ProLong Diamond , and images were acquired using a Leica SP8 confocal microscope  at 568 nm. At least 30 wing imaginal discs are analyzed for each condition. Image analysis was performed on Fiji with the macro described in de Noiron_2021 [Apoptosis was analyzed as previously described in de Noiron et al. . In brieron_2021 , whose mTM  and observed with a Leica SP8 upright confocal microscope.For tissue structure analysis, wing imaginal discs were incubated with phalloidin-ATTO 665  1 μg/mL, and Hoechst 33,342  1 μg/mL, in PBST-BSA for 1 h at room temperature. Larvae were then washed twice for 5 min in PBST. Finally, wing discs were mounted in CitifluorUAS-mCD8-GFP under the control of the vestigial-GAL4 driver were used in [w/v) of 5% (w/v)% protease (Sigma-Aldrich P8811) in 1X Ringer incubated for 20 min at 25 °C under gentle shaking (300 rpm). Dissociation was completed by gently passing the cells through a P1000 tip. After pelleting, cells were resuspended in 1X Ringer and filtered through a 40 µm sieve to retain salivary glands cells and eventual debris. Cells were then analyzed using the BD FACSAria™ III  equipped with a 488 nm laser line and the BD FACSDiva™ software . Cells were selected through forward scatter (FSC) to keep out debris and cell clusters. They were then sorted based on whether or not they expressed GFP.Third-instar larvae expressing  used in . Wing imUAS-mCD8-GFP under the control of the vestigial-GAL4 driver were used for each group. Wing imaginal discs recovered by either dissection in 1X PBS, dissection in 1X Ringer, or mass enrichment protocol in 1X Ringer were dissociated using a 0.1% (w/v) solution of protease (Sigma-Aldrich P8811) in a 1X PBS or 1X Ringer and incubated for 20 min at 25 °C under gentle shaking (300 rpm). Dissociation was completed by gently passing the cells through a P1000 tip. After pelleting, the cells were incubated for 3 min on ice in viability dye solution  diluted to 1:1000 in 1X PBS or 1X Ringer. After a wash in 1X PBS or 1X Ringer, cells were resuspended in 1X PBS or 1X Ringer and filtered through a 40 µm sieve. The fluorescence of the dye was then measured using the BD FACSARia™ III equipped with a 635 nm laser line and the BD FACSDiva™ software. Cells were selected through FSC, excluding debris and cell clusters. Data analysis was performed with FlowJo software . Graphs and statistical analysis were performed using RStudio  with a significance threshold of 5%.For the cell death assessment by viability dye, 10 third-instar larvae expressing Few protocols of mass enrichment for imaginal discs have previously been described. They are all based on the protocol developed by Fristrom and Mitchell , but the®  from Bertin. The second was the GentleMACS™ from Miltenyi, which was successfully used for such applications by the Basler lab [Drosophila larvae is not documented. The grinding process was visually monitored by assessing the integrity and location (released from the larvae or not) of fluorescent discs at the end of the process.The first step of this protocol consists of dismantling the larvae without damaging the imaginal discs. Protocols from Mitchell and Cohen laboratories used either a meat grinder that is in all likelihood not sold anymore  or a cussler lab . We expeThe development tests were mainly carried out using 2.5 mL of larvae, representing around 600 individuals. This larvae amount is collected by flushing the side of six vials. Adult flies were transferred to a fresh medium every 24 h to ensure that all offspring were the same age and obtained in sufficient quantity. We used 12 vials containing 15 virgin females and 6 males to collect the same amount (2.5 mL) of larvae from a cross where the larvae of interest represented only 50% of the progeny. These values must be adapted according to the viability of the individuals of interest.®, we used 2 mL tubes containing either 2.8 mm diameter beads or a mix of 1.4 and 2.8 mm beads with different grinding cycles. We found that 2.8 mm beads alone were more efficient in breaking cuticles and that the recovered imaginal discs were in better condition than using the bead mix. However, the yield was low since most discs remained attached to the larvae (data not shown). Furthermore, considering the quick mass enrichment protocol we wanted to set up, we did not further pursue the set-up with this device as it can only handle a limited number of larvae (100–120/tube) at once, and the recovery of the material after grinding was challenging to handle—these drawbacks are incompatible with mass enrichment.For the assays with the PrecellysThe GentleMACS™ uses tubes with spinning helix-like structures and comes with several pre-registered programs of different speeds and/or duration. Two types of tubes are available; the C tubes offer a gentler grinding and are dedicated to tissue dissociation into individual cells, whereas M tubes offer stronger grinding and are recommended for complete cell lysis as required for biomolecule extraction. We first used the “liver 1” programs  that were previously used for such purpose  or “liveThe second step of the protocol consists of isolating imaginal discs from the other organs. As per the protocol from Marty , we firsStill, to avoid gradient saturation, we added a sedimentation step. Indeed, this was a means to further improve the purity as gut and fat body pieces are much less dense than wing imaginal discs. To assess the efficiency of the sedimentation and purification steps, we needed a way to visually localize wing imaginal discs in the tubes. To do so, we took advantage of LacZ expression in those discs and added a fixation-less staining step of the beta-galactosidase  Supplem. FinallyOnce the input for the Ficoll gradient separation was cleared from most unwanted larval elements, we worked on the enrichment step. We tried many gradient configurations . We present here only a subset of them representing the milestones of the development of the method . The mosAfter centrifugation, wing imaginal discs, along with eye and antenna discs and salivary gland pieces, were found at the 15:20% interface. Some more wing imaginal discs could be recovered at the 20:25% interface, but we chose not to recover them because the interface also contains the vast majority of salivary glands and mouth hooks. For our applications, cells from imaginal discs were easy to separate from salivary gland cells, which are the primary contaminants in this case.An important difference between the protocols of Marty et al. 2014  and FrisWe used this assay to determine whether one of the buffers, PBS or Ringer, is more damaging than the other. Confirming our observation, PBS is a much more damaging agent as cells obtained by dissection in PBS were more than twice as frequently labeled compared to cells treated in Ringer solution D. In addvg-GAL4 driver. This driver induces GFP expression in a wide band at the dorso-ventral frontier, representing roughly 40% of the cells. Visual inspection of the GFP pattern readily allows the recognition of the vg pattern, confirming that discs do not undergo such stress that would lead to GFP extinction. Since salivary gland cells are bigger than wing imaginal disc cells [We continued the quality control of the material recovered by mass enrichment by analyzing the number of GFP-positive cells using flow cytometry. This analysis was carried out on larvae expressing the GFP in the wing disc under the control of the sc cells , it is eMass enrichment of discs led to a slight decrease in the proportion of GFP-positive cells compared with dissected discs (from 47.7% to 42.7%), but this decrease was expected as ground samples were not 100% pure. This fraction contains other organs (such as other discs) whose cells do not express GFP, thus increasing the GFP-negative fraction. This limited decrease indicates that the wing imaginal disc fraction recovered by mass enrichment is relatively pure. This result confirms our visual assessment and is supported by the fact that after dissociation and filtration, whatever the protocol used, the percentage of cells recovered compared to the total number of events counted by cytometry is not significantly different .vg-GAL4, mCD8-GFP, wing imaginal discs to stress the quality of the wing imaginal discs recovered by the mass enrichment protocol. Hoechst and phalloidin-ATTO 655 stained the nucleus and the plasma membrane, respectively. mCD8GFP protein is known to localize at the plasma membrane. The experiment was performed in parallel on wing discs isolated by dissection or mass enrichment protocol. The results in The tissue structure was also explored using fluorescent staining of Flushing the side of the tubes with water allows the recovery of L3 larvae from synchronized egg-laying. After rinsing in water to remove the growth medium, larvae are transferred in GentleMACS C tubes and ground in Ringer solution. The ground material is filtered through several strainers of decreasing meshes to separate organs by size; this mainly removes cuticles and fat bodies. These steps are repeated five times to ensure the good separation of organs without damaging them. The filtered organ suspension (100–200 µm) then undergoes a sedimentation step to remove gut pieces, fat body, and some other non-discs organs. Finally, a Ficoll density gradient allows the separation of imaginal discs from other organs. Salivary gland pieces constitute the primary contaminant . Wing imDrosophila imaginal discs. By studying protocols from the 1960–1970s, we set up a protocol that allows recovery of third-instar larvae wing imaginal discs without manual dissection. To do so, we had to set up every step, from the grinding of larvae to the density gradient We used around 600 larvae as an input to obtain, on average, 140 wing imaginal discs. Depending on the researcher, this amount can be dissected in about two to three hours. However, if more wing imaginal discs are needed, the dissection time increases proportionally. One of the advantages of the mass enrichment protocol is that it can process several samples simultaneously by using parallel filtration montages and doing the filtration and centrifugations (sedimentation and Ficoll density gradient) at the same time . Therefo(ii)When high amounts of organs are needed, multiple researchers usually dissect them together. As each researcher has their own method and ease of dissection, which may be influenced by the time spent dissecting, this may pose reproducibility problems. The enrichment protocol is standardized and can be performed entirely by one researcher, reducing reproducibility issues, which constitutes another advantage over manual dissection. (iii)Since dissection allows only one sample to be processed at a time, this induces a delay in the treatment of each condition. Our protocol does not present this disadvantage, as several samples can be treated almost simultaneously. Thus, it allows a more homogenous treatment between conditions. As mentioned in If larvae of interest are obtained by crossing and represent only 50% of the progeny, the number of vials must be doubled to obtain an equal amount of wing imaginal discs. Depending on the parents’ genotypes, only a fraction of the offspring may be of interest in certain conditions. Collecting these individuals based on their specific phenotypes  is possible, but this would add a time-consuming step. In this case, using a fluorescent protein expressed in the population of interest presents two advantages. Firstly, it allows the identification of discs of interest at the end of the mass enrichment without selecting the larvae before starting the protocol. Secondly, when a large amount of disc is required to isolate cells of interest, the dissociation and filtration step allows obtaining wing imaginal cells that can be sorted by flow cytometry. Depending on the experimental design, other solutions can be considered, such as tagged proteins of interest and magnetic cell sorting . Drosophila organs by modifying the filtration or the Ficoll gradient steps using the indications detailed in this specific part. Moreover, to facilitate the tracking of organs throughout the gradient, we adapted a beta-galactosidase staining protocol without fixation to color the organs blue and have a visual clue of their localization [Interestingly, this protocol can also be adapted to isolate other lization  (SupplemOlder protocols were able to graft the collected organs, which became normal adult structures . We did As another quality control, we compared the expression of GFP in wing imaginal discs in which the vestigial driver drove GFP expression. Visual examination showed no notable difference in GFP levels between discs (see In summary, this paper shows that this mass enrichment protocol allows the rapid purification of a large quantity of wing imaginal discs of equivalent quality to manually dissected discs for multiple samples simultaneously and by a single investigator. Wing imaginal discs recovered by mass enrichment allow the isolation of wing imaginal disc cells suitable for numerous applications such as cytometry analyses , transcr