Mixed Response Research Articles

Fully Synthetic TF-Based Self-Adjuvanting Vaccine Simultaneously Triggers iNKT Cells and Mincle and Protects Mice against Tumor Development.

The Thomsen-Friedenreich (TF) antigen has proven to be a promising target for developing novel therapeutic cancer vaccines. Here, a new strategy that TF antigen covalently coupled with KRN7000 and vizantin was developed. The resulting three-component vaccine KRN7000-TF-vizantin simultaneously triggers invariant natural killer T (iNKT) cells and macrophage-inducible C-type lectin (Mincle) signaling pathways, eliciting much stronger TF-specific immune responses than glycoprotein vaccine TF-KLH/alum and the corresponding two-component conjugate vaccines TF-KRN7000 and TF-vizantin. The analysis of IgG isotypes and the secretion of cytokines revealed that KRN7000-TF-vizantin induced Th1/Th2 mixed immune responses, where Th1 was dominant. In vivo experiments demonstrated that KRN7000-TF-vizantin increased the survival rate and survival time of tumor-challenged mice, and surviving mice rejected further tumor attacks without any additional treatment. This work demonstrates that covalently coupled KRN7000 and vizantin could serve as a promising TF-based vaccine carrier for antitumor immune therapy, and KRN7000-TF-vizantin features great potential to be a vaccine candidate.

Effects of GABA on Oxidative Stress and Metabolism in High-Glucose Cultured Mongolian Sheep Kidney Cells.

The Mongolian sheep, emblematic of the Inner Mongolian grasslands, is renowned for its exceptional stress resistance and adaptability to harsh environments, drawing considerable attention. Recent research has unveiled the novel role of γ-aminobutyric acid (GABA) in combating oxidative stress. This investigation examined how GABA impacts renal-cortex and medulla cells from Mongolian sheep exposed to high-glucose stress conditions, utilizing gene expression analysis and non-targeted metabolomics. Elevated glucose levels significantly reduced the viability of Mongolian sheep renal cells and increased reactive oxygen species (ROS) levels. Conversely, the introduction of GABA notably enhanced cell viability, reduced ROS production, and stimulated the expression of antioxidant genes (e.g., Gpx, SOD, CAT) in the renal cortex. In the renal medulla, CAT expression increased, while Gpx gene expression showed mixed responses. Metabolomics analysis indicated that high-glucose exposure altered various metabolites, whereas GABA alleviated the metabolic stress induced by high glucose through modulating glycolysis and the tricarboxylic acid cycle. In Mongolian sheep renal cells, GABA effectively mitigated oxidative damage triggered by high-glucose stress by upregulating antioxidant genes and regulating metabolic pathways, revealing insights into its potential mechanism for adapting to extreme environments. This finding offers a fresh perspective on understanding the stress resilience of Mongolian sheep and may provide valuable insights for research across diverse disciplines.

Abstract A004: A Phase 1B/2 trial of second line immunotherapy with pepinemab and avelumab for patients with metastatic pancreatic adenocarcinoma

Abstract Background: Pancreatic adenocarcinoma (PDAC) is a leading cause of cancer-related mortality with poor prognosis despite maximal therapy and is recalcitrant to approved immunotherapies. In PDAC, Semaphorin 4D (SEMA4D) expression is prognostic in patients who have undergone surgical resection and correlates with tumor infiltration of activated CD8+ T-cells. Furthermore, preclinical studies suggest that monoclonal blockade of SEMA4D supports CD8+T cells tumoral infiltration and activation, sensitizing PDAC to immune checkpoint blockade (ICB). Thus, we hypothesized that a humanized anti-SEMA4D antibody (pepinemab) may improve disease control and responsiveness to ICB in patients with PDAC. Methods: We designed a phase Ib/II single-arm clinical trial to evaluate the safety, tolerability, and efficacy of pepinemab with anti-PD-L1 (avelumab) in patients with chemotherapy-refractory PDAC (NCT05102721). The study follows a dose de-escalation schema starting at a dose combination of 20mg/kg pepinemab and 800mg avelumab every two weeks. Patient accrual for phase 1b utilizes the Bayesian Optimal Interval design targeting a dose-limiting toxicity rate of 30% or less. After 16 subjects receive a given combination dose, a Simon’s two stage assessment of futility will be undertaken with expansion to phase 2 if 2 or more subjects demonstrate response. Treatment response is assessed via RECIST 1.1 criteria with surveillance CT prior to enrollment and after completion of two cycles (8 weeks). Baseline and on-treatment tumor biopsies (after completion of one cycle, 4 weeks) are performed to analyze changes in immune, stromal, and genomic profiles to elucidate mechanisms of treatment response and failure. Patient reported outcomes are incorporated (FACT-Hep and FAACT subdomains) to assess disease-specific symptoms. Results: A total of 14 patients have been enrolled to date. Three subjects withdrew from the study prior to completing the evaluable period (four weeks of treatment) and nine subjects terminated treatment due to disease progression. Two subjects continue to receive combination treatment pending evaluation of tumor response. Baseline and on-treatment biopsies have been obtained and stored for correlative analysis from 8 evaluable subjects. One treatment-related adverse event occurred though the patient was able to complete treatment without further events. No treatment-related toxicities have been encountered. Conclusions: Combining pepinemab with avelumab appears to be safe and tolerable to date. Nine of eleven patients demonstrated disease progression, though 1 of these 9 demonstrated mixed response with some lesions showing radiologic partial and even complete response. Overall treatment response, survival data, and correlative science forthcoming. Citation Format: Luis I Ruffolo, Matthew Byrne, Bailey Hilty, Brian A Belt, Yatee Dave, Paul Burchard, Terrence Fisher, Elizabeth E. Evans, Crystal Mallow, Megan Boise, Darren Carpizo, Jen Jen Yeh, David C Linehan, Daniel Mulkerin. A Phase 1B/2 trial of second line immunotherapy with pepinemab and avelumab for patients with metastatic pancreatic adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A004.

Understanding mediated sports consumption by Irish children: a qualitative study exploring their exposure and understanding of gambling marketing, risks and harms

BackgroundGambling marketing communications create a public health risk by increasing the normalisation of gambling in sports. In a context where broad level studies report significant underage gambling, currently no evidence exists on how these communications are received by children in the Republic of Ireland (hereafter Ireland) and Northern Ireland. To build this evidence base and provide granular detail below broad level data, this study explored the sport consumption habits and views of Irish children (aged 14–17 years) on their exposure, awareness and perceptions of the relationship between sport and gambling.MethodsDrawing on a constructivist approach to data collection, 6 face-to-face focus groups were staged with a total of 51 youth sport consumers from both sides of the border region on the island of Ireland.ResultsFour main themes were established. First, mediated sport consumption was predominantly via mobile social media. Second, we found that their exposure to gambling marketing was high and while frequently seen through social media channels it was also prevalent in peer-to-peer conversations and on the main street. Third, we found mixed responses on their perceptions about gambling and sport. While many felt that sport and gambling were a good fit, they were aware of the financial risks involved. Few children understood the wider social risks with gambling harm. Fourth, children were sceptical of claims made in gambling communications. These findings highlight concerns about how exposure to gambling marketing is impacting children’s views on sport and on gambling. These views need to be taken into consideration when broadcasters and sport organizations are entering into commercial associations.ConclusionsGambling marketing is noticed by children watching mass and social media, and in the towns in the border regions of the island of Ireland. Our study provides children’s viewpoints on this topic which complements the larger quantitative studies in Ireland and Northern Ireland that highlight the growing prevalence of children and gambling. This study not only extends the literature on the exposure, awareness and perceptions of children on the island of Ireland but provides crucial evidence to public health advocates in this region demonstrating the pervasiveness of gambling communications in and around children’s spaces.

Joint modeling of mixed skewed longitudinal responses using convolution of normal and log-normal distributions: a Bayesian approach

This paper investigates the joint modeling of mixed ordinal and continuous longitudinal responses using a random effects model and applying a conditional approach. For the ordinal responses, a latent variable model with a logistic distribution is employed. To address skewness in the data, the model incorporates normal and log-normal convolution (NLNC) for both the error term and the random effects in the longitudinal model. Parameter estimation is carried out within a Bayesian framework using Gibbs sampling. The performance of the proposed model is evaluated through simulation studies, comparing it to joint models of mixed ordinal and continuous longitudinal responses assuming skew-normal or normal distributions. The results indicate that joint models using skew-normal or normal distributions can lead to biased parameter estimates, whereas the NLNC joint model performs better overall. Additionally, the proposed method is applied to analyze data from the British Household Panel Survey (BHPS), using life satisfaction and annual income as the correlated ordinal and continuous longitudinal responses, respectively, with annual income showing significant skewness. The results demonstrate that the proposed model provides the best fit for capturing the substantial skewness in the data.

Effects of Schistosoma haematobium infection and treatment on the systemic and mucosal immune phenotype, gene expression and microbiome: A systematic review.

Urogenital schistosomiasis caused by Schistosoma haematobium affects approximately 110 million people globally, with the majority of cases in low- and middle-income countries. Schistosome infections have been shown to impact the host immune system, gene expression, and microbiome composition. Studies have demonstrated variations in pathology between schistosome subspecies. In the case of S. haematobium, infection has been associated with HIV acquisition and bladder cancer. However, the underlying pathophysiology has been understudied compared to other schistosome species. This systematic review comprehensively investigates and assimilates the effects of S. haematobium infection on systemic and local host mucosal immunity, cellular gene expression and microbiome. We conducted a systematic review assessing the reported effects of S. haematobium infections and anthelmintic treatment on the immune system, gene expression and microbiome in humans and animal models. This review followed PRISMA guidelines and was registered prospectively in PROSPERO (CRD42022372607). Randomized clinical trials, cohort, cross-sectional, case-control, experimental ex vivo, and animal studies were included. Two reviewers performed screening independently. We screened 3,177 studies and included 94. S. haematobium was reported to lead to: (i) a mixed immune response with a predominant type 2 immune phenotype, increased T and B regulatory cells, and select pro-inflammatory cytokines; (ii) distinct molecular alterations that would compromise epithelial integrity, such as increased metalloproteinase expression, and promote immunological changes and cellular transformation, specifically upregulation of genes p53 and Bcl-2; and (iii) microbiome dysbiosis in the urinary, intestinal, and genital tracts. S. haematobium induces distinct alterations in the host's immune system, molecular profile, and microbiome. This leads to a diverse range of inflammatory and anti-inflammatory responses and impaired integrity of the local mucosal epithelial barrier, elevating the risks of secondary infections. Further, S. haematobium promotes cellular transformation with oncogenic potential and disrupts the microbiome, further influencing the immune system and genetic makeup. Understanding the pathophysiology of these interactions can improve outcomes for the sequelae of this devastating parasitic infection.

Efficacy and safety of rechallenge with [177Lu]Lu-PSMA-I&T radioligand therapy in metastatic castration resistant prostate cancer.

The purpose of this study was to evaluate the safety and outcome of rechallenge [177Lu]Lu-PSMA-I&T in newly progressed mCRPC patients after response to initial [177Lu]Lu-PSMA radioligand therapy(PRLT). We retrospectively included 18 patients who underwent rechallenge with [177Lu]Lu-PSMA-I&T. All patients presented with (i) newly progressed disease after response to initial PRLT; (ii) a [68Ga]Ga-PSMA-11 PET/CT confirming the presence of PSMA-positive metastases; iii)ECOG performance status 0-1.Adverse events were graded according to CTCAE v5.0. Response was assessed by PSA and classified according to PCWG3 recommendations. For patients who underwent restaging with [68Ga]Ga-PSMA-11 PET/CT, imaging response was categorised according to adapted PERCIST v1.0. In patients with discordant [68Ga]Ga-PSMA-11 PET/CT and PSA, other available imaging modalities were evaluated to confirm disease status. Overall survival (OS) was calculated from the first cycle of initial PRLT and rechallenge PRLT, respectively, until last patient contact or death. Patients were initially treated with a median of 5 cycles (range 4-7) and were rechallenged after a median of 9 months (range 3-13). Each patient received a median of 4 (range 2-7) rechallenge cycles (median cumulative activity 26.1GBq). None of the patients experienced life-threatening G4 adverse events during either treatment period. Grade 3 adverse events included one case of anaemia, one case of thrombocytopenia, and one case of renal failure. In 8/18 patients long-term toxicities were evaluated. Serious toxicities (≥ Grade 3) occurred in 3/8 patients (n = 1 G4 thrombocytopenia, n = 1 G4 renal failure and n = 1 pancytopenia and G4 renal failure). Best PSA50%-response was observed in 44% of patients and PSA-disease control was confirmed in 56% of patients at the last cycle. Of the 12/18 patients restaged by imaging, 6/12 (50%) patients had disease control (partial response/stable disease), 1/12 had a mixed response, and 5/12 had progression. After a median follow-up time of 25 months (range 14-44), 10 patients had died, 7 were still alive, and one patient was lost at follow-up. The median OS was 29 months (95%CI,14.3-43.7 months) for the initial treatment and 11 months (95%CI,8.1-13.8 months) for the first rechallenge course. More than half of patients benefit from rechallenge PRLT. Our analysis suggests that rechallenge may prolong survival in selected patients, with an acceptable safety profile.

Enhancing protective immunity against bacterial infection via coating nano-Rehmannia glutinosa polysaccharide with outer membrane vesicles.

With the coming of the post-antibiotic era, there is an increasingly urgent need for safe and efficient antibacterial vaccines. Bacterial outer membrane vesicles (OMVs) have received increased attention recently as a potential subunit vaccine. OMVs are non-replicative and contain the principle immunogenic bacterial antigen, which circumvents the safety concerns of live-attenuated vaccines. Here, we developed a novel nano-vaccine by coating OMVs onto PEGylated nano-Rehmannia glutinosa polysaccharide (pRL) in a structure consisting of concentric circles, resulting in a more stable vaccine with improved immunogenicity. The immunological function of the pRL-OMV formulation was evaluated in vivo and in vitro, and the underlying mechanism was studied though transcriptomic analysis. The pRL-OMV formulation significantly increased dendritic cell (DC) proliferation and cytokine secretion. Efficient phagocytosis of the formulation by DCs was accompanied by DC maturation. Further, the formulation demonstrated superior lymph node targeting, contributing to a potent mixed cellular response and bacterial-specific antibody response against Bordetella bronchiseptica infection. Specifically, transcriptomic analysis revealed that the immune protection function correlated with T-cell receptor signalling and Th1/Th2/Th17 differentiation, among other markers of enhanced immunological activity. These findings have implications for the future application of OMV-coated nano-carriers in antimicrobial immunotherapy.

A study on knowledge, awareness, and practices about eye donation in medical students

Aim: The aim of the study was to assess knowledge, awareness, and practices about eye donation in medical students of medical colleges. Setting: Tertiary Care Centre. Design: This study involves a cross-sectional study. Materials and Methods: A cross-sectional study at a tertiary care center in Western Vidarbha from August 2023 to October 2023, among medical students. Approval from the institutional ethical committee was taken. A total of 538 students participated in the study. After taking consent, data were collected in self-reported questionnaire consisting of questions about the knowledge and awareness of eye donation, the source of their information, willingness for donation, and respective reasons for willingness and nonwillingness. Data were analyzed. Results: Majority of students belonged to the age group between 21 and 30 years. Most were undergraduates, followed by interns and post-graduates. Five hundred and twenty-nine had heard about eye donation mainly through medical personnel. The majority of students believed that eye donation means giving sight to the blind. Most knew the nearest point of contact for eye donation. The answer given by most to the question “What is removed for transplantation?,” was cornea. The most common response to questions about a site for the removal of eyes was the hospital. The majority knew that no monetary benefits were given for donations. Those willing to pledge their eyes wanted to do so to help the blind, and those not willing said objection by family as the main reason among others. Conclusion: After the study, we concluded that awareness about eye donation was good in the students, while knowledge regarding eye donation varied among the students. Willingness to donate showed mixed responses.

Mixed responses of tenebrionid beetles to solar energy development in the Mojave Desert

Solar energy development causes land-use change and habitat alteration that may affect desert ecosystems. Tenebrionid beetles have evolved to exploit desert environments and heavily contribute to ecosystem functionality in aridlands, yet their species-specific, ecological responses to solar energy development are unknown. Our objective was to elucidate effects of solar energy development decisions at a solar power facility (392 MW) on a tenebrionid beetle community in the Mojave Desert. Seven years post-construction, we used pitfall traps to collect tenebrionid beetles for one month in treatments representing variably intense site preparation practices and conservation measures, including blading (i.e., bulldozing), mowing, and establishment of undeveloped patches in solar fields, replicated across three power blocks comprising the facility and in undeveloped control sites surrounding the facility. Blading caused significant, deleterious effects on tenebrionid beetle abundance, species richness, and diversity. Ordination revealed apparent overlap of non-bladed treatments and controls, suggesting that intermediate levels of disturbance from less intensive solar energy development decisions may serve to minimize negative effects of solar energy development on tenebrionid communities. Anepsius delicatulus and Triorophus laevis, the two most commonly collected tenebrionids, were significantly more abundant in non-bladed treatments in the solar facility than in undeveloped controls; thus, solar facilities with minimized disturbance to vegetation may provide preferred habitat for certain tenebrionid species. Our results demonstrate the value of understanding variable responses of insects to solar energy development decisions in deserts to inform real-world adaptive management of natural resources during the renewable energy transition.

Quitting on TikTok: Effects of Message Themes, Frames, and Sources on Engagement with Vaping Cessation Videos

This study examined how message themes, frames, and sources in vaping cessation videos on TikTok influenced positive (i.e. likes, shares, positive comments regarding quitting vaping) and negative video engagement (i.e. negative comments regarding quitting vaping). TikTok videos (N = 412) with the hashtags #quitvaping and #quittingvaping were analyzed. Aspect-based sentiment analysis was conducted to evaluate the sentiment of quitting vaping in comments. Negative binomial regression models predicted video engagement from six message themes, ratios of gain and loss frames, and message sources. Themes related to nicotine addiction and physical health effectively drove positive engagement, such as likes and shares. The theme of harmful chemicals elicited mixed responses, generating both positive and negative comments regarding quitting vaping. Videos with a higher ratio of gain frames led to more positive engagement, including likes, shares, and positive comments regarding quitting vaping. Sources with informal expertise (e.g. those who have successfully quit vaping) and current e-cigarette users were more effective in engaging the TikTok audience than non-expert and non-user sources. These findings provide insights into messaging strategies that can effectively engage TikTok audiences and encourage vaping cessation.

Potential Impacts of Heartfulness Relaxation on Interpreting Performance: A Pilot Study

This mixed-methods pilot study investigates the effects of a brief seven-minute heartfulness relaxation intervention on interpreting students’ cognitive performance, perceived stress, and consecutive interpreting skills. Seven participants completed pre- and post-intervention measures, including the Stroop test, Perceived Stress Scale (PSS-10), and consecutive interpreting tasks. Qualitative data were collected through participant interviews and analyzed using thematic analysis. The quantitative results revealed significant improvements in executive function (p<.001), perceived stress (p=.078), and Chinese to English and English to Chinese consecutive interpreting (p=.028) following the intervention. The qualitative findings indicated predominantly positive effects on interpreting performance, concentration, and focus, with mixed responses regarding relaxation, environmental factors, and effect sustainability. The convergence of the qualitative themes and quantitative improvements suggested that the heartfulness relaxation intervention enhanced the participants’ cognitive function, reduced fatigue, and optimized essential skills. These findings highlight the potential benefits of incorporating a brief relaxation practice into interpreter training and professional development. Limitations and future research directions are discussed, emphasizing the need for larger controlled trials to confirm the generalizability and long-term effects of the intervention. Overall, this pilot study provides a foundation for further investigations into the application of the brief heartfulness relaxation technique in the interpreting field.

Chemometric Modeling Revealed Oleic and Linoleic Acids as Varietal Biomarkers for Six Sesame Varieties-In Vitro and UHPLC Analyses.

Pakistan once considered self-sufficient for edible oil production now became the major importer with 88.6% imports and producing only the minor portion. Scientific negligence in oil seed crops led to a dramatic decrease in edible oil production depending mainly on only the imports. Sesamum indicum L., "Queen of Oil seeds" with 50-55% oil, is cultivated in various geographical regions of Pakistan, but farmers are not considering this crop because of insufficient knowledge, poor crop management practices, and low yielding varieties. This study was conducted to check the nutritional, biochemical, antioxidant, and yield potentials of six major varieties, i.e., TS-5, TH-6, Til-18, NIAB-Mil, NIAB-Pearl, and NS-16, and to compare the nutritionals, oil quality, and oil yield potential of these varieties. Field experiment was conducted, and various crop growth biomarkers were analyzed. Chlorophyll content and superoxide dismutase activity were found to be highest in NIAB-Mil followed by NIAB-Pearl and comparable to those of Til-18, while APX, Cat, and GPX activity was found to be highest in Til-18 with 25.6 and 5.9 and 6.02 mg/g, respectively. Seed antioxidant parameters showed a mixed response, but NIAB-Mil, NIAB pearl, and Til-18 were found to be highest in all antioxidant parameters. UHPLC analysis of seed oil resulted in a total of 14 triacylglycerols (TAGs), and principal component analysis and OPLS-Da analysis showed seven TAG biomarkers responsible for the separation of sesame varieties. Til-18 was found to be highest in oil content (53.3%) more abundant with oleic acid, while NIAB-Pearl, NIAB-Mil, and NS-16 were found to be abundant with linoleic acid, both considered as potential TAG biomarkers for sesame oil separation. This study concluded that, in general, Til-18 variety is more resistant with high nutritional status, high antioxidant activity, and oil yielding variety, followed by NIAB-Mil and NIAB-Pearl.

Taxon‐specific response of natural enemies to different flower strip mixtures

Abstract Flower strips are a prominent agri‐environmental scheme with the central objective to promote biodiversity and maintain associated ecosystem services. The promotion of natural enemies by increasing resource availability through flower strips is a promising approach for integrated pest management. In a 3‐year field study, two annual and two perennial flower strip mixtures, as well as a grass mixture, were tested regarding their attractiveness for natural enemies at three different study sites in Germany. Natural enemies were sampled annually in nine sampling rounds at a 10‐day rhythm using sweep netting and pitfall traps. To assess available floral resources, we estimated the species‐specific flower cover and classified species into flower types. Flower strip mixtures differed in their attractiveness to natural enemies. Treatment effects on arthropod activity density were most pronounced in the second and third year. Overall, perennial were more attractive than annual flower strip mixtures for most natural enemies, however, the response to flower strip mixtures varied significantly among taxa. For example, perennial flower strips showed a two‐ to fourfold increase of plant‐dwelling spider and parasitoid wasp activity density as well as higher numbers of juvenile stages of predatory bugs and rove beetles compared to annual flower strips. In contrast, annual flower strip mixtures showed the highest attractiveness for ground beetles. Moreover, different natural enemy taxa were associated with varying flower strip characteristics such as flower type proportion, herbivore availability and plant species richness. Synthesis and applications. We found taxon‐specific responses of natural enemies to different flower strips, which provide guidance for the floral composition of flower strips in order to maximize their effectiveness for natural pest control. We suggest the use of perennial mixtures with high proportion of flower heads (Asteraceae) and disk flowers (esp. Apiaceae, Rubiaceae, Brassicaceae) in particular, but also flag blossoms (Fabaceae) as these flower types were positively associated with several natural enemy groups. However, since we found mixed responses to flower types, specific flower strip composition will ultimately depends on its objective.

Environmental controls on bioluminescent dinoflagellate density, in Laguna Grande, Fajardo, Puerto Rico

Introduction: Bio bays in Puerto Rico play an important socio-economic role and declines in dominant bioluminescent dinoflagellate Pyrodinium bahamense are concerning. Studies show erratic blooms with is weak correlation to in situ environmental factors. Our study examines shorter field and longer proxy records on dinoflagellate density at Laguna Grande de Fajardo (LGF). Objectives: To quantify temporal changes in dinoflagellate density in a long-term monitoring study, understand how the marine environment modulates those changes, and determine the wider impacts of a fluctuating climate and extreme events on proxies for dinoflagellate density. Methods: Bimonthly samples were collected from 2016 to 2021 at three sampling sites in LGF. Dinoflagellates density was estimated by Sedgewick Rafter counting cells. Environmental conditions were obtained from Rio Fajardo 5007100 station and NOAA buoy 41056. Marine climate and biotic proxies were obtained from remote sensing measurements. Kruskal Wallis, Spearman correlations and cross-correlations in the shorter field and longer proxy records were used to evaluate environmental controls on LGF dinoflagellate blooms. Results: Six years of field monitoring densities found a low period in 2016-2017, frequent and intense blooms in 2018-2021 punctuated by hurricanes. Generally low values were recorded in late winter in contrast with higher values in late summer (Aug-Nov). Light winds and mixed layer response to seasonal warming in the form of high tides and low salinity, were found to sustain dinoflagellate reproduction. Conclusions: Bioluminescent dinoflagellates are vital to coastal tourism and require resource management. LGF results show that: 1) dinoflagellate counts fluctuate widely, 2) fluorescing dinoflagellates are sensitive to environmental conditions because of limited seasonality and narrow physiological range, 3) hurricanes play a role by ‘raking and refreshing’ the coastal lagoon for subsequent biotic reproduction, and 4) intra-seasonal fluctuations of density and proxies relate to air-sea thermodynamic conditions, the salinity budget and sea level.

Electroneuromyographic examination of pelvic floor muscle dysfunction in patients with descending perineum syndrome

Aim: to assess the functional state of the pelvic floor muscles and their innervation in patients with descending perineum syndrome, manifested simultaneously by obstructive defecation and chronic neurogenic pelvic pain.Materials and methods: in 2022–2023, 51 patients (40 (78.4 %) women, 11 (21.6 %) men; age 50.9 ± 14.5 years) with pelvic organ prolapse, manifested by obstructive defecation and chronic neurogenic pelvic pain were examined at the Ryzhikh National Medical Research Centre for Coloproctology. All patients underwent high-resolution anorectal manometry and stimulation electroneuromyography according to a comprehensive neurophysiological protocol using a St. Mark’s electrode.Results: manometric patterns of functional defecation disorders were detected in 40 (78.4 %) patients, in the rest they were confirmed by defecography data and a negative evacuation test according to the Rome criteria IV. With stimulation electroneuromyography, neuropathy was detected in 68.6 % of cases by increasing the latency of the M-response of the pudendal nerve on the right or left. Complex neurophysiological diagnostics made it possible to additionally identify neuropathy in another 25.5 % of patients: according to changes in the mixed reciprocating reflex response (mixed feedback-reflex response) (efferent pathway) — in 17.7 % of cases; according to changes in the bulbocavernous reflex (efferent and afferent pathways) — in 7.8 % of cases. That is, neuropathy was determined in 94.1 % of patients in total.Conclusions:Functional diagnostic methods are an effective tool for studying the state of the pelvic floor muscle structures and their innervation and make it possible to identify pathogenetic links of disorders, the clinical manifestations of which are anal incontinence, neurogenic pelvic pain and proctogenic constipation in the form of obstructive defecation.Stimulation electroneuromyography makes it possible to detect disturbed innervation of the pelvic floor muscles and anal sphincter in the standard study of the M-response in 68.6 % of cases with perineal prolapse syndrome with a combination of proctogenic disorders and neurogenic pelvic pain. The use of a comprehensive neurophysiological protocol makes it possible to detect innervation disorders in 94.1 % of cases.

Simulation-based ultrasound-guided regional anesthesia education: a national survey of Canadian anesthesiology residency training programs.

Simulation-based education in ultrasound-guided regional anesthesia (UGRA) improves knowledge, skills, and patient outcomes. Nevertheless, it is not known how simulation-based UGRA education is used across Canada. We aimed to characterize the current use of simulation-based UGRA education in Canadian anesthesiology residency training programs. We developed and distributed a structured national survey to simulation leads of all 17 Canadian anesthesiology residency training programs. The survey inquired about program demographics, simulation modalities, facilitators and barriers to simulation use, use for assessment, and beliefs around simulation-based UGRA education. We gathered data from August to November 2023 and summarized our findings descriptively. Fifteen programs (88%) responded to our survey. Eight programs (53%) used UGRA simulation for technical training and nine programs (60%) for nontechnical training. The most common simulators used were live model scanning (13 programs, 87%) and gel phantom models (7 programs, 47%). Five programs (33%) mandated simulation-based UGRA in their curriculum. We found that deliberate practice and improved patient safety were most valued in simulation training while lack of funding and faculty availability were the most common barriers to implementation. Most respondents agreed that formative simulation-based education would improve trainee skills and called for greater standardization. Nevertheless, there were mixed responses regarding summative UGRA simulation and the need for simulation proficiency before clinical practice. Our findings show significant variations in simulation implementation and views on UGRA simulation-based education among Canadian anesthesiology residency training programs. Future studies should explore avenues to overcome barriers and improve knowledge translation in UGRA.

18 Unraveling the Impact of Tumor Microenvironment on Immunotherapy Response/Resistance in Human Sarcomatoid Clear Cell Renal Cell Carcinoma: Insights from a Novel Mouse Model

Abstract Background Renal cell carcinoma (RCC) with histological sarcomatoid de-differentiation (sRCC) has a historically poor prognosis across all RCC subtypes. Recently, immunotherapy with anti-PD1 + anti-CTLA4 combo has significantly improved disease-specific survival, and complete response are seen in upwards of 20% of patients. However, half of sRCC patients still have no response to this immunotherapy, and the mechanism of response remains unclear. Methods Our preliminary bulk RNA sequencing data reveal a distinct tumor microenvironment (TME) in human sRCC tumors, characterized by heightened immune cell infiltration, particularly enriched for regulatory T cells (Tregs). Subsequently, we delineated the spatial gene expression topography within sRCC and clear cell renal cell carcinoma (ccRCC) regions, culminating in the development of the sRCC signature. Concurrently, we established an innovative immunocompetent murine model of sRCC. Leveraging CRISPR technology, we replicated the human sRCC genotype by knocking out Vhl, Bap1, and Cdkn2a/2b genes in a mouse telomerase reverse transcriptase (mTERT) overexpressing renal proximal tubule epithelial cell line. This effort yielded a successfully engineered mouse implantable sRCC cell line bearing VhlMutBap1DelCdkn2aDelCdkn2bDel (HJRCC68N). Results The syngeneic mouse sRCC model we developed faithfully replicates human sRCC histology and tumor microenvironment and recapitulates the sRCC signature identified in human sRCC regions. Importantly, like in humans, mouse sRCC tumors exhibit a heterogeneous response to anti-PD1 + anti-CTLA4 combination, and anti-CTLA4 monotherapy, reflecting the clinical variability observed in human patients. Furthermore, transcriptomic analysis from our mouse model reveals the involvement of type 1 immunity in responder, providing insights into overcoming immunotherapy resistance. Conclusions In conclusion, our study unveiled distinct changes in the TME at the transcriptomic and spatial gene expression levels in human sRCC. Leveraging our novel murine model, which faithfully replicates human sRCC histology, TME characteristics, and mixed response to immunotherapy, we elucidated the involvement of type 1 immunity in treatment response. These insights offer valuable pathways for overcoming immunotherapy resistance and pave the way for the development of tailored treatment strategies aimed at improving outcomes for sRCC patients.

Differential Age-Based Response Induced by a Commercial Probiotic Supplementation in Pastured Goats.

The potential benefit of probiotics in small ruminant production systems has largely been unexplored. We evaluated the effect of a goat commercial probiotic on health and performance indicators in pastured goats from birth until 10months. We randomly allocated 26 newborn nursing goat kids to two groups: a control group that received saline and a treatment group that received a commercial probiotic paste orally. We evaluated select observable health indicators (inappetence, diarrhea, coughing), weight, immunity (IgA, IgG, and innate immune response), total protein, hematocrit (HCT), total lactic acid bacteria (LAB), total coliforms, and prevalence of Escherichia coli (E. coli) primary virulence genes (stx1, stx2, and eae) during the experimental period. The results revealed no significant differences in the health indicators, LAB count, and total E. coli count. Prevalence of stx1 at 1week of age and both stx1 and stx2 genes 4months post-weaning was significantly (P < 0.05) higher in probiotic-supplemented goats. Probiotic supplementation significantly (P < 0.05) increased the total protein and IgA 1month post-supplementation during the pre-weaning period and innate immune markers 2days post-weaning. The HCT in probiotic-supplemented goats was significantly (P < 0.05) higher at 1 and 2months post-weaning. The growth rate was not affected by probiotic supplementation in pre- and peri-weaned goats but was significantly (P < 0.05) lowered in goats older than 4months in the supplemented group. In this pastured goat production study, there were mixed responses to a commercial probiotic in healthy goats based on age. The study suggests that early daily probiotic supplementation in pre-weaned pastured goats may have immune stimulation benefits, but in older healthy animals, post-weaning net benefits are unclear and further research is recommended.

KAP study on hand hygiene in the era post COVID-19 SARS among general population.

Background: Hand hygiene accounts one of the most effective ways of keeping diseases at bay. As of now, this practice stands as a key cornerstone post COVID-19 era, thus helping to break the chain of contact transmission of such deadly outbreak’s outcome. Aims and Objectives: To assess knowledge, attitude and practices (KAP) relating to hand hygiene among general population post COVID-19 era. Materials and Methods: A cross-sectional study was done among general population of our setup over a span of 2 months by distributing online a pre-designed, pre-validated and semi-structured Google Form survey questionnaire having questions based on knowledge (10), attitude (07) and practices (06) relating to hand hygiene post COVID-19 era. Individuals enrolled were tech-savvy with access to e-mail and internet facility, and able to understand English language. Results: Data from responses of 545 individuals among our general community was collected and analyzed, out of which majority (88.9%) responders already had a basic insight about hand hygiene. 91.2% responders clearly believed hand hygiene is important to prevent the infection spread from contact. While 25.7% responders answered 40-60 seconds as minimum time required for hand washing, comparable majority (48.9%) marked it as 20-40 seconds. 70.9% responders were strongly against use of gloves as a substitute for hand washing. Interestingly, many had a mixed say regarding time span for which an alcohol-based hand sanitizer be rubbed onto hands viz. 32.3% stating it as 10 seconds, 27.7% as 20 seconds, 21.2% as 5 seconds and 18.8% as 15 seconds, respectively. Majority (85.4%) voted hand washing as better and probably the best mode of maintaining hand hygiene. Majority (93.1%) responders felt the need of practicing hand hygiene possibly at all the times. While most of our study population practiced hand hygiene thoroughly, 77.5% responders said that it positively affected their life also. However, in contrast, only 57.4% responders washed their hands after using public transport/money exchange, while 39.3% practiced it only sometimes. Conclusion: Our study highlights that despite of a positive relation between the awareness among general masses with regard to positive attitude about hand hygiene post COVID era, there was a mixed response about practice of the same, thus stressing on the need to reinforce such practices with due clarity so that the message goes loud and clear among public at large for breaking the chain of COVID transmission till date. Further our study can aid decision-makers in forecasting the effectiveness of infection mitigation strategies or in defining valid changes based on the practices, attitudes, and understanding of the intended population.