Polycythemia vera (PV) is a myeloproliferative neoplasm primarily characterized by erythrocytosis. PV incidence is exceedingly rare in the pediatric and adolescent population. In adult patients, approximately 96% are found to have a somatic mutation in exon 14 (JAK2 V617F) and 3% display a mutation in exon 12. We present a case of a 2-year-old female with symptomatic PV secondary to a deletion in exon 12 of JAK2, initially treated with phlebotomy and switched to PEG-IFNα-2a therapy. This therapy has been effective over 15 months with resolution of symptoms, reduced phlebotomy requirements, and minimal side effects.

Background: The incidence of relapsed neuroblastoma (NB) in the central nervous system (CNS) is less than 20%. Here, we presented eleven cases with CNS relapsed NB to better understand the CNS relapsed NB. Methods: Medical records from NB patients between January 1st, 2010 and December 31st, 2017 were reviewed to examine those diagnosed with CNS relapsed NB. The following patient information was extracted, including age, gender, histological subtyping, tumor stage, risk group, biomarkers, chemotherapy regimen and dosage, surgery details, treatment outcomes and follow-up, clinical manifestations of CNS relapsed NB, treatment protocols for CNS relapsed NB, and patient outcomes (including the cause of death). Results: From January 1st, 2010 to December 31st, 2017, eleven NB patients were diagnosed with CNS relapse at our department. All the eleven patients were classified to have high risk disease. The median time from the completion of chemotherapy to CNS relapse was 5 months. Five patients did not receive any treatment for the relapsed disease and finally died of it. Curative treatments were tried in six patients. At last, two patients with isolated CNS relapsed NB survived. Conclusions: The median time from the completion of chemotherapy to CNS relapse is very short in the present retrospective study. It seems that patients with isolated CNS relapsed disease have better outcomes. We suggest the temozolomide-topotecan combination as an add-one backbone for further salvage treatment in children with CNS relapsed NB, even though this combination is not superior to others in terms of its efficacy.

Objective Pulmonary atresia (PA) is a rare type of complex cyanotic congenital heart defect mainly characterized by an undeveloped pulmonary valve or pulmonary artery. herefore, defining a disease-causing gene mutation in a pulmonary atresia family becomes a possible method of the genetic counseling, future prenatal diagnosis, and therapeutic approaches of pulmonary atresia. Methods Blood samples of six members in a PA family were collected, and the genomic DNA was extracted using the QIAamp DNA Blood Mini Kit. The gene detection was performed using the second-generation sequencing gene Panel. Results Genetic testing results indicates as follows: A heterozygous mutation originated from maternal inheritance was detected in the BMPR2 gene of the proband’s genomic DNA. The pathogenic gene was at c.2804C>T (p. A935V). The mutation was also detected in the genomic DNA of the proband’s elder brother(III-1), but not in other family members. Conclusion To our knowledge, this is the first study to report the BMPR2 variant responsible for pulmonary atresia. The frequency of c.2804C>T (p. A935V) mutation detected in this family is extremely low in the normal population (1/ 246048). The mutation was highly conservative in different species. And SIFT (sorting intorlerant from tolerant) predicts it to be a harmful mutation.

Aim: This study aimed to evaluate the effect of PFL on clinical and radiological outcomes in Lenke type 1 AIS patients. Methods: The study was retrospective and included patients diagnosed with Lenke type 1 AIS with deformity correction and spinal fusion with the right main thoracic curve. The patients were allocated into two groups according to the selection of PFL, as T2 and T4 groups. Outcome measures and classification parameters were Scoliosis Research Society - 22r (SRS - 22r) score, Oswestry disability index (ODI) Turkish version, Lenke classification and Risser sign. The data of the two groups were compared. Results: Postoperative shoulder imbalance and trunk shift values were significantly lower in the T2 group whereas SRS - 22r scores were significantly higher in the T2 group (p < 0.05 for all). While the preoperative bending angles were significantly lower in the T4 group (p < 0.05), the two groups were similar in the postoperative Cobb measurement, thoracic kyphosis and lumbar lordosis angles, sagittal imbalance, and T1 tilt values (p > 0.05 for all). Conclusions: The selection of the T2 vertebra as the PFL is more advantageous in terms of clinical and radiological results. Additionally, there is no difference between T2 and T4 fusion levels in terms of complications.

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1. The volatiles from damaged plants induce defense in neighboring plants. The phenomenon is called plant-plant communication, plant talk or plant eavesdropping. Plant-plant communication has been reported to be stronger between kin plants than genetically far plants in sagebrush. 2. Why do plants distinguish volatiles from kin or genetically far plants? We hypothesize that plants respond only to important conditions; the induced defense is not free of cost for the plant. To clarify the hypothesis, we conducted experiments and investigations using goldenrod of 4 different genotypes. 3. The arthropods community on tall goldenrods were different among 4 genotypes. The response to volatiles was stronger from genetically close plants to the emitter than from genetically distant plants from the emitter. The volatiles from each genotype of goldenrods were different; and they were categorized accordingly. Moreover, the arthropod community on each genotype of goldenrods were different. 4. Synthesis: Our results support the hypothesis: goldenrods respond to volatiles from genetically close plants because they would have similar arthropod species. These results are important clues elucidating adaptive significance of plant-plant communication.

Buck Creek is a spring-fed, cool-water tributary of the West Fork White River, Indiana. The Muncie Bureau of Water Quality sampled fishes and monitored water temperature in Buck Creek annually from 1986-2018. For this study, we utilized long-term fish data from the Bureau of Water Quality to evaluate spatial and temporal changes in the fish assemblages of Buck Creek in Delaware County, Indiana, USA. Non-metric multidimensional scaling (NMDS) was used to describe changes in the fish assemblages over space and time. Linear mixed effects models were used to evaluate the relationship between environmental factors and the fish assemblages. The spatial NMDS results were separated in distinct groups of upstream and downstream assemblages. This was characterized by a shift of headwater specialists shifting to large-river species. The temporal NMDS results were separated into distinct annual assemblages. This was characterized by a drop in pollution-tolerant species and an increase in intolerant species. Our findings indicate that the fish assemblages have improved in Buck Creek over space and time.

Background: The potential protective role of eosinophils in the COVID-19 pandemic has aroused great interest, given their potential virus clearance function and the infection resistance of asthma patients to this coronavirus. However, it is unknown whether eosinophil counts could serve as a predictor of the severity of COVID-19. Methods: A total of 1004 patients with confirmed COVID-19 who were admitted to Leishenshan Hospital in Wuhan, China, were enrolled in this study, including 905 patients in the general ward and 99 patients in the ICU. We reviewed their medical data to analyze the association between eosinophils and intensive care unit (ICU) admission and death. Results: Of our 1004 patients with COVID-19, low eosinophil counts/ratios were observed in severe cases. After adjusting for confounders that could have affected the outcome, we found that eosinophil counts might not be a predictor of intensive care unit (ICU) admission. In 99 ICU patients, 58 of whom survived and 41 of whom died, low eosinophil level was an indicator of death in patients with severe COVID-19, with a cutoff value of 0.04 × 109/L for the prediction of death. Conclusion: Our research revealed that a low eosinophil level was a predictor of death in ICU patients rather than a cause of ICU admission.

1. The dissimilarity and hierarchy of trait values that characterize niche and fitness differences, respectively, have been increasingly applied to infer mechanisms driving community assembly and to explain species co-occurrence patterns. Here, we predict that limiting similarity should result in the spatial segregation of functionally similar species, while functionally similar species will be more likely to co-occur together either due to environmental filtering or competitive exclusion of inferior competitors (hereafter hierarchical competition). 2. We used a fully mapped 50-ha subtropical forest plot in southern China to explore how pairwise spatial associations were influenced by trait dissimilarity and hierarchy between species in order to gain insight into assembly mechanisms. We assessed pairwise spatial associations using two summary statistics of spatial point patterns at different spatial scales and compared the effects of trait dissimilarity and trait hierarchy of different functional traits on the interspecific spatial associations. These comparisons allow us to disentangle the effects of limiting similarity, environmental filtering and hierarchical competition on species co-occurrence. 3. We found that trait dissimilarity was generally negatively correlated with interspecific spatial associations, meaning that species with similar trait values were more likely to co-occur together and thus supporting environmental filtering or hierarchical competition. We further found that leaf area, wood density and maximum height had stronger trait hierarchy effects on the pairwise spatial associations relative to their corresponding trait dissimilarity effects, which suggests that hierarchical competition played a more (or at least equally) important role in structuring our forest community compared to environmental filtering. 4. This study employed a novel method to disentangle the relative importance of multiple assembly mechanisms in structuring co-occurrence patterns, especially the mechanisms of environmental filtering and hierarchical competition, which lead to indistinguishable co-occurrence patterns. This study also reinforced the importance of trait hierarchy rather than trait dissimilarity in driving neighborhood competition.

Background: Expected benefits of modified ultrafiltration(MUF) include increased hematocrit, reduction of total body water & inflammatory mediators, improved left ventricular systolic function, & improved systolic blood pressure and cardiac index following cardiopulmonary bypass(CPB). This prospective randomized trial tested this hypothesis. Methods: 79 patients undergoing intracardiac repair of Tetralogy of Fallot(TOF) were randomized to MUF group(Group-M, n=39) or only conventional ultrafiltration(CUF) group(Group-C, n=40). Primary outcome was change in hematocrit. Secondary outcomes were changes in peak airway pressures, ventilatory support, blood transfusions, time to peripheral rewarming, mean arterial pressure, central venous pressure, inotrope score(IS) and cardiac index. Serum inflammatory markers were measured. Results: Following MUF, Group-M had higher hematocrit(44.3±0.98 g/dl) compared to Group-C(37.8±1.37g/dl),P=<0.001. Central venous pressure(mmHg) immediately following sternal closure was 9.27±3.12mmHg in Group-M & 10.52±2.2mmHg in Group-C(P=0.04). In the ICU, they were 11.52±2.20mmHg in Group-C and 10.84±2.78mmHg in Group-M(P=0.02). Time to peripheral rewarming was 6.30±3.91 hours in Group-M and 13.67±3.91hours in Group-C(P=0.06). Peak airway pressures in ICU were 17±2mmHg in Group-M & 20.55±2.97mmHg in Group-C, P<0.001. Duration of mechanical ventilation was 6.3±2.7 hours in Group-M compared to 14.7±3.5 hours in Group-C(P=0.002). IS was 11.52±2.20 in Group-C compared to 10.84±2.78 in Group-M. 8/39(20.5%) patients in Group-M had IS>10 compared to 22/40(55%) patients in Group-C(P=0.02). Serum Troponin-T and Interleukin-6 levels were lower in Group-M; TNF-α and CPK-MB were similar. ICU & hospital stay were similar. Conclusion: MUF group had higher post-operative hematocrit, decreased duration of mechanical ventilation, lower need for inotropes & lower Interleukin-6 & Troponin-T levels. MUF group had better post-operative outcomes.

A second-order time stepping scheme is developed for the binary fluid-surfactant phase field model coupled with hydrodynamics by using the scalar auxiliary variable approach and pressure correction method. The free energy contains a double-well potential, a nonlinear coupling entropy and a Flory-Huggins potential. By introducing one scalar auxiliary variable, the system is transformed into an equivalent form so that the nonlinear terms can be treated semi-explicitly. The scheme is linear and decoupled; thus, they can be solved eciently. We further prove that the semidiscretized scheme in time is unconditionally energy stable. Numerical experiments are performed to validate the accuracy and energy stability of the proposed scheme.

With the rapid increase of reported COVID-19 cases, German policymakers announced a 4-week “shutdown light” starting on Nov 2, 2020. Applying mathematical models, possible scenarios for the evolution of the outbreak in Germany are simulated. The results indicate that independent of the effectiveness of the current restrictive measures they might not be sufficient to mitigate the outbreak. Repeated shutdown periods or permanently applied measures over the winter could be successful alternatives.

The Lesser White-fronted Goose (Anser erythropus), smallest of the “grey” geese, is listed as Vulnerable on the IUCN Red List and protected in all range states. There are three sub-populations, with the least studied being the East Asian sub-population, shared between Russia and China. The extreme remoteness of breeding enclaves makes them largely inaccessible to researchers. As a substitute for visitation, remotely tracking birds from wintering grounds allows exploration of their summer range. Over a period of three years, and using highly accurate GPS tracking devices, eleven individuals of A. erythropus were tracked from the key wintering site of Dongting Lake, China, to breeding, molting, and staging sites in north-eastern Russia. Data obtained from that tracking, bolstered by ground survey and literature records, were used to model the summer distribution of A. erythropus. Although earlier literature suggests the summer range is patchy, the model confirms a contiguous summer range. The most suitable habitats are located along the coasts of the Laptev Sea, primarily the Lena-Delta, in the Yana-Kolyma Lowland, and smaller lowlands of Chukotka with narrow riparian extensions upstream along major rivers such as the Lena, Indigirka and Kolyma. The probability of A. erythropus presence is related to sites with altitude less than 500 m with abundant wetlands, especially riparian habitat, and a climate with precipitation of warmest quarter around 55 mm and mean temperature of wettest quarter around 14oC. Human disturbance also affects site suitability, with a gradual decrease in species presence starting around 160 km from human settlements. Remote tracking of animal species can bridge the knowledge gap required for robust estimation of species distribution patterns in remote areas. Better knowledge of species’ distribution is important in understanding the large-scale ecological consequences of rapid global change and establishing conservation management strategies.

Nontraumatic bladder rupture is a rare cause of abdominal pain. We presented a case of spontaneous bladder rupture following acute cystits.

Aim: To quantify the utility of the rate-adjusted area under the concentration curve method in increasing the probability of a correct and conclusive outcome of a bioequivalence (BE) trial for highly variable drugs when clearance (CL) varies more than volume of distribution (V). Methods. Data from a large population of subjects were generated with variability in CL and V parameters and used to simulate a two-period, crossover BE trial. The 90% confidence interval for formulation comparison was determined following BE assessment using the area under the concentration curve (AUC) ratio test, and the proposed rate-adjusted AUC ratio method. An outcome of bioequivalent, non-bioequivalent or inconclusive was then assigned in relation to predefined BE limits. Results: We illustrate the utility of the rate-adjusted AUC method for BE testing when CL varies more than V. The approach is expected to enhance the probability of correctly assigning BE or non-BE and to increase study power to further reduce the risk of an inconclusive trial. Conclusions: The rate-adjusted AUC method represents a simple and readily applicable approach to enhance the BE assessment of drug products when CL varies more than V.

Abstract Background and Purpose: Cyclic glycine-proline (cGP) regulates the function of insulin-like growth factor-1 (IGF-1), which is essential for post-natal brain development and adult cognitive function. We evaluated the efficacy of maternally administered cGP on spatial memory and the association with astrocytic plasticity, vascularization and synaptic expressions in the hippocampus of their adult offspring. Experimental Approach: Either cGP or saline was orally administered to Sprague Dawley dams from post-natal days 8-22. Spatial memory was evaluated using Morris Water Maze tests between post-natal days 70-75. Using immunohistochemistry and stereological analysis, we evaluated capillary density, astrocytic processes and expression of synaptophysin and glutamate receptor-1 (GluR-1) in the CA1 stratum-radiatum of the hippocampus. Key Results: Compared to the saline-treated offspring, cGP-treated offspring showed higher path efficiency of entry to the platform zone (p=0.03) and lower average heading errors to the platform zone (p=0.02). Astrocyte processes of cGP-treated offspring were longer and larger with more branches (p=0.03-0.0001) than saline-treated offspring. The density of capillaries (p=0.007) and GluR-1 (p=0.02) were also higher in cGP-treated offspring. The average heading error was negatively correlated with the length (r=-0.69), volume (r=-0.72) and number of astrocytic branches (r=-0.65). Independent of treatment, the changes of astrocyte processes were positively correlated with the density of capillaries (r=0.73-0.78) and expression of GluR-1 (r=0.66-0.68). Conclusion and Implications: The improved spatial memory of cGP-treated offspring after post-natal maternal administration may be mediated via promoting astrocytic plasticity, vascularization and glutamate trafficking. Therefore, cGP may have a role in regulating IGF-1 function during brain development.

As native ranges are often geographically structured, invasive species originating from a single source population only carry a fraction of the genetic diversity present in their native range. This invasion pathway is thus often associated with a drastic loss of genetic diversity resulting from a founder event. However, the fraction of diversity brought to the invasive range may vary under different invasion histories, increasing with the size of the propagule, the number of re-introduction events, and/or the total genetic diversity represented by the various source populations in a multiple-introduction scenario. In this study, we generated a SNP dataset for the invasive termite Reticulitermes flavipes from 23 native populations in the eastern United States and six introduced populations throughout the world. Using population genetic analyses and approximate Bayesian computation (ABC), we investigated its worldwide invasion history. We found a complex invasion pathway with multiple events out of the native range and bridgehead introductions from the introduced population in France. Our data suggest that extensive long-distance jump dispersal appears common in both the native and introduced ranges of this species, likely through human transportation. Overall, our results show that similar to multiple introduction events into the invasive range, admixture in the native range prior to invasion can potentially favor invasion success by increasing the genetic diversity that is later transferred to the introduced range.

Brittle cornea syndrome (BCS) is a rare autosomal recessive disorder characterized by corneal thinning and fragility, leading to corneal rupture, the main hallmark of this disorder. Non-ocular symptoms include hearing loss, but also signs of connective tissue fragility, placing it in the Ehlers-Danlos syndrome (EDS) spectrum. It is caused by biallelic pathogenic variants in ZNF469 or PRDM5, which presumably encode transcription factors for extracellular matrix components. We report the clinical and molecular features of nine novel BCS families, four of which harbor variants in ZNF469 and five in PRDM5. We also performed a genotype and phenotype-oriented literature overview of all (N=85) reported patients with ZNF469 (N=53) and PRDM5 (N=32) variants. Musculoskeletal findings may be the mean reason for referral, and often raise suspicion of another heritable connective tissue disorder such as kyphoscoliotic EDS, osteogenesis imperfecta or Marfan syndrome, especially when corneal rupture has not yet occurred. Our findings highlight the multisystemic nature of BCS and validate its inclusion in the EDS classification. Importantly, gene panels for heritable connective tissue disorders should include ZNF469 and PRDM5 to allow for timely diagnosis and appropriate preventive measures for this rare condition.

Grade Level: K-3rdSuggested Time: 60-90 minutesOverviewAfter watching a scene from the 1966 movie "How the Grinch Stole Christmas," students will use skills in engineering and measurement to "Grinch-proof" dollhouses with an array of materials. The students who design the most successful "Grinch-proof" trap will win a prize of candy canes!VocabularyDesignAnalyzeMeasureRulerForceObjectivesStudents will experiment with the strength of various materials.Students will consider various forces when designing structures.Students will learn how to plan a design.Students will work together to create a machine or that protects or catches a certain item.Students will use measurement tools to create appropriate-sized machines or traps.Students will observe the end result of their machines or traps to determine the functionality of itStudents will analyze ways to improve their machines or traps.Next Generation Science Standards2-PS1-2 Matter and Its InteractionsAnalyze data obtained from testing different materials to determine which materials have the properties that are best suited for an intended purpose.*K-2-ETS1-1 Engineering DesignAsk questions, make observations, and gather information about a situation people want to change to define a simple problem that can be solved through the development of a new or improved object or tool.K-2-ETS1-2 Engineering DesignDevelop a simple sketch, drawing, or physical model to illustrate how the shape of an object helps it function as needed to solve a given problem.3-5-ETS1-1 Engineering DesignDefine a simple design problem reflecting a need or a want that includes specified criteria for success and constraints on materials, time, or cost.3-5-ETS1-3 Engineering DesignPlan and carry out fair tests in which variables are controlled and failure points are considered to identify aspects of a model or prototype that can be improved

Aims To evaluate the effect of pre-RIRS ESWL on the efficiency and safety of RIRS in the treatment of proximal ureter stones. Methods The patients in the study population were divided into 2 groups. Group-1 was composed of patients who had undergone ESWL for proximal ureter stones before RIRS, and Group-2 was composed of patients who directly underwent RIRS without any prior ESWL. The clinical and demographic properties of the patients were analysed in the RIRSearch database. The operative outcomes, peroperative complications, postoperative complications, hospitalization time and the stone-free rates were compared between the groups. Results There were 56 patients in Group-1 and 95 patients in Group-2. The demographic and clinical properties were similar between the groups. The stone-free rates, peroperative complications and postoperative complications were also similar between the groups; however, the fluoroscopy time was significantly higher in Group-1 (p=.043). The cut-off duration of 10 weeks between ESWL and RIRS had reasonable/favourable discriminating ability, with a 51% sensitivity and 88% specificity rate for stone-free status. Conclusion Performing ESWL on the proximal ureter stones before RIRS did not change the efficacy and safety of RIRS. The time between the patient’s last ESWL session and RIRS had a predictive value for stone-free status, but did not have any effect on complications.

Background: There is limited evidence of the relationship between peripheral blood eosinophilia and clinical remission of acute exacerbations of chronic obstructive pulmonary disease (AECOPD) at different ages, especially in elderly patients, which was the objective of the present study. Methods: This retrospective study stratified patients by age (>65 or ≤65 years) and analyzed the relationship between blood eosinophilia (≥2% or <2%) and AECOPD clinical remission at observing time points of 7, 10, 14, 21, and 28 days. Results: Of 703 AECOPD cases analyzed, 616 were elderly (>65 years), 272 of whom had eosinophilic exacerbations. There were statistically significant differences in leukocyte count, high-sensitivity C-reactive protein levels (hs-CRP), and overall and daily hospital costs between eosinophilic and non-eosinophilic AECOPD patients (p<0.05, respectively). In the overall analysis, eosinophilic exacerbation was significantly associated with a higher remission rate at 7 (hazard ratio [HR]=1.457 [1.072, 1.982]), 10 (HR=1.316 [1.108, 1.562]), 14 (HR=1.334 [1.102, 1.615]), 21 (HR=1.326 [1.125, 1.562]), and 28 days (HR=1.254[1.078, 1.459]). The subgroup analysis showed that eosinophilic exacerbation yielded better clinical remission than non-eosinophilic exacerbation in elderly patients (>65 years old) at 7 (HR=1.521 [1.084, 2.136]), 10 (HR=1.319 [1.096, 1.588]), 14 (HR=1.374 [1.118, 1.689]), 21 (HR=1.326 [1.112, 1.582]), and 28 days (HR=1.234 [1.049, 1.451]), while no differences were observed in middle-aged patients (between 45 and 65 years) at all time points (all p>0.05). Conclusion: The eosinophilic phenotype was associated with better clinical remission at 7, 10, 14, 21, and 28 days among elderly but not in middle-aged patients with AECOPD.

Background: Understanding the association between water consumption and mortality is important for guiding consumers and prioritizing dietary guidelines to reduce the risk. Therefore, in the current study, we conducted a systematic review and dose-response meta-analysis of prospective cohort studies to summarize the association between total water and drinking intake and risk of mortality from all causes and CVD. Method: A comprehensive search was performed on PubMed/Medline, Scopus, and ISI Web of Science up to February 2020. The random effects model was used to calculate pooled effect size (EFs) and 95% confidence interval. Result: Seven prospective cohort studies with a total of 116816 reporting 14754 cases of all-cause mortality and 7611 of CVD mortality, were included in the present meta-analysis. No significant association was found between drinking water intake and all-cause mortality (EFs: 0.82; 95% CI: 0.63-1.08, P=0.65) and CVD mortality (EFs: 0.82; 95% CI: 0.63-1.08, P= 0.16). Total water intake was not associated with all-cause mortality (EFs: 0.95; 95% CI: 0.83-1.09, P=0.47). However, indicating a significant inverse association between total water intake and risk of CVD mortality (EFs: 0.84; 95% CI: 0.77-0.93, P<0.001). Linear dose-response meta-analysis revealed a significant inverse association between total water intake and all-cause mortality by an additional one cup per day (Pooled ES: 0.98; 95% CI: 0.97-0.99, P= 0.001). Furthermore, each additional cup of total water intake per day was associated with a 3% lower risk of death from CVD (Pooled ES: 0.97; 95% CI: 0.96-0.98, P<0.001). Conclusion: High consumption of total water is associated with a lower risk of CVD mortality. However total water intake was not associated with an increased risk of all-cause mortality.

Background and purpose: Osteoarthritis (OA) is a chronic and progressive joint disorder characterized by structural damage to one or more joints. However, drugs that could cure or at least stop the progression of this disease are still given no satisfactory outcome. The purpose of this work is to study the potential OA disease-modifying effects of atorvastatin in an experimental model of osteoarthritis and the possible underlining mechanisms if any. Experimental Approach: Seventy-six adult male Sprague-Dawley rats (250-300gms) were used throughout this study. Forty rats were used to assess the effect of atorvastatin in surgically induced OA. While 36 rats were used to assess its anti-inflammatory effect in carrageenan-induced paw edema. In the model of OA; the degree of joint stiffness was assessed by measuring the angle of knee extension besides, the histopathological changes of the OA knee joints and measurement of serum Interleukin-1beta (IL-1β), Matrix metalloproteinase-13 (MMP13), and reduced glutathione (GSH) concentration were biochemically assessed. In the carrageenan-induced paw edema, the paw thickness and pain threshold were assessed in different groups. Key Results: Atorvastatin was found to produce significant improvement of joint stiffness, the histopathological changes, a significant correction in the increased MMP13 and IL1-β, and the decreased GTH in OA rats. Also, atorvastatin showed a significant improvement in both paw thickness and pain threshold. Conclusion and Implications These results present atorvastatin as OA disease-modifying drug worse clinical trials.

Several 2-benzylbenzimidazole opioids (also referred to as ‘nitazenes’) recently emerged on the illicit market. The most frequently encountered member, isotonitazene, has been identified in multiple fatalities since its appearance in 2019. Although recent scheduling efforts targeted isotonitazene, many other analogues remain unregulated. Being structurally unrelated to fentanyl, little is known about the harm potential of these compounds. In this study, ten nitazenes and four metabolites were synthesized, analytically characterized via four different techniques, and pharmacologically evaluated using two cell-based β-arrestin2/mini-Gi recruitment assays monitoring µ-opioid receptor (MOR) activation. Based on absorption spectra and retention times, high-performance liquid chromatography coupled to diode-array detection (HPLC-DAD) allowed differentiation between most analogues. Time-of-flight mass spectrometry (LC-QTOF-MS) identified a fragment with m/z 100.11 for 12/14 compounds, which could serve as a basis for MS-based nitazene screening. MOR activity determination confirmed that nitazenes are generally highly active, with potencies and efficacies of several analogues exceeding that of fentanyl. Particularly relevant is the unexpected very high potency of the N-desethyl-isotonitazene metabolite, rivalling the potency of etonitazene and exceeding that of isotonitazene itself. Supported by its identification in fatalities, this likely has in vivo consequences. These results improve our understanding of this emerging group of opioids by laying out an analytical framework for their detection, as well as providing important new insights into their MOR activation potential.