Induction Protocol Research Articles

Agrobacterium rhizogenes mediated genetic transformation in Ficus religiosa L. and optimization of acetylcholinesterase inhibitory activity in hairy roots

The use of decoctions prepared from stem bark to treat neurodegenerative problems is amongst the widespread applications of Ficus religiosa L., a tree of great ethno-medicinal importance in the Indian sub-continent. The stem bark of the tree has been reported to have significant acetylcholinesterase inhibitory activity (AChEI). Latter is one of the most effective modalities used for the treatment of Alzheimer's disease (AD). The development of any commercial treatment modality for AD from this tree faces serious challenges for sample consistency in terms of quality (being strongly influenced by environmental and developmental stages) and quantity. Hairy root (HR) cultures are an excellent choice for such studies. Since no study has been reported for HR culture systems for F. religiosa , the present study was targeted to develop one. Three different HR lines were induced and cultured on different media. Root extracts, obtained from HRs on different media, were subjected to an AChEI assay to select the best conditions where HRs exerted higher AChEI. The wild-type strain of A. rhizogenes was used to induce hairy roots in different explants. The method of genetic transformation was optimised by changing concentrations of bacterial suspension, acetosyringone and calcium chloride in the infection and co-cultivation medium. The assay for measuring AChEI activity was modified from the assays described by Ellman et al. (1961) and Ingkaninan et al. (2000) . These methods recommend the use of IC 50 (half maximal inhibitory concentration) value for comparing AChEI activities in different samples. We developed a protocol for induction and long-term maintenance of HR cultures of F. religiosa L. In vitro conditions were optimized where hairy root extracts exerted higher AChEI activity (IC 50 value of 0.16 µg/ml) compared to that in the stem segments of the mother plants (IC 50 value of 0.49 µg/ml). The present work has been granted a patent by the Indian Patent Office, Government of India (Patent No360050, Date of grant 02/03/2021) (Siwach and Gill, 2021). The present study confirms the utility of F. religiosa in AD treatment and provides a standardized protocol for large-scale production of HR cultures (75ml medium in Erlenmeyer flask of 250 ml volume) that have higher AChEI activity; these cultures offer excellent platforms for future characterization and commercial preparation.

Ovarian Response and Fertility after Short-Term Progestagen/eCG Treatments Are Compromised in Nulliparous Sheep during Non-Breeding Season.

The objective of this investigation was to determine the ovarian response, fertility, and prolificacy of nulliparous sheep when compared to multiparous sheep after a short-term (7 days) CIDR/eCG treatment which was administered during the non-breeding season. All the multiparous sheep, whereas only 54% of the nulliparous ewes, showed signs of estrus. However, 81.8% of the multiparous sheep and 100% of the nulliparous ewes ovulated. Fertility was also low after short-term progesterone treatments during the anestrous season in maiden sheep (30.8 vs. 72.7% in multiparous ewes). Such results indicate significant differences in the response to CIDR/eCG protocols for induction and synchronization of estrus and ovulation between nulliparous and multiparous sheep during the non-breeding season.

In Vitro Evaluation of the Effects of BAP Concentration and Pre-Cooling Treatments on Morphological, Physiological, and Biochemical Traits of Different Olive (Olea euorpea L.) Cultivars

In vitro propagation of olive (Olea euorpea L.) always remained a challenging task due to its woody nature and oxidation of culture. The current study intended to optimize shoot induction and proliferation protocol for different cultivars (“Leccino”, “Gemlik”, “Moraiolo” and “Arbosana”) of olive-on-olive media (OM) provided with different concentrations (0, 0.5, 1.5, and 2.5 mgL−1) of 6-benzylaminopurine (BAP) by pre-exposing their explants (nodal segments) with different regimes (0, 24, and 48 h) of cooling. The impacts of treatments were evaluated on morphological (shoot induction percentage, primary shoot length, number of leaves shoot−1, and number of shoots per explant−1), physiological (total chlorophyll, carotenoids, CO2 assimilation, and proline), biochemical (primary and secondary metabolites) attributes of cultivars after 50 to 60 days of culture. Data recorded were subjected to statistical analysis. All traits depicted significant increases in all genotypes with increasing pre-cooling treatments and increasing supplementations of 6-benzylaminopurine (BAP). This increase was the highest for the interaction of 48 h pre-cooling and 2.5 mgL−1 BAP concentration. Moreover, correlation analysis of all traits revealed significant paired association among them in a positive direction, while principal component analysis (PCA) revealed the extent of association varied with types of treatments and the nature of genotypes. Among cultivars, Arbosana depicted more dramatic changes in morphological traits, physiological attributes, and biochemical contents due to varying interactions of pre-cooling and BAP treatments as compared to Moraiolo, Gemlik, and Leccino with in vitro systems.

Association of anti-mullerian hormone and free androgen index level on response to clomiphene citrate in PCOS infertile women

BackgroundPolycystic ovary syndrome (PCOS) is the leading cause of anovulatory infertility. Clomiphene citrate, insulin-sensitizing drugs, aromatase inhibitors, gonadotropins, or laparoscopic ovarian drilling are various methods used for ovulation induction in women with PCOS. PCOS women with high levels of anti-mullerian hormone (AMH) and free androgen index (FAI) do not respond well to ovulation induction. This prospective observational study explores the relationship between FAI and AMH levels on ovarian response to clomiphene citrate in infertile women with PCOS. MethodsThis prospective observational study included 40 infertile with PCOS who underwent ovulation induction with clomiphene citrate with dose ranging from 50 to 150 ​mg. Participants were classified into four phenotypes by NIH(National Institute of Health) consensus panel criteria. The clinical and endocrine parameters of participants who were sensitive to clomiphene were compared to those who were resistant. ResultsThe most common phenotype was A, with all three features of PCOS: hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. There was no significant difference in clinical and endocrine parameters among the different phenotypes of PCOS except AMH and FAI values. The mean FAI was 9.39 ​± ​1.11 and AMH 7.26 ​± ​0.48 (ng/ml) in clomiphene resistant and 5.31 ​± ​1.93 and 3.69 ​± ​1.84 (ng/ml) respectively in clomiphene-sensitive women. Women with FAI > 7.5 and AMH >7 ​ng/ml might be resistant to clomiphene. ConclusionFAI and AMH values were significantly higher in women resistant to clomiphene induction. AMH and FAI may help women with PCOS to tailor their ovulation induction protocol.

Failed Induction of Labor and Associated Factors Among Women Who Undergone Induction of Labor in Jimma Medical Center, Ethiopia: A Hospital-Based Cross-Sectional Study

BACKGROUNDAlthough induction of labor is intended to achieve a safe vaginal delivery, its process is not always successful. Failed induction of labor leads to an increased risk of complication for both mother and fetus. Thus, this study aimed to assess the proportion of failed induction of labor and associated factors among women who undergone induction of labor from September 1, 2019 to August 31, 2020 in Jimma Medical Center (JMC), southwest Ethiopia.METHODSA hospital-based retrospective cross-sectional study was conducted on 243 charts of women who had induction of labor in JMC selected by a simple random sampling method from January 10 to 30, 2021. The collected data were entered into Epi-data version 4.2 and analyzed by SPSS version 23. Bivariate and multivariable logistic regression analyses were done to test the association. Adjusted odds ratio at 95% confidence interval (CI) andP-value <.05 was used to ascertain statistical significance.RESULTSThe proportion of failed induction of labor in JMC was 16.9%. Maternal age ≥ 30 years (AOR 7.57; 95%CI = 2.88, 19.9) at (P-value = .000), elective induction (AOR .39; 95% CI = .16, .98) at (P-value = .045), and unfavorable bishop score (AOR 6.94; 95%CI = 2.35, 20.4) at (P-value = .000) were factors statistically significantly associated with failed induction of labor.CONCLUSIONSThe proportion of failed induction of labor was relatively low in the study area. Advanced maternal age, elective induction, and unfavorable bishop score were associated with failed induction of labor. Thus, the hospital should strictly follow the protocol for induction of labor, and candidates of emergency induction of labor should be counseled thoroughly so as to be well prepared psychologically for the labor thereby reducing the risk of failed induction.

Improved Survival of Adolescents and Young Adults (AYA) Patients Aged 14-40 Years with First Relapse of Acute Lymphoblastic Leukemia Using Pediatric-Inspired Chemotherapy Salvage Protocol (R3 induction protocol)

Background: Although survival of adolescent and young adults (AYA) with acute lymphoblastic leukemia (ALL) has improved using pediatric-inspired protocols, the outcomes of those who relapse remain poor using adult chemotherapy salvage-regimens that have historically incorporated high dose Ara-C based regimen, with long term-survival of less than 10%. The CR rate with blinatumomab for salvage in adult ALL is 34% with a median survival of 7.7 months, with an OS of less than 30% at 18 months. In contrast, pediatric salvage regimens have historically been based on multi-agent therapy and have reported superior outcomes. The ALLR3 protocol for relapsed ALL in patients up to the age of 18 years reported a CR rate of 97% and an overall survival of 72%. We therefore aimed to analyze the outcomes of young adolescents and adults treated on the R3 regimen +/- blinatumomab. Methods: Seventeen AYA patients with Philadelphia-negative ALL aged between 14 and 40 years who relapsed after first-line therapy and treated with ALLR3-protocol were identified. Relapses were classed as very early (<18 months from diagnosis) early (>18m from diagnosis and <6 months from end of therapy) and late (>6 months from end of therapy). The protocol consisted of vincristine (1.5mg/m2 IV D3,10,17,24); mitoxantrone ( 10mg/m2 IV d1 &2); PEG-asparaginase (1000 u/m2 IV d3 & 17); dexamethasone (10mg/m2 BID day d1-5 and d15-20); and intrathecal methotrexate (12mg d1 & d8) were identified. Patient aged 30-40 years received L-Asparaginase instead of PEG-Asparaginase, and 8mg/m2 of dexamethasone instead of 10mg/m2. Patients' characteristics, response rate (RR) and overall survival (OS) were analyzed. Patients who achieved minimal residual disease (MRD) negative CR proceeded with an allogeneic transplant. MRD-positive patients received blinatumomab followed by a transplant. Results: The median age at relapse was 16 years (14-40 years), 88.2% of patients were B-cell ALL and 11.8% were T-cell ALL. Most patient had standard cytogenetics on presentation (76.5%) and were CNS negative (82.4%). Relapses were very early, early or late in 2 (12%) , 5 (29%) , and 7 (41%) of cases, respectively. Relapses were isolated bone marrow (BM), isolated extramedullary (EMD) or combined BM and EMD in 14 (82%),1 (6%) and 2(12%) cases, respectively. Overall complete remission (CR) rate was 60% at the end of R3 induction; CR with negative minimal residual disease (MRD negative, <10−4 cells) at the end of R3-induction was achieved in 47.1% of patients, while 11.8% of patients had CR with positive MRD (≥10−4 cells) who received Blinatumumab thereafter with eradication of MRD. Refractory disease was seen in 41.2% of patients. Almost all patients who responded to R3 proceeded to allogeneic stem cell transplant (88.8%). Eight patients died (47%), but only one patient died because of treatment-related sepsis, while the other 7 patients died because of disease-related causes. At a median follow up of 4 years, OS was 47.3%. Conclusion: Compared to traditional adult-based chemotherapy regimens and antibody therapies used in relapsed ALL, the use of pediatric-inspired protocol (R3) in relapsed AYA-ALL patients showed favorable response rates and overall survival. Further trials are required of pediatric inspired salvage regimens, preferably incorporating antibody therapies in adult patients, especially in 1st relapse. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Impact of Additional Intensive L-Asparaginase Therapy during Consolidation Phase for High-Risk Acute Lymphoblastic Leukemia: Results of a Randomized Controlled Trial in the AIEOP-BFM ALL 2009 Protocol

Introduction Treatment of childhood Acute Lymphoblastic Leukemia (ALL) with unfavorable features remains a major challenge. An earlier intervention, aiming at lowering Minimal Residual Disease (MRD) levels, might reduce the risk of relapse and the indication to allogeneic hematopoietic stem cell transplantation (HSCT) in 1st complete remission (CR). The AIEOP-BFM ALL 2009 protocol included a randomized study for patients with high-risk (HR) features, to investigate if an intensive and continuous exposure to pegylated L-Asparaginase (Peg-L-ASP), given on top of BFM consolidation phase IB, could decrease the levels of MRD and to influence long term outcome. Patients and methods From June 1, 2010 to February 28, 2017, 6136 patients with Ph’ negative ALL, aged between 1-17 years, were enrolled in the AIEOP-BFM ALL 2009 Study (Eudract number 2007-004270-43). Participating study centers were in Australia, Austria, Czech Republic, Germany, Israel, Italy and Switzerland.1097 were classified as HR at the end of induction phase IA (day 33), on the basis of evidence ofKMT2A-AFF1 rearrangement, hypodiploidy (<45 chromosomes, or DNA-index <0.8), prednisone poor response (PPR), poor bone marrow (BM) response at day 15 (i.e. ≥10% blasts by MFC), failure to achieve morphological CR. Treatment consisted of a 7-day prephase with prednisone and 1 intrathecal dose of methotrexate (ITMTX), an induction protocol IA with prednisone or dexamethasone, vincristine, daunomycin, Peg-L-ASP (2 doses), ITMTX and, only in PPR T-ALL cyclophosphamide. Patients eligible were randomized to receive or not 4 weekly doses of Peg-L-ASP (2500 IU/sqm x 4) on top of consolidation phase IB, which included 6-MP, cyclophosphamide, cytosine arabinoside and ITMTX. Thereafter, patients received 3 blocks of non-cross-resistant drugs, 3 reinduction phases with or without cranial radiotherapy (12 Gys), and maintenance therapy till 24 months from diagnosis. The primary endpoint was PCR-MRD reduction to less than 5x10-4at the end of phase IB; secondary endpoint was Event-Free Survival (EFS). Results Overall, 1097 patients were at HR at the end of induction phase IA and 809 (79.2%) were randomized to receive (404) or not (405) 4 doses of Peg-L-Asp during consolidation phase IB. The primary endpoint could be assessed in 732 patients (90.5%). By ITT analysis, the proportion of patients with PCR MRD ≥ 5x10-4 at the end of consolidation phase IB in the experimental arm (EA) vs the control arm (CA) was 13.9% vs 17.0% with no significant difference (p-value=0.25).There were more toxic deaths during consolidation phase IB among patients randomized to Peg-L-Asp (n=14, 12 of them due to infections), vs controls (n=1): 7 of them, all in the EA, however, occurred before starting the randomized treatment. Deaths in CR after phase IB were 24 vs 24 in the EA vs CA (of whom 10 vs 12 were after HSCT in CR1, performed in 87 and 90 patients respectively). In the EA and CA the overall 5-year EFS by ITT was 70.4% (2.3) and 75.0% (2.2) (p-value=0.18), the 5-year cumulative incidence of death in CR was 9.5% (1.5) vs 5.7% (1.2) (p-value=0.08) and that of relapse was 17.7% (1.9) vs 17.2% (1.9), respectively (p-value=0.94). Non HSCT related deaths were due to infections in 23/28 in the EA, and in 10/13 in CA patients. The 5-year overall survival probability was 81.5% (2.0) and 84.0% (1.9) respectively (p-value=0.25). Discussion In this randomized study, results (ITT analysis) show a very modest, not significant decrease in the percentage of patients with high MRD in the EA, and no significant difference in EFS between the two arms. The trial results suggest that the additional administration of intensive Peg-L-Asp in phase IB is associated with higher, although not statistically significant, mortality in CR and very similar relapse incidence. However, some fatal events in phase IB occurred very early after randomization, before the start of the planned treatment with Peg-L-Asp. Importantly, in both analyses, by ITT and by treatment given, there is no evidence of benefit on reduction of relapses by the additional therapy with Peg-L-ASP in phase IB. These findings indicate that an early intensification of chemotherapy with Peg-L-Asp in patients defined as HR does not improve the outcome of children with HR ALL treated according to AIEOP-BFM regimen, thus suggesting that further improvements with conventional chemotherapy might be difficult in the context of intensive treatment protocols. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Inferior Outcome in Non-Adult Young Adolescent B-Cell Acute Lymphoblastic Leukemia Exhibiting Obesity

Background: Previously, obesity has been associated with increased risk of relapse in aggressive lymphoproliferative disorders [Teegaparavu. ASH.2021]. The mechanism is largely unknown. However, attenuated chemosensitivity driven by unbalanced adiponectin/leptin regulation and obesity specific tumor microenvironment defects represent interesting "reversible” factors. Additionally, it is possible that B-cells aberrantly proliferate under obesity induced cytokine/chemokine influence. In pediatric patients (pt), obesity correlates with suboptimal Measurable Residual Disease (MRD) after induction. High insulin levels and PI3K/ATK/mTOR activation render anti-apoptotosis and chemorefractoriness [Bresin. Cell Death & Disease. 2016]. In this study, we investigate the effect of obesity on overall survival (OS) and event free survival (EFS) in a cohort of adult B-cell Acute Lymphoblastic Leukemia (B-cell ALL) older than 40 years with and without pre-treatment obesity. Methods: After IRB approval, B-cell ALL patients (pt) treated under standard induction protocols were selected at the Instituto de Cancerologia in Mexico and Baylor College of Medicine Institutions from 2010 to 2019. Obesity was diagnosed according to standard definitions based on body mass index (BMI) to include class I, II and III subgroups (BMI>30). Detectable modification in OS and EFS were estimated by using the Kaplan Meier Method. Descriptive statistics summarized results. To assess the independent effect of intervening variables on outcomes of interest, cox regression model was performed. SAS software was used for analysis. Results: 46/71(64.7%) and 31/71 (43.6%) pt exhibited or not obesity. Median age for obese and not obese pt was 52 years (y) vs 56 y, respectively. WBC, LDH and blast at disease onset were 43.2 K/UL v 20.1 K/UL, p=0.02; 1009 U/L v 600 U/L, p= 0.2; 62.8% v 60.6% for obese vs not obese pt, respectively. Incidence of high risk genomic did not statistically differ when accounting for t (9; 22), hypodyploid karyotype, P53 and 11q23 abnormalities. OS was 779 days (d) v 6429 d, in pt with and without obesity, p=0.03 [HR=0.43, CI 0.2-0.93] [Fig 1]. When considered relapse risk, EFS was 252 d v 936 d, in pt with and without obesity, respectively, p= 0.04 [HR=0.50, CI 0.22-0.99] [Fig 2]. Univariate analysis revealed that high BMI [p=0.001] and higher WBC [p=0.04] were predictor for OS and EFS. Cox regression model demonstrated that high WBC independently predicted inferior OS and EFS, p=0.002, each. Conclusions: Obesity is associated with inferior OS and EFS in adult non- AYA pt diagnosed with B-cell ALL. High BMI >30 is associated with enhance risk of relapse, which may explain, at least in part inferior OS in our pt cohort. Higher WBC at disease onset suggests that obesity is linked with blast proliferation efficiency. Deeper understanding of obesity induced metabolic abnormalities leading to B-cell ALL chemorefractoriness is needed. Weight reduction via caloric restriction and exercise may represent interesting interventions to improve clinical outcome of leukemic pt with high BMI. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

NEUROSCIENCE OF SLEEP AND COGNITION: INVESTIGATING THE EFFECTIVENESS OF THE NEW DREAM LUCIDITY INDUCTION PROTOCOL – A PILOT STUDY

The lack of standardized, efficient, and dependable lucid dream induction methods is one of the major barriers to studying the clinical and non-clinical applications of dream lucidity. Although the interest in therapeutic usage of lucid dreaming strategies in neuroscience of consciousness studies has increased in recent decades, many of the current induction approaches appear to not be replicable reliably. In this study, a new protocol for bringing on lucid dreaming was created, implemented, analyzed, and contrasted with two distinct other approaches. Techniques to induce dream lucidity include reality testing methods, the wake-and-go-to-bed technique, mnemonic initiation, senses-initiation, and a combination of the above. Before beginning their 2 weeks of lucid dream induction new protocol study, a total of 36 participants filled out a pre-test survey, maintained a dream recall diary and sleep log for the duration of the experiment, and completed a final survey. The findings showed that both the mnemonic induction and senses-initiation techniques were mildly effective in bringing about lucid dreaming. The new hybrid protocol, combining the reality check stra- tegy with dream recall amplification, performed significantly better. For lucid dream induction to work optimally, participants needed to become more proficient at remembering their dreams after waking up, so they were requested to maintain a sleep journal for 7 days before the experiment and throughout the study to enhance their ability to dream recall. There was no correlation found between the quality of one's sleep and the frequency of dream lucidity. The results demonstrated that the new induction protocol was more successful than the other two investigated methods. There are many clinical applications for dream lucidity, such as treatment of recurring and debilitating nightmares, narcolepsy, and PTSD. Therapeu- tic potential also includes future usage for anesthesia awareness and locked-in syndrome. As such, further dream lucidity research remains important.

Establishment and optimization of a new model organism to study early land plant evolution: Germination, cultivation and oospore variation of Chara braunii Gmelin, 1826.

For studying land plant evolution, the establishment and optimization of model organisms representing streptophytic algae, sister to land plants, is essential. Long-term cultivation experiments with Chara braunii S276 were performed over 8 years, since 4 years (Nov. 2018) under constant conditions. Additionally, short-term experiments for optimization of culture conditions were performed with three strains of C. braunii (S276, NIES-1604 and Lausiger Teiche, LaT-2708). Germination success after application of sterilization agents, addition of gibberellic acid and under different incubation conditions with respect to pre-treatment, irradiance regime and substrate was investigated in order to develop protocols for generative cultivation of at least unialgal cultures. The resulting cultivation protocols for C. braunii S276, allowing maintenance of vegetative as well as generative cultures are presented in detail, including protocols for germination induction and growth of sterilized and unsterilized oospores.

Solanum tuberosum Microtuber Development under Darkness Unveiled through RNAseq Transcriptomic Analysis.

Potato microtuber (MT) development through in vitro techniques are ideal propagules for producing high quality potato plants. MT formation is influenced by several factors, i.e., photoperiod, sucrose, hormones, and osmotic stress. We have previously developed a protocol of MT induction in medium with sucrose (8% w/v), gelrite (6g/L), and 2iP as cytokinin under darkness. To understand the molecular mechanisms involved, we performed a transcriptome-wide analysis. Here we show that 1715 up- and 1624 down-regulated genes were involved in this biological process. Through the protein-protein interaction (PPI) network analyses performed in the STRING database (v11.5), we found 299 genes tightly associated in 14 clusters. Two major clusters of up-regulated proteins fundamental for life growth and development were found: 29 ribosomal proteins (RPs) interacting with 6 PEBP family members and 117 cell cycle (CC) proteins. The PPI network of up-regulated transcription factors (TFs) revealed that at least six TFs-MYB43, TSF, bZIP27, bZIP43, HAT4 and WOX9-may be involved during MTs development. The PPI network of down-regulated genes revealed a cluster of 83 proteins involved in light and photosynthesis, 110 in response to hormone, 74 in hormone mediate signaling pathway and 22 related to aging.

Central injection of abscisic acid attenuates mood disorders induced by subchronic stress in male mice.

Stressful life increases the risk of mental and psychological disorders and cognitive deficits. Abscisic acid (ABA) is a plant hormone that has been recently discovered in mammalians. ABA is produced in response to stressful stimuli and it can reduce anxiety-like behaviors and depression and improve cognitive function. This study was designed to evaluate the effects of microinjection of ABA on depression, anxiety, passive avoidance learning and memory deficits induced by subchronic stress. ABA (10 and 15 g/mouse, i.c.v.) was administered one week after recovery period for 4 consecutive days. A three-session forced swimming test (FST) protocol for induction of subchronic stress was administered to the mice. Exploratory, anxiety-like behavior, depression and cognitive function were assessed 24 h after the last swim stress session. The results indicated that ABA (15 g/mouse) could ameliorate anxiety and depression induced by FST. In addition, ABA had no effect on the subchronic stress-induced cognitive impairments. Taken together, the results suggest that ABA could improve anxiety and depression induced by subchronic stress.

Abstract 15023: Minimum Electrode Spatial Resolution for Arrhythmia Rotor Detection and Localization

Introduction: Efficacy of electrotherapy or ablation therapy for treatment of arrhythmias is dependent on accurate detection of arrhythmia focal or reentrant drivers. However, success rates have been inconsistent, with frequent arrhythmia recurrence. A likely reason is the limited current spatial resolution of mapping devices, which averages 4 mm. The goal of this study was to determine the minimum spatial resolution necessary to identify and localize arrhythmia reentrant drivers (rotors). Methods: We conducted dual-sided (epicardial and endocardial) optical mapping studies in 12 donor human right ventricular wedge preparations subjected to arrhythmia induction protocol (S1S1 restitution and/or 50Hz burst pacing). The resulting arrhythmia data (0.7mm spatial resolution) was down sampled to different spatial resolutions (1.4-7mm) and up sampled back to full resolution. Rotors were tracked for each sub-resolution and compared to the rotors in the original data. Further comparisons were made according to arrhythmia type, sex, anatomical region, and mapped surface. Results: Rotor detection accuracy for currently used mapping devices was found to be 57±4%. Localization accuracy was 61±7%. For the current average spatial resolution, clinical rotor detection and localization accuracies can be expected to fall within a confidence interval of 47-67% and 46-75%, respectively. For the spatial resolutions tested, detection accuracy was above 80% only for a resolution of 1.4 mm, while localization accuracy reached that value at 2.1 mm. Moreover, the arrhythmia type affected detection and localization accuracies, which decreased consistently with loss of spatial resolution for unstable arrhythmias and had steadier values for stable arrhythmias. When comparing mapped surfaces, rotor incidence was higher in the endocardium, likely due to the presence of adipose tissue on the epicardial surface. Differences in sex and anatomical region had no statistically significant impact. Conclusions: The spatial resolution of current devices is insufficient to map rotors accurately, especially for unstable arrhythmias. For both detection and localization accuracies to be above 80%, a resolution of at least 1.4 mm is required.

Is stress colorblind? Exploring endocrine stress responses in intergroup contexts using a virtual reality-based Trier Social Stress Test (TSST-VR)

Are social stress reactions dependent on the group identities of interaction partners? This study explored the role of ethnic context in modulating endocrine stress responses using a virtual reality (VR)-based adaptation of a standardized stress induction protocol, the Trier Social Stress Test (TSST-VR). Previous research found no clear link between endocrine stress response and ethnic context in the TSST, but conclusions remain limited due to the quasi-experimental nature of manipulating ethnic context in real-life face-to-face interactions. The VR adaptation of the TSST circumvents quasi-experimental limitations and thus provides a first, randomized-controlled investigation of the effects of ethnic context on endocrine stress responses. Forty-three men participated in the study, facing either an ingroup (“White”) or an outgroup (“Arab”) panel of interviewers. As expected, the TSST-VR produced physiological and subjective stress reactions. However, endocrine stress reactions occurred independent of interviewer ethnicity and could not be predicted based on implicit bias, explicit prejudice, or prejudice-related appearance concerns. Other physiological and subjective stress reactivity parameters also remained comparable across intergroup conditions. Implications for stress research are discussed.

Prophylactic action of ayahuasca in a non-human primate model of depressive-like behavior.

Observational studies of long-term users of ayahuasca, an Amazonian psychedelic brew, suggest an increase in resilience via improvements in emotion and cognition. Ayahuasca has also demonstrated clinical antidepressant effects in human and animal studies; however, its potential prophylactic action in depression has not been previously studied. Therefore, this experimental study sought to evaluate the potential prophylactic effects of repeated and long-term ayahuasca use, via the modulation of resilience, in a non-human primate animal model, Callithrix jacchus, subjected to a protocol for induction of depressive-like behavior. For the formation of the study groups, some juvenile marmosets were kept in their family groups (GF = 7), while for the two experimental groups, the animals were removed from the family and kept socially isolated. Then, part of the isolated animals made up the group in which ayahuasca was administered (AG, n = 6), while for others, no intervention was made (IG, n = 5). AG animals took ayahuasca (1.67 mL/300g body weight) at weeks 4 (before isolation), 8, and 12 (during isolation) of the study. More adaptive stress response was observed for the AG when compared to the IG. The AG showed higher cortisol reactivity and fecal cortisol levels than IG, while both measures were similar to FG. Moreover, AG animals showed no signs of anhedonia and no increase in chronic stress-related behaviors, which were expressed by the IG. Thus, ayahuasca seems to promote the expression of resilient responses, indicating a prophylactic action, buffering the emergence of depressive-like behaviors and cortisol alterations associated with major depression. These results are encouraging for further research on the prophylactic use of psychedelics to prevent psychopathologies associated with chronic stress.

An objective assessment in newly diagnosed multiple myeloma to avoid treatment complications and strengthen therapy adherence.

In heterogeneous multiple myeloma (MM) patients treatment decisions are challenging. The hypothesis was that adaptation of treatment intensity (dose reduction [DR] vs. none) according to an objective risk score (revised-myeloma comorbidity index [R-MCI]) rather than physician judgement alone may improve therapy efficacy and avoid toxicities. We performed this study in 250 consecutive MM patients who underwent a prospective fitness assessment at our center, after having received induction protocols based on physicians' judgement. DR, serious adverse events (SAE), response, progression-free survival (PFS) and overall survival (OS) were compared in fitness (fit, intermediate-fit, frail), age (<60, ≥70 years [y]) and therapy intensity subgroups at baseline and follow-up. Fit and <60 y patients were mostly treated with full intensity, whereas frail and ≥70 y patients usually received DR. Hematological and non-hematological SAE were more frequently seen in frail versus ≥70 y patients. Dose adaptations were mainly necessary in frail patients. OS and PFS were similar in fit and intermediate-fit but significantly worse in frail patients (P=0.0245/P<0.0001), whereas in age-based subgroups, OS and PFS differences did not reach significance (P=0.1362/P=0.0569). Non-hematological SAE were another negative predictor for impaired OS and PFS (P=0.0054/P=0.0021). In the follow-up performed at a median of 11 months after the first fitness assessment, the R-MCI improved or remained stable in 90% versus deteriorated in only 10% of patients. In conclusion, separation by R-MCI/frailty-defined subgroups was superior to age-based subgroups and can be used to improve tailored treatment. Fitter patients benefit from intensive therapies, whereas frail patients bear a need for initial DR.

Time between inhibitor detection and start of immune tolerance induction: Association with outcome in the BrazIT study

BackgroundImmune tolerance induction (ITI) is the treatment of choice for eradication of anti‐factor VIII (FVIII) neutralizing alloantibodies (inhibitors) in people with inherited hemophilia A and high‐responding inhibitor (PwHA‐HRi). The association between ITI outcome and time elapsed between inhibitor detection and start of ITI (∆tinhi‐ITI) is debatable. ObjectiveThe aim of this study was to evaluate this association among a large cohort of severe PwHA‐HRi. MethodsSevere (factor VIII activity level <1%) PwHA‐HRi on ITI (n = 142) were enrolled in 15 hemophilia treatment centers. PwHA‐HRi were treated according to the Brazilian ITI Protocol. ITI outcomes were defined as success (i.e., recovered responsiveness to exogenous FVIII) and failure (i.e., no responsiveness to exogenous FVIII and requirement of bypassing agents to control bleeding). ResultsMedian ages at inhibitor detection and at ITI start were 3.2 years (interquartile range [IQR], 1.6–8.1) and 6.9 years [IQR, 2.6–20.1), respectively. PwHA‐HRi were stratified according to ∆tinhi‐ITI quartiles: first (0.0–0.6 year), second (>0.6–1.7 year), third (>1.7–9.2 years), and fourth quartile (>9.2–24.5 years). The overall success rate was 65.5% (93/142), with no difference among first, second, third, and fourth quartiles (62.9%, 69.4%, 58.3%, and 71.4%, respectively) even after adjusting the analyses for potential confounders. ConclusionIn conclusion, delayed ITI start is not associated with failure of ITI in PwHA‐HRi. Therefore, ITI should be offered for these patients, regardless of the time elapsed between the detection of inhibitor and the ITI start.

A DISCUSSION OF THE REFRACTORY PERIOD IN VENOM IMMUNOTHERAPY

<h3>Introduction</h3> Refractory periods are inconsistently observed phenomena that follow hypersensitivity reactions wherein a reaction is followed by a period of time during which additional reactions cannot be triggered. We discuss a case that challenges this theory. <h3>Case Description</h3> A 56-year-old male with Diabetes Mellitus II, Hypertension, and HIV presented with history of two prior episodes of fire ant stings leading to anaphylaxis. His baseline tryptase level was 6.1 ug/mL and sIgE to fire ant (Solenopsis invicta) was 42.3 kU/L. The patient completed the immunotherapy ultrarush induction protocol using 1:100wt/v Solenopsis invicta whole body extract (WBE) as the maintenance (1:1v/v) concentration. One hundred minutes after completion of the protocol, he developed throat tightness, nasal congestion, diffuse pruritus, and urticaria. Symptoms resolved after two injections of 0.5mL intramuscular epinephrine. One week later, he received 0.2 mL of 1:1 v/v and developed stridor, productive cough, and injected sclera 30 minutes after injection. Symptoms resolved with intramuscular epinephrine. The following week, he received another injection (decreased to 0.5 mL of 1:10 v/v) without incident. He continued weekly buildup to maintenance dose and subsequent monthly injections without additional reactions. <h3>Discussion</h3> In this case, completion of an ultrarush induction protocol and anaphylaxis failed to result in a refractory period of unresponsiveness, allowing for a second anaphylactic reaction to S. invicta WBE one week after desensitization should have been achieved. Additional research is needed to reliably identify patients experiencing a refractory period following an anaphylactic event. This would improve the accuracy of allergy testing and has the potential to make desensitization safer.

Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols

In vivo animal models can provide worthy information on various aspects of asthma mechanism and pathogenesis. The genetic predisposition and phenotype of mice may affect the immune response itself. Here we compare the early immune response to Der p 2 or HDM allergen extract upon injection and inhalation in BALB/c and C57BL/6 mice. Female C57BL/6 and BALB/c mice were immunized with Der p 2 allergen subcutaneously followed by inhalation of Der p 2 or HDM extract. After challenge, the mice were euthanized; blood, bronchoalveolar lavage (BAL), spleens and lungs were collected. Cells from BAL were identified by May-Grünwald Giemsa staining and lung leukocyte populations were analyzed by flow cytometry. Serum antibody levels of Der p 2 specific IgE, IgG, IgG1 and IgG2a were assessed by ELISA, and cytokine secretion (IL-4, IFN-γ and IL-10) was evaluated upon stimulation with Der p 2 or HDM extract. The Th2 immune response was confirmed by elevated allergen-specific immunoglobulin E (IgE) and the allergic reaction was evidenced by infiltration of eosinophils and/or neutrophils into BAL. We found that BALB/c mice were inefficient in integrating local with systemic immune response, evidenced by almost no IgG or IgE production upon one subcutaneous injection and subsequent inhalation of Der p 2 allergen; also, the bronchoalveolar lavage infiltrate in these mice consisted of neutrophil infiltration, unlike C57BL/6 mice in which eosinophilic infiltrate predominated. The differences between BALB/c and C57BL/6 mice strains could be exploited for generating different types of responses to the Der p 2 allergen.

GnRH administration after estrus induction protocol decreases the pregnancy rate of recipient ewes following transfer of frozen-thawed embryos

The objective of this study was to investigate the effects of GnRH on the pregnancy rate and the embryo survival rate of recipient ewes following embryo transfer. Estrus was synchronized in 83 nulliparous Huyang ewes using intravaginal sponges impregnated with 45 mg flurogestone acetate. The sponge was left in the vagina for 12 days followed with an injection of 330 IU of eCG at sponge removal. The treatment group (n = 34) was subjected 48 h after sponge removal to intramuscular administration with 16 μg of the GnRH agonist triptorelin and the control group (n = 49) with 1 mL of saline solution. Embryo transfer was performed by laparoscopy on day 8 after sponge removal, which transferred frozen-thawed embryos of Suffolk ewes directly to the anterior third of the corpus luteum ipsilateral uterine horn of the recipient ewe.Each recipient with at least one normal corpus luteum received one grade I embryo. The proportion of ewes with at least one normal corpus luteum in the control group was significantly lower compared to the treatment group (73.5 % vs 91.2 %, P < 0.05). In total, 62 embryos were transferred into 62 ewes and 29 lambs were born. The pregnancy rate (59.4 % vs 33.3 %, P < 0.05) and the embryo survival rate (56.3 % vs 30.0 %, P < 0.05) in the control group were significantly higher compared to the treatment group. In conclusion, a single dose of GnRH administered 48 h after sponge removal significantly decreased the pregnancy rate and the embryo survival rate of recipient ewes following transfer of frozen-thawed embryos.