1st Trial Research Articles

Divine noni's protective impact on Swiss albino mice's short-term memory impairment caused by cyclophosphamide: A behavioral and biochemical approach

Cyclophosphamide (CYL) is a first-line cancer chemotherapeutic agent widely used for the treatment of cancer that has severe toxic effects. The primary mechanism by which CYL induces toxicity through free radical generation. Morinda citrifolia (Noni) fruit juice is an herbal remedy documented to have antioxidant properties. The aim of the current study was to investigate the protective effect of noni against CYL-induced memory impairment in Swiss albino mice. Treatment schedule: Group 1: Normal: Received vehicle; Group 2: CYL treatment: Received CYL (40.0 mg/kg b.w. i.p.) on day one; Group 3: NJ treatment: Received NJ (360 mg/b.w. p.o.) once daily for 14 days. Group 4: DNG treatment: DNG (360 mg/b.w. p.o.) once daily for 14 days, Group 5: NJ + CYL treatment: Received CYL (40.0 mg/kg b.w. i.p.) on day one and after half an hour of received NJ (360 mg/b.w. p.o.) once daily for 14 days. Group 6: DNG + CYL treatment: Received CYL (40.0 mg/kg b.w. i.p.) on day one and after half an hour received DNG (360 mg/b.w. p.o.) once daily for 14 days. Mice were subjected to the Morris water maze (MWM) challenge for two weeks as part of a behavioral study. Short-term memory impairment was observed in the behavioral activity of CYL-treated mice in the MWM test in the 1st week trial, and this effect was reversed in the 2nd week trial in the combination treatment group. The behavioral analysis proved that noni supplementation reduced the risk of memory impairment caused by CYL. Biochemical analysis revealed that CYL markedly increased the levels of AChE and MDA in brain tissue. Similarly, decreases in the levels of antioxidants, i.e., GSH, CAT, SOD and GST, were detected in the brain tissue of the mice exposed to CYL. Qualitative and quantitative examinations of histopathological examination of the mouse hippocampus supported the above findings. The results demonstrated that noni supplement therapy reversed the changes in the MDA, AChE, and antioxidant enzyme levels while improving the behavioral and histological alterations caused by CYL. Long-term hippocampal growth and memory are unaffected, suggesting that CYL is less harmful. According to our research, supplementing with noni in conjunction with CYL may be a helpful treatment strategy for treating memory impairment caused by CYL.

Training Laboratory Rabbits to Refine Routine Husbandry Procedures.

Non-aversive handling and training techniques for laboratory animals are required to facilitate experimental and routine husbandry procedures, improving both animal welfare and scientific quality. Clicker training was utilized to develop training protocols for rabbits to refine stressful routine husbandry procedures usually associated with lifting (i.e., being picked up from the floor)/restraining (i.e., being held in the arms of a human) them. Thirteen female New Zealand White rabbits were trained over three weeks. All rabbits learned the predefined goal behaviors: they followed the target stick, jumped onto the weighing scale, entered a transport box, and reared while placing their front paws onto the trainer's hand. In addition, ten animals jumped from the floor onto the sitting trainer's lap and allowed the trainer to lift their paws off the surface while sitting on the trainer's lap. For some individuals, the protocols had to be adapted by additional interim steps. At the end of the training, the rabbits reliably showed the expected goal behaviors, even after short and long training breaks. With few exceptions, a familiar person other than the trainer could elicit the goal behaviors from the rabbits (generalization), though further sessions were required for generalization. In the voluntary approach test, the rabbits preferred interacting with the trainer in the 1st trial but spent as much time with an unfamiliar person as with the trainer in the 2nd trial. The behavioral observations suggested that picking the rabbits up with the transport box, as described in the protocol, instead of restraining them with the scruff of their neck and lifting them on the arm, was less aversive. All in all, the training protocols were feasible and can serve as a refinement strategy in laboratory animal facilities. In the interest of animal welfare, the training protocols should be applied wherever possible.

Assessing the exploratory profile of two zebrafish populations: Influence of anxiety-like phenotypes and independent trials on homebase-related parameters and exploration

Anxiety is a protective behavior when animals face aversive conditions. The open field test (OFT) is used to assess the spatio-temporal dynamics of exploration, in which both homebase formation and recognition of environmental cues may reflect habituation to unfamiliar conditions. Because emotional- and affective-like states influence exploration patterns and mnemonic aspects, we aimed to verify whether the exploratory behaviors of two zebrafish populations showing distinct baselines of anxiety differ in two OFT sessions. Firstly, we assessed the baseline anxiety-like responses of short fin (SF) and leopard (LEO) populations using the novel tank test (NTT) and light-dark test (LDT) in 6-min trials. Fish were later tested in two consecutive days in the OFT, where the spatial occupancy and exploratory profile were analyzed for 30 min. In general, LEO showed pronounced diving behavior and scototaxis in the NTT and LDT, respectively, in which an “anxiety index” corroborated their exacerbated anxiety-like behavior. In the OFT, the SF population spent less time to establish the homebase in the 1st trial, while only LEO showed a markedly reduction in the latency to homebase formation in the 2nd trial. Both locomotion and homebase-related activities were decreased in the 2nd trial, in which animals also revealed increased occupancy in the center area of the apparatus. Moreover, we verified a significant percentage of homebase conservation for both populations, while only SF showed reduced the number of trips and increased the average length of trips. Principal component analyses revealed that distinct factors accounted for total variances between trials for each population tested. While homebase exploration was reduced in the 2nd trial for SF, an increased occupancy in the center area and hypolocomotion were the main factors that contribute to the effects observed in LEO during re-exposure to the OFT. In conclusion, our novel data support the homebase conservation in zebrafish subjected to independent OFT sessions, as well as corroborate a population-dependent effect on specific behavioral parameters related to exploration.

COMPARING THE EFFICIENCY OF DIFFERENT CARBON DIOXIDE SOURCES IN COLLECTING MOSQUITO VECTOR SPECIES ON THE TEMPE CAMPUS OF ARIZONA STATE UNIVERSITY.

We compared the effectiveness of 4 different carbon dioxide (CO2) sources (sugar-fermented BG-CO2, sugar-fermented Fleischmann yeast, dry ice, and compressed gas cylinders) in attracting different mosquito species in 2 separate 4 × 4 Latin square trials. The CO2 generated by dry ice and the gas cylinders collected more Culex quinquefasciatus than the sugar-fermented BG-CO2 and Fleischmann yeasts during the 1st trial (16-h surveillance periods), but there was no significant difference in Aedes aegypti numbers. There were no significant differences between the different CO2 sources in collecting Cx. quinquefasciatus and Ae. aegypti mosquitoes in the 2nd trial (24-h surveillance periods). Catches for Culiseta inornata and Cx. tarsalis were too low in both experiments for formal statistical analysis. Data can be used to inform local mosquito surveillance programs, but the selection of a CO2 source will also depend on financial and logistical constraints.

T cell CD62L expression following nivolumab therapy is associated with response to rituximab‐nivolumab in treatment naïve follicular lymphoma: the 1st FLOR study

Introduction: Inter-tumoral T cell dysfunction and efficacy of immune checkpoint inhibitors (ICI) in follicular lymphoma (FL) has been well described. However, few studies have examined peripheral blood immunity at diagnosis or the impact of frontline ICI on circulating immunity in FL. We hypothesised that immune dysregulation in peripheral blood would be present in treatment naïve FL and would correlate to response to frontline ICI. Methods: PBMC samples from 34 untreated FL patients receiving rituximab (R) and nivolumab (nivo) in the 1st FLOR trial (Hawkes JCO 2021) were collected at baseline, post 4 cycles of nivo and after 6 months of nivo±R. Immune profile was assessed using a single-tube 29 antibody FACS panel on a spectral flow cytometer and compared to the profile of 12 age matched healthy donors. Results were correlated with centrally determined PET response (Lugano criteria). Results: Compared to healthy donors, FL patients had increased proportions of TRegs (P < 0.0001) with decreased HLA-DR+ (P < 0.01) and increased PD-1+ (P < 0.01) populations. NK cells were increased (P < 0.05) with a 2-fold increase in TIM3 expression (P < 0.01). While total T cells were unchanged, expression of PD-1, TIM3, HLA-DR and 4-1BB were significantly increased in T cells from FL patients. Treatment with nivo significantly increased the populations of TRegs expressing 41BB, LAG-3, TIM3 or PD-L2 (P < 0.05) with a decreased PD-L1 positive population at PET-CT2. Expression of immune checkpoints on T cells was unchanged by therapy. To assess biomarkers of response, patients were stratified into sustained CR (CR achieved by PET-CT4 and maintained for 6 months without evidence of relapse, n = 15) vs. PR/PD (n = 21). At baseline, sustained CR was associated with a trend for increased proportions of Naïve CD4 and CD8 T cells and decreased TRegs and PD-L2 expressing TRegs. Baseline expression of PD-1 on T and NK cells did not correlate with sustained response. Most strikingly, expression of CD62L was significantly downregulated across total CD4 and CD8 T cells and TRegs at PET-CT2 in PR/PD patients (P < 0.05) and maintained at baseline levels in those patients who achieved a sustained CR. At this early timepoint final patient responses had not been established with most patients having either progressive or stable disease, suggesting that dysfunctional downregulation of CD62L in response to nivo may reflect the inability of patient’s T cells to activate and/or migrate to the tumour site and facilitate tumour clearance and long-term treatment responses. Conclusions: Grade 1-3A treatment naïve FL is associated with significant dysregulation of peripheral blood T and NK cells prior to therapy. Early downregulation of CD62L on T cells of patients treated with nivo was associated with the inability to achieve long term complete responses and may indicate aberrant T cell activation and/or function which impacts on long term response to therapy. Encore Abstract - previously submitted to EHA 2023 The research was funded by: Bristol Myers Squibb Keywords: Basic and Translational Science, Diagnostic and Prognostic Biomarkers Conflicts of interests pertinent to the abstract. A. Barraclough Honoraria: Roche, Gilead G. Chong Consultant or advisory role: Bristol Myers Squibb Research funding: Bristol Myers Squibb, Merck Serono, Pharmacyclics, Regeneron, Bayer, Astra Zeneca, Amgen, Hutch Med, Incyte E. A. Hawkes Consultant or advisory role: Roche, Merck Sharpe & Dohme, Astra Zeneca, Gilead, Antigene, Novartis, Regeneron, Janssen, Specialised Therapeutics Research funding: Bristol Myers Squibb, Roche, Merck KgA, Astra Zeneca

Enhancing the wheat growth through micronutrients enriched biochar under salt stress

Crop irrigation with brackish water having high Na+ is a common practice in developing countries like Pakistan; hence increasing the saline-sodic soils throughout the region. Therefore, two independent studies were conducted to evaluate the impact of minerals enriched contrasting biochars (MECBs) in mitigating Na+ toxicity on soil physicochemical properties, wheat growth, and physiology. In 1st trial, an incubation experiment was conducted having 32 treatments including simple biochar (SBc), manganese enriched biochar (MnBc), zinc enriched biochar (ZnBc), and iron enriched biochar (FeBc) applied at two rates (0.1% and 0.5%) with four different salinity levels such as 10, 20, 30, and 40 dSm−1. The soil was sampled after the 8th, 16th, and 24th day of incubation. Among the four sources of biochar, MnBc (0.1%) + 40 dSm−1 and FeBc (0.5%) + 40 dSm−1 showed efficient results to tolerate salinity stress in soil by efficiently reducing the availability of Na+ (60% and 30%) when compared to SBc (0.5%) + 40 dSm−1. Then, a pot study was conducted with 20 different treatments (control, SBc, MnBc, ZnBc, and FeBc) at four different salinity levels, i.e., 0, 5, 10, and 15 dSm−1. There were four replicates and the crop was harvested after 60 days of germination. The application of brackish water (control+15 dSm−1) significantly reduced the fresh weight of root (28%) and shoot (34%) of wheat (Triticum aestivum L.) as compared to FeBc+15 dSm−1. However, the soil Na+ toxicity was significantly decreased (23%) by the application of FeBc+15 dSm−1 as compared to control+15 dSm−1. Similarly, a significant increase was observed in leaf chl. a, b and photosynthetic rate (75%, 3 and 2-folds); however, a significant decrease was observed in transpiration rate (50%) in T. aestivum L. treated with FeBc+15 dSm−1 as compared to control+15 dSm−1. Interestingly, FeBc+15 dSm−1 also played a vital role to prevent oxidative damage by controlling the shoot CMP (63%) in T. aestivum L. by efficiently triggering the activities of shoot antioxidant enzymes such as APX (30%) and CAT (52%) when compared to control+15 dSm−1. In summary, it was evaluated that FeBc showed the highest adsorption capacity of Na+ ions to regulate salinity stress by improving soil health and crop physiology.

Abstract WP17: Plasma Proteins In Correlation With Lesional Iron Content And Permeability Imaging In Clinical Trial Of Cavernous Angioma With Symptomatic Hemorrhage

Background: Increases in mean lesional iron content (≥6%) measurement by QSM and vascular permeability (≥40%) assessed by DCEQP MRI have been associated with new bleeding, and are used as monitoring biomarkers in NINDS funded 1st clinical trial of pharmacotherapy (clinicaltrials.gov NCT02603328) in cavernous angiomas with symptomatic hemorrhage (CASH). Plasma protein levels previously associated with CASH in diagnostic and prognostic contexts have never been compared to lesional QSM and DCEQP in the same subjects. Methods: Plasma samples and MRI sequences were simultaneously acquired during 1 year-epochs of prospective follow-up of CASH patients. Plasma levels of 16 proteins were assayed by ELISA and correlated with lesional QSM and DCEQP during the same epochs. Univariate correlations were followed by multivariate analyses combining multiple protein levels to minimize Akaike Information Criterion (AIC) and increase R 2 . Accuracy (AUC on receiver operating curves) and sum of squared error (SSE) are reported for associations achieving statistical significance with FDR correction. Results: None of the proteins individually correlated with QSM/DCEQP (continuous or categorical). Relative change of angiopoietin2, and absolute change of the combination (endoglin+IL1B+IL16) had the highest accuracy (AUC 78.7%, SD 0.062 and 98.4%, SD 0.013 respectively) for lesional QSM increase ≥6%. Relative change of endoglin+IL1B, and absolute change of angiopoetin2+endoglin+thromobomodulin+VEGF, had the highest accuracy (AUC 64.6%, SD 0.057 and 93.2%, SD 0.040 respectively) for lesional DCEQP increase ≥40%. Relative change in angiopoetin1 had the lowest SSE (8.87) with QSM change as a continuous variable, and relative change in lipopolysaccharide binding protein had the lowest SSE (469.06) with DCEQP change as continuous variable. Conclusion: Circulating proteins reflect changes in lesional iron content and permeability in CASH during prospective follow-up. Results are a proof of concept that blood tests could replace more complex and costly imaging biomarkers in the monitoring of hemorrhage in cavernous angiomas. Additional mechanistic plasma molecules (miRNAs and metabolites) may further enhance the accuracy of these monitoring biomarkers.

Densidade populacional da lagarta-do-cartucho do outono, Spodoptera frugiperda (Smith e Lepidoptera: Noctuidae), e a sua resposta a alguns fenômenos ecológicos na cultura do milho

The fall armyworm [FAW; Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)], is considered a serious invasive pest that poses a serious threat to world food security. It can completely devastate a whole country's cereal crops. Therefore, the present work is the 1st field trial in Egypt to elucidate some ecological aspects of S. frugiperda on maize plants (Single-Hybrid 168 Yellow Corn cultivar) at Esna district, Luxor Governorate, Egypt, throughout two sequential growing seasons of maize (2021-2022). Three insect parameters were used, i.e., egg masses, number of larvae, and percentage of damaged corn plants. Effects of certain climatic conditions and corn plant ages on S. frugiperda seasonal activity and damaged plants percentage were also estimated. S. frugiperda population initiated to attack maize plants from the 3rd week of June until the harvest in every season. S. frugiperda had two seasonal activity peaks in terms of egg masses numbers and three peaks regarding the larval population density/season. Its damage percentage increased with increasing plant age weekly. The mean of S. frugiperda egg masses were 2.83 ± 0.40 and 2.96 ± 0.45 mass /10 corn plants in 2021 and 2022, respectively. While, the overall mean larval populations were 13.41 ± 0.52 and 13.03 ± 0.46 larvae/10 plants, during the two growing seasons, respectively. Corn plant damage percentages reached 68.54 ± 2.71 and 60.42 ± 2.92% in 2021 and 2022, respectively. The combined effects of both the weather conditions and maize plant ages were highly significant on egg masses, larvae population density, and damage percentage, and varied from one season to another. The mean daily maximum temperature was the most effective variable on egg masses and the larval population. Maize plant age had a clear effect on the damage percentage caused by the larvae in the field during the two years of the study. The dramatic spread of FAW and the consequent damage (that appeared in different countries of America, Africa, and Asia) mean that different management approaches must be sought for the small and large-scale producers by using available technologies for smallholder farmers will eliminate pest damage without access to reach to an economic Injury level. This information may assist the decision maker when planning the S. frugiperda IPM program for maize plants and its surveillance.

Immune activation attenuates memory acquisition and consolidation of conditioned disgust (anticipatory nausea) in rats

Anticipatory nausea is a classically conditioned response to cues (e.g. contexts) that have been previously paired with a nauseating stimulus, such as chemotherapy in humans. In rodents, anticipatory nausea can be modeled by pairing a novel context with lithium chloride (LiCl), which leads to conditioned disgust behaviours (such as gaping) when exposed to the context alone. Growing evidence suggests that selective immune activation attenuates various forms of learning and memory. The present study investigated the effects of the endotoxin lipopolysaccharide (LPS) on LiCl-induced anticipatory nausea across critical stages of associative memory including acquisition, consolidation, and extinction. Adult male Long Evans rats were subject to intraperitoneal (i.p.) LiCl (127 mg/kg) or vehicle control (NaCl) paired with a 30 min conditioning trial in a distinct context for a total of 4 trials. To study acquisition, rats were administered either LPS or NaCl (200 μg/kg, i.p.) 90 mins before the conditioning trials. To study consolidation, different rats were administered either LPS or NaCl (200 μg/kg, i.p.) immediately after the conditioning trials. These trials were followed by 4 drug-free extinction trials within the same context. LPS significantly reduced conditioned gaping behaviours by the 4th conditioning trial and on the 1st drug-free extinction trial when administered 90 mins before or immediately after the conditioning trials. LPS had no significant effect on extinction. The present study provides strong evidence for the attenuating effects of LPS exposure on the acquisition and consolidation of LiCl-induced anticipatory nausea.

ASC2ESCALATE: A Phase 2, Single-Arm, Dose-Escalation Study of Asciminib Monotherapy in Patients (Pts) with Chronic Myeloid Leukemia in Chronic Phase (CML-CP) Previously Treated with 1 Prior Tyrosine Kinase Inhibitor (TKI)

BACKGROUND: The discovery of adenosine triphosphate (ATP)-competitive TKIs has extended life expectancy in CML; despite these advances, 28% to 40% of newly diagnosed pts with CML-CP discontinue treatment by 5 years, prompting a switch to second-line (2L) therapy. Discontinuation rates increase in 2L; up to 86% of pts discontinue therapy, with 20% to 30% discontinuing 2L therapy due to disease progression. Dose-escalation studies show that some pts experiencing resistance to therapy can achieve responses with higher doses of their TKI, while use of another second-generation (2G) TKI has shown limited clinical benefit in pts experiencing resistance to a 2G TKI. Asciminib is the 1st BCR::ABL1 inhibitor that Specifically Targets the ABL Myristoyl Pocket (STAMP), allowing asciminib to have activity against most BCR::ABL1 mutations that impart resistance to ATP-competitive TKIs. Asciminib was 1st approved in the United States for pts with CML-CP after ≥2 TKIs at 80 mg once daily (QD) and 40 mg twice daily (BID) and CML-CP with the T315I mutation at 200 mg BID. In the phase 3 ASCEMBL study, asciminib 40 mg BID showed superior efficacy vs bosutinib 500 mg QD in pts with CML-CP after ≥2 prior TKIs at weeks 24 (major molecular response [MMR], 25.5% vs 13.2%) and 96 (MMR, 37.6% vs 15.8%) and favorable tolerability with >2 years’ follow-up. In a large phase 1 dose-finding trial, asciminib demonstrated safety across doses of 80 to 200 mg QD and 10 to 200 mg BID in pts with CML-CP/accelerated phase without and with the T315I mutation previously treated with ≥2 and ≥1 TKIs, respectively (maximum tolerated dose was not reached in this study). Here, we describe the phase 2 ASC2ESCALATE trial investigating the efficacy and safety of 2L asciminib with dose escalation in pts with CML-CP (NCT05384587). STUDY DESIGN AND METHODS: This is a phase 2, open-label, multicenter, single-arm, dose-escalation study in the United States (Figure). Adult pts (aged ≥18 years) with CML-CP without the T315I mutation who experienced resistance (BCR::ABL1IS >1% with 6-12 months of first-line [1L] treatment or >10% with >12 months of 1L treatment) or intolerance (BCR::ABL1IS >0.1%) with ≥6 months of treatment with 1 prior ATP-competitive TKI are eligible (Table). All pts will initiate treatment with asciminib 80 mg QD. For pts not achieving BCR::ABL1IS <1% at 6 months, dose will be escalated to 200 mg QD if pts do not have grade ≥3 toxicity or persistent grade 2 toxicity refractory to optimal management. In pts not achieving MMR at 12 months, either dose escalation from 80 to 200 mg QD or from 200 mg QD to 200 mg BID will occur or the pts will discontinue study treatment. Pts who achieve MMR at 12 months will continue asciminib at their current dose. Pts deriving clinical benefit from asciminib per investigator assessment may receive post-trial access. Pts may discontinue at any time due to unacceptable toxicity or progression or at investigator/pt discretion. The primary endpoint is the proportion of pts achieving MMR at 12 months. Secondary endpoints include MMR rates by 3, 6, 18, and 24 months; MR4.5 (BCR::ABL1IS ≤0.0032%) at 24 months; time to and duration of MMR; time to treatment failure; and safety/tolerability. To investigate progression-free and overall survival, time from the first dose to progression or death due to any cause during the study, respectively, will be measured. CONCLUSIONS: ASC2ESCALATE is the 1st clinical trial designed to assess asciminib in pts with CML-CP requiring 2L therapy and seeks to provide information on the efficacy and safety of dose escalation for pts not meeting certain response milestones. Treatment using a dose escalation strategy with asciminib in 2L could enable a greater proportion of pts with CML-CP not meeting milestones to achieve molecular responses and provide another option for those experiencing intolerance of or resistance to ATP-competitive TKIs. Recruitment for this study is planned to begin August 2022 with an approximate enrollment of 92 pts with CML-CP across ≈40 sites. This study is sponsored by Novartis. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Solar bubble dryer: alternative to sun drying for reducing drying losses in Bangladesh

The Solar Bubble Dryer (SBD) is a latest low cost drying technology and flexible alternative to sun drying. Technical performance was investigated by observing moisture content and temperature distribution at different points of the dryer. The moisture distribution was uniform and its content was reduced initially from 19.5, 20.6 and 20.3% to 14, 15.6 and 14%, respectively in the 1st, 2nd and 3rd trial of the dryer. The temperature distribution was almost same but it was sometimes lower than the ambient temperature. The drying rate, drying capacity and drying efficiency were found to be 3.9 kg/hr, 1000 kg/batch, and 12.3%, respectively. Every trial took 3 - 5 days but it was difficult to achieve desired moisture of 12.0%. The average germination rate of SBD dried paddy (89.66%) was more than that of sun-dried paddy (84%). The milling recovery was found 71% for SBD dried paddy and 72.4% for sundried paddy. The percentage of broken rice in SBD dried paddy sample was more than that of sun-dried sample because of high moisture content (14.5%) and less hardness (22.5 N). The operating cost of paddy drying in SBD dryer was found Tk. 1410.9 per ton whereas that of traditional sun drying methods was found Tk.1047.3 per ton. Although, drying by SDB is not profitable over traditional drying, it has advantage to protect drying loss of fresh harvested paddy during unpredictable weather condition in rainy season. Bangladesh J. Agri. 2022, 47(1): 1-15

OP005: AT-007 significantly reduces toxic galactitol in ACTION-galactosemia kids - the 1st therapeutic interventional clinical trial in children with classic galactosemia

Classic Galactosemia (CG) is a rare, autosomal recessive disease where galactose is not metabolized properly due to severe deficiency or absence of the GALT enzyme (galactose-1-phosphate uridyl transferase). At abnormally high levels, galactose becomes an aberrant substrate for the enzyme aldose reductase, resulting in conversion to an abnormal and toxic metabolite, galactitol. Newborn screening and implementation of a galactose-restricted diet has reduced newborn acute symptoms and significantly decreased fatalities. However, despite early dietary intervention, children continue to develop significant morbidities in speech, cognition, behavior, and motor function, which have been shown to progressively worsen with age, as well as cataracts and primary ovarian insufficiency in females. AT-007 is an oral CNS penetrant aldose reductase inhibitor, which prevents conversion of galactose to toxic galactitol, that is in development for the treatment of CG. Reduction in circulating galactitol by AT-007 has been shown to halt the progression of disease in a rat model of Galactosemia. ACTION-Galactosemia Kids is a sequential, two-part, randomized double-blind, placebo-controlled study evaluating the clinical benefit, safety, pharmacokinetics and pharmacodynamics of AT-007 in pediatric patients with CG. Patients were randomized 2:1 to AT-007 or placebo. Disease demographics, genetic variant, and patient history were assessed at baseline. Additionally, functional tests of speech, motor function, cognition and behavior were performed at baseline for all patients enrolled in the clinical study. The primary biomarker study endpoint was reduction in plasma galactitol over a minimum of 30 days of treatment with the target dose of AT-007 vs placebo. Baseline and 30-day plasma galactitol levels were drawn under fasted conditions and evaluated via a validated liquid chromatography-mass spectrometry assay. Baseline galactitol correlation with disease severity was evaluated via a linear model incorporating baseline speech, cognition, behavior and motor skills assessments. Forty-seven patients ranging in age from 2 to 17, were enrolled at 3 US clinical sites in ACTION-Galactosemia Kids across three predefined age groups (2-6 years old; 7-12 years old; 13-17 years old). The patients were predominantly White (98%), had a mean age of 9 and 51% were female/49% male. The most common baseline disease characteristics included learning disorders (∼50%), apraxia (∼40%), anxiety (∼35%), vitamin deficiency (∼30%), tremor (∼20%) and ataxia (∼20%). All the female patients older than 12 had primary ovarian insufficiency. Additionally, ∼10% of the patients suffered from seizures and ∼5% had early onset cataracts. Baseline disease and demographic characteristics were balanced across the placebo and AT-007 treatment groups. Galactitol levels at baseline correlated with total disease severity in this pediatric patient population (p=0.004). Treatment with AT-007 resulted in a 40.2% reduction in galactitol, which was significant when compared with patients on placebo (p<0.001). The reduction in toxic galactitol was consistent among age and dose groups and was not associated with changes in galactose or galacose-1-phosphate (Gal-1p). AT-007 was safe and well tolerated. The biomarker results from ACTION-Galactosemia Kids demonstrates that treatment with AT-007 resulted in a significant reduction of the toxic metabolite galactitol, a biomarker that correlated with disease severity, in pediatric patients with Classic Galactosemia. Treatment of pediatric Galactosemia patients age 2 to 17 with AT-007 was safe and well tolerated.

Potato breeding in mountainous and foothill areas

The work is to develop new competitive potato varieties. The studies were carried out in mountainous conditions on mountain meadow soils in a zone free from viral infection. According to the scheme of the breeding process, in the parent nursery, hybridization was carried out in 18 combinations, 16837 seeds were obtained. In the preliminary test nursery, 3 hybrids formed yields of over 50 t / ha. High yield of hybrids was also noted in the nursery of the main trial. Of the 7 hybrid offspring in the nursery of the 1st year competitive trial, 4 provided a yield higher than the standard variety. Hybrids 13.61 / 66 were isolated from 4 hybrids of the II year competitive test nursery. In the nursery of the third year competitive test, all tested hybrids formed a high yield (2.1-45.5% higher than the standard). The 10.11 / 770 hybrid has been submitted for state testing. Hybrid 10.11 / 765 (Osetinsky variety) is included in the State Register of Breeding Achievements.

SELECTION OF PROCESSING POTATO VARIETIES THROUGH MULTI-LOCATION TRIALS

Thirteen exotic potato varieties along with four checks were evaluated at six agro-ecological locations of Bangladesh for three generations during 2015-16 to 2017-18 in order to identify table purpose and processing quality varieties. Results indicated significant variation among the varieties. Based on the results of the 1st year multi-location trial, seven superior ones were selected for further testing in AYT and RYT in next two years. In the SYT, varieties Farida and 7four7 were the highest yielders. In the AYT, the highest average yield over location was also produced by 7four7 (38.70 t/ha). Varieties Cimega and Memphis also gave comparable yields to that of 7four7 (37.33 and 36.67 t/ha, respectively). Considering the yield of the three generations, the above four varieties were significantly better than the checks. Considering the specific qualities, Farida was found most suitable for table purpose because of its high yield, medium-sized oval and smooth tubers with good eating quality. The variety Taisiya produced tubers with good size and shape, but low in dry matter content; so not suitable for processing. Memphis might be selected for French fry as it produces maximum large sized tubers with good long oval shape. On the other hand, variety Panamera is a high yielder but its plant type was undesirable. On the whole, varieties Cimega, 7four7 and Farida are suitable for table purpose, and Memphis may be selected for French fry under Bangladesh condition. None was found quite suitable for Chips preparation.

Performance of Kampung Unggul Balitbangtan (KUB) Chicken on Household Scale Semi Intensive Maintenance In Swamp Agroecosystem In Ogan Komering Ilir District

Kampung unggul balitbangtan (KUB) Chicken is described its performance on the maintenance of household scale semi intensitively in the Ogan Komering Ilir district. This paper aims to describe the performance of KUB chickens reared semi-intensively on a household scale in the Lebak swamp diagroecosystem in the Kayuagung area, Ogan Komering Ilir distrcit. Methods The study was conducted by researchers at the Kayuagung Experimental Garden and by farmers. The study at (KP) Kayuagung was carried out twice, namely the 1st trial from February 2020 to April 2020, the 2nd trial from May 15 to December 2020. The result of the period Sensi chickens reaches an avarage 870 g on 68 days and KUB chickens 434-804 g on 74 days. Semiintensitively Chicken KUB showed first laying varies from 5,5 months to 8 months, quite high risk of disease and exhibits sedating properties. The incidence of disease is quite high at the farmer because the quantity and quality of feed is less than optimal and the weather is dominant wet. But if the quality and quantity of feed is fulfilled can produce many eggs.

Evidence of significant cognitive improvement over baseline in advanced Alzheimer’s disease (AD) patients: A regenerative therapeutic strategy

AbstractBackgroundSynaptogenix scientists have pioneered a regenerative therapeutic strategy designed to reduce (1.) a beta‐oligomers and hyperphosphorylated tau, but importantly, (2.) also restores the AD – induced loss of synaptic networks and neurons. In pre‐clinical studies (Sun and Alkon, TIPS, 2019), increased numbers of mature, mushroom spine synapses were measured with serial electron micrographs of the hippocampus with the rat spatial maze memory task, and were further significantly increased by the Bryostatin‐induced activation of the PKC epsilon‐BDNF cascade (Hongpaisan and Alkon, PNAS, 2007). Similarly, Bryostatin increased the numbers of mature synapses, normalized cognitive deficits, and prevented neuronal death in pre‐clinical models of neurodegeneration including AD (p&lt;.01, 2‐tailed), stroke, Fragile X syndrome, and traumatic brain injury. Subsequently, compassionate use trials showed marked improvements in advanced AD patients (Nelson et al., JAD, 2017).MethodThereafter, an initial pilot (13 weeks, N=150) trial of Bryostatin to treat AD, Study #202, and a 2nd (13 weeks, N=100) pilot trial, Study #203, included identical protocols: 7 i.v. doses of 20 mcg / kg Bryostatin over 11 weeks with Placebo groups in each.ResultsBryostatin Treatment (at Week #13) in the 1st trial improved Severe Impairment Battery (SIB) scores over baseline (+4.5 over baseline, p&lt;.04, 2‐tailed, pre‐specified exploratory analyses of AD patients, MMSE 14 – 4, vs. placebo, without chronic Namenda, Farlow et al., J. Alz. Dis., 2019). In a second pilot trial, Bryostatin caused significant SIB improvement (+4.12 over baseline) in the pre‐specified Moderate Stratum (MMSE 10 – 14) of advanced AD patients without Namenda (p &lt; .005, 2‐tailed). In both trials, trend analyses demonstrated increased drug benefit with increasing dosing number. In a pooled trend analysis of Studies #202, 203, Bryostatin caused highly significant benefit (p&lt;.001) and Placebo was NS.ConclusionWhile issues such as baseline imbalance (Study #203) and Namenda blockade of Bryostatin efficacy (Study #202) prevented reaching primary endpoints, these pre‐specified exploratory analyses (see above) revealed clear evidence of Bryostatin therapeutic efficacy, now being tested in an NIH‐sponsored, 6‐month trial, incorporating lessons learned from these pilot studies. This potential efficacy involved significant improvement over baseline, not only reduction in the rate of cognitive decline.

727P Phase Ib INEOV neoadjuvant trial of the anti-PDL1, durvalumab (D) +/- anti-CTLA4 tremelimumab (T) with platinum chemotherapy for patients (pts) with unresectable ovarian cancer (OC): A GINECO study

We have shown that neoadjuvant carboplatin and paclitaxel (NACP) increased tumor-infiltrating lymphocytes and PDL1 expression in OC pts. Combined PDL1/CTLA4 blockade is active in relapsed OC. INEOV is the 1st trial of neoadjuvant D +/- T with NACP in pts with unresectable OC. Main endpoints are feasibility and safety. 2° endpoints include macroscopically complete resection (CCO) and major pathological response rates after 3 cycles (C3).

Barriers to Successful Implementation of Precision Medicine to the Treatment of Refractory AML: Lessons Learned

Tremendous genomic heterogeneity has been reported for patients with acute myeloid leukemia (AML), as well as variability in gene expression. We have developed a custom high throughput drug sensitivity (HTS) assay enabling testing of ~150 drugs and combinations, both FDA approved and investigational, including chemotherapy agents and targeted inhibitors. We enrolled advanced relapsed/refractory AML patients (pts) on two successive trials, NCT01872819 “Treatment for Relapsed / Refractory AML Based on a High Throughput Drug Sensitivity Assay” and NCT NCT02551718 “Individualized Treatment for Relapsed/Refractory Acute Leukemia Based on Chemosensitivity and Genomics/Gene Expression Data.” The first trial enrolled 16 AML pts (3-5 prior regimens) of whom 14 received single agent study directed treatment, and the second trial (2-6 prior regimens) enrolled 25 AML pts of whom 14 received study directed therapy including drug combinations. We were able to choose chemotherapy drugs for all pts that had IC50's of ≤ 0.1 µM, or regimens that demonstrated synergy for drugs in combination. We have established feasibility as defined by the study, in that HTS assay results were reported at an average of 5.6 days (range 4-10), mutation analysis for 194 genes by MyAML™ reported in 12.5 days (range 9-17) with turnaround from receipt median 8 days (range 7-12) and treatment started at 7.8 (mean), 8 (median) days (range 4-11).In the first trial, 1 pt (5prior regimens), achieved complete remission with incomplete count recovery (CRi) with the single chosen drug, and 1 achieved CR, 1 CRi with subsequent regimens of additional drugs based on screening and published combinations. Median overall survival (OS) was 88 days for all patients, range 7 to 411. For the 2nd trial, where combinations were used, there were 2 CRs, 1 CRi and 3 partial responses. Median OS was 122 days, range 19-379. Of the patients in the 1st trial who had therapy that could have been directed by mutation analysis, there were 4 with FLT3, 1 KIT, 1 IDH2, 1 TP53, 1 FLT3+KIT, 2 FLT3+ IDH2 mutations (10/12 tested). For the 2nd trial there were 2 pts with FLT3D835, 2 TP53, 2 translocations involving FLT3, 1 IDH2 + FLT3-ITD, 1 FLT3-ITD + IDH1, 1 IDH1, 1 FLT3D839, 1 FLT3-ITD + TP53, 1 IDH2, 1 BCR-ABL (13/17 tested), and one patient received imatinib for BCR-ABL, 2 sorafenib for Flt3 mutations, and 1 with a TP53 mutation received decitabine. We had a number of logistical difficulties, including very advanced disease , poor performance status, inability to obtain the drugs with the lowest IC50's or as directed by mutations due to investigational status or lack of insurance approval, and inability to treat pts who returned home to their local physicians. We observed a higher response rate in pts who received drug combinations (8) vs. single agents (1). Although these were just feasibility trials, we also tried to compare their outcomes to a historical control group. We selected 2 matches for each of our pts, total of 50. They were matched to age, length of CR1, prior HCT, prior intense salvage and prior number of total salvages. One limitation was to find controls that had received ≥3 salvages like some of our studied population. Most of the patients in our clinical trial received single drug therapy (64 vs 24%) and non-intense therapy (4 vs 44%), when compared to our historical control group. Although these were very important treatment differences, the median OS did not differ statistically between both groups (88 vs 115 days, p: 0.327).We have gathered clinical data, and data on mutation analysis, gene expression, and drug sensitivity testing, that have been analyzed to determine correlations of specific features with drug resistance. Moreover, we have the ability to confirm functional drug sensitivity to the targeted inhibitors based on mutation data. All these data can contribute to future algorithms to assist in optimizing drug choices.In conclusion, our future iterations of this approach will include 1) only drug combinations, 2) study of pts with only 2 prior regimens, and 3) earlier incorporation of targeted inhibitors such as midostaurin and the new IDH2 inhibitor, enasidenib, into combination regimens. The current salvage options for relapsed/refractory AML have dismal results, as exhibited by our historical control group; only 10% were alive at 1 year. We will therefore pursue alternative approaches for this population, including the optimization of precision medicine methodology. DisclosuresWalter:ADC Therapeutics: Research Funding; Aptevo Therapeutics: Research Funding. Patay:Invivoscribe: Employment. Carson:Invivoscribe, Inc.: Employment. Radich:Amgen: Consultancy; Gilliad: Consultancy; Novartis: Consultancy, Other: lab contracts for bid and service assays; BMS: Consultancy; Ariad: Consultancy; Pfizer: Consultancy. Becker:GlycoMimetics, Inc.: Research Funding.

OP0129 BELIMUMAB AFTER RITUXIMAB SIGNIFICANTLY REDUCED IGG ANTI-DSDNA ANTIBODY LEVELS AND PROLONGED TIME TO SEVERE FLARE IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS

Background:B cell depletion with rituximab, an anti-CD20 mAb, has shown efficacy for systemic lupus erythematosus (SLE) in open-label studies but failed to meet primary endpoints in two randomised, placebo controlled trials. Rituximab increases BAFF levels which has been associated with subsequent lupus flares. We hypothesised that high BAFF levels after rituximab limit its effectiveness in SLE and that the anti-BAFF monoclonal antibody belimumab given immediately after rituximab could be a valuable therapeutic strategy.Objectives:To assess the safety and obtain preliminary evidence for efficacy of belimumab following rituximab therapy in patients with SLE.Methods:BEAT-LUPUS (Belimumab after B cell depletion in SLE) is a 52-week phase IIb, randomised, double-blind, placebo-controlled clinical trial investigating the safety and efficacy of intravenous belimumab after B cell depletion therapy (rituximab). The maximum permissible prednisolone dose throughout the trial was 20mg/day with encouragement to reduce by 50% from baseline by 6 months. The primary outcome measure was log IgG anti-dsDNA antibody serum levels at 52 weeks measured by ELISA. A linear regression ANCOVA model was fitted to evaluate the difference in 52-week anti-dsDNA levels between treatment arms adjusting for anti-dsDNA value at screening (before rituximab) and randomisation (4-8 weeks after the 1st infusion of rituximab), CD19 &gt; or &lt; 0.01x109/l at randomisation, and renal involvement at screen. Secondary outcomes included measures of disease activity and incidence of adverse events. B cell (CD19) counts were measured by flow cytometry. Intention to treat analysis was adopted. Full ethical and regulatory approval was obtained. A comprehensive description of the protocol and statistical analysis plan is available(1,2).Results:52 patients with active SLE received rituximab (2 infusions, 2 weeks apart) and then randomised to receive either belimumab (n=26) or placebo (n=26) 4-8 weeks after their 1st dose of rituximab. 32 patients completed trial treatment protocol (belimumab or placebo) through to 52 weeks, withdrawals were equally split between belimumab and placebo. There was a significant reduction in IgG anti-dsDNA antibody levels in patients treated with belimumab compared to placebo at 52 weeks (p&lt;0.001, Figure 1); 43 patients were included in the intention to treat analysis at 52 weeks.Figure 1.Serum IgG anti-dsDNA antibody levels (geometric means with 95% confidence intervals) in patients treated with rituximab, then randomised to belimumab or placebo at 1st trial infusion. An intention to treat linear regression ANCOVA model was fitted to evaluate the difference in 52-week anti-dsDNA between belimumab or placebo adjusting for baseline values and stratification factors. N= patient numbers who provided serum samples at time indicated.Kaplan-Meier curves demonstrated that belimumab reduced the risk of severe flare (BILAG A flare) compared to placebo (hazard ratio 0.27, 95% confidence interval 0.07-0.97, unadjusted log-rank p=0.03). There were 10 and 3 severe flares in the placebo and belimumab group respectively. There was no difference in cumulative steroid dose over the course of the trial between belimumab and placebo.Belimumab did not increase the incidence of infections, serious or total adverse events, nor withdrawals due to adverse events compared to placebo. Belimumab significantly suppressed B cell repopulation at 52 weeks compared to placebo (p=0.001), but not total serum IgG.Conclusion:This placebo controlled double blind trial met its primary endpoint, a significant reduction in IgG anti-dsDNA antibody levels, and demonstrated that belimumab prolongs the time to severe flare compared to placebo. These results suggest that belimumab after rituximab is a safe and effective treatment for patients with SLE and supports further development of this combination as a novel therapeutic strategy.

Development of Induced Breeding Technique for Freshwater Fish Gobi, Glossogobius giuris (Hamilton, 1822) Using Pituitary Gland (PG) Extract

This study aimed to establish induced breeding technique of freshwater fish gobi (Glossogobius giuris), using the pituitary gland (PG) extract. Six experiments were conducted at the Department of Fisheries Biology and Genetics, Faculty of Fisheries, Bangladesh Agricultural University, Mymensingh, Bangladesh. First breeding trial was conducted in June using 40, 45, and 50 mg PG kg-1 body weight of the female fish. None of the fish was ovulated in the 1st trial. Two more breeding trials were conducted in July and August, using 6 mg (T1), 8 mg (T2) and 10 mg (T3) PG kg-1 body weight of female, and 3 mg (T4), 4 mg (T5) and 5 mg (T6) PG kg-1 body weight of male fish. After treatment, the ovulation rates were recorded as 56.33±1.53, 82.67±2.52 and 75.33±1.53% in July and 58.00±2.65, 94.67±1.53 and 78.33±1.53% in August under the treatments T1, T2 and T3, respectively. Hatching rates of eggs were observed as 52.00±4.36, 81.67±3.21 and 72.33±6.03% in July, and 54.67±3.23, 91.67±3.06 and 73.67±5.13% in August under the T1, T2 and T3, respectively. Hatching time was ranged from 35 to 48 h and after the absorption of yolk sac (60-72 h), they were survived well when fed with tubificid worms and mixed zooplankton. The female treated with the dose of 8 mg PG kg-1 body weight in August showed the best performance so far as the ovulation, fertilization and hatching rates were concerned, while the single dose of 4 mg kg-1 body weight of PG was found to be effective for male fish in both months. The findings obtained from the present study reveals that induced breeding of G. giuris, using PG extract is successful for large scale production of quality seed in captive condition, which would further facilitate towards the aquaculture production and biodiversity conservation of this important fish species to a greater extent.