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In Ayurvedic samhita, it is explained that marma viddha lakshana are caused due to aghat (trauma). Sports injuries are concerned with traumatic injuries. Comparative study of marma sharira vigyan in shalyatantra of ayurveda with modern aspects of sports medicine can aware a sportsmen about the injuries. Janu sandhi (knee joint) is the mostly used joint in various sports leading to a wide range of injuries. The aim of this study was to evaluate sports injuries especially in Janusandhi Marma and structural damage occurring in it. The viddh lakshannas(traumatic injuries) of Janu Marma are limping (Khanjata) and the objective of this study is to determine the specific injury at the knee joint that might cause limping. Total 30 patients were selected as per the inclusion and exclusion criteria mention in proforma. Observation & Results were noted with the help of Objective criteria mentioned in the proforma which included sports wise distribution, types of injury, mode of trauma, structural deformities clinical examination score and final diagnosis in study group. The result was Maximum patients had Medial Collateral Ligament involvement followed by Anterior Cruciate Ligament & Medial Meniscus injuries. Patellar injuries and Lateral Collateral Ligament injuries were comparatively less. So it was concluded that with the assessment of clinical examination score it is possible to judge the severity of trauma to janu sandhi marma and the structures involved in the traumatic injuries amongst sports athletes. This study can be helpful for deciding the treatment regime of the patient.

Catalytic oxidation using non-noble metal-based catalysts is a promising approach to mitigate pollution due to VOCs in the air.

Introduction: Traditionally, teeth that are absent are substituted by partial dentures or by complete dentures. Many prefer a dental implant as it tries to imitate the natural tooth. Though the notion of immediate loading reduced the time of prosthetic treatment, debate arose whether the loading should be functional or non-functional. The objective of this study was to compare the Peri-implant Bone Level (PIBL) deviations in immediate implant placement with instantaneous functional versus non-functional loading in the mandibular posterior region. Material and Methods: Fifty patients were incorporated in the study and distributed by random sampling into two groups of equal members each. Group A had patients in whom implant was placed followed by immediate functional loading. Group B were those who had non-functional loading. After bone implant integration, temporary crowns were replaced with metal ceramic prosthesis. PIBL change (in mm) of each implant was assessed by periapical radiograph which were taken at immediate post op, after one month and then on three and six months follow up. Statistical analysis was done using software SPSS v 21.0. Comparison between the groups (Mann Whitney U test), intragroup (Friedman’s) and Wilcoxon Signed rank test for pairwise comparison were done. Results: There was PIBL gain mesially and distally of the implants in both the groups in all time intervals of one, three and six months follow up. Conclusions: To conclude immediate implants restored instantly yielded affirmative result as far as survival of the implants, bone level changes and aesthetics of the soft tissues are concerned irrespective of the type of loading.

A comprehensive study of compact flexible patch antenna, which is the suitable topology for medical telemetry operated in the industrial scientific and medical (ISM) band, is presented. For establishing efficient wireless links for assisting medical diagnostics, the antenna is required to be of a low profile, safe, and simple geometry. To achieve the compactness of the structure, Taguchi’s optimization method has been applied based on the concept of orthogonal array. The designed antenna uses two distinct flexible substrates, namely polyethylene terephthalate (PET) with dimension 43.5x36.89x0.175 mm3, and Rogers RT/Duroid 5880 with dimension 48.5x49.66x1.58 mm3 categorized under materials, namely plastic and circuit (microfiber reinforced PTFE) material respectively. The approach has been demonstrated for a simple, compact flexible antenna with reduced dimension by 5.37% for PET, and 23.96% for RT 5880 as compared to the conventional rectangular patch design parameters. The resonant frequencies of the designed antennas are 2.435 GHz and 2.45 GHz for PET, and Rogers RT/Duroid 5880, respectively, with a bandwidth of 73.2MHz for RT/Duroid, and 16.8 MHz for PET. The maximum observed directivity of the proposed antenna using PET and Rogers RT/Duroid 5880 are 5.15 dBi, and 6.117 dBi, respectively, at 2.45 GHz. Furthermore, characterization for performance variation is carried out through bending analysis for different curvatures and Specific Absorption Rate (SAR) analysis in order to demonstrate its suitability for direct integration with wearable biomedical sensing chips used for medical telemetry.

Zinc deficiency, common in children of developing countries, leads to susceptibility to infections. Relapses in nephrotic syndrome are precipitated mostly by infections. Most of morbidity and mortality in nephrotic patients is during nephrotic range proteinuria. Management of factors which reduce duration of relapse, can decrease morbidity in pediatric nephrotic patients. We aimed to study serum zinc levels, in patients of nephrotic syndrome, at onset/relapse and at remission and tried to find out correlation between serum zinc levels and time to achieve remission.Observational study was conducted in tertiary centre of North India over 12 months. Consecutive pediatric patients, with initial episode/relapse of NS were enrolled. Patients were treated as per standard protocol. Time taken for remission was noted. Serum zinc level was estimated first at confirmation of initial episode/relapse and then at remission. Data were summarized as mean ± standard deviation. Groups were compared by paired t test. Pearson correlation analysis was done to assess association between initial zinc level and time to attain remission.68 patients were screened for inclusion in the study but only 49 qualified for final analysis. Serum zinc level was statistically different in all patients at enrolment and at remission. Pearson correlation analysis showed an insignificant and inverse correlation between time to remission and serum zinc level (r=0.14, p>0.05) suggesting that, as serum zinc level at enrolment decreases, the time to remission lengthens.Zinc level at initial episode/relapse was found to have negative correlation with time to attain remission.

Introduction NICE guidance recommends that during the COVID-19 pandemic all adults are assessed for frailty, suitability for resuscitation and escalation to critical care on admission to hospital. Risks, benefits and likely outcomes should be discussed with patients/relatives. Aim To investigate impact of intervention on treatment escalation plan (TEP) use during the first wave (W1) and second wave (W2) of pandemic. Intervention W1: An ethically approved TEP and a Critical Care Decision Aid (CCDA) was added to the admission booklet. W2: The TEP update included a ‘confirm discussed with patient/relatives’ section. Method W1: We conducted retrospective review of 62 case-notes of patients admitted through COVID-19 pathway and grouped into >80 and ≤ 80 years old. Resuscitation, TEP, CCDA and communication with patient/relatives were collected. W2: 60 further case-notes of patients with COVID-19 admitted in January 2021 were analysed. Results W1: 100% patients had resuscitation decisions and 95% had TEPs in both groups. Significantly higher proportions of Frailty scores were calculated in >80s (58% vs 35% p = 0.04). On average resuscitation and TEPs were made within 13.8 hours of admission (range 1–72 hours). 53.2% of resuscitation decisions and 30% TEPs were discussed with patients of whom higher proportions were discussed in younger age group, majority were discussed at the time of deterioration. W2: 98.3% patients had resuscitation decisions made and 96.7% had TEPs across both groups. On average resuscitation decisions were made within 14.9 hours of admission. In the >80s, 86.7% of resuscitation decisions were discussed and 63.3% of the TEPs were discussed with patient/relatives. In the Conclusion We significantly improved Treatment Escalation planning during W1 and W2 of pandemic by introducing the TEP in W1 and adding prompt to improve communication with patients/relatives in W2 (p = 0.02).

Different types of scaling methods to characterize spin-glass freezing in random magnetic systems have been discussed in literature. An earlier proposed scaling method for imaginary part of ac susceptibility using a different approach is presented for 5-nm NiO particles. This work supports for the spin-glass freezing in nanoparticle systems at lower temperature.

Introduction Aspiration pneumonia is a major cause of morbidity and mortality especially in older adults. Our Trust recorded higher than expected mortality ratios in this group of patients. Aim To investigate reasons behind higher than expected mortality and improve outcomes. Intervention We developed a collaborative approach of investigating mortality in aspiration pneumonia with joint input from Speech and Language (SALT) specialists. Method We conducted structured retrospective review of annual mortality in aspiration pneumonia in 3 PDSA (plan, do, study, and act) cycles in 2015/18/20. We collected data on clinical care, diagnostic accuracy, SALT referral/input, feeding at risk discussion, communication with primary care. We monitored mortality ratios on national systems. Results We improved clinical and nursing care by auditing mouth care, bed elevation and safe feeding. We also developed electronic-SALT referral form to improve timings for the reviews (first PDSA cycle). SALT team developed “feeding at risk proforma” to formalise risk feeding where safe swallow plan was not possible (second PDSA cycle). We modified discharge summaries and made this a multidisciplinary document in the Trust so that SALT can communicate feeding plans to primary care (third PDSA cycle). Mortality ratios improved significantly in this period from Relative risk of 152 (higher than expected range) in 14/15 to 86 (within expected range) in 19/20. Conclusion We have demonstrated significant improvement in hospital mortality ratios from aspiration pneumonia and therefore improved care by collaboratively working with SALT team and bringing changes in stepwise manner. Multidisciplinary mortality reviews are key to improving outcomes for our patients.

Background Suicide is an act of deliberately killing oneself. It is a global health problem and is the second leading cause of death in 15 to 29 years. Thus, studying nursing students’ perception towards suicide attempters has paramount importance in understanding and addressing the existing gaps in healthcare delivery system. Objective To assess the nursing students’ perception towards attempted suicide and to find out the association between perception and selected variables. Method A descriptive, cross sectional design was carried out to assess the nursing students’ perception towards suicide attempters in Kathmandu University School of Medical Sciences. The respondents were 193 nursing students. A structured questionnaire was used in order to collect data. Descriptive statistics including frequency, percentage, mean and standard deviation were calculated to summarize the data and inferential statistics, Mann Whitney-U test was used for the association between perception and selected variables. Result A total 193 nursing students participated in this study. The scores between 1 and 2.4 were considered ‘positive’, ‘unsure’ or ‘uncertain’, between 2.5 and 3.4 and negative for 3.5 and above’ over all nurses showed relatively negative perception towards suicide attempters. According to results from eight domains of perception, those who did not attend the courses on psychiatry nursing presented higher score, which was found to have significant association with permissiveness, unpredictability and incomprehensibility. A younger nursing student shows significant association with incomprehensibility. Younger nursing student showed statistically significant association with duration of suicide process. Conclusion Nursing students are frequently encountered with attempted suicide patients. Therefore, they must be aware of their attitudes toward this group of patients as part of their professional and therapeutic role. Thus a nurse’s positive perception towards attempted suicide can play a key role in communicating and preventing a future suicide attempt.

TET2 is frequently mutated in myeloid neoplasms. Genetic TET2 deficiency leads to skewed myeloid differentiation and clonal expansion, but minimal residual TET activity is critical for survival of neoplastic progenitor and stem cells. Consistent with mutual exclusivity of TET2 and neomorphic IDH1/2 mutations, here we report that IDH1/2 mutant–derived 2-hydroxyglutarate is synthetically lethal to TET dioxygenase–deficient cells. In addition, a TET-selective small-molecule inhibitor decreases cytosine hydroxymethylation and restricted clonal outgrowth of TET2 mutant but not normal hematopoietic precursor cells in vitro and in vivo. Although TET inhibitor phenocopied somatic TET2 mutations, its pharmacologic effects on normal stem cells are, unlike mutations, reversible. Treatment with TET inhibitor suppresses the clonal evolution of TET2-mutant cells in murine models and TET2-mutated human leukemia xenografts. These results suggest that TET inhibitors may constitute a new class of targeted agents in TET2-mutant neoplasia. Significance: Loss-of-function somatic TET2 mutations are among the most frequent lesions in myeloid neoplasms and associated disorders. Here we report a strategy for selective targeting of residual TET dioxygenase activity in TET-deficient clones that results in restriction of clonal evolution in vitro and in vivo. See related video: https://aacrjournals.org/webinar-minimal-tet-activity-targetable-vulnerability-tet2-and-neomorphic-idh12-mutant

Eltrombopag, an FDA-approved non-peptidyl thrombopoietin receptor agonist, is clinically used for the treatment of aplastic anemia, a disease characterized by hematopoietic stem cell failure and pancytopenia, to improve platelet counts and stem cell function. Eltrombopag treatment results in a durable trilineage hematopoietic expansion in patients. Some of the eltrombopag hematopoietic activity has been attributed to its off-target effects, including iron chelation properties. However, the mechanism of action for its full spectrum of clinical effects is still poorly understood. Here, we report that eltrombopag bound to the TET2 catalytic domain and inhibited its dioxygenase activity, which was independent of its role as an iron chelator. The DNA demethylating enzyme TET2, essential for hematopoietic stem cell differentiation and lineage commitment, is frequently mutated in myeloid malignancies. Eltrombopag treatment expanded TET2-proficient normal hematopoietic stem and progenitor cells, in part because of its ability to mimic loss of TET2 with simultaneous thrombopoietin receptor activation. On the contrary, TET inhibition in TET2 mutant malignant myeloid cells prevented neoplastic clonal evolution in vitro and in vivo. This mechanism of action may offer a restorative therapeutic index and provide a scientific rationale to treat selected patients with TET2 mutant-associated or TET deficiency-associated myeloid malignancies.

We analyze the micromagnetics of short longitudinal modulations of a high-magnetization material in cylindrical nanowires made of a soft-magnetic material of lower magnetization such as permalloy, combining magnetic microscopy, analytical modeling, and micromagnetic simulations. The mismatch of magnetization induces curling of magnetization around the axis in the modulations, in an attempt to screen the interfacial magnetic charges. The curling angle increases with modulation length, until a plateau is reached with nearly full charge screening for a specific length scale~$\Delta_\mathrm{mod}$, larger than the dipolar exchange length of any of the two materials. The curling circulation can be switched by the Oersted field arising from a charge current with typical magnitude $10^{12} A/m^{2}$ for a diameter of $\sim$100 nm, and reaching a maximum for $\Delta_\mathrm{mod}$.

Medway Hospital’s ENT on-call team triages referrals according to the local ENT Emergency referral guideline. Recently, a poor referral and handover resulted in a patient with post-tonsillectomy bleeding collapsing while on an immovable trolley in ED minors. Following reflection, a referral proforma was formulated to help the on-call ENT doctor ensure safe patient management through a traffic light system. The proforma includes patient and referrer detail along with four referral outcomes or colours, depending on patient stability, each with prompts for initial management. This audit aims to measure adherence to the ENT referral proforma. We performed a retrospective cohort analysis of compliance to the new ENT referral pathway. Proformas completed between 18th October and 17th November 2020 were included. A quasi-random method of sampling was utilized, every two of three paper forms. The data was inputted on Excel. Each proforma was marked for documentation of ten aspects. Of the total seventy-seven proformas, one scored 100%. On average, proformas were 80% completed. The most documented, at 100%, was patient detail. While the least documented, at 8%, was stating if ED were informed of the plan. 5% of referrals required potential resuscitation in ED, of which none met all criteria for a safe handover. The audit shows that the referral proforma is currently not adequately completed. This provides a basis for further intervention to improve patient safety for ENT referrals. The next step would be to inform the department to discuss why adherence is poor and maximize future documentation.

Background: There are high rates of developmental delay (DD) in children with severe acute malnutrition (SAM). Vitamin B12 is mandatory for the myelination of brain during the early childhood period. Objective: The objective of the study was to find out the percentage of DD in SAM and its association with Vitamin B12 deficiency. Materials and Methods: A prospective, observational study was done in SAM children aged 0–60 months who were admitted in Nutritional Rehabilitation Centre. Developmental assessment was done by Vineland Social Maturity Scale and developmental quotient

Myelin is the protective sheath that surrounds nerves in vertebrates to protect axons, which thereby facilitates impulse conduction. Damage to myelin is associated with many neurodegenerative disea...

Simple Summary Multiple myeloma (MM) is a cancer of antibody-producing plasma cells that remains incurable. These cells heavily depend on protein disulfide isomerase, PDIA1, for folding and structural integrity of antibodies and other secretory proteins to avoid unresolvable stress caused if they remain unfolded. High PDIA1 expression confers resistance to proteasome inhibitors and other stressors due to the gain in endoplasmic reticulum (ER) function, while maintaining or increasing vulnerability to PDIA1 inhibition. Here we report the identification and characterization of an orally bioavailable novel PDIA1 inhibitor CCF642-34 that is effective against multiple myeloma in pre-clinical models. PDIA1, the ER resident enzyme essential for the folding of disulfide bond-containing proteins, is upregulated in relapse and refractory myeloma. This increase in PDIA1 confers its sensitivity to CCF642-34, a structurally optimized PDIA1 inhibitor that induces apoptosis in myeloma cells but not in normal bone-marrow-derived CD34+ hematopoietic stem and progenitor cells. Abstract Multiple myeloma is a genetically complex hematologic neoplasia in which malignant plasma cells constantly operate at the maximum limit of their unfolded protein response (UPR) due to a high secretory burden of immunoglobulins and cytokines. The endoplasmic reticulum (ER) resident protein disulfide isomerase, PDIA1 is indispensable for maintaining structural integrity of cysteine-rich antibodies and cytokines that require accurate intramolecular disulfide bond arrangement. PDIA1 expression analysis from RNA-seq of multiple myeloma patients demonstrated an inverse relationship with survival in relapsed or refractory disease, supporting its critical role in myeloma persistence. Using a structure-guided medicinal chemistry approach, we developed a potent, orally bioavailable small molecule PDIA1 inhibitor CCF642-34. The inhibition of PDIA1 overwhelms the UPR in myeloma cells, resulting in their apoptotic cell death at doses that do not affect the normal CD34+ hematopoietic stem and progenitor cells. Bortezomib resistance leads to increased PDIA1 expression and thus CCF642-34 sensitivity, suggesting that proteasome inhibitor resistance leads to PDIA1 dependence for proteostasis and survival. CCF642-34 induces acute unresolvable UPR in myeloma cells, and oral treatment increased survival of mice in the syngeneic 5TGM1 model of myeloma. Results support development of CCF642-34 to selectively target the plasma cell program and overcome the treatment-refractory state in myeloma.

COVID-19, a novel corona virus has affected the life of each and every individual worldwide. Nepal being the neighborhood country of china, though, we had a late case detection. But, since the month of July this virus has spread in an alarming manner in Nepal. Nepal being one of the developing countries, we lack in equipments, manpower resources and also in treatment centers. Looking into the devastating scenario of Covid 19 in China, Italy, New York, Brazil and our neighboring countries like India, Pakistan and Bangladesh is scary. We wondered how we were going to handle this pandemic if similar circumstances happened in our country too. At the same time being OBGYN residents, we all know pregnancy is very crucial and our patients have faced much difficulties to receive the maternal health services. In this manuscript, we have shared our experience regarding preparedness for COVID-19, management of positive cases and its effect in OBGYN trainee.

Summary This case study is from a 3D Seismic volume in the MSGBC basin covering approximately 28,000 km2 offshore Gambia-Senegal and Guinea-Bissau. The area is within the southern part of the prolific offshore MSGBC (Mauritania, Senegal, Gambia, Guinea-Bissau, Conakry) basin. The goal of this extensive 3D Seismic program is to image the prospective paleoshelf edge from shallow to deep water. We present a case study showing the importance of geologically constrained tomography to resolve the velocity variations associated with small geological features, along with the benefit of broadband data on velocity modelling and final seismic imaging.

TET2 is one of the most frequently mutated genes in myeloid neoplasms. TET2 loss-of-function perturbs myeloid differentiation and causes clonal expansion. Despite extensive knowledge regarding biochemical mechanisms underlying distorted myeloid differentiation, targeted therapies are lagging. Here we review known biochemical mechanisms and candidate therapies that emerge from this. Specifically, we discuss the potential utility of vitamin C to compensate for TET-dioxygenase deficiency, to thereby restore the biochemical function. An alternative approach exploits the TET-deficient state for synthetic lethality, exploiting the fact that a minimum level of TET-dioxygenase activity is required for cell survival, rendering TET2-mutant malignant cells selectively vulnerable to inhibitors of TET-function.

ObjectivesInforming research participants of the results of studies in which they took part is viewed as an ethical imperative. However, there is little guidance in the literature about how to do this. The Fluoxetine Or Control Under Supervision trial randomised 3127 patients with a recent acute stroke to 6 months of fluoxetine or placebo and was published in the Lancet on 5 December 2018. The trial team decided to inform the participants of the results at exactly the same time as the Lancet publication, and also whether they had been allocated fluoxetine or placebo. In this report, we describe how we informed participants of the results.DesignIn the 6-month and 12-month follow-up questionnaires, we invited participants to provide an email address if they wished to be informed of the results of the trial. We re-opened our trial telephone helpline between 5 December 2018 and 31 March 2019.SettingUK stroke services.Participants3127 participants were randomised. 2847 returned 6-month follow-up forms and 2703 returned 12-month follow-up forms; the remaining participants had died (380), withdrawn consent or did not respond.ResultsOf those returning follow-up questionnaires, a total of 1845 email addresses were provided and a further 50 people requested results to be sent by post. Results were sent to all email and postal addresses provided; 309 emails were returned unrecognised. Seventeen people replied, of whom three called the helpline and the rest responded by email.ConclusionIt is feasible to disseminate results of large trials to research participants, though only around 60% of those randomised wanted to receive the results. The system we developed was efficient and required very little resource, and could be replicated by trialists in the future.Trial registration numberISRCTN83290762; Post-results.

Role of particle size distribution on magnetization of two polydispersed samples of antiferromagnetic NiO particles is investigated. Zero field-cooled and field-cooled susceptibility as a function of temperature (10–350 K) data indicate that samples are superparamagnetic. This behavior is again confirmed from M–H loops (up to $${\pm }\,70$$ kG) at 10 and 350 K. The magnetization as a function of applied magnetic field data at 350 K is analyzed considering a lognormal distribution in particle size. The estimated particle size distributions are compared with those determined from transmission electron micrographs. We find that the particle size distribution is overestimated if magnetic anisotropy is ignored while analyzing the magnetization data.

Magnetization measurements on antiferromagnetic iron storage protein ferritin are reported. The magnetization as a function of applied magnetic field data in superparamagnetic region are analyzed considering a distribution in particle size. The estimated particle size distribution is compared with that determined from transmission electron micrograph. Effect of magnetic anisotropy on magnetization process of this nanoparticle system is also discussed.

The early detection of the cancer will dramatically improve the prognosis. There are many diagnostic aids that can be employed for screening. Hence, in our study, we aim to evaluate the efficiency of dyes in detection of cancer cells.We piloted a prospective clinical study among 30 subjects to detect the efficiency of two dyes-Lugol's iodine and toluidine blue. They were grouped equally for potentially malignant and malignant lesions. The areas were analyzed for the diagnostic accuracy of the dyes using the histopathological examination and compared using chi square test deliberatingWe observed that both the dyes showed a diagnostic accuracy of over 90% when used alone or in combination. Both the dyes had greater sensitivity while specificity was 20%.The combination of these two dyes can be used as an efficient screening aid for the identification of the cancer cells.

Background: TET-dioxygenases (TET1, TET2, and TET3) are key epigenetic regulators, which require molecular oxygen, αKG, and Fe 2+ to progressively oxidize 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxylcytosine, leading to the demethylation of mCpGs at promoter and enhancer, a key step for mounting an efficient transcription profile and thereby differentiation. Loss of function TET2 mutation (TET2 MT) is frequently observed in myeloid neoplasms (MN). Despite extensive studies of the biochemical mechanisms underlying distorted myeloid differentiation and neoplastic evolution of TET2 MT HSPCs, targeted therapies are lagging. Here, we report the therapeutic utility of preferential targeting of TET2 mutant and TET-dioxygenase deficient cells either by restoring the lost activity to induce differentiation and death or by inhibiting the residual activity to induce synthetic lethality. Methods: Genetic as well as pharmacologic in vitro and in vivo models of AML were used to validate specific targeting of TET2 mutant cells. Knockout and over-expression in the isogenic background of different cells were used to understand the pro-survival and proliferative mechanism of TET2 loss in myeloid cells. The efficacy of small molecule pharmacophore targeting TET-dioxygenase was evaluated in vitro in cell free and cell culture as well as in vivo in a mouse competitive transplant model to demonstrate the proof of therapeutic concept. Result: Ascorbate, used as an activator of αKG/Fe 2+ dependent dioxygenase including TETs, functions via maintaining the redox state of iron (II) in the dioxygenase catalytic site with a half-maximal effective concentration of 13.8 ± 1.3 µM for enhancing TET2activity as measured by an ELISA for 5hmC. However, the reported intracellular cellular concentrations of ascorbate in human remains in the millimolar range, well above the saturating concentration required for TET-dioxygenases activation. Interestingly, ascorbate failed to activate TET in primary mononuclear cells isolated from myeloid neoplasia patients with TET2 mutations. Consistent with the in-vitro observation, we found that ascorbate treatment in physiologically relevant doses has no significant effect in syngeneic murine models (p=0.1). TET-inactivation is a complex phenomenon that is controlled by context-dependent post-translational modifications including lysine acetylation by acetyltransferases and class I and II deacetylases. Therefore, the anti-leukemic effect of the supraphysiological doses of ascorbate in humans and mice leukemia models may not be dependent on TET activation by ascorbate. On the other hand, a comprehensive analysis of the configurations of TET2 MT in myeloid neoplasia (n= 2617) demonstrated a remarkable exclusivity with 2-hydroxyglutarate (2-HG) producing neomorphic IDH1/2 MT. Ectopic inducible expression of IDH1/2 MT induced synthetic lethality of malignant TET2 MT cells. Therefore, themutual exclusivity of TET2 MT and neomorphic IDH1/2 MT is due to synthetic lethality caused by 2-HG production. In addition, we demonstrate that sequential deletion of TET1 and TET3 leads to growth arrest in cellular models, which has never been observed in any myeloid malignancies. Altogether, these observations suggest that malignant TET2 mutant myeloid cells may be vulnerable to inhibition of residual TET-activity coming from TET1 and TET3. Therefore, we synthesized TET-specific inhibitor on 2HG scaffold using iterative computer-aided design, characterized in cell-free as well as cell culture model systems, and selected TETi76 with no cytotoxic effect in normal bone marrow-derived CD34 + cells. TETi76 selectively inhibits TET-dioxygenase activity with IC50 of 1.5, 9.4, and 8.8 μmol/L for TET1/2/3, and it restricts clonal outgrowth of TET2 MT both in vitro and in vivo in malignant and nonmalignant clonal hematopoiesis of indeterminate potential (CHIP). Conclusion: Taken together, here we demonstrated that anti-cancer activity of ascorbate may not come from its role as a TET activator and that a minimum level of TET-dioxygenase activity is required for cell survival, rendering TET2 -mutant malignant cells selectively vulnerable to inhibitors of TET-function. Disclosures Maciejewski: Regeneron: Consultancy; Novartis: Consultancy; Bristol Myers Squibb/Celgene: Consultancy; Alexion: Consultancy.

A set of 1,2,3-triazole incorporated quinolone antibiotic conjugates 10–15, 17–19 were synthesized via microwave assisted click chemistry technique. Some of the aryl-substituted conjugates 17–19 show promising antibacterial properties against the tested Gram-positive (S. aureus and S. pyogenes) and Gram-negative bacteria (S. typhi) with potency higher than that of the parent antibiotics 1–3. 2D-QSAR modeling supports the observed biological properties.

Undoped, 10% and 20% Al doped samples of layered hexagonal α -Co(OH)2 are synthesized. X-ray diffraction patterns show that the samples are nanocrystalline with average crystallite size 15 nm. Fourier transform infrared spectroscopy and thermogravimetric analysis are performed to establish molecular formula of samples. Analysis of susceptibility χ as a function of temperature T (5–300 K) data indicates for ferromagnetic transitions in the system at lower temperatures. Curie transition temperature T C for undoped, 10% and 20% Al doped α -Co(OH)2 are found to be 9, 7.5 and 6.0 K respectively. Fitting of χ vs. T data for T > T C to the high temperature series yields strength of intralayer and interlayer exchange interactions among Co 2 + ions. For undoped sample values of these interactions are estimated to be 1.070 and 0.240 K respectively. Strength of these interactions decrease with increasing concentration of Al 3 + ions in the lattice. Magnetization as a function of applied magnetic field (up to ± 50 kG) data confirm ferromagnetic nature of the samples at 5 K.

Magnetic behavior of 5 nm antiferromagnetic six lines ferrihydrite particles is reported. The magnetization as a function of applied magnetic field data at 100 K are fitted to the modified Langevin function without and with considering a distribution in particle magnetic moment covering different ranges of applied magnetic field strength. The resulting fit parameters are analyzed and compared. In the first case fit parameters are found to be strongly dependent on the strength of applied magnetic field. But the fit parameters are found to be independent of applied magnetic field strength in the later case.

Severe aplastic anemia (sAA) is bone marrow (BM) failure syndrome characterized by immune mediated BM hypoplasia and pancytopenia usually treated with immunosuppressive therapy (IST) or allogeneic BM transplant. Despite the successes of IST in treating acquired, idiopathic sAA, most of the available treatment options only convert severe into non-severe forms of disease. Recently, eltrombopag (Epag), a small molecule thrombopoietin receptor (TPOR) agonist has been introduced into the routine treatment of de-novo and refractory AA with robust clinical response. Epag treatment showed remarkable tri-lineage responses in AA, suggesting an effect on hematopoietic stem and progenitor cells (HSPCs) amplification. The use of hematopoietic growth factors in AA that increases HSPCs has been associated with justified fears of facilitating AA evolution to MDS. Similarly, Epag has parallel effects on the amplification of HSPCs and there has been recent signals that long term treatment of Epag may increase the MDS risk. Using an in-house developed highly stable cell free high throughput screening of bioactive small molecule chemical library, we identified Epag as one of the most potent inhibitor of TET2. TET2 is the most abundant DNA dioxygenase in HSPCs. The loss-of-function mutations of TET2 frequently occurs in myeloid neoplasia. Here using in vitro and in vivo model system we report that effect of Epag, in part, is mediated by its ability to inhibit TET-dioxygenase activity. We demonstrate that Epag directly binds to TET2 in the presence of Fe2+ and inhibits its dioxygenase function in cell free system. In cell culture model we observed that nearly 40% of Epag partitioned into to nucleus suggesting its potential TET2 inhibitory role in cells. To test if TPOR signaling activation by Epag has any impact on TET activity we engineered murine cell lines BaF3 and 32D by introducing human TPOR. Irrespective of the status of TPOR-JAK-STAT pathway activation, as reflected in STAT5 phosphorylation, Epag demonstrated a robust TET-inhibition activity. However, in similar experiments peptide agonist of TPOR or another small molecule TPOR agonist Avotrombopag did not have any TET inhibitory effects. Consistent with these observations, Epag but not TPO or Avotrombopag inhibited TET activity in murine BM. Thus, Epag mediated TET2 inhibition in these cells are specific and independent of TPOR signaling. Consistent with previous reports, we observed that Epag treatment can significantly expand murine HSPCs even in the absence of TPOR signaling activation. The hematopoietic activity of Epag in murine system is dependent on its ability to inhibit TET2 as confirmed by the lack of Epag effects on Tet2-/- HSPCs. We further confirmed that TET-inhibition by Epag is central for its ability to expand HSPCs in murine model under in vivo settings using BM transplant experiments. We transplanted a mixture of cells derived from Tet2+/+ Pep Boy (CD45.1, 95%) and Tet2-/- C57BL/6 (CD45.2, 5%) into lethally irradiated recipient PepBoy mice. Epag treatment of mice reconstituted with the mixture of BM cells increases the fraction of Tet2+/+ BM cells only. Interestingly, Epag mimic loss of Tet2 in vivo as observed in the expansion of myeloid compartments of Tet2+/+ fractions reflected in CD11b-CD11c+, CD11b+CD11c-, CD11b+CD11c-Ly6C+Ly6G- (monocytes) and CD11b+CD11c-Ly6ClowLy6G+ (neutrophils). We did not observe any significant change in B220+, CD4+ or CD8+ lymphoid populations. On the contrary, we observed a growth restrictive effect of Epag treatment on Tet2-/- myeloid cells. Consistent with our hypothesis that Epag inhibits TET-dioxygenase activity, we observed hypermethylation in the mononuclear cells derived AA patients (n= 16) after Epag treatment. This was further confirmed in ex vivo Epag treatment of BM cells derived from healthy donor. Epag treatment significant increase clonogenic expansion of normal BM cells but not TET2 mutant myeloid cells. In summary, here we demonstrate that Epag is a potent inhibitor of TET-dioxygenase and its biological consequences are independent of TPOR-JAK-STAT activation. Epag treatment mimics TET2 loss of function in AA cells, however it restricts clonal expansion of TET2 mutant myeloid cells. Disclosures Patel: Alexion: Other: educational speaker. Nazha:Jazz: Research Funding; Incyte: Speakers Bureau; MEI: Other: Data monitoring Committee; Novartis: Speakers Bureau. Saunthararajah:EpiDestiny: Consultancy, Current equity holder in private company, Patents & Royalties: University of Illinois at Chicago. Sekeres:Pfizer: Consultancy; BMS: Consultancy; Takeda/Millenium: Consultancy. Maciejewski:Alexion, BMS: Speakers Bureau; Novartis, Roche: Consultancy, Honoraria.

Multiple myeloma (MM) is a genetically complex hematological disease which is characterized by clonal proliferation of plasma cells in the bone marrow and secretion of monoclonal antibodies and cytokines that can damage bone, bone marrow, and kidney function1. MM cells constantly operate at the limit of their unfolded protein response (UPR) in the face of a secretory load of immunoglobin (Ig) and cytokines that is unparalleled by any other mammalian cell 2,3 and microenvironmental factors that aggravate the degree of physiologic misfolding that occurs during synthesis of secreted proteins. The endoplasmic reticulum (ER) resident protein disulfide isomerases (PDIs) are indispensable for folding of secreted proteins that require intramolecular disulfide-bond arrangement 4 like antibodies and many cytokines. As the main PDI family member, near-complete function of PDIA1 is essential for survival of MM cells while its inhibition should be manageable by the UPR in normal cells creating an opportunity for a large therapeutic window for PDI inhibitors in MM. Previously, we discovered and characterized an irreversible PDI inhibitor (CCF642) that induced cell death in MM cells at doses that did not affect survival of normal bone marrow cells. However, CCF642 has poor solubility and suboptimal selectivity precluding clinical translation. Using structure guided medicinal chemistry, we developed and characterized a highly potent and selective PDI inhibitor, with 10-fold higher potency (Fig 1B) and selectivity. CCF642-34 showed remarkable selectivity against PDIA1 and off-target bindings were eliminated when compared to CCF642 (Fig 1C). In addition to improved selectivity and in vitro PDI inhibition, CCF642-34 demonstrated more than 3-fold higher potency compared to CCF642 against MM1.S and bortezomib resistant MM1.S cells remained sensitive to CCF642-34. Importantly, the novel analogue CCF642-34 has 18-fold better potency in restricting the colony forming abilities of RPMI1640 cells while at no effect on the clonogenic potential of CD34+ cells derived from healthy bone marrow was observed at equivalent doses. CCF642-34 induces ER stress in MM1.S cells as observed in dose and time dependent cleavage of XBP1, IRE1α oligomerization and the profound induction of programmed cell death reflected by PARP and caspase 3 cleavage. To further analyze the modes of action of CCF642-34 and CCF642 we performed RNAseq after treatment of MM1.S cells and found exclusive induction of genes associated with UPR and downstream cell cycle and apoptotic responses for CCF642-34 while additional genes affecting were detected after CCF642 treatment. There were 362 and 568 differentially expressed genes in CCF642-34 and CCF-642 (compared to controls) treated MM1.S cells, respectively. Among these differentially expressed genes 87 down regulated and 142 upregulated were common to both, including downregulation of cell division and mitotic cell cycle process, and upregulation of response to ER stress, unfolded protein response, and apoptotic process gene sets. Results confirm that both CCF642 and CCF642-34 treatment act by inducing lethal ER-stress with greater selectivity for CCF642-34. Accordingly, hierarchical clustering showed distinct gene expression profiles in 642-34 and 642 treated MM1S cells (Fig. 2). CCF642-34 is orally bioavailable and highly efficacious in against established multiple myeloma in a syngeneic 5TGM1-luc/C57BL/KaLwRij model of myeloma. All vehicle control animals were dead by 52 days while 3 out of 6 mice lived beyond 6 months with no sign of relapse. In summary, we synthesized and characterized a novel lead PDIA1 inhibitor based on structure-guided medicinal chemistry that has improved pharmacologic properties to act as novel lead for clinical translation. References: 1. Manier S, Salem KZ, Park J, et al. Genomic complexity of multiple myeloma and its clinical implications. Nat. Rev. Clin. Oncol. 2017; 2. Fonseca R, Bergsagel PL, Drach J, et al. International Myeloma Working Group molecular classification of multiple myeloma: Spotlight review. Leukemia. 2009; 3. Wang M, Kaufman RJ. The impact of the endoplasmic reticulum protein-folding environment on cancer development. Nat. Rev. Cancer. 2014; 4. Freedman RB, Hirst TR, Tuite MF. Protein disulphide isomerase: building bridges in protein folding. Trends Biochem. Sci. 1994; Disclosures Valent: Takeda Pharmaceuticals: Other: Teaching, Speakers Bureau; Celgene: Other: Teaching, Speakers Bureau; Amgen Inc.: Other: Teaching, Speakers Bureau.

We have analyzed the magnetization data of antiferromagnetic ferritin particles. Ferritin is an ideal superparamagnetic system with lower bifurcation temperature Tbf (≈ 15.2 to 18.2 K). Therefore, the magnetization data as a function of applied magnetic field (up to 50 kOe) are measured at 150 K. The data are investigated considering the presence of magnetic anisotropy. The estimated value of magnetic anisotropy constant for ferritin is found to be ≈ 104 J/m3. This value of magnetic anisotropy constant is also verified by Neel relaxation theory. The results indicates that this method is helpful for estimation of magnetic anisotropy constant of magnetic nanoparticle systems.

Ferritin is an iron storage protein found in living organisms. It is an antiferromagnetic nanoparticle system consisting of an inorganic core surrounded by a protein shell. Ferritin is characterized by X-ray diffractometer, transmission electron microscope, atomic absorption spectrometer and thermogravimetric analyzer. We find that the ferritin core is poorly crystalline, 8 nm in size and consists of 10 wt% iron. It is believed that cores of ferritin consist of single-phase inorganic mineral ferrihydrite. Recently, we have shown that ferrihydrite decomposes directly to $$\alpha$$-$$\hbox {Fe}_{{2}}\hbox {O}_{3}$$ on heating in air at 440 $$^{\circ }$$C. In the present work, we show that ferritin cores gradually decompose to a mixture of $$\gamma$$-$$\hbox {Fe}_{{2}}\hbox {O}_{{3}}$$ and $$\alpha$$-$$\hbox {Fe}_{{2}}\hbox {O}_{{3}}$$ on heating in air. This mixture finally stabilizes to $$\alpha$$-$$\hbox {Fe}_{{2}}\hbox {O}_{{3}}$$ on further heating. The magnetic behaviour of final sample is also studied. This work confirms that the ferritin cores contain more than one phase.