Replenishment Of Stores Research Articles

Importance of ferritin research in clinical practice

Serum ferritin (SF) is typically present in serum at concentrations directly related to iron (Fe) storage and is therefore traditionally used as an indicator of Fe levels in body tissues. Reducing its level is the “gold standard” for diagnosing widespread iron deficiency conditions. No less significant is hyperferritinemia — a nonspecific syndrome that occurs when Fe reserves are overloaded, a number of immunoinflammatory, infectious, oncological diseases, liver diseases, etc. In many pathological conditions, the level of SF determines the severity and prognosis of the disease. Ferritin concentrations greater than 1000 ng/mL, regardless of cause, have been shown to be associated with higher mortality. The reason for the increase in the level of SF in liver pathologies (cancer, hepatitis, cirrhosis) is associated with the process of its release from hepatocytes during their destruction. On the other hand, excessive synthesis and/or cellular secretion of ferritin occurs under the influence of various stimuli (cytokines, oxidative stress, hypoxia, oncogenes and growth factors). The interpretation of elevated ferritin values goes far beyond the role of an indicator of replenishment of Fe stores in tissues. Only 10% of cases of hyperferritinemia are associated with iron overload; in most patients, it is defined as the result of the acute phase and a reactive increase in ferritin levels against the background of any disease. The variety of symptoms of iron deficiency syndromes and hyperferritinemia is due to the involvement of many organs and systems, which requires a thorough examination (study of complaints, anamnesis, family and concomitant diseases, as well as the necessary laboratory and instrumental studies) to search for possible causes. Systemic Fe homeostasis must be closely monitored on a regular basis. It is required to comply with the conditions for blood sampling for SF. Reference values for SF concentrations vary depending on the analytical methods used and the population studied. Age and gender play an important role. Given the range of reference values, it is important for a particular patient to focus on the initial level of his ferritin, determined against the background of health and well-being during clinical observation.

Misprogramming of glucose metabolism impairs recovery of hippocampal slices from neuronal GLT-1 knockout mice and contributes to excitotoxic injury through mitochondrial superoxide production.

We have previously reported a failure of recovery of synaptic function in the CA1 region of acute hippocampal slices from mice with a conditional neuronal knockout (KO) of GLT-1 (EAAT2, Slc1A2) driven by synapsin-Cre (synGLT-1 KO). The failure of recovery of synaptic function is due to excitotoxic injury. We hypothesized that changes in mitochondrial metabolism contribute to the heightened vulnerability to excitotoxicity in the synGLT-1 KO mice. We found impaired flux of carbon from 13C-glucose into the tricarboxylic acid cycle in synGLT-1 KO cortical and hippocampal slices compared with wild-type (WT) slices. In addition, we found downregulation of the neuronal glucose transporter GLUT3 in both genotypes. Flux of carbon from [1,2-13C]acetate, thought to be astrocyte-specific, was increased in the synGLT-KO hippocampal slices but not cortical slices. Glycogen stores, predominantly localized to astrocytes, are rapidly depleted in slices after cutting, and are replenished during exvivo incubation. In the synGLT-1 KO, replenishment of glycogen stores during exvivo incubation was compromised. These results suggest both neuronal and astrocytic metabolic perturbations in the synGLT-1 KO slices. Supplementing incubation medium during recovery with 20 mM D-glucose normalized glycogen replenishment but had no effect on recovery of synaptic function. In contrast, 20 mM non-metabolizable L-glucose substantially improved recovery of synaptic function, suggesting that D-glucose metabolism contributes to the excitotoxic injury in the synGLT-1 KO slices. L-lactate substitution for D-glucose did not promote recovery of synaptic function, implicating mitochondrial metabolism. Consistent with this hypothesis, phosphorylation of pyruvate dehydrogenase, which decreases enzyme activity, was increased in WT slices during the recovery period, but not in synGLT-1 KO slices. Since metabolism of glucose by the mitochondrial electron transport chain is associated with superoxide production, we tested the effect of drugs that scavenge and prevent superoxide production. The superoxide dismutase/catalase mimic EUK-134 conferred complete protection and full recovery of synaptic function. A site-specific inhibitor of complex III superoxide production, S3QEL-2, was also protective, but inhibitors of NADPH oxidase were not. In summary, we find that the failure of recovery of synaptic function in hippocampal slices from the synGLT-1 KO mouse, previously shown to be due to excitotoxic injury, is caused by production of superoxide by mitochondrial metabolism.

THE LOW SERUM FERRITIN AS AN INDICATOR OF INADEQUATE NUTRITION IN INTERNATIONAL-LEVEL FEMALE SWIMMER

Introduction: Iron deficiency is a common feature in athletes, and optimizing iron levels may become a contributing factor to improved athletic performance. Purpose: This study aimed to evaluate body iron stores and their relationship with sports performance. It also sought correlations between the swimmers’ hematological, hormonal, anthropometric, and functional status and their sports achievements. Methodology: The research involved 19 athletes (11 women and 8 men) from the Bulgarian national swimming team. They were tested for maximal aerobic capacity on a cycle ergometer and had blood samples taken to determine iron status and hematological and hormonal markers. Their swimming achievements (Swimming points) were evaluated as a percentage of the world records in the respective discipline. Results: There were statistically significant differences between the female and male swimmers for Unbound Iron Binding Capacity (UIBC) and ferritin. A higher UIBC in women indicated lower iron levels in them. Among female swimmers, five had ferritin levels below 40 ng/mL. In women, a highly reliable correlation between the Swimming points and ferritin concentration (r = .68) was observed. All male swimmers had ferritin values above 40 ng/mL, and there was no reliable correlation between ferritin concentration and Swimming points, probably because ferritin was in the optimal range and is no longer a significant performance factor. Conclusions: In swimmers, the optimal serum ferritin concentration is probably in the range of 40–90 ng/mL. Many female swimmers have a relative iron deficiency with sub-optimal ferritin values. In competitive female swimmers, low ferritin level is an indicator of both iron deficiency and inadequate diet. It can be expected that an adequate nutritional regime in athletes with low ferritin values will lead both to the replenishment of iron stores and to an increase in muscle mass and, ultimately, to an improvement in sports performance.

A comparative study of ferric carboxymaltose and iron sucrose as a parenteral iron treatment in iron deficiency anemia during pregnancy in a tertiary care hospital

Background: Anemia from the Greek word (anhaima) meaning “without blood” is the deficiency of red blood cell and/or hemoglobin which result in reduced oxygen-carrying capacity of blood causing tissue hypoxia. Aims and Objectives: To study the efficacy and safety of intravenous ferric carboxymaltose (FCM) versus iron sucrose in the management of iron deficiency anemia in pregnancy. Materials and Methods: A prospective, single-center, comparative interventional randomized study was carried among 180 antenatal mother admitted to antenatal ward in the Department of Obstetrics and Gynaecology of Burdwan Medical College and Hospital, from July 1st, 2020, to December 30th, 2021. Results: A total of 180 patients, 90 patients in each FCM and iron sucrose group, were included in the study. There was significant (P=0.001) mean change in hemoglobin (Hb) level in both the groups from pre-treatment to 3 and 6-week post-treatment. There was also a significant (P=0.001) mean change in ferritin level in both the groups from pre-treatment to 3 and 6-week post-treatment. The mean change in Hb level and ferritin level was higher among patients of FCM compared to iron sucrose. The adverse reactions were lower among patients of FCM than iron sucrose. Conclusions: This study found that FCM is safer than iron sucrose. Treatment with FCM resulted in rapid replenishment of iron stores in pregnant women with significantly high rise of Hb and ferritin levels over a 6-week period with lesser adverse effects.

Astroglial S100B Secretion Is Mediated by Ca2+ Mobilization from Endoplasmic Reticulum: A Study Using Forskolin and DMSO as Secretagogues

S100B, a homodimeric Ca2+-binding protein, is produced and secreted by astrocytes, and its extracellular levels have been used as a glial marker in brain damage and neurodegenerative and psychiatric diseases; however, its mechanism of secretion is elusive. We used primary astrocyte cultures and calcium measurements from real-time fluorescence microscopy to investigate the role of intracellular calcium in S100B secretion. In addition, the dimethyl sulfoxide (DMSO) effect on S100B was investigated in vitro and in vivo using Wistar rats. We found that DMSO, a widely used vehicle in biological assays, is a powerful S100B secretagogue, which caused a biphasic response of Ca2+ mobilization. Our data show that astroglial S100B secretion is triggered by the increase in intracellular Ca2+ and indicate that this increase is due to Ca2+ mobilization from the endoplasmic reticulum. Also, blocking plasma membrane Ca2+ channels involved in the Ca2+ replenishment of internal stores decreased S100B secretion. The DMSO-induced S100B secretion was confirmed in vivo and in ex vivo hippocampal slices. Our data support a nonclassic vesicular export of S100B modulated by Ca2+, and the results might contribute to understanding the mechanism underlying the astroglial release of S100B.

Intravenous ferric carboxymaltose and ferric derisomaltose alter the intestinal microbiome in female iron-deficient anemic mice.

Iron deficiency anemia (IDA) is a leading global health concern affecting approximately 30% of the population. Treatment for IDA consists of replenishment of iron stores, either by oral or intravenous (IV) supplementation. There is a complex bidirectional interplay between the gut microbiota, the host's iron status, and dietary iron availability. Dietary iron deficiency and supplementation can influence the gut microbiome; however, the effect of IV iron on the gut microbiome is unknown. We studied how commonly used IV iron preparations, ferric carboxymaltose (FCM) and ferric derisomaltose (FDI), affected the gut microbiome in female iron-deficient anemic mice. At the phylum level, vehicle-treated mice showed an expansion in Verrucomicrobia, mostly because of the increased abundance of Akkermansia muciniphila, along with contraction in Firmicutes, resulting in a lower Firmicutes/Bacteroidetes ratio (indicator of dysbiosis). Treatment with either FCM or FDI restored the microbiome such that Firmicutes and Bacteroidetes were the dominant phyla. Interestingly, the phyla Proteobacteria and several members of Bacteroidetes (e.g., Alistipes) were expanded in mice treated with FCM compared with those treated with FDI. In contrast, several Clostridia class members were expanded in mice treated with FDI compared with FCM (e.g., Dorea spp., Eubacterium). Our data demonstrate that IV iron increases gut microbiome diversity independently of the iron preparation used; however, differences exist between FCM and FDI treatments. In conclusion, replenishing iron stores with IV iron preparations in clinical conditions, such as inflammatory bowel disease or chronic kidney disease, could affect gut microbiome composition and consequently contribute to an altered disease outcome.

Diving behaviour of southern elephant seals: new models of behavioural and ecophysiological adjustments of oxygen store management.

Among pinnipeds, southern elephant seals (SESs, Mirounga leonina) are extreme divers that dive deeply and continuously along foraging trips to restore their body stores after fasting on land during breeding or moulting. Their replenishment of body stores influences their energy expenditure during dives and their oxygen (O2) reserves (via muscular mass), yet how they manage their O2 stores during their dives is not fully understood. In this study, 63 female SESs from Kerguelen Island were equipped with accelerometers and time-depth recorders to investigate changes in diving parameters through their foraging trips. Two categories of dive behaviour were identified and related to the body size of individuals, with smaller SESs performing shallower and shorter dives requiring greater mean stroke amplitude compared with larger individuals. In relation to body size, the larger seals had lower estimated oxygen consumption levels for a given buoyancy (i.e. body density) compared with smaller individuals. However, both groups were estimated to have the same oxygen consumption of 0.079±0.001 ml O2 stroke-1 kg-1 for a given dive duration and at neutral buoyancy when the cost of transport was minimal. Based on these relationships, we built two models that estimate changes in oxygen consumption according to dive duration and body density. The study highlights that replenishing body stores improves SES foraging efficiency, as indicated by increased time spent at the bottom of the ocean. Thus, prey-capture attempts increase as SES buoyancy approaches the neutral buoyancy point.

ACUTE EFFECTS OF CLUSTER SET AND TRADITIONAL SET POST-ACTIVATION POTENTIATION PROTOCOLS ON VERTICAL JUMP PERFORMANCE

The aim of this study was to investigate the acute effects of different post-activation potential (PAP) protocols on Countermovement Jump (CMJ) performance in female athletes. Twelve elite female taekwondo athletes (age 15.17±.718 years, height 168.66±4.81 cm and body weight 49.25±2.37 kg) participated in the study voluntarily. The resistance training methods of the traditional set (1RM 75% x 3 sets x 12 repetitions and 180 s rest between sets) and cluster set (1RM 75% x 3 sets x 4+4+4 (total 12) repetitions, 30 s rest between sets and 180 s rest between sets) were applied to the research group. CMJ test performances were recorded for 30 s, 4 min and 8 min before and after both PAP protocols. Paired sample t-test was applied for pre-posttest comparison. In the traditional set structure, there was a statistically significant difference between CMJ pre test and CMJ 4 min test. In the cluster set structure, a statistically significant difference was found between CMJ pre test and CMJ30 sec, between CMJ pre test and CMJ 4 min and between CMJ pre test and CMJ 8 min. As a result, the PAP effect is better with the cluster set structure than with the traditional set due to the intermittent rest that will be given with the cluster set structure due to the sudden replenishment of energy stores.<p> </p><p><strong> Article visualizations:</strong></p><p><img src="/-counters-/soc/0017/a.php" alt="Hit counter" /></p>

Practical Clinical Consensus Guidelines for the Management of Cancer Associated Anemia in Low- and Middle-Income Countries.

Purvish M. ParikhCancer-associated anemia (CAA) remains a major unmet need that compromises overall survival (OS) and quality of life (QoL). Currently, available guidelines do not take into consideration the unique challenges in low- and middle-income countries (LMIC). Our CAA patients have to battle preexisting impaired nutritional status, depleted body iron stores, financial limitations, and difficulty in having easily accessible affordable healthcare. Hence, we fulfilled the need of guidelines for LMIC. A group of subject experts were put together, given background literature, met in a face-to-face discussion, voted using Delphi process, and finally agreed on the contents of this guideline document. As many as 50% of cancer patients will have significant anemia (hemoglobin < 10 g/dL) at initial diagnosis. It is most commonly seen with gastrointestinal malignancies, head and neck cancers, and acute leukemias. The hemoglobin falls further after initiation of cancer directed therapy, due to chemotherapy itself or heightened nutritional deficiency. Its evaluation should include tests for complete blood count, red blood cell morphology, reticulocyte count, Coombs test, and levels of vitamin B12 and folic acid. Iron status should be monitored using test to measure serum iron, total iron binding capacity, transferring saturation, and serum ferritin levels. A minimum of 50% of cancer patients with anemia require iron supplements. The preferred mode of therapy is with intravenous (IV) iron using ferric carboxymaltose (FCM). Most patients respond satisfactorily to single dose of 1000 mg. It is also safe and does not require use of a test dose. Significant anemia is found in at least half of all cancer patients in India, South Asian Association for Regional Cooperation region, and other LMIC countries. Its awareness among healthcare professionals will prevent it from remaining undiagnosed (in up to 70% of all cancer patients) and adversely affecting OS and QoL. The benefits of treating them with IV iron therapy are quick replenishment of iron stores, hemoglobin returning to normal, better QoL, and avoiding risk of infections/reactions with blood transfusions. Many publications have proven the value of single-dose FCM in such clinical situations. CAA has been proven to be an independent prognostic factor that adversely affects both QoL and OS in cancer patients. Use of FCM as single IV dose of 1000 mg is safe and effective in the majority of patients with CAA.

Obstetric Determinants of Birth Weight Status of Babies Born at Jaramogi Oginga Odinga Teaching and Referral Hospital, Kenya

Background: Low birth weight (LBW) is a serious public health problem, especially in developing countries. Globally, 15.5% of all births are born LBW and 95.6% of them are in developing countries. In Kenya, 8% of babies are born LBW and at Jaramogi Oginga Odinga Teaching and Referral Hospital (JOOTRH), it is 11.1%. Despite several efforts such as antenatal care services put in place to improve the quality of maternal and child health, the rates of LBW are still high. LBW is a major cause of mortality and morbidity in infants. Objective: The objective of this study was to assess obstetric determinants of the birth weight status of babies born at JOOTRH, Kenya. Materials and Methods: This was a cross-sectional study with a sample size of 131 babies plus their mothers from a total population of 538 deliveries during the study period. Systematic random sampling was used to select the respondents. Data was collected using a questionnaire and record review checklist. Data analysis was done using descriptive statistics and inferential statistics, specifically logistic regression. Result: The study results showed that the prevalence of LBW was 13.7%. LBW was strongly associated with a birth interval of &lt; 24 months (AOR = 13.722), hypertension (AOR = 11.753), previous history of LBW (COR = 14.0), and gestation at birth (COR = 3.75). Gravidity, previous history of miscarriage and gestation at which antenatal visits started had no association with LBW. Conclusion: The prevalence of LBW is 13.7%, higher than the national rate of 8% and statistically significant with a Z score of 2.398, p = 0.016. The birth interval of &lt; 24 months, hypertension, previous history of LBW, and gestation at birth influenced the occurrence of LBW. Focused antenatal care to ensure early detection and management of high-risk pregnancies and educate pregnant mothers on the birth spacing of more than two years between two successive pregnancies to allow replenishment of nutrient stores is important in reducing LBW.

Fe-Armacist Run Iron Deficiency Management Clinic to Optimize Intravenous Iron Infusions in an Ambulatory Oncology Clinic

Background At Boston Medical Center, patients across disciplines requiring IV iron are referred to a hematologist in the oncology infusion center. The high demand for patient appointments resulted in wait times up to 8 weeks from referral to visit across all hematologic indications. In addition, the absence of a standardized procedure led to increased drug denials, lack of reimbursement, treatment delays, and increased chair time. Pharmacist run clinics are becoming increasingly common and have offset time for general practitioners, improve access, and increase patient satisfaction. This quality improvement (QI) project aimed to increase the number of patients with iron deficiency anemia (IDA) managed by a pharmacist to improve physician access. Methods Patients 18 years of age and older requiring IV iron for IDA were referred for management under a pharmacy run protocol in an outpatient clinic. Providers placed referrals through the electronic medical record (EMR), Epic, that were reviewed daily by pharmacists. Key inclusion criteria were established IDA inadequately controlled by oral iron due to poor absorption, inability to take oral iron, or requirement of rapid replenishment of iron stores. Under a scope of practice with hematologists, pharmacists assessed symptoms of IDA, ordered and interpreted iron laboratory results, selected appropriate IV iron formulation, created a treatment plan within the EMR, and scheduled follow-up. A standard operating procedure (SOP) was developed outlining a treatment algorithm with all available IV iron. Standard documentation was utilized and documented in the hospital EMR for each visit. Per the IHI model, Plan-Do-Study-Act (PDSA) cycles were implemented to assess change over time. PDSA cycle 1 was implementation of a pilot clinic for referrals restricted to hematologists. Outcome measures included total number of referred patients and provider time offset. Process metrics included percentage of visits with appropriate labs as outlined in the SOP. Balancing metrics included number of infusion-related reactions, incidence of hypophosphatemia, and number of pharmacist hours. Results A total of 109 patients were referred to the pharmacy service from February 1, 2022 through June 17, 2022. Of 109 patients referred, 101 (92.6%) patients met inclusion criteria with an average of 5 patients referred weekly (Figure 1). Eight patients (7.3%) were excluded due to: complicated IDA that required further physician workup (4/8, 50%), the need to establish hematology care per the pilot protocol (2/8, 25%), adequate control on oral iron without previous IV therapy (1/8, 12.5%), or the absence of IDA (1, 12.5%). By June 17th, a total of 36 patients (35.6%) had an encounter with a pharmacist. The remaining patients (N=65) received an iron infusion at time of referral and did not require immediate follow-up. Time to next available appointment for patients requiring IV iron was < 7 days. Of patients seen by a pharmacist (N=36), 100% of patients had all per protocol iron studies completed. Six (16%) patients had phosphorus collected during their visit. Hypophosphatemia (phosphate < 2.7 mg/dL) was reported in 3 patients at time of initial pharmacist visit. There were no infusion-related reactions reported or documented in the patients evaluated. The total time spent by a pharmacist on all aspects of the protocol was 51.7 hours since clinic go-live for an average of 2.6 hours per week. More than 150 visits were projected to offset provider visits within 6 months based on the return to clinic time at referral, with a large majority of patients requiring follow-up at 6 months (37%, 37/101) or 3 months (33%, 33/101) (figure 2). Using the average reimbursement for an ambulatory oncology pharmacist visit, it was projected that this service would generate approximately $22,200 in visit revenue during the 6-month time period. Conclusion The pharmacist driven iron clinic was successfully implemented in our ambulatory infusion center. Pharmacists served as an ancillary service under a scope of practice to help improve clinic efficiency by standardizing IV iron and facilitating timely access for patients requiring treatment. Planned PDSA cycles include expansion of referring providers to general internal medicine practitioners. This QI project will continue to streamline workflow for patients with IDA and will aid in future development of similar pharmacist driven practices. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Mutations in proteins involved in E-C coupling and SOCE and congenital myopathies.

In skeletal muscle, Ca2+ necessary for muscle contraction is stored and released from the sarcoplasmic reticulum (SR), a specialized form of endoplasmic reticulum through the mechanism known as excitation-contraction (E-C) coupling. Following activation of skeletal muscle contraction by the E-C coupling mechanism, replenishment of intracellular stores requires reuptake of cytosolic Ca2+ into the SR by the activity of SR Ca2+-ATPases, but also Ca2+ entry from the extracellular space, through a mechanism called store-operated calcium entry (SOCE). The fine orchestration of these processes requires several proteins, including Ca2+ channels, Ca2+ sensors, and Ca2+ buffers, as well as the active involvement of mitochondria. Mutations in genes coding for proteins participating in E-C coupling and SOCE are causative of several myopathies characterized by a wide spectrum of clinical phenotypes, a variety of histological features, and alterations in intracellular Ca2+ balance. This review summarizes current knowledge on these myopathies and discusses available knowledge on the pathogenic mechanisms of disease.

A Systematic Review, Meta-Analysis, and Indirect Comparison of Blindly Adjudicated Cardiovascular Event Incidence with Ferric Derisomaltose, Ferric Carboxymaltose, and Iron Sucrose.

IntroductionIntravenous (IV) iron is the preferred treatment for patients with iron deficiency anemia (IDA) who require rapid replenishment of iron stores or in whom oral iron is not tolerated or effective. Data from two large-scale randomized controlled trials (RCTs) have recently been published reporting the incidence of adjudicated cardiovascular events after ferric derisomaltose (FDI) and iron sucrose (IS). The objective was to calculate the relative incidence of cardiovascular events with FDI and IS, and to conduct an indirect comparison with ferric carboxymaltose (FCM) based on previously published studies of cardiovascular risk.MethodsRCTs reporting the incidence of blindly adjudicated cardiovascular events in IDA patients treated with IV iron were identified by systematic literature review (SLR). Pairwise random effects meta-analyses of FDI versus IS, and FCM versus IS were conducted for the pre-specified adjudicated composite cardiovascular endpoint of: death due to any cause, nonfatal myocardial infarction, nonfatal stroke, unstable angina requiring hospitalization, congestive heart failure, arrhythmia, and protocol-defined hypertensive and hypotensive events. Analyses were also conducted for the composite endpoint excluding blood pressure events. Meta-analysis results were combined in an adjusted indirect comparison to provide an indirect estimate of cardiovascular risk with FDI versus FCM.ResultsThe SLR retrieved 694 unique articles, of which four were RCTs reporting the incidence of the composite cardiovascular endpoint; two studies comparing FCM (N = 1529) with IS (N = 1505), and two studies comparing FDI (N = 2008) with IS (N = 1000). The odds ratios of the composite CV endpoint were 0.59 (95% confidence interval: 0.39–0.90) for FDI versus IS, 1.12 (95% CI 0.90–1.40) for FCM versus IS, and the indirect OR for FDI versus FCM was 0.53 (95% CI 0.33–0.85).ConclusionsPooling data from four large-scale RCTs suggested that FDI was associated with significantly lower incidence of cardiovascular adverse events compared to both FCM and IS.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12325-022-02242-x.

CIC-39Na reverses the thrombocytopenia that characterizes tubular aggregate myopathy

AbstractStore-operated Ca2+-entry is a cellular mechanism that governs the replenishment of intracellular stores of Ca2+ upon depletion caused by the opening of intracellular Ca2+-channels. Gain-of-function mutations of the 2 key proteins of store-operated Ca2+-entry, STIM1 and ORAI1, are associated with several ultra-rare diseases clustered as tubular aggregate myopathies. Our group has previously demonstrated that a mouse model bearing the STIM1 p.I115F mutation recapitulates the main features of the STIM1 gain-of-function disorders: muscle weakness and thrombocytopenia. Similar findings have been found in other mice bearing different mutations on STIM1. At present, no valid treatment is available for these patients. In the present contribution, we report that CIC-39Na, a store-operated Ca2+-entry inhibitor, restores platelet number and counteracts the abnormal bleeding that characterizes these mice. Subtle differences in thrombopoiesis were observed in STIM1 p.I115F mice, but the main difference between wild-type and STIM1 p.I115F mice was in platelet clearance and in the levels of platelet cytosolic basal Ca2+. Both were restored on treatment of animals with CIC-39Na. This finding paves the way to a pharmacological treatment strategy for thrombocytopenia in tubular aggregate myopathy patients.

Loss of Estrogen‐Related Receptor Alpha (ESRRA) Impairs Neuronal Mitochondrial Function and Results in Disturbed Sleep and Neural Oscillatory Patterns

ESRRA is an orphan nuclear receptor enriched in metabolically active tissues and has been known to affect the transcription of a set of genes that regulate mitochondrial bioenergetics in peripheral organs. Of note, ESRRA is also abundantly expressed in the brain, but the functional role of brain ESRRA remains poorly understood. To test the role of ESRRA on energy metabolism in neurons, we performed the Seahorse assay in mouse primary cortical neurons with or without ESRRA expression and found that neurons lacking ESRRA have significantly reduced cellular respiration compared to control neurons, indicating that loss of ESRRA decreases mitochondrial oxidative capacity and thus neuronal energy production. We have previously reported that global ESRRA null (KO) displayed markedly reduced locomotor activity (home cage vibration plate p&lt;0.05, wheel running p&lt;0.001), suggesting that these mice do display an energy deficit as a result of this manipulation. Based on the commonly appreciated role of sleep in the replenishment of energy stores for normal brain function, we hypothesized that reduced brain energy levels caused by genetic loss of ESRRA would lead to hypersomnia and diminished neural oscillations, which may underlie hypolocomotion seen in ESRRA KO mice. To test this hypothesis, wireless electroencephalography/electromyography (EEG/EMG) devices were implanted in a cohort of ESRRA KO and wildtype (WT) mice and 24hr sleep state was recorded and scored. Sleep architecture analysis from 24‐hour wireless EEG/EMG recording revealed that ESRRA KO mice exhibit significantly increased non‐rapid eye movement (NREM) (p&lt;0.05) sleep as well as disrupted circadian pattern of rapid eye movement (REM) (p&lt;0.01) sleep compared to WT littermates. Power spectral analysis further revealed that high range gamma (40‐100Hz) (p&lt;0.001) and alpha (9‐13Hz) (p&lt;0.05) oscillations are significantly decreased while low range gamma (25‐40Hz) (p&lt;0.05) and beta (13‐25Hz) (p&lt;0.05) oscillations are increased in ESRRA KO mice. Our findings not only reveal a previously unappreciated role of ESRRA in regulating neuronal mitochondrial function, sleep‐wake cycle, and neural oscillations, but also support a novel concept that a single molecular cause of diminished brain energy levels can prolong sleep duration and results in aberrant REM sleep pattern.

Comparative study of efficacy of oral iron versus parenteral iron in the treatment of iron deficiency anaemia

Background: Iron deficiency anaemia (IDA) is one of the most widespread nutritional deficiency and common medical conditions seen in everyday clinical practice. IDA has a substantial haematological complication with potentially serious clinical consequences that often may require iron therapy. In most patients, body stores of iron can be restored by oral iron therapy but parenteral iron therapy results rapid correction of haemoglobin levels, faster and higher replenishment of iron stores with better compliance. The study was to compare the efficacy of oral iron (ferrous sulphate) with parenteral iron (iron sucrose) to treat iron deficiency anaemia. Methods: 221 patients were included in the study. 101 patients were given oral iron in the form of ferrous sulphate containing 67 mg of elemental iron three times daily. 120 patients were treated with iron sucrose. After a 25 mg test dose on the first infusion only, this was given at a dose of 300 mg by intravenous infusion diluted in 250 ml of normal saline, every alternate day. Haemoglobin level and serum ferritin of both groups were done before iron therapy, 3 weeks and 12 weeks after iron therapy. Results: The mean±SD rise of haemoglobin concentration 3 weeks after iron therapy in iron sucrose group was 11.2±0.60 g/dL, while in ferrous sulphate group was 9.0±0.58 g/dL. The mean±SD ferritin 3 weeks after iron therapy in iron sucrose group was 81.0±15.18 ng/mL, while in ferrous sulphate group was 27.0±12.22 ng/mL. The mean±SD haemoglobin concentration 12 weeks after iron therapy in iron sucrose group was 12.2±0.60 g/dL, while in ferrous sulphate group was 11.0±0.58 g/dL. The mean±SD ferritin 12 weeks after iron therapy in iron sucrose group was 82.0±16.17 ng/mL, while in ferrous sulphate group was 52.0±12.22 ng/mL. No serious adverse events were reported in either the ferrous sulphate group or iron sucrose group. Conclusion: Iron sucrose causes higher rise in haemoglobin level and serum ferritin as compared to oral iron therapy.

Computer-Based Drug Design of Positive Modulators of Store-Operated Calcium Channels to Prevent Synaptic Dysfunction in Alzheimer's Disease.

Store-operated calcium entry (SOCE) constitutes a fine-tuning mechanism responsible for the replenishment of intracellular stores. Hippocampal SOCE is regulated by store-operated channels (SOC) organized in tripartite complex TRPC6/ORAI2/STIM2. It is suggested that in neurons, SOCE maintains intracellular homeostatic Ca2+ concentration at resting conditions and is needed to support the structure of dendritic spines. Recent evidence suggests that positive modulators of SOC are prospective drug candidates to treat Alzheimer’s disease (AD) at early stages. Although STIM2 and ORAI2 are definitely involved in the regulation of nSOC amplitude and a play major role in AD pathogenesis, growing evidence suggest that it is not easy to target these proteins pharmacologically. Existing positive modulators of TRPC6 are unsuitable for drug development due to either bad pharmacokinetics or side effects. Thus, we concentrate the review on perspectives to develop specific nSOC modulators based on available 3D structures of TRPC6, ORAI2, and STIM2. We shortly describe the structural features of existing models and the methods used to prepare them. We provide commonly used steps applied for drug design based on 3D structures of target proteins that might be used to develop novel AD preventing therapy.

Multigravida Women With Moderate to Severe Anaemia in Third Trimester: Fetomaternal Outcomes.

Background & Objectives: Anaemia has been reported to be associated with adverse pregnancy outcomes, especially when presenting in the last trimester. In addition to prevalent common causes of anaemia in pregnant women, poor replenishment of iron stores after a pregnancy event is specific to women with higher birth orders. Anaemic women presenting in the third trimester are more prone to maternal complications such as infections, toxaemia, antepartum haemorrhage, cardiac failure, pre-eclampsia as well as fetal hazards too such as low birth weight, pre-term deliveries, developmental anomalies, and even neonatal death. When presented near term there are higher chances of feto-maternal morbidity and mortality. In the current study, analysis is done of feto-maternal outcomes, routes of delivery of women, causes of anaemia in multigravida women in the third trimester suffering from moderate to severe anaemia in a tertiary care centre of Western Rajasthan of India.Methods: A prospective observational clinical study was conducted on patients attending Geetanjali Hospital over a period of 18 months. A total of 70 consecutive multigravida pregnant women having moderate to severe anaemia in the third trimester were selected. Statistical analysis of the data collected was done and a p-value <0.05 was taken as significant.Results: Moderate and severe anaemia in the study population were 44.28% and 55.71%, respectively. The mean haemoglobin level of all study groups was 7.0 gm%. Pre-eclampsia, placenta praevia, postpartum haemorrhage (PPH), congestive cardiac failure (CHF), neonatal intensive care unit (NICU) admission, preterm birth (PTB), low birth weight (LBW), intrauterine death (IUD), low Appearance, Pulse, Grimace, Activity, and Respiration (APGAR) score, and birth asphyxia records were investigated. Of the patients studied, 18.57% had PPH, 15.71% had pre-eclampsia, 8.57% had IUD, and 37.14% newborns were LBW.Interpretations and Conclusion: Multiparity itself is a major risk factor of anemia. Anemia presenting in the third trimester of pregnancy is a proxy indicator of care received by gravid women in the early antenatal period. In combination, a multigravida in the third trimester with less time to restock iron and vitamin stores may result in considerable maternal as well as perinatal mortality and morbidity.

Towards Clinical Translation of Hematopoietic Cell Gene Editing for Treating Hyper-IgM Type 1

Hyper-IgM Type 1 (HIGM1) is caused by mutations of CD40L, whose absence in CD4 T cells impairs signaling for B cell activation and Ig class-switching. Since unregulated CD40L expression leads to lymphoproliferations/lymphomas in the mouse model of the disease, gene correction must preserve the physiological regulation of the gene. Gene editing of either autologous T cells or hematopoietic stem cells (HSC) held promise for treating HIGM1. We developed a “one size fits all” editing strategy to insert a 5'-truncated corrective CD40L cDNA in the first intron of the native human gene, effectively making expression conditional to targeted insertion in the intended locus. By exploiting a protocol that preserves T stem memory cells (TSCM), we reproducibly obtained ~40% of editing efficiency in healthy donor and patients derived T cells, restoring regulated, although partial, CD40L surface expression. The reconstituted level of expression, however, was sufficient to fully restore helper function to B cells. In order to select, track and potentially deplete edited T cells, we coupled the corrective cDNA with a clinically compatible selector gene and confirmed that enriched T cells preserved their engraftment capacity in NSG mice. Unexpectedly, the presence of an IRES-linked downstream coding frame counteracted the shorter half-life of transcript from the edited locus, allowing replenishment of intracellular stores and surface translocation of physiological amounts of CD40L upon activation. We also tailored the CD40L editing strategy to human HSC, reaching up to 15-30% editing in HSC long term engrafting NSG mice, depending on the HSC source. We then modelled the therapeutic potential of both T cell and HSC gene therapy by infusing increasing proportions of WT murine cells, as surrogates of edited cells, in HIGM1 mice. Administration of functional T cells at clinically relevant doses in HIGM1 mice, preconditioned or not with different lymphodepleting regimens, achieved long term stable T cell engraftment and partial rescue of antigen specific IgG response and germinal center formation in splenic follicles after vaccination with a thymus dependent antigen. Remarkably, infusion of T cells from mice pre-exposed to the antigen, mimicking treatment of chronically infected patients, was effective even in absence of conditioning and protected the mice from a disease relevant infection induced by the opportunistic pathogen Pneumocystis murina. Transplantation of functional T cells admixed with an equal number of HIGM1 T cells resulted in lower vaccination response, indicating competition between WT and HIGM1 cells and implying that increasing the fraction of corrected cells in the graft by selection would improve immune reconstitution. Concerning HSC gene therapy, transplanting 25% WT cells along with HIGM1 ones in HIGM1 mice - mirroring the editing efficiencies achieved in human HSC - rescued antigen specific IgG response and established protection from pathogen comparably to T cell therapy. These findings suggest that autologous edited T cells can provide immediate and substantial benefits to HIGM1 patients and position T cell as competitive strategy to HSC gene therapy, because of more straightforward translation, lower safety challenges and potentially comparable clinical benefits. We thus embarked in assessing GMP compliant reagents and protocols for T cell activation, culture and editing and developed a scalable manufacturing process. Optimization of clinical grade culture conditions allowed further increasing editing efficiency, total cellular yield and maintenance of TSCM thus paving the way to the design of a clinical trial. DisclosuresNaldini: Genenta Science: Consultancy, Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees, Other: Founder.

Protocol for a multicentre, parallel-group, open-label randomised controlled trial comparing ferric carboxymaltose with the standard of care in anaemic Malawian pregnant women: the REVAMP trial

IntroductionAnaemia in pregnancy remains a critical global health problem, affecting 46% of pregnant women in Africa and 49% in Asia. Oral iron therapy requires extended adherence to achieve correction of...