Background: The genetic causes of atrial fibrillation (AF) with slow conduction are unknown. Methods: Eight kindreds with familial AF and slow conduction, including a family affected by early onset AF, heart block and incompletely penetrant non-ischemic cardiomyopathy (NICM) underwent whole exome sequencing. Results: A known pathogenic mutation in the desmin (DES) gene resulting in S13F substitution at a PKC phosphorylation site was identified in all four members of the kindred with early-onset AF and heart block, while only two developed NICM. Higher penetrance of the mutation for AF and heart block prompted the screening for DES modifier(s). A second deleterious mutation in the phosphodiesterase 4D interacting-protein (PDE4DIP) gene resulting in A123T substitution segregated with early onset AF, heart block and the DES mutation. Three additional novel deleterious PDE4DIP mutations were identified in four other unrelated kindreds. Characterization of PDE4DIPA123T in vitro suggested impaired compartmentalization of PKA and PDE4D characterized by reduced colocalization with PDE4D, increased cAMP activation leading to higher PKA phosphorylation of the β2-adrenergic-receptor, and decreased PKA phosphorylation of Desmin in response to isoproterenol stimulation compared to wildtype PDE4DIP. Conclusion: Our findings identify an epistatic interaction between DES and PDE4DIP variants, increasing the penetrance for conduction disease and arrhythmia.

The hypercoagulable state observed in COVID-19 could be responsible for morbidity and mortality. In this retrospective study we investigated whether therapeutic anticoagulation prior to infection has a beneficial effect in hospitalized COVID-19 patients. 1154 COVID-19 patients admitted to 6 hospitals in the Netherlands between March and May 2020 were included. We applied 1:3 propensity score matching to evaluate the association between prior therapeutic anticoagulation use and clinical outcome, with in hospital mortality as primary endpoint. 190 (16%) patients used therapeutic anticoagulation prior to admission. In the propensity score matched analyses, we observed no associations between prior use of therapeutic anticoagulation and overall mortality (RR 1.02 (95% CI; 0.80-1.30) or length of hospital stay (7.0 [4-12] vs 7.0 {4-12] days, p=0.69), although we observed a lower risk of pulmonary embolism (RR 0.19 (95% CI; 0.05-0.80). This study shows that prior use of therapeutic anticoagulation is not associated with improved clinical outcome in hospitalized COVID-19 patients.

Introduction: Posterior fossa ependymomas in children are associated with a poor prognosis. This retrospective study tries to determine prognostic factors for the outcomes and the quality of life. Material and Methods: Thirty-three patients with posterior fossa ependymomas were treated from 2002 to 2018. All patients had a cranio-spinal MRI and 3.12% were metastatic. Removal was complete in 29 patients (90.62%). All patients received a complementary treatment: 11patients chemotherapy, radiotherapy in 35 patients, 24 cases of neoadjuvant type and for recurrence in 11. Results: Average age was 5.8 years with a range from 9 months to 18 years. The sex ratio was 2.3 (M/F: 23/10). Sixteen patients had one recurrence (3 metastatic). 90% of the EpPCF were of the PFA group. Overall Survival was 65% with a mean follow-up of 8.4 and a median of 9 years. Twenty-one patients were alive (63.63%) at last follow-up. All were tumour-free according the MRI except 2 patients, 1 with a stable residue for 5 years and 1 in palliative treatment for metastatic recurrence for 5 years. Post-operative complications were as follow, facial nerve palsy in four cases (12.12%), swallowing disorders in three cases (9.09%), transient cerebellar syndrome in 4 cases (12.12%). Fifteen patients had normal schooling. Conclusion: Posterior fossa ependymomas are aggressive tumors. Complete surgical removal remains the most important prognostic factor even if responsible of sequels. Despite molecular studies and the expression of different genes, no obvious therapeutic target has yet emerged.

Improved sample preparation has the potential to address a huge unmet need for fast turnaround sepsis tests that enable early administration of appropriate antimicrobial therapy. In recent years, inertial and elasto-inertial microfluidics-based sample preparation has gained substantial interest for bioparticle separation applications. However, for applications in blood stream infections the throughput and bacteria separation efficiency has thus far been limited. In this work, for the first time we report elasto-inertial microfluidics-based bacteria isolation from blood at throughputs and efficiencies unparalleled with current microfluidics-based state of the art. In the method, bacteria-spiked blood sample is prepositioned close to the outer wall of a spiral microchannel using a viscoelastic sheath buffer. The blood cells will remain fully focused throughout the length of the spiral channel while bacteria migrate to the inner wall for effective separation. Initially, microparticles were used to investigate particle focusing and the separation performance of the spiral device. A separation efficiency of 96% for the 1 µm particles was achieved, while 100% of 3 µm particles were recovered at the desired outlet at a throughput (sample + sheath) of 1 mL/min. Following, processing blood samples revealed a minimum of 1:2 dilution was necessary to keep the blood cells fully focus at the outer wall. In experiments involving bacteria spiked in diluted blood, viable E.coli were continuously separated at a total flow rate of 1 mL/min, with a separation efficiency between 82 to 90% depending on the blood dilution. Using a single spiral, it takes 40 minutes to process 1 mL of whole blood at a separation efficiency of 82% and 3 hr at 90% efficiency. To the best of our knowledge, this is the highest bacteria separation efficiency from blood sample reported using inertial and elasto-inertial methods. As such, the label-free, passive high efficiency and high throughput of bacteria isolation method has a great potential for speeding up downstream phenotypic and molecular analysis of bacteria.

The presence of iron in circulating monocytes is well known as they play an essential role in iron recycling. It has been demonstrated that the iron content of blood cells can be measured through their magnetic behavior; however, the magnetic properties of different monocyte subtypes remain unknown. In this study, we report for the first time, the magnetic behavior of classical, intermediate and non-classical monocytes, which is related to their iron storage capacity. The magnetic properties of monocytes were compared to other blood cells, such as lymphocytes and red blood cells in the oxyhemoglobin and methemoglobin states, and a cancer cell type. For this analysis, we used an instrument referred to as Cell Tracking Velocimetry (CTV), which quantitatively characterizes the magnetic behavior of biological entities. Our results demonstrate that significant fractions of the intermediate and non-classical monocytes have high magnetophoretic mobilities, equivalent to methemoglobin red blood cells and higher than the classical subset, suggesting their higher iron storage capacities. Moreover, our findings have implications for the immunomagnetic separation industry; we demonstrate that negative magnetic isolation techniques for recovering monocytes from blood should be used with caution, as it is possible to lose magnetic monocytes when using this technique.

Background The long-term impact of childhood cancer treatment on dietary intake is likely to be complex and the length of time dietary behaviours are affected after childhood cancer treatment is unknown. Aim The aim of this study was to determine the diet quality in childhood cancer survivors recently off treatment and identify possible contributing factors that may affect diet quality in this population. Methods Participants were 65 parents and/or carers of childhood cancer survivors (CCS) (aged 2-18 years), recently off treatment and 81 age-matched controls. Methods Participants completed two self-administered dietary intake and eating behaviour questionnaires. Study data was explored to determine between group differences, bivariate analysis using Spearman’s correlations was used to determine the relationship between diet quality and identified variables, and hierarchical cluster analysis was completed to characterise specific variables into clusters. Results CCS had a significantly poorer diet quality score than the age-matched controls (t=-2.226, p=0.028). Childhood cancer survivors had significantly higher parent-reported rates of ‘picky eating’ behaviour than the control group (t=0.106 p=0.044). Factors such as picky eating, emotional overeating and Body Mass Index z-score appeared to drive diet quality in survivors. Conclusions A CCS with picky eating behaviours could avoid complete food groups, have strong food preferences/aversions and over- consume high energy foods to maintain their energy intake, possibly affecting diet quality. The outcomes highlighted the need for a tailored intervention aimed at improving healthy eating behaviours in CCS after treatment for cancer.

The occurrence of alternative morphs within populations is common but the underlying molecular mechanisms remain poorly understood. Many animals, for example, exhibit facultative migration, where two or more alternative migratory tactics (AMTs) coexist within populations. In certain salmonid species, some individuals remain in natal rivers all their lives, whilst others (in particular, females) migrate to sea for a period of marine growth. Here we performed transcriptional profiling (“RNA-seq”) of the brain and liver of male and female brown trout to understand the genes and processes that differentiate migratory and residency morphs (AMT-associated genes) and how they may differ in expression between the sexes. We found tissue-specific differences with greater number of genes expressed differentially in the liver (n = 867 genes) compared to the brain (n = 10) between the morphs. Genes with increased expression in resident livers were enriched for Gene Ontology terms associated with metabolic processes, highlighting key molecular-genetic pathways underlying the energetic requirements associated with divergent migratory tactics. In contrast, smolt-biased genes were enriched for biological processes such as response to cytokines, suggestive of possible immune function differences between smolts and residents. Finally, we identified evidence of sex-biased gene expression for AMT-associated genes in the liver (n = 18) but not the brain. Collectively, our results provide insights into tissue-specific gene expression underlying the production of alternative life-histories within and between the sexes, and point towards a key role for metabolic processes in the liver in mediating divergent physiological trajectories of migrants versus residents.

Abstract Continuous intravenous (IV) sildenafil may avoid the acute systemic vasodilatory effects of bolus dosing in infants with pulmonary hypertension (PH). We aimed to examine the tolerability of different methods of IV sildenafil administration. Methods: We retrospectively evaluated subjects less than 12 months old with PH, who had been started on IV sildenafil. Vital signs, oxygen requirement, and vasoactive-inotropic score (VIS) before and after sildenafil initiation were noted, as was the need for discontinuation due to side effects. Results: Forty-three subjects were identified (23 continuous, 20 intermittent dosing). There were no statistically significant differences between groups in gender or gestational age, but higher baseline inspired oxygen (FiO2) and VIS in the continuous group suggested a higher baseline severity of illness (p=0.012). After sildenafil initiation, there were no significant differences in the change in blood pressure, oxygen saturation, FiO2, or VIS between groups, and no difference in the number of subjects requiring discontinuation due to side effects (4 in the continuous group, 1 intermittent, p=0.35). Eight subjects (34.8%) in the continuous group and 3 (15%) in the intermittent group died (p=0.024). Conclusions: In this small cohort of infants with PH treated with continuous or intermittent IV sildenafil, there were no statistically significant differences between groups in the change in vital signs, VIS, and oxygen requirement, or the need for discontinuation of therapy due to side effects. A higher mortality rate in the continuous infusion group may be explained by higher baseline illness severity.

Aim The present study aimed to evaluate the effect of quercetin in COVID-19 treatment. Methods This was a single-centre, prospective randomised controlled cohort study. Routine care versus QCB (quercetin, vitamin C, bromelain) supplementation was compared between 447 patients with at least one chronic disease and moderate-to-severe respiratory symptoms. Demographic features, signs, laboratory results and drug administration data of patients were recorded. The endpoint was that QCB supplementation was continued throughout the follow-up period from study baseline to discharge, intubation, or death. Results The most common complaints at presentation were fatigue (62.4%), cough (61.1%), anorexia (57%), thirst (53.7%), respiratory distress (51%) and chills (48.3%). The decrease in CRP, procalcitonin and ferritin levels was higher in the QCB group (all Ps were <0.05). In the QCB group, an increase in platelet and lymphocyte counts were higher (all Ps were <0.05). QCB did not reduce the risk of events during follow-up. Adjustments for statistically significant parameters, including the lung stage, use of favipiravir and presence of comorbidity did not change the results. While there was no difference between the groups in terms of event frequency, QCB group had more advanced pulmonary findings. QCB supplement is shown to have a positive effect on laboratory recovery. Conclusion We suggest that suboptimal bioavailability of QCB may explain this. So, we conclude that if a stable blood level can be achieved for QCB, it may make a difference in the treatment of COVID-19.

Over the past fifteen years, the number of papers focused on “eco-evo dynamics” has increased exponentially (Figure 1). This pattern suggests the rapid growth of a new, integrative discipline. We argue that this overstates the case. First, the terms “eco-evo dynamics” and “eco-evo interactions” are used too imprecisely. As a result, many studies that claim to describe eco-evo dynamics are actually describing basic ecological or evolutionary processes. Second, these terms are often used as if the study of how ecological and evolutionary processes are intertwined is novel when, in fact, it is not. The result is confusion over what the term “eco-evolution” and its derivatives describe, a loss of appreciation for the history of genuine eco-evolutionary studies, and a loss of appreciation for the novelty associated with the original rise of the term. We advocate a more precise definition of eco-evolution that is more useful in our effort to understand and characterize the diversity of ecological and evolutionary processes and that focuses attention on the subset of those processes that offer novel results.

Global influenza virus circulation declined and has been below traditional seasonal levels during the COVID-19 pandemic.1-3 We reviewed WHO influenza surveillance outputs from May 1-December 31, 2020 (epidemiologic weeks 18-53) from tropical Asian countries. For each country we report influenza surveillance specimens tested, and the percentage positive for influenza, by type and subtype. We compared current data to historical data from 2015-2019 in order to place the current season in historical context. Twelve included countries tested 17,407 surveillance specimens, with 592 (3.4%) testing positive for influenza viruses. From April 27-July 26, 2020 (epidemiologic weeks 18-30), specimens tested decreased from an average of 14,102 per year in 2015-2019 to 3,969 (71.9% decrease) and influenza positivity from 22% to <1%. During weeks 31-53, specimens tested decreased from an average of 24,782 per year in 2015-2019 to 13,438 (45.8% decrease) and influenza positivity from 18% to 4%. In six countries that maintained testing of surveillance specimens for >90% of weeks, influenza circulation was unseasonably low, or absent, during weeks 18-30, 2020. However, during weeks 31-53, the percentage of surveillance specimens testing positive for influenza approached or reached positivity rates of 2015-2019 in Bangladesh and Cambodia; and increased but remained lower than historical positivity in Lao PDR and Viet Nam. The data presented here are a reminder that the low levels of influenza circulation in the northern hemisphere in summer 2020 may not necessarily persist into the upcoming influenza season, and influenza surveillance and prevention strategies should continue as planned and not be delayed.

A document by Kefei Rao, written on Authorea.

Aims: It is generally accepted that geriatric patients are more sensitive to propofol than adults; thus, a dose-adjusted propofol is recommended for these patients during the induction of anesthesia. However, for patients aged 75 years and over, established guidelines do not provide dose references for the anesthesiologists. To this end, we observed 80 surgical patients (female 39, male 41, American Society of Anesthesiologists physical status score Ⅰ ~ Ⅱ) to access the appropriate dose of propofol for inducing loss of consciousness (LOC). Methods: Patients were subdivided into group A (20 patients, 45~64 yr), group B (20 patients, 65~74 yr), group C (20 patients, 75~84 yr), and group D (20 patients, ≥ 85 yr). All patients received propofol (at a rate of 0.3 mg/kg/min) alone for inducing LOC, which was defined by loss of both eyelash reflex and verbal response. Results: Compared with group A, the propofol requirement for LOC in Group B, C and D decreased by 14.8%, 25.2% and 38.5%, respectively. Bivariate linear correlation analysis showed that propofol requirement was negatively correlated with age. After adjusting for potential confounders, age was still an independent factor affecting propofol requirement. Conclusion: The propofol requirement for inducing LOC decreased significantly in elderly patients. We demonstrated that age was an independent factor impacting propofol requirement for LOC during the induction of general anesthesia, implying that the propofol dose for anesthesia induction should be further reduced in elderly surgical patients, especially those aged 75 years and over.

Wind is a fundamental driver of the distribution and energy expenditure of birds at sea. Wind can also influence mortality at the nest. Yet airflows have never been fully integrated into models of breeding habitat selection. We use computational fluid dynamics to provide the first assessment of whether and how airflows predict the distribution of seabird colonies, taking common guillemots (Uria aalge) breeding on Skomer island as our study system. We reveal that air pressure predicts occupancy, demonstrating the importance of exposure (rather than wind speed) in habitat selection. Our simple model with pressure and slope correctly identified 80% of the largest colonies and 93% of avoided sites. While previous approaches have not predicted space use in novel sites, our model predicted 73% of the largest colonies on a neighbouring island. This suggests generality in the mechanisms linking airflows and breeding distributions, and highlights a novel route by which seabirds may be affected by global change.       

In this study, we have looked the periodic table from the Barut’s point of view and discussed the deviations from the Madelung rule. Expected, observed and computed total energies (Hartree-Fock and Gaussian) are given for two different (one for expected and the other one is observed) configurations of the Cr atom. The data shows that preferred electronic configuration for the Cr is 4s13d5 not 4s23d4 as dictated by the Madelung rule. This event may be due to the spin correlation effect which is closely related to the Hund’s rule.

Human granzyme B (hGzmB), which is present in various immune cells, has attracted much attention due to its role in various pathophysiological conditions. The hGzmB activity is triggered at a catalytic triad (His59, Asp103, Ser198), cleaving its specific substrates. To date, the drug design strategy against hGzmB mainly targets the catalytic triad, which causes the non-specificity problem of inhibitors due to the highly conserved active site in serine proteases. In the present work, microsecond classical molecular dynamics simulations are devoted to exploring the structural dynamics of the hGzmB catalytic cycle in the presence of Ac-IEPD-AMC, a known substrate (active hGzmB), and Ac-IEPD-CHO, a known inhibitor (inactive hGzmB). By comparing active and inactive forms of hGzmB in the six different stages of the hGzmB catalytic cycle, we revealed, for the very first time, an additional network of interactions involving Arg216, a residue located outside the conventional binding site. Upon activation, the His59∙∙∙Asp103 hydrogen bond is broken due to the formation of the Asp103∙∙∙Arg216 salt bridge, expanding the active site to facilitate the substrate-binding. On the contrary, the binding of inhibitor Ac-IEPD-CHO to hGzmB prevents the Arg216-mediated interactions within the catalytic triad, thus preventing hGzmB activity. In silico Arg216Ala mutation confirms the role of Arg216 in enzyme activity, as the substrate Ac-IEPD-AMC failed to bind to the mutated hGzmB. Importantly, as Arg216 is not conserved amongst the various granzymes, the current findings can be a major step to guide the design of hGzmB specific therapeutics.

The effective and reasonable construction of the low impact development (LID) facilities in loess area depend on the functionality of typical LID facilities and the safety of surrounding structures in areas. A full-scale field test on rainwater concentrated infiltration of bioretentions in a collapsible loess site was conducted in this study. The water content and deformation law of the site were analyzed, and the water movement law of the rainwater-concentrated infiltration at bioretention facilities in the loess site was determined. The site settlements were calculated as per the wetting deformation curve and infiltration depths were calculated on an improved infiltration depth model tailored to the loess area. The rainwater infiltration rules of different bioretention structural forms are different in the collapsible loess field. The diffusion rate of the retaining wall type in loess decreases over time, while that on a sloping type does not. Within the same infiltration time, the retaining wall has a stronger influence on the site than the sloping type. When the water is concentrated in the site, its influence on the subgrade settlement is small (generally less than 1.5 mm) enough to satisfy the relevant engineering requirements. A modified Green-Ampt model based on assumed loess saturated unsaturated stratification can be used to predict the infiltration depth of facility water at the site. The adverse effects of water infiltration related to stagnant bioretentions can be mitigated by adjusting the initial water content and saturated water content at the loess site.

The increased frequency of flood events has motivated interest in natural flood management (NFM), in particular the potential for woodlands to reduce flooding. Woodlands can reduce the risk of rainfall-generated flooding through increased interception, soil infiltration, and available storage. Despite growing evidence, there is still low confidence in woodlands as a flood mitigation method due to limited empirical data available, particularly for semi-natural woodlands. We established a correlation catchment study in Haweswater, Cumbria, UK. Nine small upland catchments, each less than 0.2 km2 in area, were established on semi-natural broadleaf woodland sites where no stock grazing occurs or pasture with varied grazing intensity. At each site soil characteristics were investigated, namely soil moisture, permeability and bulk density. In addition, a v-notch weir was installed within in each catchment to calculate flow. The specific peak discharge (SPD), peak runoff coefficient, volume runoff coefficient and time taken to flow response was determined at each site for 28 storm events, of up to 205 mm, identified over a 13-month period. We found that semi-natural woodland reduced SPD by 33-52 % compared with pasture, reducing SPD by 36 % during larger storms (> 1 mm/hr peak discharge). Woodland reduced the peak runoff coefficient by 31-52 % and the volume runoff coefficient by 13-22 % compared to pasture. Additionally, response to storm events took 1-4 hours longer in woodland. These differences in flood response can be somewhat explained by the more permeable woodland soils, 4.6 times greater than pasture soil. Our analysis strengthens the argument that woodlands can reduce rainfall-generated flooding as a land use management method of NFM. Data collected here should be used to inform the parameters in flood prediction models and contribute to the evidence base for NFM.

A coupled atmospheric-hydrologic system models the complex interactions between the land surface and the atmospheric boundary layer, and the water-energy cycle from groundwater across the land surface to the top of the atmosphere. A regional climate model called WRF (Weather Research Forecasting) was coupled with a land surface scheme (Noah) to simulate intensive storms in central Alberta, Canada. Accounting for the land-atmosphere feedback enhances the predictability of the fine-tuned WRF-Noah system. Soil moisture, vegetation, and land surface temperature influence latent and sensible heat fluxes, and modulate both thermal and dynamical characteristics of land and lower atmosphere. WRF was set up in a two-way, three-domain nested framework so that the output of the outermost domain (D1) was used to run the second domain (D2) and the output of D2 was used to run the innermost domain (D3). In two-way nesting, D3 and D2 provide the feedback to their outer domains (D2 and D1), respectively. D3 was set at a 3-km resolution adequate to simulate convective storms. WRF-Noah was forced with climate outputs from Global Climate Models (GCMs) for the baseline period 1980–2005. A quantile-quantile bias correction method and a regional frequency analysis were applied to develop intensity-duration-frequency (IDF) curves from precipitation simulated by WRF-Noah. The simulated baseline precipitation of central Alberta agreed well with observed rain gauge data of Edmonton. The 5th‐generation NCAR mesoscale atmospheric model (MM5) was also set up in a 3-domain, but one-way nesting configuration. As expected, after bias correction, precipitation simulated by MM5 was less accurate than that simulated by WRF-Noah. For storms of short durations and return periods of more than 25 years, both MM5 driven by SRES climate scenarios of CMIP3 and WRF-Noah driven by RCP climate scenarios of CMIP5 projected storm intensities in central Alberta to increase from the base period to the 2050s, and to the 2080s.

Protein structure networks (PSNs) have long been used to provide a coarse yet meaningful representation of protein structure, dynamics, and internal communication pathways. An important question is what criteria should be applied to construct the network so that to include relevant interresidue contacts while avoiding unnecessary connections. To address this issue we systematically considered varying residue distance cutoff length and the probability threshold for contact formation to construct PSNs based on atomistic molecular dynamics in order to assess the amount of mutual information within the resulting representations. We found that the minimum in mutual information is universally achieved at the cutoff length of 5 Å, irrespective of the applied contact formation probability threshold in all considered, distinct proteins. Assuming that the optimal PSNs should be characterised by the least amount of redundancy, which corresponds to the minimum in mutual information, this finding suggests an objective criterion for cutoff distance and supports the existing preference towards its customary selection around 5 Å length, typically based to date on heuristic criteria.

The hyper-diverse order Coleoptera comprises a staggering ~25% of known species on Earth. Despite recent breakthroughs in next generation sequencing, there remains a limited representation of beetle diversity in assembled genomes. Most notably, the ground beetle family Carabidae, comprising more than 40,000 described species, has not been studied in a comparative genomics framework using whole genome data. Here we generate a high-quality genome assembly for Nebria riversi, to examine sources of novelty in the genome evolution of beetles, as well as genetic changes associated with specialization to high elevation alpine habitats. In particular, this genome resource provides a foundation for expanding comparative molecular research into mechanisms of insect cold adaptation. Comparison to other beetles shows a strong signature of genome compaction, with N. riversi possessing a relatively small genome (~147 Mb) compared to other beetles, with associated reductions in repeat element content and intron length. Small genome size is not, however, associated with fewer protein-coding genes, and an analysis of gene family diversity shows significant expansions of genes associated with cellular membranes and membrane transport, as well as protein phosphorylation and muscle filament structure. Finally, our genomic analyses show that these high elevation beetles have endosymbiotic Spiroplasma, with several metabolic pathways (e.g. propanoate biosynthesis) that might complement N. riversi, although its role as a beneficial symbiont or as a reproductive parasite remains equivocal.

Background: Iron deficiency is the most common micronutrient deficiency affecting nearly one-third of the population and is the leading cause of anemia worldwide. n this study, we evaluated the effectiveness of ferrous gluconate and ferrous sulfate supplements to identify the best iron supplement with the most effective and the least side effects in toddlers 6-24 months old. Methods: A randomized, single-blind clinical trial was performed. A total of 120 healthy toddlers aged 6 to 24 months old (two groups of 60) entered the study. Toddlers receiving ferrous sulfate (FS group) and ferrous gluconate (FG group) supplements. Blood indices such as hemoglobin & ferritin levels were evaluated at baseline and 6 months post- supplementation. Results: The FG group that received ferrous gluconate chelate iron showed approximately 2.4 g/dl higher Hb level in comparison to the FS group with ferrous sulfate supplementation 6 months post-supplementation (12.51±0.58 g/dL vs. 10.10±0.83, p = 0.045). Side effects were significantly more common in the FS group than the FG group (43.3 % vs. 16.7 %, P ≤ 0.001. Conclusion: The present study shows that educating mothers to feed toddlers with breast milk and iron supplements, including ferrous sulfate and ferrous gluconate, can be helpful in the prophylaxis of iron deficiency. Our results show that ferrous gluconate can be used in cases where ferrous sulfate causes unacceptable side effects.

Aim Although the fludarabine (F-araA)-treosulfan based toxicity reduced conditioning regimen has improved hematopoietic cell transplantation (HCT) outcome in patients with high-risk beta-thalassemia major (TM), rejection and regimen related toxicities (RRT) are still of major concern. This study aims to assess the role of F-araA pharmacokinetics (PK) and pharmacogenetics (PG) in a uniform cohort of patients with TM. Methods All patients with TM who receiving F-araA based regimen prior to HCT between September 2010 and 2019 were enrolled in this study. F-araA plasma levels were analyzed using LC-MS/MS. Selected polymorphisms in genes encoding for the enzymes (NT5E (Ecto-5’-nucleotidase) and DCK (Deoxycytidine kinase) involved in the metabolism of F-araA were screened. The influence of F-araA PK and PG on clinical outcomes were evaluated. Results F-araA PK showed wide inter-individual variation (27 and 19 fold in F-araA AUC and CL) which was explained by a promoter polymorphism (rs2295890) in the NT5E gene. Patients carrying the NT5E promoter variant showed no graft rejection (0% vs 7.7%, p=0.07) or Sinusoidal Obstruction Syndrome (0% Vs 19%, p=0.0007) and a trend to better EFS (87.5% vs 75.7%, p=0.1). F-araA systemic exposure was not associated with HCT outcome. Conclusion Our results suggest that the NT5E promoter polymorphism could be a predictive biomarker in F-araA based HCT setting in TM, however extensive functional studies are warranted to validate the clinical utility of this finding.

CHO cells have been recently shown to produce amino acid catabolism derived byproducts, which accumulate in fed-batch cultures to growth-inhibitory levels. Residual amino acid limitation or genetic engineering strategies have been successfully employed to suppress production of these novel growth inhibitory metabolic byproducts. However, the growth advantage attained due to suppression of these metabolic byproducts in fed-batch cultures is more pronounced when lactate accumulation is also controlled. BCAT1 knock-out (KO) CHO cells, which produce negligible levels of the metabolic byproducts isovalerate, isobutyrate and 2-methylbutyrate, grow to significantly higher peak cell densities in fed-batch cultures with lactate control (HiPDOG) as compared to cultures without lactate control. Henceforth, strategies involving novel metabolic byproduct control should preferably include lactate control to more easily assess the enhanced cell growth and productivities attainable.