Relative Excess Research Articles

Bruno Schulz's 1930 article "The Hereditary Relationships of Old-Age Paranoid Psychosis".

In the 1899 6th edition of his influential textbook, Kraepelin proposed a diagnostic category of "Old-Age Paranoid Psychosis." In this 1930 article, Bruno Schulz studied the morbid risk (MR) of several disorders and traits in the parents, siblings, offspring, and nieces/nephews of 51 probands with "Old-Age Paranoid Psychosis." His results permitted an evaluation of the validity of Kraepelin's category of Old-Age Paranoid Psychosis, in particular, whether it was a form of psychosis resulting from "senile changes" or late-onset schizophrenia. The MR of schizophrenia in these four groups of relatives varied from 0 to 2.4% with 3 of 4 somewhat higher than population expectations but much lower than parallel results in relatives of schizophrenics. By contrast, the rates of eccentricity in these relatives were uniformly elevated over population rates, sometimes approaching those seen in relatives of schizophrenics. Schulz concluded, from his study, that Old-Age Paranoid Psychosis was a distinct disorder not closely related to schizophrenia. However, he suggested that a family history and/or a premorbid trait of eccentricity increases the risk of developing a paranoid psychosis in old age, particularly when associated with physical or mental decline. He was uncertain about whether the trait of eccentricity he found in this study was very similar or distinct from that observed in excess in relatives of schizophrenics. This study was the first, to the best of our knowledge, to use a family study design explicitly to address a nosologic question-in this case the familial relationship between Old-Age Paranoid Psychosis and schizophrenia.

Antivenom supply and demand: An analysis of antivenom availability and utilization in South Africa

IntroductionSnakebites are a neglected tropical disease. In many areas, envenoming incidence and antivenom administration rates are unknown. This study compared antivenom (AV) availability to rates of envenoming and recommendations to treat (RTT) in South Africa. MethodsThis retrospective study identified, extracted, and reviewed all cases of envenoming (snake bites and spits) reported to the Poisons Information Helpline of the Western Cape of South Africa (PIHWC) from June 1, 2015 to May 31, 2020 by public hospitals in the Western Cape. A standardized interview was administered to the pharmacies of the 40 hospitals in winter and summer to determine how many vials of monovalent and polyvalent AV they had on hand at the time of the call and their expiration dates. Descriptive analysis was used to compare rates of envenoming and recommendations to treat to antivenom stock in winter and summer and by hospital type and location. ResultsPublic hospitals reported 300 envenomings, 122 from snakes. The PIHWC recommended antivenom administration in 26% of cases (N = 32). All hospital pharmacies queried answered our questions. Our study demonstrates urban district hospitals have higher ratios of AV vials compared to mean annual rates of envenoming and RTT than rural district hospitals at both the winter and summer timepoints. ConclusionThis study evaluates antivenom supply and demand in a province of South Africa. The findings suggest South African urban hospitals have a relative excess of antivenom, and thus more capacity to meet demand, than their rural counterparts. It supports consideration of a redistribution of antivenom supply chains to match seasonal and local rates of envenoming. It indicates a need for higher quality, prospective data characterizing envenoming incidence and treatment.

Dissociation of Arousal Index Between REM and NREM Sleep in Elderly Adults with Cognitive Impairment, No Dementia: A Pilot Study.

Sleep disruption in elderly has been associated with an increased risk of cognitive impairment and its transition into Alzheimer's disease (AD). High arousal indices (AIs) during sleep may serve as an early-stage biomarker of cognitive impairment non-dementia (CIND). Using full-night polysomnography (PSG), we investigated whether CIND is related to different AIs between NREM and REM sleep stages. Fourteen older adults voluntarily participated in this population-based study that included Mini-Mental State Examination, Neuropsi battery, Katz Index of Independence in Activities of Daily Living, and single-night PSG. Subjects were divided into two groups (n = 7 each) according to their results in Neuropsi memory and attention subtests: cognitively unimpaired (CU), with normal results; and CIND, with -2.5 standard deviations in memory and/or attention subtests. AIs per hour of sleep during N1, N2, N3, and REM stages were obtained and correlated with Neuropsi total score (NTS). AI(REM) was significantly higher in CU group than in CIND group. For the total sample, a positive correlation between AI(REM) and NTS was found (r = 0.68, p = 0.006), which remained significant when controlling for the effect of age and education. In CIND group, the AI(N2) was significantly higher than the AI(REM). In CIND older adults, this attenuation of normal arousal mechanisms in REM sleep are dissociated from the relative excess of arousals observed in stage N2. We propose as probable etiology an early hypoactivity at the locus coeruleus noradrenergic system, associated to its early pathological damage, present in the AD continuum.

P25-11: The effect of the introduction of European Union drinking water standard on arsenic exposure and related lifetime excess cancer risk in Hungary – a population based estimation of public health and economic benefits

P25-11: The effect of the introduction of European Union drinking water standard on arsenic exposure and related lifetime excess cancer risk in Hungary – a population based estimation of public health and economic benefits

Postmarketing observational study on the safety of 2021/2022 and 2022/2023 influenza vaccination campaigns in Italy: TheShinISS-Vax|Flu study protocol

IntroductionThe purpose of TheShinISS-Vax|Flu study is to examine the association between influenza vaccines and adverse events requiring hospital admission or emergency care during the influenza vaccination campaigns 2021/2022 and 2022/2023...

Th17 / Treg ratio: A prospective study in a group of pregnant women with preeclampsia and fetal growth restriction

IntroductionDuring pregnancy, the maternal immune system is challenged to tolerate a semi-allogenic fetus. A proinflammatory profile has been linked to adverse pregnancy outcomes and poor placental development. In this study, the authors evaluated the number of circulating Tregs and Th17 cells in a group of patients diagnosed with preeclampsia(PE) and fetal growth restriction(FGR). MethodsProspective longitudinal observational study where peripheral blood lymphocyte subsets were analyzed in a cohort of pregnant patients with PE, FGR, and a control group of healthy pregnant women. ResultsThe diagnosis of PE was associated with a significative higher number of circulating Th17 cells and a significative relative reduction in the Treg cell count. This proinflammatory profile was also expressed in the evolution of the Th17/ CD4+CD25highFOXP3+ Treg ratio. In the FGR group, the Th17 cell count was significantly higher during the third trimester of pregnancy. This proinflammatory profile was also expressed in the evolution of the Th17/ CD4+CD25highFOXP3+ Treg ratio. When we compare the immunological profiles of patients with PE and FGR we observed a higher number of proinflammatory Th17 cells and a significative lower number of Treg cells in PE patients. This is particularly expressed in the differences found between the Th17/ CD4+CD25highFOXP3+ Treg ratios of these two groups.Discussion/ConclusionOur data showed a that a proinflammatory profile and a relative excess of Th17 cells was associated with the diagnosis of PE and FGR. A more exuberant systemic proinflammatory profile present in the PE patients is absent in patients with FGR without preeclampsia

Application of multi-method-multi-model inference to radiation related solid cancer excess risks models for astronaut risk assessment

The impact of including model-averaged excess radiation risks (ER) into a measure of radiation attributed decrease of survival (RADS) for the outcome all solid cancer incidence and the impact on the uncertainties is demonstrated. It is shown that RADS applying weighted model averaged ER based on AIC weights result in smaller risk estimates with narrower 95% CI than RADS using ER based on BIC weights. Further a multi-method-multi-model inference approach is introduced that allows calculating one general RADS estimate providing a weighted average risk estimate for a lunar and a Mars mission. For males the general RADS estimate is found to be 0.42% (95% CI: 0.38%; 0.45%) and for females 0.67% (95% CI: 0.59%; 0.75%) for a lunar mission and 2.45% (95% CI: 2.23%; 2.67%) for males and 3.91% (95% CI: 3.44%; 4.39%) for females for a Mars mission considering an age at exposure of 40 years and an attained age of 65 years. It is recommended to include these types of uncertainties and to include model-averaged excess risks in astronaut risk assessment.

Nearby active galactic nuclei and starburst galaxies as sources of the measured UHECRs anisotropy signal

The Pierre Auger and the Telescope Array observatories have measured independent and statistical significant anisotropy in the arrival direction of ultra-high-energy cosmic rays (UHECR). Three hotspot regions with relative excess of events and a dipole signal have been identified in different regions of the sky and energy ranges. In this paper, we investigate the conditions under which these anisotropy signal could be generated by nearby (<23 Mpc) active galactic nuclei (AGN) and/or starburst galaxies (SBG). We studied a wide range of possibilities including injected nuclei (p, He, N, Si, and Fe), three UHECR luminosity proxies and three extragalactic magnetic field models. The results shows that both local AGN and SBG are needed to describe all the anisotropy signal. The contribution of AGN to hotspots and to the generation of the dipole is dominant in most cases. SBG is required only to explain the hotspot measured by the Telescope Array Observatory.

A Receptor Story: Insulin Resistance Pathophysiology and Physiologic Insulin Resensitization's Role as a Treatment Modality.

Physiologic insulin secretion consists of an oscillating pattern of secretion followed by distinct trough periods that stimulate ligand and receptor activation. Apart from the large postprandial bolus release of insulin, β cells also secrete small amounts of insulin every 4-8 min independent of a meal. Insulin resistance is associated with a disruption in the normal cyclical pattern of insulin secretion. In the case of type-2 diabetes, β-cell mass is reduced due to apoptosis and β cells secrete insulin asynchronously. When ligand/receptors are constantly exposed to insulin, a negative feedback loop down regulates insulin receptor availability to insulin, creating a relative hyperinsulinemia. The relative excess of insulin leads to insulin resistance (IR) due to decreased receptor availability. Over time, progressive insulin resistance compromises carbohydrate metabolism, and may progress to type-2 diabetes (T2D). In this review, we discuss insulin resistance pathophysiology and the use of dynamic exogenous insulin administration in a manner consistent with more normal insulin secretion periodicity to reverse insulin resistance. Administration of insulin in such a physiologic manner appears to improve insulin sensitivity, lower HgbA1c, and, in some instances, has been associated with the reversal of end-organ damage that leads to complications of diabetes. This review outlines the rationale for how the physiologic secretion of insulin orchestrates glucose metabolism, and how mimicking this secretion profile may serve to improve glycemic control, reduce cellular inflammation, and potentially improve outcomes in patients with diabetes.

Tide drives nutrients variation and exchange flux in the semi-enclosed Shuidong Bay coastal water in winter, South China Sea

Nutrients play a key role in coastal ecosystems. Coastal tide is an important driver of the daily nutrient variability in semi-enclosed bay under the intensive human activities and climate change. However, the extent of tidal effects on the nutrients variation and exchange flux in the semi-enclosed bay in winter is still scarcely. In this study, seawater samples were collected by synchronous continuous observation during winter in 2021, to investigate tide drives nutrients variation and exchange flux between Shuidong Bay (SDB) and South China Sea (SCS). The results showed most nutrients were significantly impacted by the tidal variation in the SDB with the semi-diurnal cycle. Coastal flood water can increase seawater exchange capacity due to strong dilution influenced by the outer SCS water, resulting mitigate adverse effects of nutrient enrichment in the semi-enclosed bay. In addition, the concentrations of total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and silicate (DSi) were 146.75 μmol/L, 1.06 μmol/L, and 6.15 μmol/L. Besides, there was a relative excess of nitrogen, phosphorus and silicate were limiting factors in the SDB, and nutrients significantly impacted the Chlorophyll-a in coastal water. Moreover, both TDP and DSi were transported from the SDB to the SCS with a net exchange flux of 0.23 t and 1.99 t, while TDN was transported from the SCS to the SDB with a net exchange flux of 0.14 t, resulting in nutrients ratio increase in the SDB coastal water. Statistical analysis identified the tide induced hydrological variations and terrestrial inputs dominated nutrient dynamics in the SDB coastal water. This study provided a new understanding of the variation and transport of nutrients driven by tide in the SDB, which is implications for understanding the biogeochemistry of nutrients in daily scale and water quality management in the semi-enclosed bay in future.

Modified H-plasty in large basal cell carcinoma defect reconstruction on the forehead.

Basal cell carcinoma (BCC) is the most common malignancy worldwide. Surgical removal is considered the gold standard treatment. However, large defects following excisional surgery can pose a significant challenge for reconstruction, especially in cases where primary closure is not possible. In these cases, skin flaps may be used. Most traditional skin flaps are limited by design to take advantage of only one region of relative skin excess. This paper reports a case of a large forehead defect following BCC excision that was successfully reconstructed by a modified Hplasty involving a U-plasty and rotation-advancement flap. Maintenance of aesthetics without impaired function is the aim of closure defects with combined flaps.

Yoga as secondary prevention and cardiac rehabilitation in myocardial infarction

Abstract Funding Acknowledgements Type of funding sources: None. Introduction The prevalence of coronary artery disease is racing at a fast pace. The risk factors of myocardial infarction are old age, male sex, previous MI , diabetes, hypercholesterolemia, hypertension, unhealthy diet , smoking , physical inactivity and psychosocial stress. Interventional and pharmacological approaches are accessible these days for reduction of mortality and morbidity in myocardial infarction. Yoga as a compliment is inexpensive and devoid of side effects which has been shown to reduce body weight, blood pressure, cholesterol, blood sugar &amp; improve physical and mental well being due to relative parasympathetic excess. Many studies performed earlier had small sample size and duration. Therefore, yoga is affordable and can be recommended along with conventional treatment to prevent and improve the outcome in MI cases. Purpose 1. To study the effect of yoga on risk factors following myocardial infarction. 2. To study the effect of yoga on Beck's Depression Inventory score. Methods Our study that comprised of 300 patients were randomized into test group (n=150) who were subjected to conventional treatment and yoga and control group (n=150) subjected to conventional treatment only. The study included patients diagnosed as myocardial infarction above 18 years with their consent to participate. Patients with hemodynamic instability, heart failure, pregnancy, prior yoga practice, arrhythmia ,cardiac pacemaker, autonomic neuropathy and musculoskeletal disease were excluded. Initially, assessment of clinical &amp; biochemical parameters and routine investigations were performed. Beck's depression score was calculated. Yoga was started on 4th day after MI (test group) and there were sessions of 10 minutes of pranayam , 15 minutes of shavasan and 15 minutes of transcendental meditation. The patients were followed after 24 weeks for assessment of clinical &amp; biochemical parameters. Carotid doppler &amp; depression score was evaluated. The primary outcome included change in risk factors from baseline to the end of study period and secondary outcome was change in Beck's Depression score. Results After 24 weeks of intervention, a significant improvement was noted among yoga group in comparison to control group for BMI (p=0.04), heart rate (p=0.01), systolic &amp; diastolic blood pressure (p&amp;lt;0.05), fasting &amp; post prandial blood sugar (p&amp;lt;0.05), HbA1c (p=0.03), Beck's depression score (p=0.001) and lipid profile (p&amp;lt;0.05).In contrast, no improvement was seen for carotid intima media thickness. Conclusion On the basis of the study, it was concluded that conventional therapy with yoga has a positive impact on risk factors of MI and psychosocial stress. Therefore, yoga can be recommended to reduce morbidity and mortality after MI. Large trials with more number of cases and long term follow up are needed so that yoga as an intervention, can be added for secondary prevention and rehabilitation in myocardial infarction cases.

Плотность древесины сосны обыкновенной в осушаемом сосняке сфагновом

The task of increasing forest productivity has been and remains one of the current interests of the European North of Russia, especially after the period of primary development of the forest areas. In the European Northeast, excessive humidity is observed at rather low average temperatures, which prevents active surface evaporation. In the twentieth century, the hydrotechnical reclamations were conducted on the large areas of swampy forests that increased the total productivity. The ecologists have an ambivalent attitude concerning forest draining. The supporters, who try to protect the undisturbed forest territories, counteract such activities. At the same time, the cultivation of highly productive forests of the right composition and quality requires measures for natural afforestation, or artificial forests must be created. Besides the total productivity of the stands, wood quality is an important factor in the hydroamelioration. It is mainly determined by density and the ratio between early and late wood. The influence of the landscape peculiarities of the drained forests on the quality of the wood was examined by the devotion of the areas according to the most typical groups of microrelief: flat areas, elevations, transitional areas. In cases of extensive humidity, the death of seedlings and artificial forest plantings was observed (soaking, frost extrusion, etc.). The creation of micro-elevations and the drainage of the excessive moisture guarantee the regeneration and establishment of the highly productive forests. The differences in the density of pine wood before and after the hydrotechnical reclamation were detected according to the landscape. The largest changes in the indicator were determined at the elevations and the flat areas, with the increase and the decrease, respectively. The trees that grow on the flat terrain after the reclamation show a relative excess of early wood over late wood, while at the elevations the presence of the late wood is predominant. For citation: Zagorodski M.A., Tretyakov S.V., Koptev S.V. Density of Scots Pine Wood in Drained Sphagnum Pine Forest. Lesnoy Zhurnal = Russian Forestry Journal, 2023, no. 2, pp. 160–171. (In Russ.). https://doi.org/10.37482/0536-1036-2023-2-160-171

Optimal bond holding dynamics with hedging against real exchange rate risks

This study examines optimal bond holding dynamics using a two-country DSGE model with two bonds. Unlike previous studies focusing on steady-state properties of optimal bond holdings, we solve the model by employing a third-order approximation method and show how optimal bond holdings respond when macro fundamentals change. In particular, we derive impulse responses of optimal home and foreign bond holdings to productivity shocks. Simulation results show that with a positive productivity shock at home, optimizing home agents decrease holdings of home bonds and increase holdings of foreign bonds. Correlation between relative price and excess bond returns following a positive productivity shock makes home (foreign) bond a less (more) attractive hedging tool against real exchange rate risks. In addition, we use a SVAR method and verify that the optimal bond holding dynamics derived from the model are observed in the data.

The miR-20a/miR-92b Profile Is Associated with Circulating γδ T-Cell Perturbations in Mild Psoriasis.

Psoriasis vulgaris (PV) is an autoinflammatory dermatosis of unknown etiology. Current evidence suggests a pathogenic role of γδT cells, but the growing complexity of this population has made the offending subset difficult to pinpoint. The work on γδTCRint and γδTCRhi subsets, which express intermediate and high levels of γδTCR at their surface, respectively, is particularly scarce, leaving their inner workings in PV essentially unresolved. We have shown here that the γδTCRint/γδTCRhi cell composition and their transcriptom are related to the differential miRNA expression by performing a targeted miRNA and mRNA quantification (RT-qPCR) in multiplexed, flow-sorted γδ blood T cells from healthy controls (n = 14) and patients with PV (n = 13). A significant loss of miR-20a in bulk γδT cells (~fourfold decrease, PV vs. controls) largely mirrored increasing Vδ1-Vδ2- and γδintVδ1-Vδ2- cell densities in the bloodstream, culminating in a relative excess of γδintVδ1-Vδ2- cells for PV. Transcripts encoding DNA-binding factors (ZBTB16), cytokine receptors (IL18R1), and cell adhesion molecules (SELPLG) were depleted in the process, closely tracking miR-20a availability in bulk γδ T-cell RNA. Compared to controls, PV was also associated with enhanced miR-92b expression (~13-fold) in bulk γδT cells that lacked association with the γδT cell composition. The miR-29a and let-7c expressions remained unaltered in case-control comparisons. Overall, our data expand the current landscape of the peripheral γδT cell composition, underlining changes in its mRNA/miRNA transcriptional circuits that may inform PV pathogenesis.

Different post-pancreatectomy glucagon responses to a meal test between surgical approaches.

Residual pancreatic endocrine function is important for maintaining metabolic status after pancreatectomy and is closely related to patient nutritional status and prognosis. In contrast to insulin secretion, the significance of glucagon secretion following pancreatectomy remains unclear. In this study, we assessed the changes in pancreatic glucagon secretion during pancreatectomy to determine their pathophysiological significance. We evaluated glucagon and insulin secretion using a liquid meal tolerance test before and after pancreatectomy in patients scheduled to undergo pancreaticoduodenectomy (PD) or distal pancreatectomy (DP). After pancreatectomy, fasting plasma glucagon levels were significantly decreased in both the PD (n = 10) and DP (n = 5) groups (PD: from 18.4 to 10.5 pg/mL, p = 0.037; DP: from 21.0 to 12.1 pg/mL, p = 0.043), whereas postprandial plasma glucagon levels were not changed. In the liquid meal tolerance test after pancreatectomy, 60-min plasma glucagon levels and the area under the curve (AUC) for 0-120 min of PD were significantly higher than those for DP (60-min plasma glucagon: PD 49.0 vs. DP 21.7 pg/mL, p = 0.040; AUC0-120min: PD 4,749 vs. DP 3,564 μg min/mL, p = 0.028). Postoperative plasma glucose, serum insulin, and serum C-peptide levels during the liquid meal tolerance test were not significantly different between the two groups. Although fasting plasma glucagon levels decreased, postprandial glucagon responses were maintained after both PD and DP. The difference in residual meal-stimulated glucagon response between PD and DP suggests that a relative excess of postprandial glucagon is involved in the postoperative nutritional status after PD through its impact on systemic metabolic status.

Enhanced detoxification of Cr6+ by Shewanella oneidensis via adsorption on spherical and flower-like manganese ferrite nanostructures.

Maximizing the safe removal of hexavalent chromium (Cr6+) from waste streams is an increasing demand due to the environmental, economic and health benefits. The integrated adsorption and bio-reduction method can be applied for the elimination of the highly toxic Cr6+ and its detoxification. This work describes a synthetic method for achieving the best chemical composition of spherical and flower-like manganese ferrite (MnxFe3-xO4) nanostructures (NS) for Cr6+ adsorption. We selected NS with the highest adsorption performance to study its efficiency in the extracellular reduction of Cr6+ into a trivalent state (Cr3+) by Shewanella oneidensis (S. oneidensis) MR-1. MnxFe3-xO4 NS were prepared by a polyol solvothermal synthesis process. They were characterised by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectrometry (XPS), dynamic light scattering (DLS) and Fourier transform-infrared (FTIR) spectroscopy. The elemental composition of MnxFe3-xO4 was evaluated by inductively coupled plasma atomic emission spectroscopy. Our results reveal that the oxidation state of the manganese precursor significantly affects the Cr6+ adsorption efficiency of MnxFe3-xO4 NS. The best adsorption capacity for Cr6+ is 16.8 ± 1.6 mg Cr6+/g by the spherical Mn0.22+Fe2.83+O4 nanoparticles at pH 7, which is 1.4 times higher than that of Mn0.8Fe2.2O4 nanoflowers. This was attributed to the relative excess of divalent manganese in Mn0.22+Fe2.83+O4 based on our XPS analysis. The lethal concentration of Cr6+ for S. oneidensis MR-1 was 60 mg L-1 (determined by flow cytometry). The addition of Mn0.22+Fe2.83+O4 nanoparticles to S. oneidensis MR-1 enhanced the bio-reduction of Cr6+ 2.66 times compared to the presence of the bacteria alone. This work provides a cost-effective method for the removal of Cr6+ with a minimum amount of sludge production.

The Relationship between Serum Adiponectin, Urinary Albumin/Creatinine Ratio and Type 2 Diabetes: A Population-Based Cross-Sectional Study.

The relationship between serum adiponectin concentration (S-Adipo) and various diseases, such as type 2 diabetes (T2D) is conflicting. We hypothesized that the extent of kidney damage in patients with T2D may be responsible for this inconsistency and, thus, examined association between S-Adipo and T2D after consideration for the extent of kidney damage present. Of the 1816 participants in the population-based Iwaki study of Japanese people, 1751 participants with a complete dataset were included. Multivariate logistic regression analyses revealed that low S-Adipo was independently associated with T2D (&lt;0.001), as was high urinary albumin to creatinine ratio (uACR) (&lt;0.001). Principal components analysis showed that the relative value of S-Adipo to uACR (adiponectin relative excess) was significantly associated with T2D (odds ratio: 0.49, p &lt; 0.001). Receiver operating curve analyses revealed that an index of adiponectin relative excess the ratio of S-Adipo to uACR was superior to S-Adipo per se as a marker of T2D (area under the curve: 0.746 vs. 0.579, p &lt; 0.001). This finding indicates that the relationship between S-Adipo and T2D should be evaluated according to the extent of kidney damage present and may warrant similar analyses of the relationships between S-Adipo and other medicalconditions, such as cardiovascular disease.

Autoimmune Diseases Affecting Hemostasis: A Narrative Review

Hemostasis reflects a homeostatic mechanism that aims to balance out pro-coagulant and anti-coagulant forces to maintain blood flow within the circulation. Simplistically, a relative excess of procoagulant forces can lead to thrombosis, and a relative excess of anticoagulant forces can lead to bleeding. There are a wide variety of congenital disorders associated with bleeding or thrombosis. In addition, there exist a vast array of autoimmune diseases that can also lead to either bleeding or thrombosis. For example, autoantibodies generated against clotting factors can lead to bleeding, of which acquired hemophilia A is the most common. As another example, autoimmune-mediated antibodies against phospholipids can generate a prothrombotic milieu in a condition known as antiphospholipid (antibody) syndrome (APS). Moreover, there exist various autoimmunity promoting environments that can lead to a variety of antibodies that affect hemostasis. Coronavirus disease 2019 (COVID-19) represents perhaps the contemporary example of such a state, with potential development of a kaleidoscope of such antibodies that primarily drive thrombosis, but may also lead to bleeding on rarer occasions. We provide here a narrative review to discuss the interaction between various autoimmune diseases and hemostasis.

Characterization of Hemoglobin Variants Due to in-Frame Deletions in the β-Globin Gene from CRISPR/Cas9 Gene Correction for Sickle Cell Disease

Introduction: Sickle cell disease (SCD) is caused by a single nucleotide mutation in the β-globin gene (HBB). CRISPR/Cas9 based gene correction approaches rely on inducing a double-strand break (DSB) at or near the sickle mutation, which subsequently results in gene correction via the homology-directed repair (HDR) or insertions/deletions (Indels) via non-homologous end joining (NHEJ). Frameshift Indels induce a premature stop codon and disrupt HBB production. Indels that preserve the reading frame may yield structurally abnormal β-globin chain with altered amino acid sequences and protein structures that can affect the globin chain stability, leading to thalassemia of uncertain pathogenic significance. The high rate and diversity of in-frame Indels as an unintended by-product of gene correction therapy confers a challenge for predicting therapeutic outcomes when it is not possible to frame the clinical behavior of hemoglobin (Hb) variants. Here we provided the molecular and structural characterization of the most commonly identified Hb variants due to CRISPR-induced in-frame deletions and proposed a putative pathogenic mechanism for the observed phenotype using human umbilical cord-derived erythroid progenitor cells (HUDEP2). Methods: HUDEP2 clonal cell lines carrying mutated Hb variants from in-frame deletions on HBB exon1 introduced by R-66S and R-02 CRISPR sgRNAs currently in use in preclinical and clinical studies for correcting sickle mutation, respectively, were established, and clonal genotypes validated by next-generation sequencing and droplet digital PCR based copy number assay. The Hb variants derived from R-66S include p.Val7_Ser10del (-12bp), p.Val7del (-3bp), p.Glut8del (-3bp), p.Glut8_Lys9del (-6bp), pVal7Thr13del (-21bp), p.His3_Lys9delinsGln (-18bp), and p.Thr5_Lys18del (-42bp), in the order of allelic abundance observed in the bulk population of edited cells. The Hb variants derived from R-02 include p.Val12_Ala14del (-9bp), p.Val12_Thr13del (-6bp), p.Ala14del (-3bp), p.Thr13_Gly17del (-15bp), and p.Thr13_Val19delinsMet (-18bp). Selected ten clonal cell lines carrying a unique combination of genotypes and the parent HUDEP2 were cultured for expansion in serum-free media for one week. All clones were subsequently subjected to erythroid differentiation media for ten days. Samples were collected on days 4, 7, and 10 of differentiation for analysis via high-performance liquid chromatography (HPLC) using an ionic column to access the Hb profile and spectrophotometry via a Hb assay kit to assess the total Hb quantity. Results: HPLC analysis demonstrated a significant qualitative variation in Hb profile in HUDEP2 clones carrying different Hb variants. p.Val7del is Hb Leiden, and the rest of Hb variants are not listed in the HbVar database. In clones carrying Hb Leiden in homozygous or heterozygous states, Hb Leiden eluted in the position of sickle Hb (HbS). A low percentage of multiple abnormal Hb X fractions was observed in all clones carrying Hb variants compared to parent HUDEP2, which hinted at the presence of unstable Hb variants or alpha precipitates. Levels of fetal Hb (HbF) increased in mutant clones, suggesting globin chain imbalance and hematopoietic stress. Clones missing multiple amino acids showed ineffective erythropoiesis and cell death by day 4 of differentiation, most likely exacerbated by the relative excess of the α-chains. Discussion: We found that unintended gene modifications can significantly impact Hb quality and quantity, reinforcing concerns about the safety of gene-editing therapy for SCD. In patients, cells carrying in-frame deletions in the β-globin chain could produce an unstable Hb and may contribute to a β-thalassemia-like phenotype. More work is required to further assess the consequences of the diverse Hb variants in combination with other unintended modifications that disrupt HBB in a bulk population of gene-edited cells. Detecting, profiling, and reducing these unintended modifications remains a priority to increase the efficacy and safety of gene-editing therapy for treating SCD or β-thalassemia. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal