Association of the HALP Score with Dyslipidemia: A Large, Nationwide Retrospective Study.

Background and Objectives: Dyslipidemia is a major risk factor for cardiovascular disease (CVD). The identification of new biomarkers that may enhance the risk assessment of lipid abnormalities is a promising approach in improving risk prediction of CVD. There is no information on the association of the hemoglobin, albumin, lymphocyte, and platelet (HALP) score with dyslipidemia. The aim of this study was to investigate the clinical utility of the HALP score in light of dyslipidemia. Materials and Methods: A retrospective analysis of 7192 subjects was initiated to assess the association between the HALP score and disturbed lipid markers. Medians were compared by Mann-Whitney U or Kruskal-Wallis tests and the diagnostic performance and risk assessment were calculated. Results: Median HALP score among all subjects was 53.3, with varying values between males and females. Notably, median HALP was significantly elevated in all forms of dyslipidemia and among males and females irrespective of age. The odds of having elevated HALP score values were significantly higher in all lipid abnormalities. Moreover, HALP score was significantly yet weakly correlated with lipid markers, while the highest diagnostic accuracy of the HALP score was observed with an elevated ratio of total cholesterol to high-density lipoprotein (TC/HDL) (area under the curve, AUC = 0.6411, p < 0.0001). The decision curve analysis (DCA) showed that the HALP score can reliably predict the presence of dyslipidemia. Conclusions: This study demonstrates that the HALP score is a novel, cost-effective index that is associated with a disturbed lipid profile. Further investigation of the nature of this association is needed.

Geraniin inhibits whole blood IFN-γ and IL-6 and promotes IL-1ß and IL-8, and stimulates calcium-dependent and sucrose-sensitive erythrocyte death.

Correlations between circulating cytokine levels and disease states are well established, and pharmacological modulation of the immune response is thus an important aspect of the assessment of investigational new drugs. Moreover, chemotherapy-related anemia is a major obstacle in cancer treatment. Geraniin (GRN), a tannin extracted from Geranium and other plants, possesses promising antitumor potential. However, the effect of GRN on whole blood (WB) cytokine response and RBC physiology remains unexplored. Heparinized blood from consented, healthy adults was challenged with 100 ng/mL of lipopolysaccharide (LPS) with and without pretreatment with 10 µM of GRN for 24 h at 37 °C, and tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-1ß (IL-1ß), IL-6, IL-8, and IL-10 were assayed by ELISA. Moreover, single-cell RBC suspensions were treated with 5-100 µM of GRN for 24 or 48 h at 37 °C and cytotoxicity and canonical eryptotic markers were examined by flow cytometry. It was revealed that GRN significantly attenuated LPS-induced IFN-γ levels, increased IL-1ß, decreased IL-6 only in absence of LPS, and aggravated LPS-induced IL-8 while together with LPS significantly diminished IL-10. Furthermore, GRN induced dose-responsive, Ca2+-dependent, and sucrose-sensitive hemolysis, along with phosphatidylserine exposure and Ca2+ accumulation with no appreciable cell shrinkage or oxidative damage. GRN was also selectively toxic to platelets, significantly delayed reticulocyte maturation, and significantly disrupted leukocyte proportions. In conclusion, GRN regulates the WB cytokine response and promotes premature hemolysis and eryptosis. This study provides insights into the therapeutic utility of GRN in a highly relevant cellular model system.

Breast Cancer Incidence Patterns in the Saudi Female Population: A 17-Year Retrospective Analysis.

Background and Objectives: Breast cancer is considered the most commonly diagnosed type of cancer among women globally and in Saudi Arabia. This study aimed to assess breast cancer incidence patterns and trends among the Saudi female population. Materials and Methods: Breast cancer incidence parameters were obtained from the Saudi Cancer Registry (SCR). The data were retrospectively analyzed for the period from 2001 to 2017 to investigate changes in incidence rates. Temporal trends were also analyzed through joinpoint regression analysis and were dissected by age groups and administrative regions. Results: During the specified period, breast cancer jumped by 55% to constitute 30.9% of all cancer cases among Saudi females. The median age at diagnosis increased to reach 51 years at the end of that period, with an overall increase of 6.3%. The overall Age-Standardized Incidence Rate (ASR) escalated by 151.7% from 11.8/100,000 to 29.7/100,000 population for that period. The Eastern region noticeably had the highest ASR and peaked at 52.2/100,000 population. The joinpoint analysis of the ASR showed increased trends, with an annual percent change (APC) of 5.13% (p < 0.05, [95% CI 4−6.3]). An age-specific analysis was also performed and showed that the age group 70−74 years had the highest trend (APC 10.2%, [95% CI 7.2−13.4], p < 0.05). Region-specific analysis revealed that the Jouf region had the highest trend among the regions (APC 8.8%, [95% CI 3.7−14.2], p < 0.05). Conclusions: Our analysis indicates increased breast cancer incidence in Saudi Arabia with an alarming pace. With the existing trend, it is expected that Saudi Arabia will continue to display an increase in breast cancer incidence. Long-term preventive measures and more effective screening strategies are warranted to alleviate the burden of the disease.

Isolated and Combined Effect of Age and Gender on Neutrophil-Lymphocyte Ratio in the Hyperglycemic Saudi Population.

Inflammation is pivotal to the pathogenesis of diabetes mellitus (DM), but pathological alterations of the neutrophil−lymphocyte ratio (NLR), an emerging inflammatory index in DM management, remains understudied. The aim of this study is to examine the relationship between NLR and glycemic control in the Saudi population. Gender, age, WBC count, and fasting blood glucose (FBG) were obtained from Al-Borg Medical Laboratories for 14,205 subjects. Means, prevalence, risk measures, and the diagnostic accuracy of elevated NLR and hyperglycemia (HG) were evaluated. Subjects with elevated NLR (>3) had significantly higher FBG (105.10 ± 0.33 vs. 114.0 ± 2.81) and NLR was significantly elevated in impaired fasting glycemia (IFG; 1.21 ± 0.01 vs. 1.25 ± 0.01) and HG (1.21 ± 0.01 vs. 1.39 ± 0.02). Elevations of NLR in HG but not in IFG persisted across all age groups except young males and elderly females. The prevalence of elevated NLR in hyperglycemic subjects was 4.12% compared to 2.16% in subjects with normal FBG. HG was more prevalent in subjects with elevated NLR (17.33% vs. 12.46%) who had a relative risk (RR) of 1.68 (95% CI = 1.38−2.06, p < 0.0001) and an odds ratio (OR) of 1.94 (95% CI = 1.48−2.56, p < 0.0001) to be hyperglycemic. Nevertheless, NLR failed to discriminate individuals with normal FBG from those with HG based on ROC curve analysis. Pathological fluctuations in NLR may serve as supportive evidence in DM management.

Physcion Induces Hemolysis and Premature Phosphatidylserine Externalization in Human Erythrocytes.

The prevalence of cancer-associated anemia (CIA) is high, and the mechanisms governing its development remain poorly understood. Eryptosis, the suicidal cell death of red blood cells (RBCs), may account for CIA as it is triggered by clinically approved chemotherapeutics including cisplatin and paclitaxel. Physcion (PSN), an anthraquinone extracted from rhubarb and other plants, has shown great promise as an anticancer agent. However, the potential toxicity of PSN to RBCs remains elusive. RBCs were isolated from heparinized blood, and incubated with 10-100 µM of PSN for 24 h at 37 °C. Hemolysis was photometrically calculated from hemoglobin concentration in the medium at 405 nm, while flow cytometry was employed to investigate cardinal markers of eryptosis. Phosphatidylserine (PS) exposure was detected by Annexin-V-fluorescein isothiocyanate (FITC), intracellular calcium by Fluo4/AM, cellular volume from forward scatter (FSC), and oxidative stress by 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA). PSN induced overt hemolysis at 50 and 100 µM which was not mediated through calcium influx, protein kinase C, casein kinase 1α, or receptor-interacting protein 1. Moreover, PSN caused significant increase in Annexin-V-FITC and Fluo4 fluorescence with no appreciable influence on FSC or DCF values. Accordingly, PSN stimulates premature eryptosis characterized by PS externalization and intracellular calcium overload without cell shrinkage or oxidative damage. In conclusion, this report shows, for the first time, that PSN is cytotoxic to RBCs by inducing hemolysis and programmed cell death which may limit its success as a chemotherapeutic agent.

SNAIL is induced by tamoxifen and leads to growth inhibition in invasive lobular breast carcinoma.

PURPOSE: Invasive lobular carcinoma (ILC) is a histological subtype of breast cancer that is predominantly estrogen receptor alpha (ER)-positive (+) and is thus treated with endocrine therapies. Herein, we sought to understand the molecular underpinnings of the 4-hydroxytamoxifen (4OHT) resistance in ILC by assessing the potential role of the epithelial-to-mesenchymal transition transcription factor (EMT-TF) SNAIL (SNAI1). METHODS: Using a series of breast cancer cell lines, we measured the basal, estrogen and 4OHT-induced expression of SNAIL and other EMT-TF family members by quantitative reverse transcription-polymerase chain reaction and immunoblotting. Chromatin immunoprecipitation experiments were performed to assess ER binding to the SNAIL promoter. Cell proliferation, cell cycle and apoptosis were assessed in 2D cultures. 3D growth was assessed in Matrigel and Collagen I cultures. RESULTS: Estrogen and 4OHT induced SNAIL expression, but not that of the other EMT-TF family members SLUG (SNAI2) and SMUC (SNAI3), with the 4OHT effect being specific to the lobular but not the ductal subtype. We observed estrogen and 4OHT-induced ER recruitment to the SNAI1 promoter and high endogenous basal levels of SNAIL and several EMT-TFs in ILC cell lines. While SNAIL knockdown had a minor impact on the 4OHT partial agonism in estrogen-depleted conditions, it led to a surprising increase in cell proliferation in full serum. In complementary experiments, inducible SNAI1 overexpression caused decreased proliferation, associated with a cell cycle arrest in G0/G1. Additionally, apoptosis was observed in BCK4 cells. CONCLUSION: These data suggest a previously unrecognized role for SNAIL in ILC, substantiating a context-dependent behavior for this EMT-TF.

Novel nuclear localization of fatty acid synthase correlates with prostate cancer aggressiveness.

Fatty acid synthase is up-regulated in a variety of cancers, including prostate cancer. Up-regulation of fatty acid synthase not only increases production of fatty acids in tumors but also contributes to the transformed phenotype by conferring growth and survival advantages. In addition, increased fatty acid synthase expression in prostate cancer correlates with poor prognosis, although the mechanism(s) by which this occurs are not completely understood. Because fatty acid synthase is expressed at low levels in normal cells, it is currently a major target for anticancer drug design. Fatty acid synthase is normally found in the cytosol; however, we have discovered that it also localizes to the nucleus in a subset of prostate cancer cells. Analysis of the fatty acid synthase protein sequence indicated the presence of a nuclear localization signal, and subcellular fractionation of LNCaP prostate cancer cells, as well as immunofluorescent confocal microscopy of patient prostate tumor tissue and LNCaPs confirmed nuclear localization of this protein. Finally, immunohistochemical analysis of prostate cancer tissue indicated that nuclear localization of fatty acid synthase correlates with Gleason grade, implicating a potentially novel role in prostate cancer progression. Possible clinical implications include improving the accuracy of prostate biopsies in the diagnosis of low- versus intermediate-risk prostate cancer and the uncovering of novel metabolic pathways for the therapeutic targeting of androgen-independent prostate cancer.

A Comprehensive Two-Decade Analysis of Lymphoma Incidence Patterns in Saudi Arabia.

Background: Lymphomas account for approximately 10% of all cancer cases among the Saudi population. Even when separated, Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) are in the top ten most commonly diagnosed cancers among Saudi men and women. Despite the substantial cost of HL and NHL to public health, the resources to assess their impact are insufficient. This study provides a two-decade detailed assessment of lymphoma incidence trends in the Saudi population. Methods: Analysis of the Saudi Cancer Registry (SCR) data for various incidence metrics from 2001 to 2020 was conducted. Joinpoint regression analysis was further performed to investigate temporal trends globally and by age group, gender, and administrative region. Results: HL cases grew by 174.1%, whereas NHL cases increased by only 80% for that time period. The HL overall Age-Standardized Incidence Rate (ASR) increased by 100% for both genders combined but remained unchanged for NHL. The median age at diagnosis for HL (20-30 years) and NHL (46-57 years) was lower than in many other nations. Our model identified increasing trends for HL with annual percentage changes (APCs) of 2.94% (CI: 2.2-3.7) and 3.67% (CI: 2.6-4.7) for males and females, respectively. The rise was mainly among young groups under 40. On the contrary, the NHL cohort revealed notable declining tendencies. We discovered alarming rates of HL in Saudi Arabia's APC (2.23% for males and 3.88% for females) and ASR compared to other Western countries. Overall, the majority of the patients presented with advanced-stage disease at a younger age and with slight male predominance. Conclusions: The overall incidence of lymphoma (especially HL) has been rising among Saudis. Implementation of secondary and tertiary prevention measures, as well as management of modifiable risk factors, is warranted.

Comprehensive Retrospective Analysis of Colorectal Cancer Incidence Patterns in Saudi Arabia.

Colorectal cancer (CRC) is the commonest cancer in Saudi males and the third most common in Saudi females. Although CRC represents a major public health challenge, the resources to evaluate its burden are inadequate. This study aims to elucidate the magnitude of CRC incidence trends in the Saudi population by age, gender, and administrative region. Data for multiple incidence measures were analyzed from the Saudi Cancer Registry (SCR) retrospectively from 2001 to 2018. Temporal trends were further analyzed by age group, gender, administrative region, and globally using joinpoint regression analysis. The number of CRC cases climbed by 335.6% and the disease increased by 56.4% to comprise 12.2% of all cancers cases. The age-standardized incidence rate (ASR) increased by 152% overall, and the median age at diagnosis peaked at 60 and 58 years for males and females, respectively. Riyadh and the Eastern Region had the highest ASR for both genders, peaking at 21.8 and 19.2 for males and 17.4 and 16.5 for females per 100 K population. Our prediction model identified growing trends with annual percentage changes (APCs) of 4.59% in males (CI: 3.1-6.1) and 3.91% among females (CI: 2.4-5.5). Males above 75 years had the highest APC (7.9%, CI: 5.3-10.7), whereas the highest APC among females was found in the age group 70-74 (5.4%, CI: 2.8-8). Globally, APC was the highest for both genders compared to selected countries. CRC incidence is increasing alarmingly in Saudi Arabia and is projected to continue. There is a need for better screening strategies, preventative measures, and awareness-building.

Blood indices of omega-3 and omega-6 polyunsaturated fatty acids are altered in hyperglycemia.

Polyunsaturated fatty acids (PUFAs) may favorably influence the risk and clinical course of diabetes mellitus (DM). In particular, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and arachidonic acid (AA) alleviate oxidative injury and insulin resistance characteristic of DM. Uncertainty still remains, however, as to the composition and proportions of blood PUFAs in relation to fasting blood glucose levels. This study, thus, aims to examine the patterns of blood PUFA indices in normoglycemic (NG) and hyperglycemic (HG) Saudi subjects. Age, gender, FA profiles, and laboratory records of 143 subjects collected from September 2014 to March 2018 were retrospectively analyzed. Means, prevalence rates, associations, risk measures, and the diagnostic accuracy of PUFAs were determined. HG subjects had significantly lower AA (0.70%, 95% CI: 0.59-0.80% vs 0.46%, 95% CI: 0.38-0.53%) and higher EPA/AA ratio (0.36, 95% CI: 0.30-0.42 vs 0.69, 95% CI: 0.61-0.77). Gender-wise comparisons revealed that ώ-6/ώ-3 ratio was the only PUFA index significantly elevated in HG males (0.36, 95% CI: 0.26-0.45 vs 5.68, 95% CI: 4.98-6.38) while both DHA (2.91%, 95% CI: 2.54-3.29% vs 3.37%, 95% CI: 3.13-3.60%) and ώ-3 index (3.1%, 95% CI: 2.70-3.49% vs 3.63%, 95% CI: 3.38-3.88%) were significantly elevated in HG females. Furthermore, reduced AA and elevated EPA/AA ratio were more prevalent in HG subjects (26.53 vs 28.72 and 30.61 vs 38.29, respectively) and exhibited the highest diagnostic accuracy for HG among all PUFA indices. Altogether, our study revealed that distinct, gender-specific blood PUFA indices are differentially regulated in HG subjects which may be valuable for DM management.

WCRC-25: A novel luminal Invasive Lobular Carcinoma cell line model.

Breast cancer is categorized by the molecular and histologic presentation of the tumor, with the major histologic subtypes being No Special Type (NST) and Invasive Lobular Carcinoma (ILC). ILC are characterized by growth in a single file discohesive manner with stromal infiltration attributed to their hallmark pathognomonic loss of E-cadherin ( CDH1 ). Few ILC cell line models are available to researchers. Here we report the successful establishment and characterization of a novel ILC cell line, WCRC-25, from a metastatic pleural effusion from a postmenopausal Caucasian woman with metastatic ILC. WCRC-25 is an ER-negative luminal epithelial ILC cell line with both luminal and Her2-like features. It exhibits anchorage independent growth and haptotactic migration towards Collagen I. Sequencing revealed a CDH1 Q706* truncating mutation, together with mutations in FOXA1, CTCF, BRCA2 and TP53 , which were also seen in a series of metastatic lesions from the patient. Copy number analyses revealed amplification and deletion of genes frequently altered in ILC while optical genome mapping revealed novel structural rearrangements. RNA-seq analysis comparing the primary tumor, metastases and the cell line revealed signatures for cell cycle progression and receptor tyrosine kinase signaling. To assess targetability, we treated WCRC-25 with AZD5363 and Alpelisib confirming WCRC-25 as susceptible to PI3K/AKT signaling inhibition as predicted by our RNA sequencing analysis. In conclusion, we report WCRC-25 as a novel ILC cell line with promise as a valuable research tool to advance our understanding of ILC and its therapeutic vulnerabilities. Financial support: The work was in part supported by a Susan G Komen Leadership Grant to SO (SAC160073) and NCI R01 CA252378 (SO/AVL). AVL and SO are Komen Scholars, Hillman Foundation Fellows and supported by BCRF. This project used the UPMC Hillman Cancer Center and Tissue and Research Pathology/Pitt Biospecimen Core shared resource which is supported in part by award P30CA047904. This research was also supported in part by the University of Pittsburgh Center for Research Computing, RRID:SCR_022735, through the resources provided. Specifically, this work used the HTC cluster, which is supported by NIH award number S10OD028483. Finally, partial support was provided by the Magee-Womens Research Institute and Foundation, The Shear Family Foundation, and The Metastatic Breast Cancer Network.

Regioselective formation of acrolein-derived cyclic 1,N(2)-propanodeoxyguanosine adducts mediated by amino acids, proteins, and cell lysates.

Acrolein (Acr) is a major component in cigarette smoke and a ubiquitous environmental pollutant. It is also formed as a product of lipid peroxidation. Following ring closure via the Michael addition, Acr modifies deoxyguanosine (dG) in DNA by forming cyclic 1,N(2)-propanodeoxyguanosine adducts (OHPdG). The reactions of Acr with dG yield, depending on the direction of ring closure, two regioisomers, α- and γ-OHPdG, in approximately equal amounts. However, previous (32)P-postlabeling studies showed that the γ isomers were detected predominantly in the DNA of rodent and human tissues. Because of the potential differential biological activity of the isomeric OHPdG adducts, it is important to confirm and study the chemical basis of the regioselective formation of γ isomers in vivo. In this study, it is confirmed that γ-OHPdG adducts are indeed the major isomers formed in vivo as evidenced by a LC-MS/MS method specifically developed for Acr-derived dG adducts. Furthermore, we have shown that the formation of γ-isomers is increased in the presence of amino-containing compounds, including amino acids, proteins, and cell lysates. A product of Acr and arginine that appears to mediate the regioselective formation of γ isomers was identified, but its structure was not fully characterized due to its instability. This study demonstrates that intracellular amino-containing compounds may influence the regiochemistry of the formation of OHPdG adducts and reveals a mechanism for the preferential formation of γ-OHPdG by Acr in vivo.

The Role of Immune Checkpoint Inhibitors in Cancer Therapy.

Over the years, immune checkpoint inhibitors (CPIs) have become a powerful treatment strategy in the field of cancer immunotherapy. In the last decade, the number of FDA-approved CPIs has been increasing prominently, opening new horizons for the treatment of a wide range of tumor types. Pointedly, three immune checkpoint molecules have been under extensive research, which include cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein-1 (PD-1) and its ligand-1 (PD-L1). Despite remarkable success, not all patients respond positively to therapy, which highlights the complexity of the tumor microenvironment (TME) and immune system. This has led to the identification of molecular biomarkers to predict response and toxicity. In addition, there has been an emerging focus on developing new delivery and targeting approaches for better drug efficacy and potency. In this review, we highlight the mechanism of action of major CPIs, their clinical impact, variation in effectiveness, response prediction, updated clinical indications, current challenges and limitations, promising novel approaches, and future directions.

Induction of hemolysis and eryptosis by occupational pollutant nickel chloride is mediated through calcium influx and p38 MAP kinase signaling.

OBJECTIVES: Nickel (Ni) is an abundant environmental hazard and an occupational pollutant. Exposure to Ni compounds is prevalent in electroplating workers and in the printing industry, among others. The toxicity of Ni manifests as dermatological, gastrointestinal, respiratory, allergic, and cardiovascular symptoms. In particular, hyperbilirubinemia and reticulocytosis have been detected in intoxicated subjects; an observation possibly implicating selective red blood cell (RBC) toxicity. Herein, the interaction of nickel chloride (NiCl2) with human RBCs and associated molecular mechanisms are described. MATERIAL AND METHODS: Cells from healthy donors were incubated for 24 h at 37°C in the presence or absence of 0.5‒10 mM of NiCl2, and cytotoxicity was determined through hemoglobin leakage by colorimetry under different experimental conditions. Eryptotic markers were also identified by flow cytofluorometry using Annexin-V-FITC tagging for phosphatidylserine (PS) exposure, light scatter properties for cellular dimensions, Fluo4/AM labeling for intracellular calcium, and H2DCFDA staining for reactive oxygen species (ROS). Additionally, small molecule inhibitors were used to probe the signaling pathways involved. RESULTS: It was found that NiCl2 at 10 mM caused profound intracellular calcium overload and significant calcium-dependent hemolysis. Also, NiCl2 reduced forward scatter and increased side scatter, Annexin- positive cells, and ROS levels. Importantly, NiCl2-induced hemolysis was significantly attenuated by the exclusion of extracellular calcium, and in the presence of p38 MAP kinase (MAPK) inhibitor SB203580. CONCLUSIONS: It is concluded that NiCl2 induces p38 MAPK-dependent hemolysis, and stimulates the canonical features of premature eryptosis. This report presents the first description of the molecular mechanisms underlying the hemolytic and eryptotic potential of NiCl2 and, thus, may explain changes in hematological parameters observed in poisoning victims. Int J Occup Med Environ Health. 2022;35(1):1-11.

Calcium-oxidative stress signaling axis and casein kinase 1α mediate eryptosis and hemolysis elicited by novel p53 agonist inauhzin.

Inauhzin (INZ) is a novel p53 agonist with antitumor activity. Anemia is a common side effect of chemotherapy and may arise from red blood cell (RBC) hemolysis or eryptosis. In this study, we investigate the mechanisms of INZ toxicity in human RBCs. RBCs were isolated from healthy donors and treated with antitumor concentrations of INZ (5-500 µM) for 24 h at 37 °C. Hemoglobin was photometrically measured, and cells were stained with Annexin-V-FITC for phosphatidylserine (PS), Fluo4/AM for calcium, and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) for oxidative stress. INZ caused significant dose-responsive, calcium-dependent hemolysis starting at 40 µM. Furthermore, INZ significantly increased Annexin-positive cells and Fluo4 and DCF fluorescence. The cytotoxicity of INZ was also significantly mitigated in presence of D4476. INZ possesses hemolytic and eryptotic potential characterized by cell membrane scrambling, intracellular calcium overload, cell shrinkage, and oxidative stress secondary to calcium influx from the extracellular space.

Patterns of 25-Hydroxyvitamin D3, Calcium Status, and Anemia in the Saudi Population: A Cross-Sectional Study.

BACKGROUND: Emerging evidence suggests an intricate relationship between vitamin D, Ca2+, and inflammation-driven anemia. We, thus, investigated the patterns of serum 25(OH)D3, Ca2+, ferritin, and iron in healthy and anemic members of the Saudi population. METHODS: A population-based, retrospective, cross-sectional study was designed to analyze data for 14,229 subjects, aged 3-110 years, obtained from Al-Borg Medical Laboratories, over a six-year period (2014-2020). Gender and age differences were analyzed for 25(OH)D3, Ca2+, hemoglobin, ferritin, and iron. RESULTS: Vitamin D deficiency was extremely prevalent (98.47%) irrespective of age or gender, despite an increasing trend with age, in clear contrast to serum Ca2+. Ferritin was significantly lower in young adult and adult females, compared to elderly females, whereas iron was significantly reduced in females; in particular, adult females compared to young adults or elderly adults. Only anemic adult males had significantly lower 25(OH)D3, while Ca2+ was consistently significantly diminished in anemics of all age groups, independent of gender. Notably, hypocalcemic subjects were 2.36 times more likely to be anemic. Moreover, ferritin, but not iron, was significantly diminished in anemics, which was only evident in young adults and adults. However, both ferritin and iron showed positive correlation with hematocrit, hemoglobin, MCH, MCHC, and MCV. CONCLUSIONS: Despite being significantly lower in anemics, 25(OH)D3 is not particularly associated with anemia, while hypocalcemia is associated with an increased risk for anemia. Assessment of vitamin D and Ca2+ status may be valuable in the clinical management of anemia in the Saudi population.

Monocyte-Lymphocyte Ratio and Dysglycemia: A Retrospective, Cross-Sectional Study of the Saudi Population.

Background: Abnormalities in fasting blood glucose (FBG) resulting in hypoglycemia (OG), impaired fasting glycemia (IFG), or hyperglycemia (HG) arise from disordered metabolic regulation caused in part by inflammation. To date, there is a dearth of evidence regarding the clinical utility of the monocyte−lymphocyte ratio (MLR), an emerging inflammatory index, in the management of dysglycemia. Methods: This retrospective, cross-sectional study explored MLR fluctuations as a function of glycemic control in 14,173 Saudi subjects. Data collected from 11 August 2014 to 18 July 2020 were retrieved from Al-Borg Medical Laboratories. Medians were compared by Mann−Whitney U or Kruskal−Wallis tests and the prevalence, relative risk (RR), and odds ratio (OR) were calculated. Results: MLR was significantly elevated in IFG (p < 0.0001) and HG (p < 0.05) groups compared to the normoglycemia (NG) group, and individuals with elevated MLR (>0.191) had significantly increased FBG (p < 0.001). The risk of IFG (RR = 1.12, 95% CI: 1.06−1.19, p < 0.0002) and HG (RR = 1.10, 95% CI: 1.01−1.20, p < 0.0216) was significantly increased if MLR was elevated, and individuals with elevated MLR were 1.17 times more likely to have IFG (OR = 1.17, 95% CI: 1.08−1.26, p < 0.0002) and 1.13 times more likely to have HG (OR = 1.13, 95% CI: 1.02−1.24, p < 0.0216). Conclusion: Elevated MLR is correlated with and carries a greater risk for IFG and HG. However, large prospective cohort studies are needed to establish the temporal relationship between MLR and FBG and to examine the prognostic value of this novel marker.

Patterns of Dyslipidemia in the Anemic and Nonanemic Hypertensive Saudi Population: A Cross-Sectional Study.

Background: Risk factors of cardiovascular disease include dyslipidemia, hypertension (HTN), and anemia. Our objective is to assess the patterns of dyslipidemia in the anemic and non-anemic hypertensive Saudi population. Methods: A retrospective, cross-sectional study of the gender, blood pressure, lipid markers, and CBC parameters of 3111 subjects, which were retrieved from the database of Al-Borg Medical Laboratories over a six-year period (2014-2019), was carried out. Means were compared among study groups and the prevalence, association, and diagnostic accuracy of lipid markers for HTN were evaluated. Results: TG, LDL/HDL, and TG/HDL were significantly higher (P < 0.0001) in hypertensives. Anemia reduces TC and LDL (P < 0.0001) in both genders, and reduces all markers and increases HDL (P < 0.01) in male hypertensives. HTN was more prevalent in anemics with high TC than normal TC (38.23% vs 11.17%, P < 0.001) and in non-anemics with high TG than normal TG (56.31% vs 21.22%, P < 0.001). Furthermore, non-anemics with high TG/HDL had the highest risk for HTN (RR = 1.20, 95% CI = 1.1551-1.2473, P < 0.0001). Elevated TC (P = 0.0142), TG (P < 0.0001), TC/HDL (P < 0.0001), LDL/HDL (P < 0.0001), and TG/HDL (P < 0.0001), and low HDL (P < 0.0001) were risk factors for HTN as shown by ORs. In anemics, high TC/HDL, LDL/HDL, and TG/HDL were not. Importantly, only TG and TG/HDL had a discriminating capacity for HTN. Conclusion: The anemic state of hypertensive Saudi patients influences dyslipidemia which warrants further investigation.

Epidemic dropsy toxin, sanguinarine chloride, stimulates sucrose-sensitive hemolysis and breakdown of membrane phospholipid asymmetry in human erythrocytes.

Sanguinarine (SGN) is a benzophenathridine alkaloid extracted from Sanguinaria canadensis plant. SGN is incriminated in epidemic dropsy (ED) characterized by multiple-organ failure and anemia. Nevertheless, how SGN leads to anemia of ED remains poorly understood. This study was thus initiated to investigate the interaction of SGN with human red blood cells (RBCs) and to delineate associated molecular mechanisms. Heparin- and EDTA-anticoagulated blood was collected from healthy participants and whole blood was analyzed for a complete blood count, while isolated RBCs were examined for hemolytic and eryptotic markers following exposure to 1-100 µM SGN for 24 h at 37 °C. Calcium was measured by Fluo4/AM, hemolysis by hemoglobin leakage, membrane scrambling by Annexin V-FITC, cell size by forward scatter (FSC), cell granularity by side scatter (SSC), and oxidative stress by H2DCFDA. SGN led to increased Fluo4 fluorescence and dose-dependent hemolysis which was not ameliorated by exclusion of extracellular Ca2+ but was nevertheless sensitive to hyperosmotic conditions and to the presence of aspirin. SGN also caused significant increase in Annexin V-positive cells, decreased FSC and SSC values, and elevated DCF fluorescence. Moreover, significantly reduced lymphocyte and basophil percentages along with selective toxicity to platelets was noted. Collectively, SGN possesses sucrose- and cyclooxygenase-sensitive hemolytic potential and elicits eryptosis characterized by Ca2+ accumulation, phosphatidylserine externalization, morphological alterations including cell shrinkage and loss of granularity, and oxidative stress. In conclusion, this report reveals a novel activity of SGN against human RBCs and informs prospective policies in ED prevention and management.

Antiproliferative Wnt inhibitor wogonin prevents eryptosis following ionophoric challenge, hyperosmotic shock, oxidative stress, and metabolic deprivation.

Anemia is a common complication of chemotherapy and may arise due to premature or suicidal death of red blood cells (RBCs). Prevention of RBC death thus lends itself as a promising strategy to counteract anemia. Wogonin (WGN; 5,7-dihydroxy-8-methoxyflavone) is a Wnt inhibitor derived from Scutellaria baicalensis plant with potent cytotoxic and antitumor potential. However, the nature of interaction of WGN with human RBCs is unknown. RBCs from healthy participants were exposed to different hemolytic and eryptotic stimuli for 24 or 48 hr at 37°C in the presence and absence of 100 µM WGN. Calcium overload was induced by 2 µM ionomycin, hyperosmotic shock with excessive sucrose, oxidative stress by 2-phenethyl isothiocyanate (PEITC), and metabolic deprivation by exclusion of glucose. Hemolysis was estimated from extracellular hemoglobin, phosphatidylserine (PS) exposure by Annexin V-FITC, intracellular calcium by Fluo4/AM, and oxidative stress by 2',7'-dichlorodihydrofluorescein diacetate (H2 DCFDA). While WGN did not rescue the cells from the hemolytic activity of ionomycin, it reduced PS externalization by interfering with calcium influx under both ionomycin treatment and metabolic exhaustion. WGN also significantly inhibited PS exposure upon hyperosmotic shock, but the effect was independent of calcium entry. Moreover, WGN exhibited antihemolytic and anti-eryptotic activities against PEITC without appreciable reduction in ROS levels. Altogether, WGN prevents PEITC-induced hemolysis and suppresses eryptosis due to calcium accumulation, hyperosmotic shock, oxidative stress, and metabolic exhaustion. These novel insights may open new avenues into the therapeutic application of WGN for conditions in which anemia is commonly encountered, most notably cancer. PRACTICAL APPLICATIONS: The herbal supplement Sho-Saiko-To (Xiaochaihu-tang) is commonly prescribed to relieve symptoms of liver disease. Flavonoids from the herbal constituents of Sho-Saiko-To have demonstrated considerable anti-inflammatory, antioxidant, antimicrobial, antitumor, and immunomodulatory properties. In this work, we identify WGN, a major flavonoid in Sho-Saiko-To, as a novel inhibitor of hemolysis and eryptosis of human erythrocytes. Since inordinate erythrocyte death is a major obstacle in therapeutic drug development, our findings argue for the use of WGN as a natural alternative, either as a primary or an adjuvant drug, for a wide assortment of pathological conditions including cancer.