Whole Blood Hemoglobin Research Articles

A-142 Comparison of Alinity c Hemoglobin A1c Immunoassay with Premier Hb9210 Automated HPLC Assay: Very Few Patients May be Outside Reportable Range Using Alinity c.

Abstract Background Hemoglobin A1c (HbA1c; glycosylated hemoglobin) is used for diagnosis and monitoring of patients with diabetes. In the clinical laboratory both immunoassays and high-performance liquid chromatography (HPLC) based tests are available for such measurement. We use Premier Hb9210 analyzer (HPLC method; Trinity Biotech, Jamestown, NY) for measuring HBA1c in whole blood. As our laboratory is transitioning to Abbott chemistry systems, we compared HbA1c values obtained by the Alinity c and Premier Hb9210. Methods The Premier Hb9210 analyzer is based on boronate affinity high performance liquid chromatography with output in 66 sec. The analytical measurement range is 3.8 to 18.5% although values as low as 2% can be reported by the analyzer but result may not be accurate. The Alinity c Hemoglobin A1c assay measures both total hemoglobin and HbA1c (enzymatic assay) in whole blood and then calculates %HbA1c. The analytical measurement range of this assay is 4 to 14% of HbA1c. Specimens cannot be diluted for Alinity c HbA1c measurement. Results Alinity c HbA1c has excellent total precision (0.4% at HbA1c of 5.1% and 0.25% at Hb A1c of 9.4%). In contrast, Premier HB9210 has total precision of 1.89% at HbA1c of 5.5% and 1.81% at HbA1c of 8.4%. However, analytical measurement range of Premier HB9219 is slightly wider compared Alinity c assay. Plotting HbA1c results obtained by the Premier Hb9210 analyzer in the x-axis and the corresponding values obtained by the Alinity c assay, we observed the following regression equation: y = 0.9499 x + 0.1108 ( n = 50, 0.99). The 95% confidence interval of the slope was 0.9241 to 0.9757. Rarely, we get specimen where HbA1c is between 3.8 and 4.0% (42 specimens out of 52 803 between 2/7/22 to 2/7/23; 0.08%) or above 14% (174 specimens out of 52 802; same period; 0.33%) and these specimens would not produce any result with Alinity c assay. We recently observed HbA1c result of 2.4 % in a sickle cell patient (reported as <3.8% per our lab policy). Discussion Our result indicates that HbA1c immunoassay on the Alinity c analyzer showed values comparable to HPLC method. However, for very few patients HbA1c may exceed 14% due to uncontrollable diabetes. Because specimen cannot be diluted, real value could not be measured using Alinity c but can be measured by the HPLC method. Conclusions We conclude that HbA1c assay on the Alinity c analyzer is a viable alternative to HPLC for measuring HbA1c in clinical laboratory. However, for very few patients (sickle cell trait, uncontrolled diabetes), HbA1c may be outside the range of Alinity c, requiring HPLC analysis.

Selected biomarkers of oxidative stress in healthy Beagle dogs: A preliminary study.

While oxidative stress has been studied in pathologic conditions in dogs, data in presumably healthy dogs and standardized protocols are lacking. This work purposed to bridge the gap by presenting provisional physiological ranges for oxidative stress biomarkers in a group of Beagle dogs. Based on our long-standing clinical expertise in the field of oxidative stress, nine plasma biomarkers of oxidative stress were evaluated for their concentrations (mean ± SD) in 14 healthy adult Beagle dogs. Selected biomarkers were: vitamins C (7.90 ± 1.36 μg/mL) and E (34.1 ± 6.63 μg/mL), zinc (0.80 ± 0.17 mg/L), copper (0.54 ± 0.048 mg/L), selenium (256 ± 25.7 μg/L), total and oxidized glutathione (822 ± 108 μM and 3.56 ± 1.76 μM), myeloperoxidase (67.4 ± 56.2 ng/mL), and isoprostanes (340 ± 95.3 ng/mL). Glutathione peroxidase activity and superoxide anion production in whole blood were also measured. Glutathione peroxidase activity was 473 ± 34.0 IU/g of hemoglobin and superoxide anion production in whole blood was 18,930 ± 12,742 counts per 30 min. Reduced glutathione/oxidized glutathione and copper/zinc ratios were, respectively, 280 ± 139 and 0.70 ± 0.15. Sex-related differences were recorded for zinc (p = 0.0081), copper/zinc ratio (p = 0.0036) and plasma isoprostanes (p = 0.0045). Provisional physiological norms covering 95% of our group were proposed for each biomarker and should be of interest for future studies of canine oxidative stress.

Absolute Quantification of Total Hemoglobin in Whole Blood by High-Performance Liquid Chromatography Isotope Dilution Inductively Coupled Plasma-Mass Spectrometry.

Accurate and traceable measurement of hemoglobin (HGB) is of great importance in clinical testing. Although the HiCN method is the internationally accepted conventional reference method for this biomarker and frequently used in clinical routine diagnostics, the HiCN method cannot be traceable to the International System of Units (SI) and thus does not meet highest metrological demands. In this study, an absolute quantitative approach for total HGB in a whole blood sample is proposed based on the determination of natural Fe and S present in the heme-group of HGB by HPLC isotope dilution ICP-MS. IRMM/IFCC-467 is used for method validation, and then clinical blood samples are measured by the established strategy and HiCN method. The measurable ranges of total HGB were 10.0-240.0 g L-1. Limits of detection via Fe and S were 0.01 and 0.07 g L-1, respectively. The intra-assay imprecision CVs via Fe and S were 0.89-1.35 and 0.99-1.56%, and the interassay CVs were 1.19-2.15 and 1.55-2.55%, respectively. Good agreement was achieved in the method validation. In the comparison with HiCN experiments, the ID-ICP-MS assays via Fe and S showed correlations of r2 = 0.991 and 0.970 against HiCN methods. Moreover, the concentration of transferrin (Tf) was also simultaneously measured. This strategy has potential to serve as a reference measurement procedure for total HGB in whole blood, which could be traceable to SI and does not require toxic derivation reagent.

Selective Adsorption of Hemoglobin in Human Whole Blood with a Nickel Monosubstituted Silicotungstic Acid Hybrid

A nickel monosubstituted polyoxometalate (POM)/polyaniline organic–inorganic hybrid SiW11Ni/PANI was synthesized using the liquid-phase method at room temperature to achieve the solidification of water-soluble POMs. The SiW11Ni/PANI hybrid was characterized by scanning electron microscopy, thermogravimetric analysis, Fourier transform infrared spectroscopy, X-ray diffraction, and zeta potential. On the basis of π–π stacking and affinity interaction between the SiW11Ni/PANI hybrid and proteins, the SiW11Ni/PANI hybrid showed good adsorption selectivity to hemoglobin (Hb). At pH 7.0, 0.5 mg of SiW11Ni/PANI resulted in an adsorption efficiency of 92.4% for 1 mL of 100 μg mL–1 Hb. The adsorption behavior of Hb on the surface of the hybrid fitted with the Langmuir model, and the maximum adsorption capacity was 692 mg g–1. The adsorbed Hb was eluted by BR (0.04 mol L–1, pH 9), providing a recovery of 94%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis assay results indicated that the Hb in human whole blood could selectively be adsorbed by the SiW11Ni/PANI hybrid, and the obtained Hb was with high purity. It could expand the application of POMs in life science due to the application of the POM hybrid in protein isolation/purification.

Development and validation of an in-house handheld ELISA kit to detect glycosylated hemoglobin in human whole blood

Background: Diabetes mellitus (DM) is known as one the oldest disease known to a human being and it is defined as a group of metabolic disorders characterized by the presence of hyperglycemia. Aims and Objectives: The present study describes the development and evaluation of a cost-effective and straightforward in-house handheld enzyme-linked immunosorbent assay (ELISA) kit for the determination of HbA1c in human whole blood of diabetic patients and is suitable for resource-poor settings. Materials and Methods: A total of 1056 specimens were collected for assay validation purposes. To check the proficiency of our diagnostic strategy, we compared our test results with high-performance liquid chromatography (HPLC) and a commercially available ELISA kit for HbA1c. The Institutional Ethical Committee approved this study (IEC). SPSS 16 statistical software analyzed all the data. The correlation regression was done to compare the assay. The level of significance of this study was P<0.001. Results: The sensitivity, specificity, and efficiency of the inhouse HbA1c ELISA kit range from 98.8%, 100%, and 99.24%. Conclusion: The newly developed in-house handheld HbA1c ELISA test kit is not only cost-effective, accurate, and straightforward but also gives good correlation with optimized methods such as HPLC and commercially available techniques in deciding the glycemic status of patients. Therefore, highly recommended for use in resource-poor settings in the management and diagnosis of DM.

Suppression of fertility by <em>Capparis spinosa</em> root extract through antioxidant alteration in reproductive system of male rats

Aim: To evaluate the suppression of fertility by oral mode of Capparis spinosa root extract through alteration in antioxidant defence of the reproductive system of male albino rats. Methods: The methanol extract of Capparis spinosa root (100mg/kg/day) was fed for 60 days to male, Wistar strain, albino rats. The fertility and testicular functions were assessed by mating tests, antioxidant defence indices, and spermatogenesis, histometry, and biochemical estimations for fertility suppression along with serum biochemistry and hematology for assessment of toxicology. Results: Capparis spinosa root extract administration for a period of 60 days did not cause body weight loss significantly, while the weight of testes, Cauda epididymis, seminal vesicle, and ventral prostate were significantly reduced. The lipid peroxidation and antioxidants levels i.e. superoxide dismutase, catalase, glutathione reduced, glutathione peroxidase, vitamin C, and Vitamin E in testes, Cauda epididymis, seminal vesicle, and ventral prostate were altered significantly (P<0.001) after the 60 days treatment of Capparis spinosa extract when compared to the controls. The root extract feeding caused a marked reduction in the number of primary spermatocytes, secondary spermatocytes, and spermatids however spermatogonia population showed non-significant change. Sertoli cell population was also affected. The number of mature Leydig cells was decreased and degenerating cells increased proportionately. The seminiferous tubule and Leydig cell nuclear area were reduced significantly (P<0.001) when compared to the controls. The protein and sialic acid content of the testes, Cauda epididymis, seminal vesicle, and ventral prostate were significantly (P<0.001) decreased. The glycogen in testes and fructose in seminal vesicle was also significantly reduced, whereas the testicular cholesterol was elevated as compared with the controls. Reduced sperm count and motility resulted in a total suppression of fertility. The RBC and WBC count, hemoglobin, and hematocrit in whole blood whereas marker enzymes and glucose in serum were found to be within the normal range after the 60 days of Capparis spinosa extract treatment. Conclusion: Capparis spinosa root extract may selectively act on developing germ cells mediated via Sertoli cells through increased oxidative stress or impairment of antioxidant defence system in reproductive organs of male rats, leading to suppression of fertility and suggests the feasibility of developing non-toxic, herbal male contraceptive drug.

An investigation of the effectiveness of oral cyclosporine on perioperative myocardial injury (PMI) in patients who undergo the surgical procedure of coronary artery bypass graft (CABG): A Randomized Controlled Clinical Trial.

Background:Routine clinical strategies for the prevention of myocardial infarction (MI) during the surgical procedure of CABG include cross-clamp fibrillation and cardioplegia have failed to decrease the risk of perioperative myocardial injury (PMI). Cyclosporine-A (CsA) might be able to prevent mitochondrial dysfunction and PMI.Methods:In the present clinical trial, patients were divided into two groups (Case receive 2.5 mg/kg CsA and Control receive a placebo) randomly. Moreover, patients were controlled by placebo through a double-blind, single-center trial 4-12 h before anesthesia. Perioperative blood tests include bilirubin, complete blood count, the amount of hemoglobin in whole blood, liver transaminases, and glomerular filtration rate (GFR). Blood samples were taken before surgery and at 24, 48, and 72 h after surgery and serum Troponin-I and CK-MB levels were determined in all blood samples using ELISA.Results:There were no significant differences between the two groups in the results of routine pre-operative blood results, intraoperative variables, and baseline characteristics (P > 0.05). There are significant correlations between cross clamp time and cTnI and CKMB levels in patients taking CsA. In patients with both diabetes and hypertension, postsurgical cTnI and CKMB levels decrease significantly in CsA compared to placebo group on 24, 48, and 72 h (P < 0.05). Moreover, patients with old MI, both postsurgical cTnI and CKMB levels decrease significantly in CsA compared to placebo group on 24 h and 48 h (P < 0.05).Conclusions:In patients with a long cross-clamping period, using an oral CSA single dose before conducting CABG surgery, the risk of PMI could be decreased. Also, oral CsA has protective effect for CABG in diabetic patients with hypertension.

Absolute Quantification of Haemoglobin in Whole Blood by Species-unspecific Isotope-dilution Liquid Chromatography-inductively Coupled Plasma Mass Spectrometry

Absolute Quantification of Haemoglobin in Whole Blood by Species-unspecific Isotope-dilution Liquid Chromatography-inductively Coupled Plasma Mass Spectrometry

Associations between selected perfluoroalkyl acids in serum and hemoglobin in whole blood, a biomarker of anemia: Impact of deteriorating kidney function

Data (N = 11251) from National Health and Nutrition Examination Survey (NHANES) for 2003–2016 for US adults aged ≥20 years were stratified by gender and anemia and analyzed to evaluate the associations between the concentrations of whole blood hemoglobin (WBHGB) and selected perfluoroalkyl acids (PFAAs) in serum by stages of glomerular filtration (GF). Investigated PFAAs were perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), perfluorodecanoic acid (PFDA), perfluorohexane sulfonic acid (PFHxS), and perfluorononanoic acid (PFNA). Females with whole blood hemoglobin concentrations <12 g/dL and males with whole blood hemoglobin concentrations <13 g/dL were classified as being anemic. Regression models with log10 transformed concentrations of whole blood hemoglobin as dependent variable and age, poverty income ratio, body mass index, serum cotinine, daily alcohol intake, survey year, and log10 concentrations of one of the PFAA as independent variables were fitted. For anemic females, association between WBHGB and PFAA concentrations were uniformly positive across worsening stages of renal failure and percent increases for 10% increases in PFAAs varied between 0.03% and 0.39%. For anemic males, association between WBHGB and PFAA concentrations were positive except at GF-3A (45 ≤ eGFR<60 mL/min/1.73 m2) and percent increases for 10% increases in PFAAs varied between 0.02% and 0.53%. Thus, more often than not, presence of positive associations between WBHGB and PFAA among anemics imply elevated levels of PFAA are associated with higher levels of WBHGB. Similar results were observed for non-anemic males and females, however strengths of associations between whole blood hemoglobin and PFAAs were several fold higher among anemic compared to non-anemic participants. Hemoglobin is consistently associated with serum PFAAs.

Parental Males of the Plainfin Midshipman Are Physiologically Resilient to the Challenges of the Intertidal Zone.

The decision of where to rear young is influenced by both the needs of offspring and the costs parents incur in certain rearing environments. Plainfin midshipman fish (Porichthys notatus) provide extended paternal care in rocky intertidal zones, where they experience regular bouts of aquatic hypoxia and air exposure during low-tide events. We investigated the physiological responses of plainfin midshipman males to three conditions for 6 h that simulate what these fish naturally experience during tidal cycles while nesting: normoxia, progressive hypoxia, or air exposure. Hypoxia- and air-exposed fish exhibited shifts in energy metabolites, driven largely by elevated lactate and glucose content and reduced glycogen content in several tissues (muscle, heart, liver, and brain), but the magnitude of these changes was relatively modest. Hematocrit increased most in air-exposed fish relative to normoxia-exposed fish, contributing to an increase in whole-blood hemoglobin concentration. Air exposure reduced swim bladder oxygen content, suggesting that internal O2 stores are drawn on during air exposure. In a second experiment, we found that aquatic surface respiration and gill ventilation frequency increased in hypoxia-exposed fish relative to normoxia-exposed fish. Overall, our results suggest that plainfin midshipman overcome the challenges of the intertidal environment through a variety of physiological strategies and exhibit little physiological disturbance in response to the fluctuating and extreme conditions created by regular low tides.

Association between milk consumption and the nutritional status of poor rural Chinese students in 2016.

Milk promotes the growth of children's height. However, the relationship between milk consumption and anemia or obesity remain unclear. We explored the association between milk consumption and the nutritional status of poor rural Chinese students, including anemia, height, weight, malnutrition, and overweight/obesity. A total of 22,315 students aged 8-16 years were recruited. The frequency of milk consumption and other information were investigated using the questionnaire. Students' morning fasting height, weight, and whole-blood hemoglobin were measured. The children were classified as malnutrition, normal weight, and overweight/obesity according to their age-specific height and BMI. Multivariate linear and multiple logistic regression were used to analyze the association between milk consumption and nutritional status. In total, 10.6% of students drank milk at least once a day. Compared to students who drank milk <1 time/week, The heights and weights of students who consumed milk 1-3 times/week, 4-6 times/week, and at least once per day were 0.8, 0.9, and 1.3 cm greater and 0.8, 0.6, and 1.0 kg heavier. Students who drank milk at least once a day (OR=0.817), and 1-3 times/week (OR=0.868) had a significantly lower prevalence of malnutrition. Students who drank milk 4-6 times/week (OR=0.472) had a significantly lower prevalence of anemia. However, no significant association was identified between milk consumption and overweight/obesity. Frequent consumption of milk was associated with these students having a lower risk of malnutrition and anemia, being taller, and being heavier.

Productive traits, selenium status and blood metabolic status in fattening lambs affected by selenium biofortified corn

The widespread selenium (Se) deficiency in soil and in feedstuffs from such soil, needs to be prevented by various methods. Recently, biofortification of Se has been carried out by soil fertilization and foliar application of crops. The aim of this study was evaluate the productive traits, Se status and blood metabolic profile in fattening lambs affected by Se biofortified corn. The research was conducted with 20 Merinolandschaf lambs of the average age of 70 days, during 30 days of fattening. The feed mixture of the control group contained corn (0.014 mg Se/kg dry matter, DM); in the experimental group (Se-BC), corn was biofortified (0.278 mg Se/kg DM). The feed mixture, hay and water were offered to lambs ad libitum. Blood was collected from the jugular vein on days 0 and 30 of experiment. Haematological indices were determined in whole blood, whereas concentrations of biochemical indices, enzyme activities, and concentrations of metabolic hormones were determined in serum. After the slaughter, samples of musculus semimembranosus, liver, kidney, lungs, spleen, peritoneum and heart tissues were taken. No significant differences were obtained in productive traits when feeding lambs with Se-BC. Higher concentrations of Se in lungs and liver (control: 0.26 and 0.75 mg/kg, respectively; Se-BC: 0.33 and 0.92 mg/kg, respectively) was determined in Se-BC compared to control. Higher activity of glutathione peroxidase enzyme in serum, higher erythrocyte count, higher content of haemoglobin and haematocrit in whole blood of Se-BC lambs were determined. The results of the study indicate the posibility of using Se-BC in lambs’ diets.

Peculiarities of Metabolism of Rats with Fructose-induced Insulin Resistance Against the Background of Congenital and Acquired Iodine Deficiency

Animals following a high-fructose diet during eight weeks, have experienced changes in metabolism and the signs of insulin resistance have developed. Under such conditions, moderate hyperglycemia, hyperinsulinemia, hyperuricemia, an increase of the level of glycosylated hemoglobin in whole blood were observed. The significant role of the HOMA-IR index, as an early marker of carbohydrate metabolism disorders at the stage of pre-diabetes, has been confirmed. In experimental animals against the background of the high-fructose diet, the changes in the lipid spectrum of the blood were revealed: an increase of the total cholesterol level, low-density lipoproteins, triglycerides against the background of a high-density lipoproteins decrease. These disorders and a significant increase of the atherogenicity reflect the development of secondary dyslipidemia. In this case, the disorders of carbohydrate metabolism were combined with the degree of dyslipidemia. Males were found to have at increased risk of development the insulin resistance and comorbid pathology. Iodine deficiency, especially of congenital nature, is an aggravating factor of metabolic disorders. The obtained data can serve as a basis for extend of preventive measures and identification of the priority treatment schemes for type 2 diabetes mellitus in residents of endemic regions.

Fast and Sensitive Detection of Paramagnetic Species Using Coupled Charge and Spin Dynamics in Strongly Fluorescent Nanodiamonds.

Sensing of a few unpaired electron spins, such as in metal ions and radicals, is a useful but difficult task in nanoscale physics, biology, and chemistry. Single negatively charged nitrogen-vacancy (NV-) centers in diamond offer high sensitivity and spatial resolution in the optical detection of weak magnetic fields produced by a spin bath but often require long acquisition times on the order of seconds. Here, we present an approach based on coupled spin and charge dynamics in dense NV ensembles in strongly fluorescent nanodiamonds (NDs) to sense external magnetic dipoles. We apply this approach to various paramagnetic species, including gadolinium complexes, magnetite nanoparticles, and hemoglobin in whole blood. Taking advantage of the high NV density, we demonstrate a dramatic reduction in acquisition time (down to tens of milliseconds) while maintaining high sensitivity to paramagnetic centers. Strong luminescence, high sensitivity, and short acquisition time make dense NV- ensembles in NDs a potentially promising tool for biosensing and bioimaging applications.

Burnout Syndrome in Physicians-Psychological Assessment and Biomarker Research.

Background and objectives: Burnout is a syndrome typically occurring in work environments with continuous and chronic stress. Physicians are at increased risk for burnout, as a result of 24-h work, delayed work–life balance gratification, and the challenges associated with patient care. The aim of the present study was to evaluate the psychological parameters of burnout symptoms in relation to biomarkers of stress among physicians with different medical specialties. Materials and methods: A total of 303 physicians were contacted as potential participants. A comparison group of 111 individuals working outside medicine was used as a control to verify the results. The physicians were specialists in internal medicine, general surgery, pathology, and primary care. Serum cortisol, salivary cortisol, adrenocorticotropic hormone (ACTH), insulin (IRI), and prolactin levels were analyzed by chemiluminescence enzyme immunoassay (Access 2, Beckman Coulter). Fasting glucose in serum and glycated hemoglobin (HbA1C) in whole blood were measured using the automatic analyzer AU 480 Beckman Coulter system. Symptoms of burnout were measured with the Maslach Burnout Inventory (MBI). Results: The group with burnout presented significantly higher levels of serum and saliva cortisol, ACTH, prolactin, fasting glucose, and HbA1C compared with the control group. The correlation analysis between biomarkers showed a positive correlation with moderate strength between serum and saliva cortisol (r = 0.516, p = 0.01),as well as serum and saliva cortisol with ACTH (r = 0.418; r = 0.412, p = 0.01) and HbA1C (r = 0.382; r = 0.395, p = 0.01). A weak positive correlation was found between serum and saliva cortisol with prolactin (r = 0.236; r = 0.267, p < 0.01) and glucose (r = 0.271; r = 0.297, p < 0.01). In the multiple logistic regression model, saliva cortisol, HbA1C, and age were significantly associated with burnout (chi-square = 16.848, p < 0.032). Conclusion: Our findings demonstrated the interest of exploring biomarkers of stress related to burnout in health professionals.

Capturing hemoglobin on graphene sheet from sub-microliter whole blood for quantitative characterization by internal extractive electrospray ionization mass spectrometry

A disposable blood sampler, which is consisted of a sub-microliter whole blood collector and a graphene filter, loading graphene sheet to selectively capture hemoglobin from sub-microliter whole blood, was developed for both qualitative and quantitative characterization hemoglobin by internal extractive electrospray ionization mass spectrometry (iEESI-MS). The blood collector was elegantly fabricated in syringe-like fashion for precisely sampling tiny amounts (1.0 μL - 2%) of whole blood, which was immediately diluted by water inside the syringe and was then pressed through the graphene filter placed between the waste outlet and the syringe reservoir to capture the hemoglobin in the blood sample. Then the graphene with hemoglobin was directly eluted by a charged (+2.5 kV) solution (mathanol/water/formic acid, 48/48/4, v/v/v) to produce the hemoglobin ions for mass spectrometric analysis. Low detection-of-limit (19.3 mg L−1 (89.5 picomol)), acceptable linear response range (300–1500 mg L−1, R2 = 0.998), relative standard deviation (0.5–6.5%, n = 3), low sample consumption (≤1.0 μL) and a relatively high speed (≤4 min per sample, including the sample loading) were achieved, demonstrating that the graphene based iEESI-MS was an alternative choice for direct detection of hemoglobin in whole blood with minimal sample consumption.

Characterization of suicide involving insulin analog injection: Interest of injection site analysis by LC/HRMS

To provide evidence of the role of insulin in the occurrence of death is always challenging: glycemic state can be estimated only in vitreous matrix, and postmortem blood sample detection of insulin or its analogs is known to be problematic. This case report is of a death involving insulin analog injection characterized at the injection site on liquid chromatography coupled to an exactive high-resolution/high accuracy mass spectrometry (LC/HRMS). A 56-year-old nurse with suicidal tendencies was found dead in her car 24 hours after disappearing. An empty bottle of whiskey, empty alprazolam blisters and insulin pens were found near the corpse. Autopsy showed common signs of toxic death. Three needle marks were found on the left arm, and were sampled along with the usual toxicological matrices. General toxicological screening was performed to screen for ethanol, carbon monoxide, medical drugs and drugs of abuse. Glucose and lactate in vitreous humor (VH) and glycated hemoglobin (HBA1c) in whole blood were quantified by an enzymatic method on a Cobas ® analyzer. Immunoassay (Cobas ® ) was performed on femoral blood (FB) serum to screen for insulin and C-peptide. For insulin assay in injection site tissue, 1 g of tissue was pretreated by grinding and ultrasonic bath in an acid ethanol solution; then solid phase extraction by anion exchange was performed before analysis on LC/HRMS. Ethanol was detected only in urine, at 0.11 g/L. Ethylglucuronide was quantified at 0.06 μg/mL and 133 μg/mL in FB and urine, respectively. Alprazolam (45 ng/mL) and tramadol (270 ng/mL) were found in FB. Morphine and codeine were quantified in urine at 102 ng/mL and 290 ng/mL respectively, but not detected in FB. HBA1c level was 4.5%. Glucose assay was negative (< 0.10 mmol/L) in both VH samples. Lactate was quantified at 10.3 and 9.9 mmol/L in right and left VH, respectively. Serum immunoassays were negative for insulin (< 1 mU/L) and C-peptide (< 0.03 nmol/L). Analysis of suspected injection site tissue identified aspart, an insulin analog. Alprazolam and tramadol concentrations were subtoxic in blood; opiates and ethanol were detected only in urine. Toxic death was not demonstrated from these Results. The values of glucose and lactate in vitreous humor suggested a potentially lethal hypoglycemic state, according to the reference values proposed by Karlovsek [1] . However, despite the insulin pens found in the car, insulin immunoassay was negative in blood. Injection site analysis by LC/HRMS allowed identification of the insulin analog, aspart. This finding of insulin analog intake in a non-diabetic subject, associated with the glucose and lactate concentrations in VH, suggested severe hypoglycemia induced by insulin analog injection. Various hypotheses could explain the failure to detect aspart in serum: the poor ability of immunoassay to detect analogs, metabolism during the time between injection and death, and/or degradation by hemolysis during the postmortem interval. Demonstration of death by insulin injection is difficult and should not be limited to blood immunoassay. Estimation of glycemic status at time of death and identification of insulin or analogs by injection site analysis by LC/HRMS are fundamental to characterizing death involving insulin injection. In the present case, the medicolegal examiner diagnosed a hypoglycemic state with fatal outcome secondary to aspart injection as cause of death, and suicide as the manner.

Micronutrient intakes and status assessed by probability approach among the urban adult population of Hyderabad city in South India.

To assess the dietary inadequacies of micronutrients and the associated factors among the apparently healthy urban adults. This community-based cross-sectional study involved 300 urban adults (distributed into age groups: 21-40, 41-60, and > 60years) residing in Hyderabad city, South India. Hemoglobin in whole blood, ferritin, folate, and vitamin B12 (B12) in plasma was estimated. Dietary intakes were assessed by three 24-h dietary recalls and calculated the probability of adequacy (PA) using estimated average requirement. The prevalence of anemia (30%), iron deficiency (ID, 23%), and iron deficiency anemia (IDA, 14.3%) was independent of age but higher in women. While folate deficiency (32.2%) was independent of age and gender, B12 deficiency (35.5%) varied by both age and gender. The PA of iron (89%) was higher, while that of folate, B12, and zinc (1-11%) were noticeably low. The mean PA (MPA) across the ten micronutrients was 38%, independent of age and gender, but associated with the educational status. Energy intake was a strong predictor of the MPA. Cereals and millets predominantly contributed to the intake of thiamine, niacin, zinc, and iron; green leafy vegetables and fruits to vitamins A, C, folate, and iron; animal foods to B12; and milk and milk products to calcium, vitamin A, riboflavin, and B12. The unadjusted and adjusted logistic regression models revealed that micronutrient inadequacy was associated with greater risk of IDA and folate deficiency. These results indicate a higher prevalence of micronutrient deficiencies among the healthy urban adults possibly due to the inadequacy of multiple micronutrients.

Recent developments and innovations in red blood cells diagnostics

Although the diagnosis of anemia is a relatively simple and straightforward endeavor, since it is mostly based on the measurement of hemoglobin in whole blood, the accurate characterization of red blood cell (RBC) disorders has emerged as a mainstay in the era of precision (laboratory) medicine. The many technological advances occurred in laboratory medicine over recent times have enabled the introduction of a vast array of innovations also for RBC diagnostics, mostly represented by modular automation, digital hematology, innovative erythrocyte parameters and adaptation of disruptive technologies to laboratory hematology, especially mass spectrometry and molecular biology. These important breakthroughs have enormously contributed to broadening the diagnostic armamentarium and making more efficient and sustainable the diagnostics of RBC disorders. Some on these important innovations will be discussed in this article.

Apoptosis in pancreatic β-cells is induced by arsenic and atorvastatin in Wistar rats with diabetes mellitus type 2

IntroductionDiabetes Mellitus type 2 (T2D) is a multifactorial disease. However, it is known that there is an important effect in pancreatic β-cells caused by apoptosis of pro-apoptotic proteins, possibly related to arsenic exposure and atorvastatin treatment. ObjectiveThe goal of this study was to evaluate the effects of atorvastatin treatment on apoptosis of pancreatic β-cells in Wistar rats with induced diabetes type 2 exposed to arsenic. Material & methodsT2D in Wistar rats was induced by administration of Streptozotocin. The plasmatic glucose concentrations were measured using the glucose oxidase method, and the concentration of glycated hemoglobin (HbA1c) in whole blood was determined. Exposure to arsenic was measured from urine using atomic absorption with hydride generation, and pro-apoptotic proteins in pancreatic β-cells were observed using the Western blotting technique. ResultsCaspase-3 was present in rats that were treated with 10 mg/kg of oral atorvastatin and exposed to 0.01 and 0.025 mg/L of arsenic, but no others proteins were present, such as pro Caspase-8, bcl-2, and Fas. The glycemic levels were 129.2 ± 7.0 mg/dL in the control group and 161.8 ± 14.6 mg/dL and 198.3 ± 18.2 mg/dL (p < .05) in the study groups. HbA1c increased from 2.53% to 3.64% (p < .05) in the control and study groups. ConclusionsAtorvastatin treatment and arsenic exposure alone are capable of generating apoptosis in pancreatic β-cells of Wistar rats with T2D. Together, all of these factors induce apoptosis in pancreatic cells.