Antisickling effect of chrysin is associated with modulation of oxygenated and deoxygenated haemoglobin via alteration of functional chemistry and metabolic pathways of human sickle erythrocytes.

Sickle cell disease (SCD) treatment and management remain a challenging puzzle especially among developing Nations. Chrysin's sickling-suppressive properties in human sickle (SS) erythrocytes in addition to its effect on AA-genotype erythrocytes were evaluated. Sickling was induced (76%) with 2% sodium metabisulphite at 3 h. Chrysin prevented (81.19%) the sickling and reversed same (84.63%) with strong IC50s (0.0257 µM and 0.00275 µM, respectively). The levels of oxygenated haemoglobin in the two groups (before and after induction approaches) were similar but significantly (P < 0.05) higher than that of SS erythrocytes (the 'induced' control), with chrysin-treated AA-genotype showing no effects relative to the untreated. The level of deoxygenated haemoglobin in the 'induced' control group was significantly (P < 0.05) higher than those of the chrysin-treated SS erythrocytes. Normal and chrysin-untreated erythrocytes (AA-untreated) were significantly more resistant to osmotic fragility than the SS-untreated. However, treatment with chrysin significantly reduced the osmotic fragility of the cells relative to the untreated cells. Furthermore, chrysin treatment significantly lowers the high level of 2,3-diphosphoglycerate (2,3-DPG) observed in the sickle erythrocytes, with no effects on AA-genotype erythrocytes. Based on functional chemistry, chrysin treatment alters the functional groups in favour of its antisickling effects judging from the observed bends and shifts. From metabolomics analysis, it was observed that chrysin treatment favors fatty acid alkyl monoesters (FAMEs) production with concomitant shutting down-effects on selenocompound metabolism. Thus, sickling-suppressive effects of chrysin could potentially be associated with modulation of oxygenated and deoxygenated haemoglobin via alteration of human sickle erythrocyte's functional chemistry and metabolic pathways implicated in SCD crisis.

Sickling-suppressive effects of chrysin may be associated with sequestration of deoxy-haemoglobin, 2,3-bisphosphoglycerate mutase, alteration of redox homeostasis and functional chemistry of sickle erythrocytes.

Sickle cell disease (SCD) is a medical condition caused by mutation in a single nucleotide in the ß-globin gene. It is a health problem for people in sub-Saharan Africa, the Middle East and India. Orthodox drugs developed so far for SCD focus largely on symptomatic respite of pain and crisis mitigation. We investigated the antisickling effects of chrysin via modulation of deoxy-haemoglobin, 2,3-bisphosphoglycerate mutase, redox homeostasis and alteration of functional chemistry in human sickle erythrocytes. In silico and in vitro methods were adopted for the studies. Chrysin was docked against deoxy-haemoglobin and 2,3-bisphosphoglycerate mutase, with binding energies (-24.064 and -18.171 kcal/mol) and inhibition constant (K i) of 0.990 µM and 0.993 µM at their active sites through strong hydrophobic and hydrogen bond interactions. Sickling was induced with 2% metabisulphite at 3 h. Chrysin was able to prevent sickling maximally at 2.5 µg/mL and reversed the same at 12.5 µg/mL, by 66.5% and 69.6%, respectively. Treatment with chrysin significantly (p < 0.05) re-established the integrity of erythrocytes membrane as evident from the observed percentage of haemolysis relative to induced erythrocytes. Chrysin also significantly (p < 0.05) prevented and reversed lipid peroxidation. Similarly, glutathione and catalase levels were observed to significantly (p < 0.05) increase with concomitant significant (p < 0.05) decrease in superoxide dismutase activity relative to untreated. From Fourier-transform infrared results, treatment with chrysin was able to favourably alter the functional chemistry, judging from the shifts and functional groups observed. Sickling-suppressive effects of chrysin may therefore be associated with sequestration of deoxy-haemoglobin, 2,3-bisphosphoglycerate mutase, alteration of redox homeostasis and functional chemistry of sickle erythrocytes.

Burden and Correlates of Anaemia among Patients with Diabetes Mellitus in a Tertiary Health Facility in Northwestern Nigeria.

BACKGROUND: Anaemia is a common disorder occurring in about 33% of the global population. It is an important cardiovascular risk factor and a key indicator of some chronic complications of Diabetes Mellitus (DM). This study aimed to determine the burden of anaemia and its correlation with some clinical and biochemical parameters among patients with DM attending a tertiary health facility in Zaria, Northwestern Nigeria. SUBJECTS, MATERIALS AND METHODS: This was a case-control study in which 168 participants were enrolled (84 DM patients, 84 controls). It was conducted in the Endocrinology and Metabolic clinics of Ahmadu Bello University Teaching Hospital, Zaria. Consenting DM patients were enrolled consecutively and subsequently, sex- and age-matched with non-diabetic controls. Data on age, gender and Haemoglobin (Hb) concentrations were collated for all study participants. Additional data on type of DM, duration of DM once diagnosis, treatment, type of treatment, history of hypertension, chronic kidney disease, peripheral neuropathy, and Fasting Blood Sugar (FBS) were collated for all cases. Data were collated and analyzed using SPSS version 21. Level of significance was set at <0.05. Ethical approval for the study was obtained from the Institutional Health Research Ethics Committee and informed consent was obtained from the all the participants. RESULTS: Females constituted 39/84(46.4%) of each arm of the study. The mean ± SD of age for both cases and controls was 53.7 ± 8.9 years. The mean ± SD duration of DM, treatment for DM and FBS were 8.4 ± 5.7 years, 5.0 ± 3.6 years and 6.1 ± 2.5mmol/L respectively. Cases had significantly lower Hb concentration compared to controls (12.1±2.2g/dl vs. 13.1 ± 1.4g/dl, t= -3.446, p = 0.001). Overall prevalence of anaemia among cases and controls was 36/84(42.9%) vs. 26/84(31.0%) Z = 1.6, p = 0.110. Among cases, haemoglobin concentration had very weak, inverse and non-statistically significant relationships with age, duration of DM diagnosis, duration of therapy and FBS levels. There was a significant relationship between anaemia on one hand and type of DM and treatment on the other. The odds of DM patients with history of CKD or uncontrolled FBS having anaemia were OR= 0.600 (95% CI 0.196, 1.836) and OR=1.755 (95% CI 0.737, 4.181) respectively. CONCLUSION: The burden of anaemia amongst patients with DM is high in Zaria, Northwestern Nigeria, and it is associated with poor glycaemic control. Hence, the need to include haematological assessment as part of routine care of patients with DM.

Erythropoietic Response to Anaemia of Dialysis Naïve Patients with Chronic Kidney Disease in Zaria, North-West Nigeria.

INTRODUCTION: Chronic kidney disease (CKD) is a global health problem with an increasing prevalence worldwide. Anemia is one of its consistent and severe hematological complications although its mechanism is not fully elucidated. The primary defect could manifest as serum erythropoietin (sEPO) deficiency or EPO resistance. We set out to determine the erythropoietic response to anemia of patients with CKD and its relationship with their iron status in a cross-sectional descriptive study of 91 patients in various stages of CKD. Materials and Methods: Soluble transferrin receptor (sTfR), sEpo, and serum ferritin levels were determined using ELISA method (Diagnostic Automation Inc and WKEA med supplies corp.). Data generated were analyzed using Epi Info version 3.5.3 and level of statistical significance was set at ≤0.05. Results: Participants comprised of 50 females (54.9%) and 41 (45.1%) males with an overall mean age of 47 ± 15 years. The major causes of CKD were hypertension (HTN) (50.54%), diabetes mellitus (DM) (6.59%), and HTN + DM (19.78%). The mean hemoglobin (Hb) concentration of the participants was 10.97 ± 2.28 g/dl; the red cell indices were within normal ranges except for Red cell distribution width-Coefficient of variation (%) which was elevated (16.29%). The mean serum ferritin, sTfR, and sEpo were 70.58 ± 46.44 ng/ml (interquartile range [IQR] 82.00), 22.9 ± 49.7 ng/ml (IQR 15.00), and 12.49 ± 33.47 IU/L (IQR 6.00), respectively, with a high variance. Serum ferritin and sTfR are consistently low across the stages of CKD (range between 54.54 ng/ml and 88.64 ng/ml), but sEPO for stage 3 and 4 showed a 2-fold increase when compared to normal level at Hb 10.97 g/dl (29.54 IU/L and 38.83 IU/L, respectively). Correlation between sTfR and sEpo (r2 = 0.96, P = 0.001), while between sEpo and serum ferritin (r2 = 0.02, P = 0.185), and between Hb and stage of CKD undulating (r2 = 0.41, P = 0.001). CONCLUSION: In contrast to some existing literature, this study has demonstrated that EPO resistance and iron deficiency contributes to anemia in CKD and serum ferritin can be used to assess the iron level of dialysis naïve CKD patients at every stage of the disease.

Variations in the ß-globin genes of sickle cell anaemia patients in Zaria, Northwestern, Nigeria.

CONTEXT: Sickle Cell Anaemia (SCA) is a genetic disorder with a life-long disability, which is of public health importance. The diversity in its clinico-pathologic and laboratory presentations may be due to the interplay between additional genetic differences and environmental factors. The genetic factors may be within the ß-globin gene itself, the ß-globin gene cluster or elsewhere in the genome. AIM: To characterize the ß-globin gene for variations associated with the Sickle Cell mutation. SETTINGS AND DESIGN: A cross-sectional descriptive study involving 51 adult SCA patients attending Sickle Cell Clinic of Haematology Department Ahmadu Bello University (ABUTH) Zaria, Kaduna State, Nigeria. METHODS AND MATERIAL: The buccal swab specimens were collected and ß-globin gene DNA sequencing was done. The sequences obtained were compared with a Genbank Reference ß-globin gene (NC_000011.9) using Basic Local Alignment Search Tool (BLAST), and variations noted. Data generated were analyzed using SPSS Version 20.0. STATISTICAL ANALYSIS USED: Data generated was summarized by using charts, means±2SD, and 95% confidence intervals. RESULTS: There were 40 (78.43%) females and 11 (21.57%) males. The mean age of the participants was 25.35 ± 7.67 years, 95% CI (23.20, 27.51). The classic sickle cell mutation A T was present in all participants. The mean number of ß-Globin gene variations was 8.61±11.30, 95% CI (5.43, 11.78). The number of Substitutions were 122 (27.79%), insertions 184 (41.91%), and deletions 133 (30.30%). These occurred in various combinations. The mean number of substitutions, insertions, and deletions were 2.39±3.23, 3.61±7.66, and 2.60±2.46 with 95% CIs of (1.48, 3.30), (1.45, 5.76), and (1.92, 3.30) respectively. CONCLUSIONS: There are ß-globin gene variations in SCA patients in Zaria, and locally relevant genetic database of the SCA population will be the cornerstone in understanding genotype-phenotype interactions in this disorder.

Fournier's Gangrene as Seen in University of Maiduguri Teaching Hospital.

Background. Fournier's gangrene is uncommon but increasingly being seen over the last two decades probably due to increasing socioeconomic problems including an upsurge in HIV infection especially in the tropics. Patients and Methods. The study retrospectively reviewed all patients with Fournier's gangrene managed in UMTH between January 2007 and December 2012. Results. Thirty-eight males aged 2 weeks to 80 years (mean 37.82) were reviewed, with most aged 30-39 years (13 (34.21%)). Clinical features were scrotal pain and swelling, 36 (94.74%), fever, 19 (50.00%), and discharging scrotal wound, 19 (50.00%). The predisposing conditions were UTI secondary to obstructive uropathy in 11 (28.95%), perianal suppuration, and HIV, in 8 (21.05%) patients each. Wound biopsy culture revealed mixed organisms in 27 (71.05%). Twenty-six (68.42%) had blood transfusions. Thirty-seven (97.37%) patients had wound debridement. Twenty (52.63%) had flap rotation for skin cover. There were 6 (15.79%) mortalities, of which 4 (10.53%) were HIV positive, 1 (2.63%) was diabetic, and 1 (2.63%) was both diabetic and HIV positive. Conclusion. Fournier's gangrene is a fulminant synergistic necrotising fasciitis of the perineum and genitalia with poor prognosis especially when associated with HIV and diabetes, requiringprompt and aggressive management for good outcome.

Characterization and sequencing of prototypic human T-lymphotropic virus type 1 (HTLV-1) from an HTLV-1/2 seroindeterminate patient.

Serological screening for human T-lymphotropic virus type 1 (HTLV-1) parallels the standard screening process for human immunodeficiency virus (HIV), in which samples found positive by enzyme-linked immunosorbent assay (ELISA) are confirmed with a modified Western blot procedure. There are a significant number of cases in which HTLV-1/2 ELISA-positive specimens demonstrate an incomplete banding pattern on this Western blot. Individuals providing these atypical antibody responses are categorized as seroindeterminate for HTLV-1/2. Although HTLV-1 genomic sequences are readily detectable in the peripheral blood lymphocytes (PBL) of seropositive individuals, previous studies have repeatedly demonstrated that PBL from the vast majority of HTLV-1/2 seroindeterminate individuals are PCR negative for HTLV-1. As a result, identification of the agent responsible for this indeterminate reactivity has been of interest. We have generated an HTLV-1-positive B-cell line (SI-1 B) from one of these seroindeterminate individuals. Previous screening for HTLV-1 in PBL from this patient had been routinely negative by primary PCR; however, HTLV-1 tax had been periodically detected by nested PCR. DNA sequence data generated with genomic DNA from the SI-1 B cell line and HTLV-1-specific primers demonstrated the presence of a full-length viral genome with >97% homology to the Cosmopolitan form of HTLV-1. A 12-bp deletion was identified in the 3'-gag/5'-prot region, which would predict translation of altered or nonfunctional proteins from these genes. We propose that this HTLV-1/2-seroindeterminate patient is infected with a prototypic form of HTLV-1 at an extremely low viral load and that this finding may explain HTLV-1/2 seroindeterminate reactivity in at least a subset of these individuals.

HTLV-I/II seroindeterminate Western blot reactivity in a cohort of patients with neurological disease.

The human T-cell lymphotropic virus type I (HTLV-I) is associated with a chronic, progressive neurological disease known as HTLV-I-associated myelopathy/tropical spastic paraparesis. Screening for HTLV-I involves the detection of virus-specific serum antibodies by EIA and confirmation by Western blot. HTLV-I/II seroindeterminate Western blot patterns have been described worldwide. However, the significance of this blot pattern is unclear. We identified 8 patients with neurological disease and an HTLV-I/II seroindeterminate Western blot pattern, none of whom demonstrated increased spontaneous proliferation and HTLV-I-specific cytotoxic T lymphocyte activity. However, HTLV-I tax sequence was amplified from the peripheral blood lymphocytes of 4 of them. These data suggest that patients with chronic progressive neurological disease and HTLV-I/II Western blot seroindeterminate reactivity may harbor either defective HTLV-I, novel retrovirus with partial homology to HTLV-I, or HTLV-I in low copy number.

Implicit and explicit learning in an auditory serial reaction time task.

OBJECTIVE: To explore the role of the motor cortex during implicit and explicit learning. MATERIALS AND METHODS: EEG signals were recorded from 30 channels by measuring task-related desynchronization (TRD) when 10 right-handed naive volunteers performed a variation of the serial reaction task. Stimuli, consisting of 4 pure tones of 500, 1000, 1500, and 2000 HZ, lasting 200 ms, were presented binaurally through a pair of tubephones at 60 dB with a 2-s constant interstimulus interval. A series of 10 repetitive tones represented the test sequence; the random sequence was the control. RESULTS: All subjects developed implicit and explicit knowledge reflected by decreased response time, increased accuracy, and the ability to generate the sequence. Six of 10 subjects demonstrated implicit learning without explicit learning during the first 3 blocks. When subjects acquired full explicit learning, 10 Hz TRD at C3 reached a peak amplitude, declining thereafter. CONCLUSIONS: Properties of the sensorimotor cortex change during learning and these changes are independent of stimulus modality.

Stereotactic neurosurgery planning with 3-D spiral CT-angiography.

OBJECTIVE: To assess the feasibility and value of spiral CT angiography of the brain vessels for the planning of neurosurgical stereotactic interventions. MATERIAL AND METHODS: Fourty-two patients harboring cerebral lesions underwent spiral CT angiography prior to stereotactic biopsy. Thin spiral CT slices with a collimator slice thickness of 1 mm and a pitch of 1 were used. Multiplanar reconstructions and maximum intensity projections (MIP) were obtained as well as 3-D tissue definition. RESULTS: There was a sufficient visualization of vessels and of their relationship to the lesion. Tumor neovascularization was clearly demonstrated. Arteries could be shown separately. Stereotactic coordinates of targets were chosen at a safe distance from the vessels and the simulation of tarjectories using the cine loop was made possible. In three cases the presence of a pathological vascularization warned against a stereotactic biopsy. CONCLUSION: Spiral CT angiography seems to yield enough topographical information for the accurate planning of stereotactic surgery for brain lesions. CT angiography with the helical technique is rapid and less invasive than digital subtraction angiography.

[Value of CT-angiography in diagnosis of cerebral sinus and venous thromboses]. / Stellenwert der CT-Angiographie in der Diagnostik von zerebralen Sinus-und Venenthrombosen.

Dural sinus thrombosis is not uncommon. Due to the nonspecific symptomatology, as well as the manifold etiology, clinical diagnosis may be difficult. In these cases imaging procedures are frequently crucial in deciding how to proceed and how to treat. The aim of our study was to evaluate the diagnostic utility of helical CT in the detection of dural sinus thrombosis. In 20 patients with clinically suspected thrombosis CT angiography was performed. In 6 patients dural sinus thrombosis was diagnosed. In order to acquire also arterial vessels, a short delay of about 22 s after the onset of the application of contrast medium was selected. By this method we found an occlusion of the MCA in two patients with clinically suspected sinus thrombosis. In all patients the transverse slices and the multiplanar reconstructions showed filling defects or an "empty delta" sign. With irregular outlines the thrombus could be depicted over the complete course of the sinus. The MIP reconstructions were particularly helpful in the evaluation of the vessel anatomy and the pathological collateral venous drainage. In three patients MR angiograms were available for comparison. The smaller veins, such as the v. vermis inferior, were less clearly depicted than in CT angiography. CT angiography is a fast and reliable method to exclude or verify a sinus thrombosis. It can be performed immediately after non-enhanced CT. According to our present experience CT angiography is sufficient for the diagnosis of a sinus thrombosis.

[CT-angiography in planning stereotaxic biopsies]. / CT-Angiographie in der Planung stereotaktischer Biopsien.

Twenty-one patients referred for stereotactic biopsy were studied by CT angiography. Helical CT with 1 mm collimation was obtained (pitch of 1:1). Multiplanar reconstructions were performed; maximum intensity projections and shaded-surface displays were generated by connectivity-based editing tools. The visualization of cerebral vessels was excellent. No further conventional angiography was needed. Improved information was obtained about localization of the intracranial lesion and its relationship to neighboring vessels. No bleeding complications were detected by CT after stereotactic biopsy.

Low-dose radiotherapy for stage I seminoma: early results.

One hundred and one patients with stage I seminoma were irradiated with total doses of 30, 25.5 and 20 Gy to gradually reduced target volumes (paraaortic, pelvic, and inguinal regions to paraaortic only). Low doses and small target volumes resulted in excellent survival and freedom of recurrence but in more frequent nausea.