Criterion for patient selection and demographic comparison between the two groups, the numbers in individual groups (normal union or defective union) how the observers were blinded for the samples and control group measurements in the graphs were not mentioned. No data on actual measurement levels is given. No where it is mentioned like it is an average of all normal union or defective union in tables or diagrams. The treatment methodology maybe still focused including either conservative or surgical treatment since a displaced fracture cannot be compared with an osteotomy. The cases shall be followed up for a longer period. Some suggestions on how to blind the observers is given. In the one year period there was neither mention of the failure of treatment or complications of any of neither these 36 cases nor any drop out for follow-ups. There was also no mention of any case which initially put on conservation was changed to surgical management. The authors could have one group of patients taking this food stuff and another group who refused this food stuff. The statistical test used to compare the levels of factor is not mentioned. Mere statement that ‘p’ values were significant will not benefit the reader. Failure to produce X-rays even for a single case weakens the study. X-rays are needed to confirm the diagnosis of a fracture and confirm the position of implants and fracture fragments. The remaining period after confirming the fracture/implant position the case shall be followed only with marker estimation. Once the desired levels of increase or tapering of marker level achieved then X-rays can be taken to correlate with clinical findings and radiology. Union as one group and the second group shall be called as non-union or delayed union. The second group (the poor callus group) is mentioned as malunion possibly by over-sight. Probably they were meaning the non union or delayed union group or defective union as malunion. This should be preferably be written non union or delayed union. Malunion means the fracture actually unites and union process is completed. In a group of fractures (hypertrophic non unions) the callus formation is excessive, still the fracture is ununited. Thus the enzyme or markers alone cannot disclose the details of the completion of union they can herald bone formation.
Blinding; Marker estimation; X-rays; Non-union; Malunion
Iron is an essential trace element and plays a number of vital roles in biological system. It also leads the chains of pathological actions if present in excess and/or present in free form. Major portion of iron in circulation is associated with transferrin, a classical iron transporter, which prevent the existence of free iron. The fraction of iron which is free of transferrin is known as “non transferrin bound iron”. Along with the incidence in iron over loaded patient non transferrin bound iron has been indicated in patients without iron overload. It has been suggested as cause as well as consequence in a number of pathological conditions. The major organs influenced by iron toxicity are heart, pancreas, kidney, organs involved in hematopoiesis etc. The most commonly suggested way for iron mediated pathogenesis is through increased oxidative stress and their secondary effects. Generation of free oxygen radicals by iron has been well documented in Fenton chemistry and Haber–Weiss reaction. Non transferrin bound iron has obvious chance to generate the free reactive radicals as it is not been shielded by the protective carrier protein apo transferrin. The nature of non transferrin bound iron is not clear at present time but it is definitely a group of heterogenous iron forms free from transferrin and ferritin. A variety of analytical approaches like colorimetry, chromatography, fluorimetry etc. have been experimented in different research laboratories for estimation of non transferrin bound iron. However the universally accepted gold standard method which can be operated in pathological laboratories is still to be developed.
Non transferrin bound iron; Oxidative stress; Haber–Weiss reaction; Fenton chemistry; Transferrin; Apo transferrin
Cinnamon has been used as an anti-diabetic agent for centuries but only in recent few years its mechanism of action has been under investigation. Previous studies showed that cinnamon might exert its anti-diabetic effect via increasing glucose transporter isotype-4 (GLUT4) gene and glycoprotein contents in fat cells. To study if hydro-alcoholic cinnamon extract (HACE) enhances GLUT4 translocation from intracellular compartments of nuclear or endoplasmic reticulum membranes (N/ER) into the cytoplasmic membrane (CM). C2C12 myoblastic cell line were seeded in DMEM plus 20 % FBS and differentiated to myotubes using 2 % horse serum. After myotubes formation, 100 or 1,000 μg/ml HACE, as intervention, and as control 1 % DMSO were added for 3 h. Cells were washed and homogenized followed by ultracentrifuge fractionation, protein separation by SDS-PAGE and GLUT4 detection using semi-quantitative Western blotting. Data analysis was done by two-independent samples t test for comparison of mean ± SD of GLUT4 percent in categories. GLUT4 contents were higher in CM of groups 100 and 1,000 μg/ml HACE and lower in 1 % DMSO treated myotubes (CI = 0.95, P < 0.05). For N/ER reverse results were obtained (CI = 0.95, P < 0.05). As our results have shown HACE induces GLUT4 translocation from intra-cell into cell surface. We conclude that cinnamon maybe a choice of type-2 diabetes mellitus treatment because its extract enhances GLUT4 contents in CM where it facilitates glucose entrance into the cell. However it is necessary to trace the signaling pathways which are activated by HACE in muscular tissue.
Anti-diabetic; Cinnamon; Diabetes type-2; GLUT4; Muscle
Recent studies have demonstrated the presence of prostate specific antigen (PSA) in cord blood of male as well as female babies. The placental progesterone and estradiol up-regulate the synthesis and secretion of PSA in Placenta. This PSA is presumed to play a role in intrauterine growth of fetus by virtue of its proteolytic action on several substrates including insulin-like-growth-factor-binding-protein-3, insulin chains and Interleukin-2. This study was planned with the objective of correlating the levels of PSA in cord blood to gestation at delivery, the type of delivery and gender of the fetus. Fifty-seven cord blood samples were collected from the umbilical cord during delivery or mid-trimester abortion and analyzed for PSA using ‘Active PSA DSL-9700 ultra sensitive’ kit employing two-site immuno-radiometric assay principle and having a detection limit of 0.001 ng/ml. Mean PSA levels in cord blood were found to be 0.112 ± 0.027 ng/ml. The concentration of PSA in cord blood was found to be higher in case of higher gestational age, male baby and operative delivery. 50 % of cord bloods for female babies had PSA below detection limit (range <0.001–0.460 ng/ml), while all the male samples had detectable PSA (range 0.11–0.973 ng/ml). Higher Progesterone levels found in prenatal maternal blood in case of male babies may be responsible for the higher cord blood PSA. Mean cord blood PSA was 0.150 ± 0.150 ng/ml in forceps delivery and 0.078 ± 0.012 ng/ml in normal vaginal delivery. Forceps delivery causes much more stress and strain as compared to a normal vaginal delivery, resulting in increased levels of adrenal glucocorticoids, and therefore, higher cord blood PSA.
Prostate specific antigen; Cord blood; Gestation; Mode of delivery; Gender of baby
Abnormal glomerular permeability is the primary step towards the glomerulosclerosis. The progression rate of glomerulosclerosis is proportionate to abundance and severity of lesions created at incipient stage, which is reflected as proteinuria even though eGFR remains in the normal range. Therefore, there is a current need to find out the association between relative risks for the factors leading to proteinuria. The relations could be more informative, if it is with respect to the macromolecules like “IgG” excretion in urine. Type 2 diabetic patients were selected for this study with eGFR > 75 ml/min/1.73 m2 and grouped into four quartiles based on UIgGCR. The markers of key factors affecting progression of proteinuria were estimated through biochemical tests. The impact of these markers on proteinuria was accessed by applying multinomial logistic regression. The adjusted odds ratio for the UGAGCR was 1.186 (95 % CI: 1.061–1.327) P < 0.003 in highest quartiles of UIgGCR, followed by odds ratio for markers of collagen catabolism 1.051 (95 % CI: 1.025–1.079) P < 0.001, and USACR 1.044 (95 % CI: 1.013–1.077) P < 0.006 respectively. The marker of glycation, i.e., glycated hemoglobin showed the highest odds ratio 5.449 (95 % CI: 1.132–26.236) P < 0.035. In addition, odds for the systolic blood pressure was observed 1.387 (95 % CI: 1.124–1.712) P < 0.002. The higher odds inform and could help to discriminate the diabetic patients with fast progressive diabetic nephropathy. The study describes critical relationship between the urinary excretion of IgG and factors leading to proteinuria in type 2 diabetic patients.
Diabetic nephropathy; Urinary glycosaminoglycans; Urinary sialic acid; Proteinuria; Glomerular basement membrane
Mucopolysaccharidoses (MPS) are a group of genetic diseases and its diagnosis is a challenging task due to multiple differential diagnosis. We had combined clinical findings, radiological and ophthalmological features. Biochemical test for urine glycosaminoglycans (GAG) was done for confirmation of diagnosis in the patient. The case of Sanfilippo disease was characterized by slowly progressive, severe CNS involvement with mild somatic disease. Radiological features were suggestive of Sanfilippo disease and urine GAG test for MPS was positive in the case. With the clinical features we had multiple differential diagnoses. The radiological investigations minimized the list and the biochemical test confirmed GAG in urine. In this case the combination of clinical, radiological and biochemical findings confirmed the diagnosis of Sanfilippo disease.
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The online version of this article (doi:10.1007/s12291-012-0211-1) contains supplementary material, which is available to authorized users.
Sanfilippo disease; Diagnosis; Clinical; Radiological; Biochemical
The Antarctic continent on the planet Earth is full of environmental extremes. It is considered as natural stress model. Therefore, the present study examined the effect of harsh environment on the certain salivary markers of 28th Indian Antarctic expeditioners. Thirty healthy men and women (median age 36 year; range 22–61 year) participated in this study. Parameters measured were salivary IgA (SIgA), IgM (SIgM), TGF-β and cortisol level at three different time points: (I) before leaving India on 26th October 2008 for base line level; (II) after 1 month on-board journey on 31st January 2009 in Southern Ocean and (III) after 1 month staying at Maitri, Antarctica on 3rd March 2009. Our observation indicated that Ship borne journey and Antarctic environment increased the SIgA levels while that of SIgM level was not altered on-board but decreased by staying for 1 month at Antarctica. No significant alteration was found in the TGF-β and cortisol level at any point of time. The present study concluded that ship borne journey and Antarctic environment may induce the SIgA level while SIgM level decreased in environmental extremes of Antarctica.
Cytokine; Immunoglobulins; Antarctica
Hemoglobin J-Rajappen (alpha)90 Lys → Thr is an alpha chain variant found in heterozygous state and presents normal hematological blood picture. Due to the ambiguity in results obtained while analyzing by HPLC and alkaline gel electrophoresis, we report this rare case of HbJ-Rajappen using non denaturing gel electrophoresis and matrix assisted laser desorption ionization mass spectrometry. Though HbJ-Rajappen has earlier been reported using different techniques, this is the first report being validated using mass spectrometry technique.
Hemoglobin J-Rajappen; Mass spectrometry; High performance liquid chromatography; Electrophoresis
The present study was conducted on North Indian population to observe rpoB gene mutation profile in multidrug resistant Mycobacterium tuberculosis. This was an observational study. 30 cases of MDR-TB proven by culture and drug sensitivity were selected. DNA sequencing of 81 bp (codon 507–533) long RRDR of Mycobacterium tuberculosis was done to detect the sites of mutation. Out of 30 cases, 24 showed a single mutation in the RRDR region of rpoB gene in which 16 (53.33 %) showed mutation in codon 531(TCG→TTG), 5 cases (16.66 %) showed mutation in codon 526(CAC→TAC), mutation in codon 516(GAC→GTC, AAC) was present in 3 cases (10 %). It was also observed that mutation in more than one codon was present in 4 cases (13.33 %), which included deletion at codon 509(AGC→–GC), mutation at 513(CAA→CTA), 516, 526, 529(CGA→CTA) and 531. No mutation was detected in RRDR in 2 cases (6.66 %). Our finding of 13.33 % cases with multiple sites of mutation in RRDR region is in contrast to earlier studies done in North India which showed single mutation detected in RRDR of rpoB gene that highlights the emerging change in the trend of mutation profile of rpoB gene in rifampicin resistant Mycobacterium tuberculosis.
Multiple mutations; Rifampicin resistance; Rifampicin resistance determining region
The objective of this study to evaluate heme oxygenase (COHb), leptin and coenzyme Q10 (CoQ10) in pre-eclamptic women. Also Zinc, copper, Iron, total iron binding capacity, Ferritin and uric acid were assessed. 120 female subjects were included in this study. They were divided into, 60 female with normal pregnancy attending the outpatient clinic, 60 pre-eclamptic patients were recruited from obstetrics and gynaecology department El-kasr El-Aini hospital. The results showed that in pre-eclampatic group, leptin level was significantly increased while COHb and CoQ10 was significantly decreased. It is concluded that hemeoxygenase, leptin and coenzyme CoQ10 can be considered as new markers for prediction of pre-eclampsia.
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The online version of this article (doi:10.1007/s12291-012-0226-7) contains supplementary material, which is available to authorized users.
Pre-eclampsia; Hemeoxygenase; Carbon monoxide; Leptin; Coenzyme Q10
Most cytokine receptors including interleukin (IL)-9 have soluble counterparts in body fluids. We planned to investigate the pathophysiological significance of the serum soluble IL-9 receptor (sIL-9R) level. We determined the serum sIL-9Rα chain (sIL-9Rα) levels in 96 healthy Japanese individuals to establish a control value by means of specific human sIL-9Rα ELISA, followed by a preliminary application in a patient with diarrhea positive hemolytic uremic syndrome. Age was negatively correlated with the sIL-9Rα level (Spearman r = −0.241, n = 96, p = 0.0180). The serum sIL-9Rα level showed a progressive decline to the normal adult level by the age of 30. The serum sIL-9Rα level of the patient with HUS was markedly higher than those of the age-matched control from the onset of the disease. Because of the remarkable age-dependent variability of sIL-9Rα in healthy subjects, disease-related changes, as well as therapy-dependent alterations, should be considered with caution. Thus, it is recommended that when the serum sIL-9Rα levels of patients are evaluated, the values should be compared with those of age-matched controls. The established control value will be used to discriminate between normal and the pathological conditions in our future studies.
Soluble interleukin 9 receptor α chain (sIL-9Rα); Hemolytic uremic syndrome (HUS); ELISA
The nature of the dyslipidemia associated with diabetes mellitus is complex and is the major risk factor for atherosclerosis and coronary artery disease. Aim of this study was to assess the effect of glycemic control, achieved by metformin, glibenclamide and insulin, on lipid profile in type 2 diabetic patients. One hundred and sixty-five type 2 diabetes mellitus patients were classified into good glycemic control (Group I) and poor glycemic control (Group II) on the basis of their blood HbA1c values. The Group II was characterized with high serum triglyceride (190.46 ± 15.20 mg/dl), total cholesterol (175.3 ± 6.31 mg/dl) as well as high LDL-cholesterol (109.0 ± 5.88 mg/dl). Significant correlations were evident between HbA1c and dyslipidemia, particularly serum TG (r = 0.28, P < 0.05), and between HbA1c and total cholesterol (r = 0.310, P < 0.05). Better glycemic control and improved dyslipidemia were observed in patients on combination therapy of metformin plus glibenclamide.
Diabetes mellitus; Glycemic control; HBA1c; Dyslipidemia; Metformin; Glibenclamide
Multiple myeloma is a disseminated malignancy of monoclonal plasma cells that accounts for 15 % of all hematological cancers. The present study was conducted to evaluate the role of inflammation and oxidant-antioxidant dynamics in the etiology of this disease. The study population comprised of 20 cases of multiple myeloma and 20 healthy controls. The parameters evaluated were serum malondialdehyde (MDA), superoxide dismutase (SOD) and ferritin levels. The serum MDA levels were 1.9 ± 0.96 nmol/ml in cases as compared to 0.98 ± 0.55 nmol/ml in the controls. Similarly, a statistically significant difference was noted in the SOD and ferritin levels between the cases and controls (93.2 ± 23.8 vs. 210.1 ± 190.5 U/ml and 285.8 ± 216.4 vs. 131.8 ± 30.1 ng/ml respectively). Our study highlights the imbalance in the oxidant-anti oxidant mechanism and the role of smoldering inflammation in the etiology of multiple myeloma.
Multiple myeloma; Oxidative stress; Thiobarbitone reactive substances (TBARS); Antioxidants; Superoxide dismutase (SOD); Inflammation; Ferritin
Malondialdehyde (MDA) is widely used as oxidative stress biomarker in biomedical research. Plasma is stored in deep freezers generally till analysis. Effect of such storage on MDA values, which may be variable and prolong, was incidentally observed in the ongoing study which is to estimate oxidative stress with oral iron. Plasma from blood samples of pregnant women (20–30 years age) in third trimester of singleton pregnancy (n = 139), consuming oral iron tablets was stored at −20 °C with intention of MDA estimation, as soon as possible. However logistic problems led this storage for prolonged and variable period (1–708 days). When values of MDA estimated using “Ohkawa” 79 method and readings were plotted against time to check the temporal effect, it showed a hyperbolic curve. Standard deviation (SD) was lowest when samples were tested within 3 weeks time. The samples analyzed within 3 weeks had mean ± SD value of 31.59 ± 26.11 μmol/L, while 123.7 ± 93.97 and 366.5 ± 189.8 μmol/L for samples stored for 1–3 and 4 months to 1 year respectively. Mean ± SD were 539.9 ± 196.8 in the samples store for more than a year. Rate of change in values was also lowest (0.0433 μmol/L/day) in the samples tested within first 3 weeks, which rose to 1.2 μmol/L/day during 3 month’s storage. This rate peaked at storage of 120 days (1.87 μmol/L/day) and fell to 0.502 μmol/L/day in the second year of storage. It is concluded that at −20 °C, only 3 weeks of storage time should be considered valid for fairly acceptable stability in MDA values.
Malondialdehyde; Storage; Temporal; Oxidative stress
The present study was designed to test the hypothesis of a circadian variation in circulating levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in women with fibromyalgia syndrome (FMS). Serum levels of IL-6 and TNF-α were measured at 4 h intervals of the day in 50 women with FMS satisfying American College of Rheumatology criteria for FMS (age 36.68 ± 9.89) as well as 50 healthy control women (age 32.82 ± 10.53). Serum TNF-α levels were substantially increased in patients with FMS but showed no circadian variation. In contrast, no difference in the levels of IL-6 was found. Moreover, there was also no circadian variation in both the groups of patients and controls. We conclude that no circadian pattern exists in the circulating levels of serum IL-6 and TNF-α in patients with FMS, although TNF-α levels are found raised in patients with FMS.
Fibromyalgia syndrome; Circadian rhythm; Interleukin-6; Tumor necrosis factor
This study was undertaken to determine the association of inflammatory biomarker, oxidative stress and antioxidant capacity marker with fetal haemoglobin (HbF) level among sickle cell trait and sickle cell disease (SCD) patients in Chattisgarh. The study group consisted of 51 SCD (SS) patients with painful episode, 49 SCD (SS) patients with steady state, 50 sickle cell trait (AS) and 50 controls. Malondialdehyde (MDA), CRP, total antioxidant power (FARP), total thiol and HbF levels were quantified. We found a significant positive (p < 0.0001) association between CRP and MDA levels and its inverse association with HbF level in SS patients. We also observed that antioxidant capacity had significantly positively (p < 0.0001) associated with HbF level. The protective effect of HbF was found, because the increase in HbF levels resulted in decrease in lipid peroxidation and inflammation in SCD patients. A decrease in the HbF level and its antioxidant capacity has been associated with the pathogenesis of SCD. These finding may explain the high level of HbF is ameliorating oxidative stress and inflammation in SCD patients.
C-reactive protein; Oxidative stress; Antioxidant capacity; HbF level; Sickle cell disease
In the present study, oxidative stress and lymphocytic DNA damage in both pre-op and post-op benign prostrate hyperplasia (BPH) patients with age >50 years was evaluated and compared with normal healthy subjects (controls- without any evidence of disease) of the same sex and age group. From December 2007 to November 2009, oxidative stress in 45 BPH patients were evaluated both before (pre-op patients) and after 7 days of surgery (post-op patients) in terms of measurements of plasma levels of (1) various anti-oxidative enzymes, (2) non-enzymatic antioxidants and (3) malondialdehyde which is a product of lipid peroxidation. The lymphocyte DNA damage was also evaluated by single cell alkaline gel electrophoresis in terms of tail length migration in these patients. These values were compared with their respective control subjects of similar sex and age group. The activities of antioxidant enzymes and the levels of antioxidant, reduced glutathione were found significantly decreased (p < 0.05) in serum samples of pre-operative group of BPH patients as compared to the controls. These altered parameters increased significantly (p < 0.05) and returned to their near normal control values, but not up to baseline values, in post operative patients i.e. after the cancer load was decreased by surgery. Lymphocytic DNA damage was found to be significantly increased in pre-op group as compared to controls and was reduced after surgery in post-op group. The present study therefore, shows significantly increased levels of oxidative stress and DNA damage in BPH patients which were reduced after removal of tumour load. Thus oxidative damage plays an important role in prostate tumourogenesis and timely management of oxidative stress can be of importance in preventing the occurrence of BPH.
BPH; Oxidative stress; Antioxidants; Lipid peroxidation; DNA damage
Thalassemia has been recognized by the World Health Organization as important inherited disorders principally impacting on the populations of low income countries. In this report, the prevalence of common β-thalassemia mutations in India was defined in 126 β-thalassemia carrier subjects in a western Indian population mainly from the south-western Maharashtra. The six most common β-thalassemia mutations were detected, which included IVS I-5 (G–C), IVS I-1 (G–T), codon 8–9 (+G), codon 41/42 (–TCTT), Codon 15 (G–A), and 619 bp deletion at 3′ end of β-globin gene. These mutations accounted for 93.66 % in 126 β-thalassemia carrier subjects and 6.34 % remained uncharacterized. Out of 126, 82 (65.07 %) showed the most common (prevalent) type of mutation, IVS I-5 (G–C), followed by IVS I-1 (G–T) showed by 12 (9.52 %) subjects. Three (2.38 %) subjects showed 619 bp deletion, codon 8/9 (+G) and codon 15 (G–A) mutations were present in eight subjects each (6.34 %). Only five (3.96 %) subjects showed codon 41/42 (–TCTT). There were eight (6.34 %) subjects where mutation was not any of the six mutations studied. This study provides the pattern of β thalassemia mutations from south-western Maharashtra, which will help to prevent β-thalassemia using prenatal diagnosis and proper counseling.
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The online version of this article (doi:10.1007/s12291-012-0230-y) contains supplementary material, which is available to authorized users.
β-Thalassemia; South-western Maharashtra; Mutations; β-Thalassemia carrier; Prenatal diagnosis
The main objective of the current study is to examine the role of the statistical relation between BCL2 gene (Ala43Thr) single nucleotide polymorphism and growth hormone (GH1) levels in Egyptian HCV genotype-4 patients before and after treatment with pegylated interferon plus ribavirin. Eighty patients with HCV genotype-4 and 40 healthy volunteers as controls were enrolled in the prospective study. Gene polymorphism of BCL2 (Ala43Thr) using PCR-RFLP technique and GH1 concentrations using ELISA procedure were measured for all patients and controls. The present study resulted that Responder HCV genotype-4 Patients, with BCL2 43Ala genotype, have high significant increase in pre-treatment GH1 levels (>1 ng/ml); which represent normal levels, as compared to non-responders pre-treatment GH1 levels (<1 ng/ml); which represent low concentrations. We concluded that HCV genotype-4 patients who have normal GH1 concentrations and BCL-2 43Ala genotype can successfully achieve response to interferon based therapy.
BCL2; SNP; GH1; HCV; PEG-IFN-α/RBV
Hepatocellular carcinoma (HCC) is one of the commonest tumors worldwide. The treatment of HCC is vital for disease diagnosis and prognosis, as the liver is the most important organ controlling metabolic functions. Now-a-days, western folklore medicines are largely dependent on the phyto compounds which are highly effective in therapy and with low side effects. Luteolin is a flavonoid (3,4,5,7-Tetrahydro flavones) possess anti-inflammatory, anticancer and anti allergic property. The present study evaluates the efficacy of luteolin against N-nitrosodiethylamine (DEN) induced HCC in albino rats. In the highlight of the above, luteolin was evaluated for its efficacy against DEN induced HCC in male Wistar albino rats. The Biochemical parameters such as tissue damaging enzymes viz., AST, ALP, LDH and γ-GT, enzymatic antioxidants viz., SOD, CAT, GSH and GPx and histopathological changes have been estimated. The tissue damaging enzymes were found to be high in DEN alone treated group whereas the enzymatic antioxidants decreased destructively. Severe lesions and cirrhosis were observed in the toxin (DEN alone) treated group. The luteolin treated DEN group altered the tissue damaging enzymes and the enzymatic antioxidants. The damaged lesion in the histoarchitecture of DEN treated rat liver was almost completely restored. Finally this study strongly demonstrates that luteolin has potent curative property against HCC in albino rats.
Hepatocellular carcinoma; Luteolin; DEN; Enzymatic/Non-enzymatic Antioxidants; Ultrastructural study
The purpose of this study is to find out the frequency of TP53 mutations in acute myeloid leukemia (AML) patients and correlate sensitivity of drug response with TP53 mutations. In AML more than 90 % of cases comprise of wild type TP53. 94.2 % of TP53 mutations are found within exon 5–8 of which 73 % are point mutations. TP53 mutations were analysed with high resolution melting curve analysis. We analysed 106 AML samples of which we found nine mutations which represents 8.5 % mutation rate and found one rare SNP. The effect of TP53 mutations were studied on the chemosensitivity of two new drugs AZD115 and RHPS4, an Aurora Kinase B inhibitor and Telomerase inhibitor respectively. Four mutations were found out of 17 for RHPS4 stating significant (p = 0.002) increase in sensitivity and no mutation found in AZD1152 database, but need more study to get definite conclusion.
TP53; Acute myeloid leukemia; High resolution melting curve analysis; Drug sensitivity
Malaria infection is known to cause severe hemolysis due to production of abnormal RBCs and enhanced RBC destruction through apoptosis. Infected RBC lysis exposes uninfected RBC to the large amount of pro-oxidant molecules such as methemoglobin. Methemoglobin (MetHb) exposure dose dependently makes RBCs susceptible to osmotic stress and causes hemolysis. MetHb mediated oxidative stress in RBC correlated well with osmotic fragility and hemolysis. Interestingly, a reactive oxygen species (ROS) spike at 15 min was responsible for the observed effects on RBC cells. Two natural antioxidants N-acetyl cysteine and mannitol protected the RBC from MetHb-mediated defects, which clearly indicated involvement of oxidative stress in the process. MetHb due to its pseudo-peroxidase activity produces ROS in the external microenvironment. Therefore, classical peroxidase inhibitors were tested to probe peroxidase activity mediated ROS production with defects in RBCs. Clotrimazole (CLT), which irreversibly inactivates the MetHb (CLT-MetHb) and abolishes peroxidase activity, did not produce significant ROS outside RBC and was inefficient to cause osmotic fragility and hemolysis. Hence, initiating a chain reaction, MetHb released from ruptured RBC produces significant ROS in the external microenvironment to make RBC membrane leaky and enhanced hemolysis. Together data presented in the current work explored the role of MetHb in accelerated humorless during malaria which could be responsible for severe outcomes of pathological disorders.
RBC; Methemoglobin; Reactive-oxygen species; Hemolysis; Antioxidant; Malaria