Determination Of Adenosine Deaminase Activity Research Articles

Evaluation of serum adenosine deaminase activity and autoantibodies in systemic lupus erythematosus

Introduction: Systemic lupus erythematosus (SLE) is a complex autoimmune disease with no single specific and sensitive test available for its diagnosis. Adenosine deaminase (ADA) is an enzyme that can act as an indicator of cellular immunity reflecting the extent of inflammation. The aim of this study was to investigate the role of serum adenosine deaminase activity and its association with antinuclear antibodies (ANA) and anti-dsDNA (anti-double stranded deoxyribonucleic acid) in SLE patients. Material and methods: In this study, we included 36 diagnosed case of SLE as per the American Rheumatology Association (ARA) criteria and 30 healthy control. Serum ADA activity was measured by a spectrophotometric technique based on Giusti and Gallanti. ANA and Anti ds-DNA were measured by indirect Enzyme-Linked Immune Sorbent Assay (ELISA). Normal values of serum ADA activity, ANA and anti ds-DNA was < 25 IU/L, < 10 U/mL and < 25 IU/mL respectively. Results: The mean ADA activity (37.03 ± 13.03 IU/L) in SLE patients was higher as compared to control (18.23 ± 10.12 IU/L), and the difference was statistically significant (p < 0.001). Median values of ANA and anti-dsDNA in SLE patients was 16.9 (8.5, 42.8) and 24.7 (20.5, 45.0) respectively. Serum ANA was positive in 25 out of 36 cases, anti-dsDNA was positive in 17 out of 36 cases and ADA was positive in 29 out of 36 cases. We observed a weak positive correlation between ANA and anti-dsDNA (r = 0.46, p = 0.005), whereas a moderate correlation between ANA and ADA (r = 0.525, p = 0.001) in SLE patients. Conclusion: Serum ADA was significantly higher in SLE patients as compared to control and more specific than autoantibodies. Determination of ADA activity is a reliable, cost-effective, and easy test that can be used as an alternative parameter for diagnosing and evaluating disease activity in SLE patients.

Changes in the activity of adenosine deaminase and the content of polyamines in the blood of cats with breast cancer

In this work, the clinical significance of changes in the activity of adenosine deaminase (ADA) and the content of polyamines in the blood plasma of cats with breast cancer was studied. There were examined 30 cats aged 1 to 20 years, operated on in veterinary clinics due to spontaneous neoplasms. Histological studies of surgical material were performed to establish nosological forms of the tumor. The results of the measurements showed that there were no significant differences between ADA activity in the blood plasma of cats diagnosed with invasive non-specific carcinoma and ADA activity in the blood of healthy animals. In the plasma of animals diagnosed with fibroadenomatous breast hyperplasia, ADA activity was reduced by 35% compared with the control group. The content of putrescine in the red blood cells of cats with benign neoplasms of the mammary gland (fibroepithelial hyperplasia) was statistically significantly increased 5 times compared with the control group. In malignant neoplasms of the mammary gland (invasive non-specific carcinoma, tubular cancer and fibrosarcoma), the content of putrescine and spermine in the blood significantly exceeded the control values by 60 and 10 times, respectively, compared with the control group. Thus, the data obtained indicate the possibility of using polyamines in order to improve the accuracy of diagnosis and treatment of cats with breast cancer. The determination of ADA activity in the blood plasma of cats with breast tumors in the early stages of cancer is not an informationally significant indicator as a tumor biomarker.

Changes in the activity of adenosine deaminase and the content of polyamines in the blood of cats with breast cancer

In this work, the clinical significance of changes in the activity of adenosine deaminase (ADA) and the content of polyamines in the blood plasma of cats with breast cancer was studied. There were examined 30 cats aged 1 to 20 years, operated on in veterinary clinics due to spontaneous neoplasms. Histological studies of surgical material were performed to establish nosological forms of the tumor. The results of the measurements showed that there were no significant differences between ADA activity in the blood plasma of cats diagnosed with invasive nonspecific carcinoma and ADA activity in the blood of healthy animals. In the plasma of animals diagnosed with fibroadenomatous breast hyperplasia, ADA activity was reduced by 35% compared with the control group. The content of putrescine in the red blood cells of cats with benign neoplasms of the mammary gland (fibroepithelial hyperplasia) was statistically significantly increased 5 times compared with the control group. In malignant neoplasms of the mammary gland (invasive non-specific carcinoma, tubular cancer and fibrosarcoma), the content of putrescine and spermine in the blood significantly exceeded the control values by 60 and 10 times, respectively, compared with the control group. Thus, the data obtained indicate the possibility of using polyamines in order to improve the accuracy of diagnosis and treatment of cats with breast cancer. The determination of ADA activity in the blood plasma of cats with breast tumors in the early stages of cancer is not an informationally significant indicator as a tumor biomarker.

Serum Adenosine Deaminase (ADA) Activity: A Novel Screening Test to Differentiate HIV Monoinfection From HIV-HBV and HIV-HCV Coinfections.

CD4(+) cell count, the common HIV infection screening test, is costly and unable to differentiate HIV monoinfection from its concurrent infection with hepatitis B or C virus. We aimed to ascertain diagnostic value of serum adenosine deaminase (ADA) activity as a useful tool to differentiate HIV mono- and co-infection. Blood samples were collected from 30 HIV-HBV and 30 HIV-HCV coinfected patients, 33 HIV positive subjects, and 72 controls. CD4(+) cell count, serum total ADA (tADA), and ADA1, and ADA2 isoenzyme activities were determined and their sensitivity and specificity were computed. tADA and ADA2 activities were significantly higher and CD4(+) counts were markedly lower in all patients compared with controls. Strong inverse agreements between CD4(+) cell counts and both tADA and ADA2 activities were observed. Serum tADA and ADA1 activities showed the highest specificity and the highest sensitivity, respectively, for differentiating HIV monoinfection from HIV-HBV and HIV-HCV coinfections. We showed strong agreement and correlation between CD4(+) cell count and ADA enzyme activity. Based on high ADA sensitivity and specificity, it is concluded that determination of ADA activity might be a novel diagnostic tool to distinguish of HIV monoinfection from its coinfection with HBV or HCV.

Influence of hemodialysis on the plasma concentration of adenosine deaminase in patients with chronic kidney disease

Introduction: Over the past years there has been a significant increase in hospitalizations and treatments due to kidney complications that eventually resulted in the increased number of patients on dialysis. The adenosine deaminase (ADA) enzyme mediates the formation of some defense cells of the organism and is therefore a marker of inflammation. Objective: The objective of this study was to evaluate biomarkers of renal function and serum ADA of hemodialysis patients. Materials and methods: Blood samples were collected from 80 patients – 40 women and 40 men – between 19 and 60 years old, before and after the completion of hemodialysis. Results: There was a significant difference in levels of creatinine, urea and ADA in pre- and post-hemodialysis periods (p < 0.0001). There was a significant increase in post-dialysis ADA regardless of sex; however there was a significantly greater increase in men. Conclusion: The results showed a reduction in urea and creatinine parameters, evidencing the main purpose of hemodialysis. This study suggests that the determination of ADA activity could be used to monitor inflammation in hemodialysis patients, however wider and more specific studies are needed to show the effectiveness of serum ADA activity as an inflammatory marker in patients with chronic kidney disease.

Syzygium cumini is more effective in preventing the increase of erythrocytic ADA activity than phenolic compounds under hyperglycemic conditions in vitro

Syzygium cumini (S. cumini) is a plant known for its antidiabetic properties. The aim of this study was to evaluate the effect of Sc aqueous leaf extract (ASc) on adenosine deaminase (ADA) activity in erythrocytes (RBCs) exposed to high glucose concentrations (30mM) in vitro. We also investigated the effects of the main phenolic compounds found in ASc (gallic acid, rutin, and chlorogenic acid) and the effects of insulin, caffeine, and dipyridamole, which are substances involved in the adenosine metabolism, on ADA activity in vitro. Blood samples were obtained from healthy volunteers and a suspension of RBCs was used for the determination of ADA activity. The results showed that: (1) the effect of ASc on ADA activity was more significant than the combination of phenolic compounds; (2) insulin, caffeine, or dipyridamole prevented high glucose increase of ADA activity at doses as low as 50μU/mL, 25μM, and 1μM, respectively; (3) the inhibitory effect caused by ASc on erythrocyte ADA activity remained practically the same after the combination of the extract with insulin or caffeine; (4) when RBCs were exposed to ASc plus dipyridamole, this chemical attenuated the effect of ASc on ADA activity, suggesting an antagonism or a competition with ASc by the same site of action. Therefore, ASc was more effective in preventing the increase in ADA activity than phenolic compounds, suggesting that ASc may collaborate to improve endothelial dysfunction, antioxidant, anti-inflammatory, and antithrombotic properties of adenosine by affecting its metabolism. The results of this study help to provide evidence of the empirically supported benefits of the use of S. cumini in diabetes.

G-Quadruplex-Based DNAzyme as a Sensing Platform for Ultrasensitive Colorimetric Adenosine Deaminase Detection

Adenosine deaminase plays a crucial role in the development and maintenance of the normal immune system. Although the determination of adenosine deaminase in serum may serve as a noninvasive diagnostic tool in evaluation of the active phase of systemic lupus erythematosus and the severity of this disease, detection of this analyte in a biologically complex mixture remains a major challenge. In this report, a colorimetric biosensor is reported for the determination of adenosine deaminase activity. The sensor employed DNAzyme as the signal transformation element which does not have a complicated label process. The proposed method was simple, rapid, and economical, allowing monitoring of adenosine deaminase to levels less than 0.1 U L−1. The simplicity of this device suggests it may be a candidate for the rapid screening of proteins in human serum and have wide applications in the field of clinical diagnosis.

Avaliação da atividade da adenosina deaminase no derrame pleural tuberculoso

Tuberculose pleural; Adenosina desaminase; Biomarcador

Serum Adenosine deaminase (ADA) levels in surgically treated hydatid cyst patients

Adenosine deaminase (ADA) is an essential enzyme in the monocytes - macrophage system and in the proliferation and differentiation of lymphocytes. ADA activity is routinely used in the differentiation of other etiologies of tuberculosis pleurisy in clinical practice and elevated ADA levels are stated to indicate the severity of the disease. In this study the purpose was to compare the serum ADA levels between patients operated due to hydatid cyst and healthy individuals (control group). The serum ADA levels of surgically treated patient with hydatid cyst were analyzed and a significant decrease was observed in comparison with the control group (10.98±6.53 vs 21.11±14.07, p<0.05). Decreasing of ADA levels on surgical treated hydatid cyst patients can be interpreted as the inhibition of tissue damage or cessation of lymphocyte proliferation as a result of the elimination of the parasite by means of surgical treatment and breaking the relation between the parasite and immune cells of the host. Determination of ADA activity in hydatid cyst patients might be a diagnostic tool if further clinical trials are carried out. Key words: Adenosine deaminase, hydatid cyst, operated patients.

Determination of Adenosine Deaminase Activity in Dried Blood Spots by a Nonradiochemical Assay Using Reversed-Phase High-Performance Liquid Chromatography

Adenosine deaminase (ADA) deficiency is a rare metabolic disease causing severe combined immunodeficiency (SCID). An assay to determine ADA activity in dried blood spots was developed using reversed-phase HPLC. The assay was linear with reaction times up to at least 4 hours, and protein concentrations up to at least 2.2 mg/ml. The intra-assay CV and the inter-assay CV for the complete assay was 3.5 and 8.4%, respectively. The ADA activity in a control blood spot, stored at 4°C, remained stable for at least one year. Only a slightly decreased ADA activity (35 ± 13 nmol/mg/h, n = 4) was observed in heterozygotes for a c.704G > A mutation in the ADA gene when compared to that observed in controls (41 ± 13 nmol/mg/h, n = 108). In addition, increased ADA activity as found in a rare form of congenital anemia can be assessed, as observed in a bloodspot from a patient diagnosed with Diamond Blackfan anemia (ADA activity 150 nmol/mg/h).

Determination of adenosine deaminase activity in the pleural effusion and its clinical significance--a study using experimental pleuritis in rabbits

Determination of adenosine deaminase activity in the pleural effusion and its clinical significance--a study using experimental pleuritis in rabbits

Determination of adenosine deaminase activity in cattle naturally infected with Theileria annulata

The purpose of this study was to determine serum ADA activity in cattle naturally infected with Theileria annulata. In this study, a total of 37 cross-bred cattle which 27 of it showing clinical signs of theileriosis constituted infected group and 10 healthy cattle as control group were used as animal materials. Infected group divided into three groups according to their PCV values. Cattle with PCV > or = 25 were put on group I (n = 9), those with PCV 13-24 were put on group II (n = 11) and those with PCV < or = 12 were put on group III (n = 7). Microscopical diagnosis of the disease was also made. Hematological parameters, serum enzyme activities (ADA, AST, ALT and ALP) were determined in all cattle. Hematological results revealed that significant progressive decreases in HGB, PLT, PBML counts and ratios from group I onwards to group III, whereas the WBC, PBPL counts and ratios showed an increase from group I onwards to group III. The serum ADA, AST, ALT and ALP activity increased significantly in all infected groups compared to control group. However, these parameters were also observed to decrease progressively from group I to group III. Furthermore, the highest increase in enzyme activities observed in the infected group I. But, these enzyme's activities started to decrease in infected group II and III in parallel with PBML and PLT counts. Eventhough, this decrease did not reach to the values obtained from control group. On the contrary, PBPL counts and ratios increased in infected group II and III in contrast to decrease in PCV. As a result, increased serum ADA activity in tropical theileriosis may reflect the involvement of the cellular immune responses.

Adenosine deaminase activity in cerebrospinal fluid of HIV-infected patients: limited value for diagnosis of tuberculous meningitis

Adenosine deaminase activity (ADA) determination in cerebrospinal fluid (CSF) is considered a specific test for the diagnosis of tuberculous meningitis. In order to study the variability of this marker in patients with different neurological disorders associated with HIV infection, and its utility for the diagnosis of tuberculous meningitis in these patients, the ADA levels in 417 CSF samples from HIV-infected patients with neurological symptoms were reviewed. HIV infection, HIV-associated neurological disorders, and progressive multifocal leukoencephalopathy were not associated with elevated ADA in CSF. Among patients with meningitis, receiver operating characteristic curve analysis gave an optimal ADA cut-off point of 8.5 IU/l for the diagnosis of tuberculous meningitis, with 57% sensitivity, 87% specificity, and an area under the curve of 0.747 (similar to that for CSF glucose concentration). False-positive results were found in patients with neurological CMV disease and cryptococcal, lymphomatous, and probable candidal meningitis. The results of this study indicate that ADA determination in CSF has limited utility for the diagnosis of tuberculous meningitis in HIV-infected patients.

Combined Use of the Polymerase Chain Reaction and Detection of Adenosine Deaminase Activity on Pleural Fluid Improves the Rate of Diagnosis of Pleural Tuberculosis

Evaluation of the combined use of polymerase chain reaction (PCR) and adenosine deaminase (ADA) activity on the diagnosis of pleural tuberculosis (pTB) in a region of high prevalence of tuberculosis. PCR and determination of ADA activity were performed on the pleural fluid of every patient presenting with pleural effusion suspected to be associated with tuberculosis. The case definition of pTB involved parameters including the combination of clinical and radiologic findings; biochemical, microbiologic, and cytologic examination of the pleural fluid; and the histopathologic findings of pleural fragments obtained by biopsy. The diagnosis of pTB was confirmed in any patient presenting with positive culture findings of Mycobacterium tuberculosis, either on the pleural fluid or other biological material, or the presence of histopathologic findings suggestive of pTB on pleural biopsy, and also, in the absence of negative laboratory results, those patients with clinical improvement after empirical treatment. We studied 45 patients with pleural effusion. Of these, 16 patients met the diagnosis of pTB by our broad case definition. PCR findings were positive in six patients. The reaction was also positive in a patient whose diagnosis of tuberculosis could not be confirmed. ADA activity was considered positive in 11 patients with pTB. The combined use of PCR and ADA activity confirmed pTB in 14 patients; however, when analyzed in combination with the conventional methods, diagnosis of pTB was achieved in all 16 patients. Our results show that, even in a highly endemic area, neither PCR nor ADA activity should be relied on as a single test that substitutes for the diagnostic methods already available, but rather they should be used as an extra tool in the diagnosis of pTB. The combined analysis of PCR and ADA activity, however, is a very useful diagnostic approach to achieve a more rapid and precise diagnosis in the cases of pTB.

Investigation of the relationship between nitric oxide metabolites' levels and adenosine deaminase activity in 7,12-dimethylbenz[a]anthracene induced mouse liver.

7,12-Dimethylbenz[a]anthracene (7,12-DMBA) is a member of the polycyclic aromatic hydrocarbons with a severe carcinogenic effect. In this study, nitrate levels and ADA (Adenosine deaminase) activity in the liver homogenates of mice were measured and the effect of free radicals induced by 7,12-DMBA on inducible nitric oxide synthase (iNOS) and ADA activity were investigated. Antioxidant effects of melatonin were also compared. Three groups of mice were included in the study. The first served as control, the second was treated only with 7,12-DMBA and the third was treated with 7,12-DMBA + melatonin. Spectrophotometric methods were used at all measurements. Data were analyzed using Kruskal-Wallis Variance Analysis Test and Mann-Whitney U Test that were applied to the groups. The nitrate concentrations of mouse liver were as follows: 4.98 +/- 0.63 micro mol/L in the control group (n = 10), 8.23 +/- 1.58 micro mol/L (higher than control group, p < 0.05) in the 7,12-DMBA-treated group (n = 10), and 6.43 +/- 0.57 micro mol/L (lower than 7,12-DMBA-treated group, p < 0.05) in the 7,12-DMBA + melatonin-treated group (n = 10). Liver ADA activities were measured to be 4.14 +/- 0.674 U/L in the control group, 6.25 +/- 1.261 U/L (higher than control group, p < 0.05) in the 7,12-DMBA-treated group, and 4.93 +/- 0.916 U/L (lower than 7,12-DMBA-treated group, p < 0.05) in the 7,12-DMBA+melatonin-treated group. Differences between free nitrite levels were no significantly. Results demonstrated that nitrate levels and ADA activities were increased by means of free radicals induced by 7,12-DMBA. Melatonin inhibited the 7,12-DMBA induced increase that was observed in the activities of ADA enzyme and nitrate levels. It is concluded that determination of ADA activity and nitrate levels can be useful in the assessment of liver damage caused by toxic chemicals.

Determination of adenosine deaminase activity and its isoenzymes for diagnosis of pleural effusions *

Determination of adenosine deaminase activity and its isoenzymes for diagnosis of pleural effusions *

Determination of adenosine deaminase activity and its isoenzymes for diagnosis of pleural effusions*

We aimed to investigate adenosine deaminase (ADA) activity and the activities of its ADA1 and ADA2 isoenzymes in pleural effusions and also sera with different aetiological origins. The pleural effusions of 87 patients were examined. The patients were separated into four groups: transudates, parapneumonic, malignant, and tuberculous effusions. The cases were also designated as tuberculous or non-tuberculous group. Adenosine deaminase activity was determined by the colorimetric method described by Giusti and Galanti. The intermean differences were statistically significant for total ADA, ADA1 and ADA2, except for pleural fluid ADA1 in the malignant group when compared to the tuberculous effusion group. The intermean differences between the tuberculous and non-tuberculous group were statistically significant for all three parameters except for pleural fluid and serum ADA1 activity. The sensitivities of total ADA, ADA1 and ADA2 activities for tuberculosis were 91, 57 and 93%, respectively; their specificities 89, 88 and 92%, respectively; their positive predictive values 82, 70 and 86%; and their diagnostic accuracies 89, 76 and 92%, respectively, in pleural fluid. Determination of ADA and its isoenzymes can help to differentiate the causes of pleural effusion. Increased ADA2 activity is a striking marker of tuberculous effusions. In contrast, increased ADA1 activity was significantly elevated in parapneumonic effusions.

Determination of adenosine deaminase activity in human erythrocytes by on-column capillary isotachophoresis-capillary zone electrophoresis in the presence of electroosmotic flow.

Transient capillary isotachophoresis (CITP)-capillary zone electrophoresis (CZE) in presence of electroosmotic flow (EOF) was utilized for the measurement of adenosine deaminase activity in human erythrocytes. Phosphates, dominant anions of the sample matrix, were used as leading ions for transient isotachophoresis, and borates (0.3 M, pH 10) were used as terminating ions and background electrolyte for CZE. Final experimental conditions made it possible to inject 70% of the total capillary volume (1.45 microL) with the sample. Enzymatic conversion products (inosine and hypoxanthine), present in the sample in the low-micromolar range, were determined using optimized conditions. The limit of detection was 28 nM using UV detection at 202 nm. The presented data shows that CITP-CZE can be performed in uncoated capillaries in the presence of strong EOF.

Determination of adenosine deaminase activity in human erythrocytes by capillary eletrohoresis

ZAM is an LTR-retrotransposon from Drosophila melanogaster that belongs to the genus errantivirus, viruses similar in structure and replication cycle to vertebrate retroviruses. A key component to its lifecycle is its reverse transcriptase which copies single-stranded genomic RNA into DNA. Here, we provide a detailed characterization of the enzymatic activities of the reverse transcriptase encoded by ZAM. When expressed in vitro, the reverse transcriptase domain associated with the RNase H domain encoded by the ZAM pol gene forms homodimers and displays an efficient RNA-dependent DNA-polymerase activity. It requires either Mg2+ or Mn2+ divalent cations, and works in basic pH, with a peak at around pH9. The so-called [RT-RH] polypeptide displays an optimal activity at 22 °C, a property that makes it well-adapted to the temperature of its host. This study contributes to our understanding of the general structures and functions of retroviral reverse transcriptases, a necessary process in the search for novel inhibitors.

Rapid diagnosis of pleural tuberculosis by polymerase chain reaction.

We have investigated the use of polymerase chain reaction (PCR) for the rapid diagnosis of pleural tuberculosis. The study was composed of 21 patients with pleural tuberculosis, confirmed by culture or pleural biopsy, and 86 control subjects. The PCR assay was based on detecting a 123-bp DNA segment belonging to the insertion sequence IS6110, specific of Mycobacterium tuberculosis complex. In 21 patients diagnosed with pleural tuberculosis, Ziehl-Neelsen staining was positive in three (14%) (95% CI, 7 to 21%) and pleural fluid culture in 11 (52%) (95% CI, 43 to 61%). Pleural biopsy revealed granulomas with caseous necrosis in 72%, and the culture was positive in 67% of the patients. Adenosine deaminase activity determination was positive (> 45 IU/L) in 86% (95% CI, 79 to 93%). The sensitivity and specificity for PCR was 81% (95% CI, 74 to 88%) and 100% (95% CI, 95 to 100%), respectively. All culture-positive specimens were PCR positive. We conclude that PCR is a rapid, sensitive, and specific method for the diagnosis of pleural tuberculosis. However, further prospective studies are required to properly evaluate the yield of the technique.