How Reliable Is Automated Urinalysis in Acute Kidney Injury?

OBJECTIVE: Examination of urine sediment is crucial in acute kidney injury (AKI). In such renal injury, tubular epithelial cells, epithelial cell casts, and dysmorphic red cells may provide clues to etiology. The aim of this study was to compare automated urinalysis findings with manual microscopic analysis in AKI. METHODS: Samples from patients diagnosed with AKI and control patients were included in the study. Red blood cells, white blood cells, renal tubular epithelial cells/small round cells, casts, and pathologic (path) cast counts obtained microscopically and by a UF1000i cytometer were compared by Spearman test. Logistic regression analysis was used to assess the ability to predict AKI from parameters obtained from the UF1000i. RESULTS: There was poor correlation between manual and automated analysis in AKI. None of the parameters could predict AKI using logistic regression analysis. However, the increment in the automated path cast count increased the odds of AKI 93 times. CONCLUSION: Automated urinalysis parameters are poor predictors of AKI, and there is no agreement with manual microscopy.

Recurrent genetic abnormalities detected by FISH in adult B ALL and association with hematological parameters.

BACKGROUND: Recurrent genetic abnormalities influence prognosis in B lymphoblastic leukemia. BCR-ABL rearrangement is associated with higher leukocyte counts and older age at presentation. Among adults, BCR -ABL - is the commonest recurrent abnormality whereas, IgH rearrangements are rare. AIM: Aim of this study was to identify common recurrent genetic abnormalities in adult B ALL and study their association with hematological findings. METHODS: Bone marrow and peripheral blood from patients with B acute lymphoblastic leukemia were analyzed for complete blood counts, bone marrow morphology and cytogenetic abnormalities. The study group was divided into smaller groups based on cytogenetic abnormalities. Hematological parameters and presence of recurrent genetic abnormalities was compared across age groups and gender by non parametric tests. RESULTS: BCR-ABL positive group had a higher leukocyte count than BCR-ABL negative group. Among groups 1 to 5, group 1 with gains of chromosomes was associated with leucopenia and higher age at presentation. BCR-ABL is commonest recurrent abnormality followed by IgH rearrangements. CONCLUSION: Patients with gains of chromosomes alone have low total leukocyte counts at presentation.

EGFR mutations and ROS1 and ALK rearrangements in a large series of non-small cell lung cancer in South India.

BACKGROUND: Driver mutations are seen in 80% of lung adenocarcinomas, and they influence prognosis and choice of therapy. AIM: Aim of this study was to analyse the frequency of epidermal growth factor receptor (EGFR) mutations, ALK and ROS1 rearrangements and their association with age and gender in non-small cell lung cancer reported from a tertiary care center in South India. METHODS: Tumors from patients with non-small cell carcinoma of lung were evaluated for EGFR mutations, ALK and ROS1 rearrangements and their association with age and gender were studied. RESULTS: Two thirds of non-small cell carcinomas had driver mutations or rearrangements. EGFR mutation was common and seen in 34.1%, whereas ALK rearrangement was seen in 11.1% and ROS1 rearrangement in 2% patients. Among EGFR mutations, most common were Exon 19 deletion and L858R seen in 21.3% and 11% of patients, respectively. Adenocarcinoma was the histologic diagnosis in 81% to 85% of patients with exon 19 deletion and L858R mutation, respectively. EGFR mutation frequency in patients less than 36 years was 13.6%, whereas in older patients, it varied from 34% to 36%. Exon 19 deletion was seen in 29.8% females and 17.2% of males. CONCLUSION: EGFR mutations are more common than ALK and ROS1 rearrangements. They are more common in females. Patients less than 36 years have reduced frequency of EGFR mutations. Exon 19 deletion and L858R are most common and are more prevalent in lung adenocarcinomas. Rare EGFR mutations are seen in patients aged more than 50 years.

Can Platelet Distribution Width Be Used to Predict the Possibility of Chronic Myeloproliferative Neoplasms?

Platelet distribution width (PDW) and mean platelet volume are markers of platelet activation and have prognostic value in coronary heart diseases, as well as in cancers of solid organs. In this study, we evaluated the possibility of using PDW to predict chronic myeloproliferative neoplasms by comparing platelet indices obtained by automated analyzers in chronic myeloproliferative neoplasms with those in control specimens. We found that PDW greater than 66.4% has specificity of 99% and likelihood ratio of 19.5 for predicting chronic myeloproliferative neoplasms. Also, the area under curve (AUC) for platelet distribution width is 0.68.

Improved contrast microscopy: modification of bright field for urine sediment visualisation.

BACKGROUND: Phase contrast microscopy is the recommended technique for urine sediment examination. Bright field microscopy does not differentiate objects with slight changes in the refractive index and hence phase contrast is a superior alternative. METHODS: In this article, we describe a novel method to improve contrast in bright field microscopy. A strategically placed disc of specific dimensions enhances the diffraction of rays by Fresnel principle causing a shift in wavelength in the rays which are perceived as differences in contrast by the eye due to constructive and destructive interference. RESULTS: Epithelial cells, red blood cells (RBCs), dysmorphic red blood cells, casts, bacteria and crystals are easily seen and differentiated by this technique. CONCLUSIONS: The images obtained are similar to those obtained by phase contrast microscopy.

Cytogenetic Profiling of Myelomas, Association With Complete Blood Count: Study of 180 Patients.

OBJECTIVES: To analyze the most common primary and secondary cytogenetic events in myelomas using a probe panel designed in our laboratory, and to associate those events with hematological and biochemical findings. METHODS: Blood specimens from patients diagnosed with myeloma were processed to determine complete blood count and levels of albumin, creatinine, and beta-2 microglobulin. We evaluated bone-marrow specimens for plasma-cell percentage by light microscopy and for cytogenetic abnormalities by fluorescence in situ hybridization (FISH). The Mann-Whitney U test was used to compare hematological and biochemical parameters. RESULTS: We observed immunoglobulin heavy chain (IgH) gene translocations in 43.3% and t(4;14) in 21% of specimens; t(11;14) was observed in 7.7% of specimens. Gain of chromosomes was observed in 67.2% and loss observed in 16.6% of specimens. CONCLUSIONS: Gains of chromosomes were observed in two-thirds of patients with myeloma. The most common IgH translocation was t(4;14); del13/monosomy13 was the most common secondary cytogenetic abnormality. Partial or complete tetrasomies were associated with higher beta-2 microglobulin levels.

Simplified Method for Local Correction of System International Normalized Ratio.

BACKGROUND: International normalized ratio (INR) derivation is dependent on the international sensitivity index (ISI) of thromboplastin. It varies with instrument and reagents used. OBJECTIVE: To evaluate the role of a correction factor in the derivation of INR. METHODS: We studied prothrombin time (PT) and INR from patients using 3 thromboplastins of varying ISI values. The correction factor was applied to the observed INR to obtain a corrected INR. RESULTS: The difference between corrected INR and observed INR values varies from -0.8 through 0.96. CONCLUSIONS: Corrected INR is dependent on PT only. It can be applied to all patients irrespective of cause for elevated PT.

Hb A1c Separation by High Performance Liquid Chromatography in Hemoglobinopathies.

Hb A1c measurement is subject to interference by hemoglobin traits and this is dependent on the method used for determination. In this paper we studied the difference between Hb A1c measured by HPLC in hemoglobin traits and normal chromatograms. We also studied the correlation of Hb A1c with age. Hemoglobin analysis was carried out by high performance liquid chromatography. Spearman's rank correlation was used to study correlation between A1c levels and age. Mann-Whitney U test was used to study the difference in Hb A1c between patients with normal hemoglobin and hemoglobin traits. A total of 431 patients were studied. There was positive correlation with age in patients with normal chromatograms only. No correlation was seen in Hb E trait or beta thalassemia trait. No significant difference in Hb A1c of patients with normal chromatograms and patients with hemoglobin traits was seen. There is no interference by abnormal hemoglobin in the detection of A1c by high performance liquid chromatography. This method cannot be used for detection of A1c in compound heterozygous and homozygous disorders.

Dilute Russell's viper venom and activated partial thromboplastin time in lupus anticoagulant diagnosis: is mixing essential?

Dilute Russell's viper venom (DRVV) testing and activated partial thromboplastin time (APTT) have been effectively used in combination for lupus anticoagulant testing. The purpose of our study was to evaluate the role of mixing in activated partial thromboplastin and dilute Russell's viper venom testing for evaluation of lupus anticoagulants. Citrated blood from patients who were not on oral anticoagulant therapy was studied. Mixing study with 1 : 1 normal plasma for elevated APTT and also few samples with elevated screen time was carried out. Elevated APTT was seen in only 48.1% of patients with lupus anticoagulant. Correction of APTT was seen in 27.8% of lupus anticoagulant-positive patients. DRVV test on mixing resulted in 83.8% false-negative values. Integrated DRVV test could be a standalone test for testing lupus anticoagulant. Mixing study may be restricted for patients on oral anticoagulants or patients with strong lupus anticoagulant.

Does platelet count in platelet-rich plasma influence slope, maximal amplitude and lag phase in healthy individuals? Results of light transmission aggregometry.

Light transmission aggregometry lacks in standardisation and normal reference values are not widely available. The aims of our study were to establish reference ranges for aggregation, slope and lag phase in healthy controls with platelet counts between 150 and 450 × 10(9)/l in platelet-rich plasma (PRP) as well as evaluate the influence of platelet count. Ninety-nine subjects were evaluated with four agonists and divided into two groups based on platelet count and the groups were compared by Student's t-test. There was no difference between the means of the two groups for amplitude and slope barring the lag phase for collagen. Platelet counts between 150 and 450 × 10(9)/l have no effects on light transmission aggregometry and hence adjustment of platelet count is not necessary.

Minor hemoglobins other than A2 in iron deficiency.

OBJECTIVE: To determine any difference in levels of minor hemoglobins, other than hemoglobin A2 (HbA2), in patients with iron deficiency. METHODS: We divided ethylenediaminetetraacetic acid (EDTA)-anticoagulated blood specimens from 188 patients into 2 groups, of which 94 displayed a normal or high ferritin level and the remaining 94 had a low ferritin level. We correlated the red cell indices, hemoglobin levels, and ferritin levels with the percentage of various minor hemoglobins. The Student's t-test was used to assess the statistical significance of the difference between the mean concentrations of minor hemoglobins in the 2 groups. RESULTS: Only hemoglobin HbA1a had a positive correlation (0.33 in iron-deficient and 0.35 in iron-replete specimens) with serum ferritin concentration. Among the minor hemoglobins, HbA1a (P =.008) and HbA2 (P <.001) were significantly different between the 2 groups. CONCLUSION: HbA1a may be low in iron deficiency, independent of HbA2. This information may be used to predict coexistent iron deficiency in ß-thalassemia traits, in which concomitant infection or inflammation leads to normal ferritin levels.

Florid erythrophagocytosis on the peripheral smear.

Erythrophagocytosis is a relatively rare observation on blood smears. It has been reported in auto immune hemolytic anemias and sporadically in few other conditions. Here, we report a case of florid erythrophagocytosis with severe anemia following a viral infection in an 18-year-old girl. Her complete blood count (CBC) revealed hemoglobin of 3.6 gm/dl and a hematocrit of 10%. The peripheral smear showed erythrophagocytosis by neutrophils and rosetting of erythrocytes around neutrophils. The direct Coombs test and direct Donath- Landsteiner tests were positive.

DIC Score: Statistical Relationship with PT, APTT, and Simplified Scoring Systems with Combinations of PT and APTT.

We looked into the statistical association of prothrombin time (PT) and activated partial thromboplastin time (APTT) with disseminated intravascular coagulation (DIC) score calculated using the International society for thrombosis and haemostasis (ISTH) scoring system. The PT, APTT, PT + APTT, and PT/APTT ratios were evaluated against the DIC score by linear regression analysis in fifty inpatients with suspected DIC. The PT, PT + APTT, and PT/APTT ratios were all found to be statistically significant in predicting DIC scores with P values of 0.02, 0.03, and 0.02, respectively. The APTT alone was not found to be statistically significant in predicting DIC score and had a P value of 0.09. This scoring system does not need d-dimer levels and the platelet count.

Hemoglobin disorders in South India.

Cation exchange-high performance liquid chromatography (CE-HPLC) is increasingly being used as a first line of investigation for hemoglobinopathies and thalassemias. Together with a complete blood count, the CE-HPLC is effective in categorizing hemoglobinopathies as traits, homozygous disorders and compound heterozygous disorders. We carried out a one year study in Apollo Hospitals, Chennai (Tamil Nadu, South India) during which 543 abnormal chromatogram patterns were seen. The commonest disorder we encountered was ß-thalassemia trait (37.9%), followed by HbE trait (23.2%), homozygous HbE disease (18.9%), HbS trait (5.3%), HbE ß-thalassemia (4.6%), HbS ß-thalassemia (2.5%), ß-thalassemia major (2.3%), HbH (1.6%), homozygous HbS (1.4%), HbD trait (0.7%). The average value of HbA2 in ß-thalassemia minor was 5.4%. ß-thalassemia major had an average HbF of 88% and in HbH the mean A2 was 1.4%. Among the HbE disorders the HbA2 + HbE was 30.1% in the heterozygous state, 90.8% in the homozygous state and 54.8% in HbE ß-thalassemia. In the sickle cell disorders, HbS varied from 30.9% in the trait to 79.9% in the homozygous state to 65.6% in HbS ß-thalassemia.