Showing papers in "Journal of Clinical Laboratory Analysis in 2014"

Neutrophil to Lymphocyte Ratio Predicts Poor Prognosis in Ischemic Cerebrovascular Disease

Abstract: Background Inflammation plays a role in the pathogenesis of carotid atherosclerosis. Although previous data demonstrated an association between inflammatory biomarkers and stroke, there is no publication reporting the relation of neutrophil to lymphocyte (N/L) ratio with ischemic stroke. We aimed to analyze the predictive ability of N/L ratio in acute ischemic cerebrovascular disease. Methods A total of 190 patients including 70 patients with first-ever atherothrombotic acute ischemic stroke (AAIS), 50 patients with transient ischemic attack, and 70 healthy subjects were enrolled in this study. We analyzed the values of white blood cell (WBC), N/L ratio, C-reactive protein (CRP), gamma-glutamyltransferase (GGT), homocysteine (HCY), mean platelet volume (MPV) in patients with ischemic group and compared those with control individuals. Results WBC, CRP, HCY, N/L ratio were found to increase significantly in AAIS patients than the controls (P 0.05). A cut-off value of 4.1 for N/L ratio was detected in predicting mortality with a sensitivity of 66.7% and a specificity of 74.1% (κ = 0.299, P = 0.006). Conclusions These findings support the role of N/L ratio as a simple inexpensive and readily available marker of prognosis in acute ischemic stroke.

Iron Metabolism: From Health to Disease

Abstract: Background: Iron is vital for almost all living organisms by participating in a wide range of metabolic processes. However, iron concentration in body tissues must be tightly regulated since excessive iron may lead to microbial infections or cause tissue damage. Disorders of iron metabolism are among the most common human diseases and cover several conditions with varied clinical manifestations. Methods: An extensive literature review on the basic aspects of iron metabolism was performed, and the most recent findings on this field were highlighted as well. Results: New insights on iron metabolism have shed light into its real complexity, and its role in both healthy and pathological states has been recognized. Important discoveries about the iron regulatory machine and imbalances in its regulation have been made, which may lead in a near future to the development of new therapeutic strategies against iron disorders. Besides, the toxicity of free iron and its association with several pathologies has been addressed, although it requires further investigations. Conclusion: This review will provide students in the fields of biochemistry and health sciences a brief and clear overview of iron physiology and toxicity, as well as imbalances in the iron homeostasis and associated pathological conditions. J. Clin. Lab. Anal. 28:210‐218, 2014. C � 2014 Wiley Periodicals, Inc.

Changes in platelet count and mean platelet volume during infectious and inflammatory disease and their correlation with ESR and CRP.

Abstract: Background The measurements of platelet count and mean platelet volume (MPV) are routinely available nowadays. The aim of this study was to evaluate the platelet count and MPV trend in infectious and inflammatory processes. We also investigated whether these parameters were associated with the known markers of disease activity, erythrocyte sedimentation rate (ESR) and C- reactive protein (CRP). Methods This cross-sectional study was conducted on 100 children with diagnosis of infectious and inflammatory diseases. Platelet count, MPV, ESR, and CRP were measured at the time of hospitalization and thereafter in the recovery phase. Results Mean platelet count increased in the patients at the time of admission in the hospital compared to the recovery and discharge time (mean 430,820 ± 134,643/μl vs. 350,970 ± 99,374/μl, P < 0.001). However, MPV decreased significantly during the same period (8.2 ± 1.1 fl vs. 8.7 ± 0.9, P < 0.001). Platelet count was directly correlated with CRP (mean 6.4 ± 0.3 mg/l), (r = 0.49, P < 0.001) and ESR (mean 10.9 ± 1.1 mm/hr), (r = 0.32, P = 0.003). On the other hand, MPV was inversely correlated with CRP (r = 0.39, P < 0.001) and ESR (r = −0.24, P = 0.034). Conclusions This study demonstrated a higher level of platelet count and lower MPV in the patients with active disease compared to the recovered patients. These parameters were well correlated with the known disease activity markers. We propose that platelet parameters can be considered as reliable markers for assessment of disease activity and response to treatment.

Association of neutrophil to lymphocyte ratio with presence and severity of gastritis due to Helicobacter pylori infection.

Abstract: Objective Infection with the bacterial pathogen Helicobacter pylori (HP) clearly results in chronic mucosal inflammation in the stomach and duodenum, which, in turn, might lead to abnormalities in gastroduodenal motility and sensitivity and is the most frequent cause of dyspepsia and peptic disease. Some studies have shown that there was a correlation between low-grade inflammation as C-reactive protein (CRP) and HP infection. The aim of this study was to investigate the relationship between the presence of gastritis due to HP infection and neutrophil/lymphocyte ratio (NLR), a simple and reliable indicator of inflammation. Design Fifty patients met the HP criteria and half of them have had severe symptoms and upper endoscopy showed atrophic gastritis, and fifty age- and sex-matched control subjects with gastritis without HP infection were included in this randomized controlled trial. Patients were diagnosed to have HP according to the use of urea breath testing (UBT) and multiple biopsies. NLR was calculated from complete blood count at the time of diagnosis and before initiating the treatment to all groups. Results Patients with HP infection had significantly higher NLR compared to those without HP. Moreover, the patients with symptomatic HP and grade 4 gastritis had higher NLR than those asymptomatic with past history of peptic disease (P 0.007 and P 0.068, respectively). Although NLR increased as the severity of gastritis and HP symptoms increased (r = 0.564, P < 0.001), Receiver operating characteristic (ROC) Curve analysis was performed. The cut-off level for NLR with optimal sensitivity and specificity was calculated as 1.82 (area under curve [AUC] = 0.825 [0.753–0.884], P < 0.001). Conclusion The present study indicated, for the first time, a significant correlation between HP infection and inflammation on the basis of NLR, a simple and reliable indicator of inflammation. Furthermore, there is an increase in NLR as the severity of gastritis with HP increases. This elevated ratio gets normalized with treatment.

Performance evaluation of platelet counting by novel fluorescent dye staining in the XN-series automated hematology analyzers.

Abstract: Background Conventional automated hematology analyzers have limitations in platelet measurements such as poor accuracy and precision in the low count range and interference by nonplatelet particles. In order to improve it, the newly developed XN-Series automated hematology analyzers (Sysmex Corporation, Kobe, Japan) have been installed with a new dedicated channel for platelet analysis (PLT-F), which is based on a fluorescence flow cytometry method with uses of a novel fluorescent dye specifically staining platelets. We evaluated the basic performance of this new PLT-F channel. Methods Basic performance of the PLT-F channel in within-run reproducibility and assay linearity was studied using standard methods. Correlation was studied between PLT-F and a conventional automated hematology analyzer (XE-2100) and immunoplatelet analysis using anti-CD61 monoclonal antibody (Cell-Dyn Sapphire; Abbott Laboratories). The assay interference by nonplatelet particles such as fragmented red and white blood cells was evaluated by using clinical samples, respectively, from burn injury and acute leukemia. Results Basic performance of the PLT-F platelet counting was satisfactory in within-run reproducibility, linearity and correlation with the conventional analyzer. The correlation was satisfactory also with the immunoplatelet analysis, even for samples from a patient with burn injury, and those with white blood cell fragments displayed, platelet abnormal flag and low platelet counts (<50 × 109/l). Conclusion The platelet counting performance of the PLT-F channel of the XN Series had improved accuracy and precision in the low range and in abnormal samples, avoiding the interference by nonplatelet particles.

The Development and Organization of Newborn Screening Programs in Turkey

Abstract: Background Newborn screening tests have been designed to identify infants with severe disorders that are relatively prevalent and treatable or controllable. Comparing to other countries, the incidence of these diseases are very high in Turkey where the rate of consanguineous marriage is high. Methods In this article, it is aimed to evaluate the development and organization of newborn screening programs in Turkey which include phenylketonuria, congenital hypothyroidism and biotinidase deficiency screenings. The point reached today, limitations of the program, expectations and projects for the future are discussed. Results Today, the point reached in screening programs of the country is appreciable. While the screening rate of the live born babies was 4,7% in 1987, this rate reached to 95% by 2008. Predicted target for newborn screening program at the strategic plan of Ministry of Health for 2010–2014 was to enhance this rate above 95% by the end of 2012. It seems that the envisaged goal has been reached. Conclusion National newborn screening program appears to be conducted successfully and extensively as a result of political determination and performance of health care workers who are in charge of this program. Nevertheless, limited numbers of the nutrition and metabolism clinics and specialists on these branches have caused some access difficulties, waste of time, and financial loss. Therefore, special planning to improve quality and the number of the clinics would be useful.

Serum Vascular Endothelial Growth Factor, Transforming Growth Factor β1, and Nitric Oxide Levels in Patients With Psoriasis Vulgaris: Their Correlation to Disease Severity

Abstract: Background Vascular endothelial growth factor (VEGF), transforming growth factor β1 (TGF-β1), and nitric oxide (NO) have been reported to be contributory factors to the pathogenesis of psoriasis vulgaris. In the current study, we aimed to investigate the association between the levels of VEGF, TGF-β1, and NO and psoriasis severity (as expressed by psoriasis area severity index, PASI). Methods Fifty-eight patients with psoriasis vulgaris and twenty-two controls were included in the study. The serum levels of VEGF and TGF-β1 were estimated by ELISA technique. The serum levels of NO were determined by colorimetric method. Results The serum levels of VEGF, TGF-β1, and NO were significantly higher in patients than controls. Moreover, the serum levels of the studied biochemical variables in patients with severe disease activity were significantly higher than mild cases. The duration of disease showed significant positive correlations with each VEGF (r = 0.35, P < 0.01) and TGF-β1 (r = 0.41, P < 0.05). In addition, the PASI score was significantly positively correlated with VEGF (r = 0.65, P < 0.001), TGF-β1 (r = 0.31, P < 0.05), and NO (r = 0.51, P < 0.001). Conclusion These findings suggest an association between psoriasis disease severity and serum levels of VEGF, TGF-β1, and NO, which can be recognized as markers of the psoriasis severity. The modulation of their production may represent a therapeutic potential strategy for psoriasis.

Significant increase of prostaglandin E-major urinary metabolite in male smokers: a screening study of age and gender differences using a simple radioimmunoassay.

Abstract: Background For the assessment of inflammatory status, we have developed a simple, reliable radioimmunoassay (RIA) of prostaglandin E-major urinary metabolite (PGE-MUM), which remains stable in urine after it is metabolized. Using this method, we conducted a screening study to compare standard values of PGE-MUM to serum C-reactive protein (CRP) levels in health check volunteers. Methods PGE-MUM (micrograms per gram creatinine) was measured in normal urine samples obtained from 797 samples in volunteers for health check, using a newly developed RIA, and analyzed in relation to age, gender, smoking, and drinking habits. Results were compared to serum CRP. Results PGE-MUM was significantly higher in males than in females. It was significantly higher in smoking males, compared to males who had never smoked (nonsmokers), particularly in those above 40 years of age. In nonsmokers, PGE-MUM declined in males with advancing age, while it rose in females. Although PGE-MUM reflected current smoking status, independent of smoking index (SI), serum CRP indicated both current and former smoking condition, rather dependent upon SI. Conclusions PGE-MUM increases in smokers, as suggested by possible inflammatory injury of pulmonary tissue. This RIA method for PGE-MUM may be thus a sensitive and reliable biomarker for current inflammation, different from serum CRP.

Restriction Analysis of β-Tubulin Gene for Differentiation of the Common Pathogenic Dermatophytes

Abstract: Background: Identification of dermatophytes at the species level, relying on macro- and microscopic properties of the colonies is time-consuming, questioned in many circumstances, and requires considerable expertise. In this study, we examined the potency of a new genetic marker, β-tubulin (BT2) gene, for differentiation of dermatophytes in an in silico and experimental restriction fragment length polymorphism (RFLP) profile. Methods: The BT2 sequences of dermatophyte species were retrieved from GenBank and analyzed using bioinformatics softwares to choose suitable restriction enzyme(s). Forty reference culture collections and 100 clinical isolates were PCR-amplified using the primers T1 and Bt2b and consequently subjected to virtual RFLP analysis. The dermatophytes were identified according to specific lengths of bands in agarose gel electrophoresis. Results: After digestion of partially amplified β-tubulin gene with the restriction enzyme FatI, three dermatophyte species, that is, Microsporum gypseum, M. canis, and Trichophyton verrucosum yielded unique restriction maps while the remaining species including T. interdigitale, T. rubrum, T. tonsurans, T. schoenleinii, and T. violaceum, were identified by further restriction digestion by Alw21I, MwoI, and HpyCH4V endonucleases. The length of RFLP products was same as of those expected by computer analysis. Conclusion: The two-step BT2 restriction mapping used in this study is an effective tool for reliable differentiation of the clinically relevant species of dermatophytes. © 2014 Wiley Periodicals, Inc.

Is neutrophil lymphocyte ratio an indicator for proteinuria in chronic kidney disease

Abstract: Background Recent studies have shown that neutrophil lymphocyte ratio (NLR) is a strong indicator in determining inflammation in cardiac and non-cardiac diseases. We aimed to evaluate the relationship between proteinuria and NLR in chronic kidney disease (CKD) patients without diabetes mellitus (DM). Methods Between 2011 and 2012 files of a total of 1000 CKD patients attending outpatient clinic were retrospectively scanned. Patients with DM, chronic disease, malignancy or stage 5 CKD were excluded. After these patients were excluded, a total of 69 patients with stage 3 and 4 CKD were evaluated. Results The study comprised 27 patients with CKD without proteinuria (Group 1), 42 patients with CKD and proteinuria (Group 2) and 30 healthy volunteers (Group 3). NLR was highest in Group 2 and this was statistically significant compared with the control group (p = 0.012). The platelet lymphocyte ratio (PLR) in Group 2 was higher than the control group at a significant level (p = 0.004). There was a moderate positive correlation found between proteinuria and NLR (p = 0.013, r = 0.3). There was a positive correlation found between proteinuria and PLR (p = 0.002, r = 0.306). Conclusion In conclusion, NLR, a parameter easily found in routine blood counts of CKD patients, is a marker with prognostic value for the presence and degree of proteinuria.

Ischemia-Modified Albumin May be a Novel Marker for the Diagnosis and Follow-up of Necrotizing Enterocolitis

Abstract: Aim We investigate the efficacy of serial ischemia-modified albumin (IMA) measurements in diagnosis and follow-up of necrotizing enterocolitis (NEC), and compare its effectiveness with C-reactive protein (CRP), interleukin-6 (IL-6), in NEC. Methods Preterm infants, whose gestational age and weight matched each other, were grouped as control (n = 36) and NEC (n = 37). IMA, CRP, IL-6 levels were measured on the third day of life for the control group and on the day of diagnosis (first day), third, and seventh days of NEC. Results IMA, CRP, and IL-6 levels were significantly increased in NEC patients compared to the control group (P < 0.001) on the follow-up. IMA levels were significantly higher in infants with stage-III NEC than those in infants with stage-II NEC on the first, third, and seventh days (P < 0.001). The area under curve for IMA (0.815 at diagnosis, 0.933 at the third day, 0.935 at the seventh day) were significantly higher than CRP and IL-6 at all days for predicting perforation in infants with NEC (P < 0.001). Similarly, the area under curve for IMA (0.952 at diagnosis, 0.929 at the third day, 0.971 at the seventh day) was significantly higher than CRP and IL-6 at all consequent days of diagnosis for predicting mortality in infants with NEC (P < 0.001). Conclusion Ischemia-modified albumin was found to be superior to CRP and IL-6 in both diagnosis and follow-up of NEC.

Distribution of lymphocyte subpopulations in thyroid glands of human autoimmune thyroid disease.

Abstract: Background The autoimmune thyroid disease (AITD) is an organ-specific autoimmune disease characterized by the breakdown of self-tolerance to thyroid antigens. Some lymphocytes have been identified to be related notably to the pathogenesis of AITD. This article evaluated the distribution of the lymphocytic subpopulation in thyroid glands in order to develop the immunospecific forms of therapy for AITD. Methods Damaged thyroid specimens were obtained from 18 Graves' disease (GD) and 17 Hashimoto's thyroiditis (HT) patients. Normal thyroid specimens were obtained from unaffected glands of 17 patients who underwent parathyroidectomy. We evaluated the distribution of lymphocytic subpopulation by analyzing the expression difference and correlationship among CD4+ T lymphocyte, CD8+ T lymphocyte, CD20+ B lymphocyte as well as regulatory T cells(Tregs)' marker FoxP3 in the thyroid tissues via immunohistochemistry. Results Our research uncovered that no distinct lymphocyte infiltrated in the normal thyroid specimens. Scarcely any lymphocyte infiltration could be found in half of the totally 18 GD thyroid specimens. For the rest 9 GD specimens, CD8+ T cells and CD20+ B cells were expressed more or less in all of them, FoxP3+ Tregs were detected in 7 of them and CD4+ T cells were weakly expressed in only 2 of them. For the 17 HT thyroid specimens, CD20+ B cells were stained strongly in all of them, CD4+, CD8+ T cells were expressed more or less in most of them and FoxP3+ Tregs could be detected in 9 of them. Conclusion Based on CD20+ B cells predominantly infiltrating in all HT thyroid tissues we suggested CD20 antibody might be of help for HT treatment. Furthermore based on FoxP3+ Tregs abundantly infiltrating in some of the AITD thyroid specimens, we considered that activating the Tregs' function in comparison to increasing the Tregs' number only, may be a more effective approach to the treatment of AITD in some cases.

A comparison of whole blood and plasma osmolality and osmolarity.

Abstract: Background Substituting whole blood osmolality for plasma osmolality could expedite treatments otherwise delayed by the time required to separate erythrocytes from plasma. The purpose of this study was to compare the measured osmolality (mmol/kg) and calculated osmolarity (mmol/l) of whole blood and plasma. Methods The osmolality of whole blood and plasma was measured using freezing point depression by micro-osmometer and osmolarity calculated from biosensor measures of sodium, glucose, and blood urea nitrogen. The influence of sample volume was also investigated post hoc by comparing measured osmolality at 20 and 250 μl. Results Sixty-two volunteers provided 168 paired whole blood and plasma samples for analysis. The mean difference (whole blood − plasma; ±standard deviation) in osmolality was 10 ± 3 mmol/kg. Whole blood was greater than plasma in 168 of 168 cases (100%) and data distributions overlapped by 27%. The mean difference in osmolarity was 0 ± 2 mmol/l. Whole blood was greater than plasma in 90 of 168 cases (56%) and data distributions overlapped by 90%. The osmol gap (osmolality − osmolarity) was 16 ± 6 mmol for whole blood and 7 ± 5 mmol for plasma. Ten volunteers were tested on one occasion post hoc to investigate the potential effects of sample volume. The difference between whole blood and plasma was reduced to 3 ± 2 mmol/kg with a larger (250 μl vs. 20 μl) sample volume. Conclusions This investigation provides strong evidence that whole blood and plasma osmolality are not interchangeable measurements when a 20 μl sample is used.

Clinical relevance of plasma miR-21 in new-onset systemic lupus erythematosus patients.

Abstract: Background Plasma miR-21 is widely investigated as biomarker in many diseases. Recent studies show that miR-21 participates in the development of systemic lupus erythematosus (SLE). The aim of this study was to evaluate the expression profile of miR-21 in the plasma of SLE patients. Methods Relative quantities of plasma miR-21 both in SLE patients and healthy controls were determined by relative qRT-PCR under endogenous and exogenous controls. The diagnostic value of plasma miR-21 was evaluated in SLE patients. Data of some SLE-associated clinical parameters were collected. Results Eighty participants from Central China were recruited. Forty-four participants were new-onset SLE patients and the others were healthy controls. Plasma miR-21 level in SLE patients was higher than that of healthy controls (P = 0.031). Receiver operating characteristic analysis of plasma miR-21 revealed an Area Under Curve (AUC) of 0.64 ± 0.06 (95% CI: 0.51–0.76, P = 0.03854) when differentiating SLE from healthy controls. The level of plasma miR-21 was not associated with the level of white blood cells (P = 0.4284), red blood cells (P = 0.4079), and platelets (P = 0.4961), but significantly correlated with the level of plasma complement C3 (r = −0.5297, P = 0.0004), C4 (r = −0.4732, P = 0.0020), and serum uric acid (r = 0.3932, P = 0.0121) in SLE patients. Conclusions Plasma miR-21 in SLE patients from Central China is overexpressed. Since circulating miR-21 is aberrantly expressed in many diseases, the applying of it as a disease biomarker should be considered carefully.

Comparison between screening and confirmatory serological assays in blood donors in a region of South Italy.

Abstract: Background Screening assays are needed in order to guarantee safety of donated blood, but a significant number of safe donations are removed from blood supply because of reactive screening results. It is important to evaluate the positive predictive value (PPV) of screening assays in order to modulate confirmatory algorithm and implement an adequate counseling. Methods An analysis of 17,912 blood donations has been conducted at Transfusion Medicine at Second University Naples, Italy, in 2009–2012. Serological screening for syphilis, hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) was performed by ARCHITECT (Abbott Diagnostics, Wiesbaden, Germany); repeatedly reactive (RR) samples were checked by respective confirmatory tests. The relationship between sample/cutoff and confirmed seropositivity were analyzed. Results RR rates were low as expected in blood donors: 0.47% for syphilis, 0.42% for HBV, 0.50% for HCV, and 0.15% for HIV. The specificity on RR + gray zone (GZ) was 99.67%, 99.79%, 99.77%, and 99.88%, respectively; due to the low prevalence, PPV value was 30.6% for syphilis, 50.7% for HBV, 42.2% for HCV, and 18.5% for HIV. These values increased substantially reaching a plateau of 89.3% for syphilis, 94.6% for HBV, 85.7% for HCV, and 100% for HIV at the threshold established by receiver operating characteristics curve analysis. Conclusions Supplemental testing on samples with high signal by screening assays seems to add little information. GZ settings and confirmatory testing for positive screening results should be designed taking in account several factors, including difference in the natural history among blood-borne infections, the characteristics of first- and second-level tests, and, when available, the results of nucleic acid amplification testing.

Novel parameters of extended complete blood cell count under fluorescence flow cytometry in patients with sepsis.

Abstract: Background: We first describe a patient who developed urosepsis from an ordinary urinary tract infection. In this case, the new hematological parameters of immature leukocytes, that is, the high-fluorescence lymphocyte cell (HFLC) and immature granulocyte (IG) counts peaked early, whereas the established infection parameters, that is, C-reactive protein (CRP) and total white blood cell count showed less dynamic regarding infection and therapy. Methods: To investigate this phenomenon in greater detail, the novel parameters HFLC and IG counts are investigated retrospectively in a cohort of 38 patients who were admitted to the anesthesia intensive care unit. Three groups of patients have been analyzed and compared: patients without signs of infection, patients with limited infections, and patients with sepsis. Data were collected with a Sysmex XE-5000 hematological analyzer. Results: In patients (n = 22) without any signs of infection, both values are very low. In patients with limited local infections (n = 10), moderate elevations of the IG and HFLC counts are seen. In patients with sepsis (n = 6), the IG and HFLC counts are significantly higher.Conclusion: The total IG countseemstobeusefulfordistinguishinga

Clinical utility of serum cystatin C in predicting coronary artery disease in patients without chronic kidney disease.

Abstract: Background Cystatin C has been proposed as a novel marker of renal function and predictor of cardiovascular risk. The aim of this study was to investigate the role of cystatin C level as a predictor of cardiovascular events in patients with coronary artery disease (CAD). Methods Three hundred and five coronary artery patients were included in this study. Serum cystatin C levels, high-sensitive C-reactive protein (hs-CRP), and oxidative stress were measured. Estimated glomerular filtration rate (eGFR) and the CAD severity score were calculated. Results Cystatin C was correlated with the CAD severity score (r = 0.631, P 50. Every 0.1 mg/l increase in cystatin C, 2 mg/l increase in hs-CRP, 0.2 mmol/l decrease in high-density lipoprotein cholesterol, 13.7 ml/min decrease in eGFR, and 1.51 μmol/l increase in homocysteine caused a 34, 12, 5, and 22% increase in the risk of having CAD, respectively. Conclusion Cystatin C could be a useful laboratory biochemical marker in predicting the severity of CAD. Cystatin C is associated with biochemical atherosclerosis markers such as CRP and homocysteine.

Point‐of‐Care Fluorescence Immunoassay for Cardiac Panel Biomarkers

Abstract: Background Myoglobin, creatine kinase-MB isoenzyme (CK-MB), and cardiac troponin I (cTnI) are cardiac biomarkers that are widely used to assist in the early and late diagnoses of acute myocardial infarction (AMI). Here, we present a clinically applicable fluorescence (FL) immunoassay for cardiac biomarkers. Methods Whole blood was mixed with FL-labeled detector Ab (antibody) and then loaded onto a capture Ab-immobilized strip in a test cartridge. The FL intensities at test and control line on the strip were obtained and converted in a laser FL scanner to determine the concentration of biomarker. The analytical performance of immunoassay system was evaluated by linearity and imprecision tests. The comparability of the FL immunoassay method was examined with a reference method. Results FL intensities and the levels of myoglobin, CK-MB, and cTnI displayed good linearity and high correlations (r = 0.999, 0.998, and 0.989, respectively). The coefficient of variations (CVs) of imprecision for all cardiac biomarkers were less than 8% in both intra- and interassays. When the results from the developed method and bioMerieux VIDAS assay were analyzed by Bland–Altman plot and Passing–Bablok plot, the two assay methods were in good agreement. Conclusion The FL immunoassay system can provide a platform for point-of-care testing (POCT), and it is an easy, fast, and reliable method for the quantification of cardiac biomarkers.

Association of FOS-like antigen 1 promoter polymorphism with podocyte foot process effacement in immunoglobulin A nephropathy patients.

Abstract: Background FOS has been implicated in the progression of renal disease including IgAN. In this study, we investigated whether polymorphisms of FOS family genes [FOS, FOSB, FOS-like antigen 1 (FOSL1), and FOSL2] were associated with immunoglobulin A nephropathy (IgAN) and the clinical phenotypes of IgAN patients. Methods We genotyped single nucleotide polymorphisms (SNPs) of FOS family genes (rs2239615 and rs7101 for FOS, rs12373539 and rs2282695 for FOSB, rs637571 for FOSL1, and rs925255 for FOSL2) using direct sequencing in 198 IgAN patients and 290 control subjects. Results No SNPs were associated with IgAN; however, in the analysis of clinical phenotypes, we found that rs637571 of FOSL1 was associated with podocyte foot process effacement of IgAN in additive (CT vs. TT vs. CC, P = 0.0031, OR = 2.08, 95% CI = 1.27–3.40) and dominant models (CT/TT vs. CC, P = 0.0034, OR = 2.50, 95% CI = 1.35–4.64). The frequency of genotypes containing the T allele was increased in IgAN patients with podocyte foot process effacement, compared to those without podocyte foot process effacement. Conclusion These results suggest that FOSL1 may be related to IgAN severity.

Comparison of Methods, Storage Conditions, and Time to Analysis of Serum and Urine Creatinine Measured from Microsamples by Liquid Chromatography Mass Spectrometery (LC/MS) vs. Jaffe

Abstract: Background Measurement of serum creatinine (SCr) and urine creatinine (UCr) is regularly used in clinical and research settings. For small animal experiments and for studies in which sample collection is spare (i.e. neonatal cohorts), measuring SCr and UCr using tiny amounts of sample (as low as 10 mcl) would maximize exploration and minimize iatrogenic blood loss. Methods We performed an evaluation in six healthy adults to determine differences between SCr and UCr values in different methodologies and storage environments and time. Study was conducted using 20 mcl of sample. Analyses were done using two-way repeated measures of ANOVA. Results Scr values showed no significant differences between LC/MS vs. Jaffe. However, the SCr using LC/MS method was lowest when measured immediately compared to other time points (F = 7.2; P< 0.001). Similarly, Jaffe measurements showed changes in the mean differences over time; however, these were not significant. UCr values were consistently higher using LC/MS than Jaffe (F = 19; P< 0.01), and UCr changed over time (F = 8.7; P < 0.02). In addition, the interaction term for method and time was also significant (F = 5.8; P < 0.04) which reflects the stability of the Jaffe measurements over time whereas the LC/MS measurements declined; especially after being frozen for 1 year (P < 0.001). Conclusion UCr measured by Jaffe is lower than samples measured by LC/MS. UCr measurements by LC/MS vary more over time, mostly due to the sample measured after 1 year; therefore, storage of urine for more than 90 days measured by LC/MS may provide altered results.

Indexing Glomerular Filtration Rate to Body Surface Area: Clinical Consequences

Abstract: Background Kidney function is mostly expressed in terms of glomerular filtration rate (GFR). A common feature is the expression as ml/min per 1.73 m2, which represents the adjustment of the individual kidney function to a standard body surface area (BSA) to allow comparison between individuals. We investigated the impact of indexing GFR to BSA in cancer patients, as this BSA indexation might affect the reported individual kidney function. Methods Cross-sectional study of 895 adults who had their kidney function measured with 51chrome ethylene diamine tetraacetic acid. Mean values of BSA-indexed GFR vs. mean absolute GFR were analyzed with a t-test for paired data. Bland–Altman plot was used to analyze agreement between the indexed and absolute GFR values. Results and Conclusion BSA-GFR in patients with a BSA 2 m2 with a bias up to −20.76 ml/min (−23.59%). BSA is not a good normalization index (NI) in patients with extreme body sizes. Therefore, until a better NI is found, we recommend clinicians to use the absolute GFR to calculate individual drug chemotherapy dosage as well as express individual kidney function.

Rapid and Sensitive Headspace Gas Chromatography–Mass Spectrometry Method for the Analysis of Ethanol in the Whole Blood

Abstract: Background For the forensic aim, a sensitive and specific method using headspace gas chromatography coupled with mass spectrometry (GC/MS) has been developed for the quantitative determination of ethanol in blood using n-propanol as internal standard. GC was performed in isothermal mode with a GC run-time of 5.0 min. Methods The quantification was performed using selected ions monitoring mode adopting a quantitative ion and qualifier ion for ethanol and the internal standard. Results The method was linear (r2 = 0.999, in the concentration range of 39.5–1,262.9 μg/ml), specific, sensitive (limit of quantification and limit of detection of 39.5 and 0.4 μg/ml, respectively), and robust. A slightly modified method was also developed for the quantification of 50 commonly abused drunken in blood. The method used an isothermal GC program with a run-time of 5.0 min. The quantification was performed using selected ions monitoring mode and integrating the area under the peak using n-propanol as an internal standard. The method was linear 40–1,263 μg/ml and sensitive. Conclusions The method was proved superior in speed and selectivity to previously reported methods and was successfully applied to the pharmacokinetic study of ethanol.

Detection and Assessment of Clarithromycin Inducible Resistant Strains Among Korean Mycobacterium abscessus Clinical Strains: PCR Methods

Abstract: Background Mycobacterium abscessus group belongs to a group of rapidly growing mycobacteria (RGM) and, following Mycobacterium avium complex, is the second most common pathogen responsible for lung disease caused by nontuberculous mycobacteria (NTM). Clarithromycin is known to be the key drug in the treatment of M. abscessus group disease, but a high failure rate of treatment response is reported due to clarithromycin inducible resistance. Methods Using the results from a clarithromycin susceptibility test we examined the proportion of clarithromycin inducible resistant M. abscessus (sensu stricto; hereafter referred to as M. abscessus) clinical strains. Also, we attempted to detect the clarithromycin resistant strains, using the amplification refractory mutation system-PCR (ARMS-PCR) and real-time PCR methods for rapid detection of single-nucleotide polymorphisms (SNPs) at position 28 (T or C) of the erm(41) gene of M. abscessus leading to resistance to clarithromycin. Results Of the 157 M. abscessus clinical strains, clarithromycin susceptible, resistant, and inducible resistant strains accounted for 10.83% (n = 17), 22.29% (n = 35), and 66.88% (n = 105), respectively. Clarithromycin resistant strains were able to separate from clarithromycin susceptible strains by ARMS-PCR and real-time PCR identical to DNA sequence analysis. Conclusion Most M. abscessus clinical strains in Korea are resistant to clarithromycin, and ARMS-PCR and real-time PCR are useful tools for the rapid detection of single-nucleotide polymorphisms (SNPs) at position 28 of the erm(41) gene.

Comparison of creatinine and specific gravity for hydration corrections on measurement of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine.

Abstract: Background Tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) was measured in all participants aged 6 years and older from the Centers for Disease Control and Prevention's National Health and Nutrition Examination Survey 2007–2008. The suitability of using creatinine or specific gravity for urinary NNAL correction in exposure assessment is examined in this study. Methods Effects of both specific gravity and creatinine correction on urinary NNAL among smokers were investigated with multiple linear regression models using either normalization or the fitting of creatinine and specific gravity in the model as covariates. Results When log-scaled NNAL was normalized by either creatinine or specific gravity, R2 was slightly higher for creatinine than for specific gravity (R2 = 0.1694 and 0.1439, for creatinine and specific gravity, respectively). When log-scaled NNAL was normalized by both factors, the R2 was improved (R2 = 0.2068). When specific gravity or creatinine was included as a covariate separately in the models, they were highly significant factors (P < 0.001, R2 = 0.2226 and 0.1681 for creatinine and specific gravity, respectively). However, when both were included in the model as covariates, creatinine remained highly significant (P < 0.001), whereas the significance of specific gravity was eliminated (P = 0.4294). Conclusion This study confirms significant relationships between NNAL concentrations and both urine creatinine and specific gravity. We conclude that creatinine is the more influential and preferred variable to account for urine dilution in tobacco-specific nitrosamine exposure assessment.

Storage at -80°C preserves the antioxidant capacity of preterm human milk.

Abstract: Background and Objective: It is essential to establish optimum parameters for maintaining the quality of stored milk until the moment of consumption with minimal deterioration of its properties The aim of the study was to evaluate total antioxidant capacity (TAC) and total oxidation status (TOS) of fresh and freeze-stored samples (at −80 ◦ C) of preterm human milk (HM) Methods: Samples of colostrum were collected from 98 healthy women within the first 4 days after delivery The total milk volume collected (6 ml) was divided in two aliquot parts: 3 ml for the fresh analysis which was done immediately after the extraction and 3 ml for storage under freezing conditions at −80 ◦ C for three months The

Serum 25-hydroxyvitamin D level and extent and complexity of coronary artery disease.

Abstract: Background There are limited number of studies about relationship between serum vitamin D level and presence and severity of coronary artery disease (CAD). We assessed the relationship between the extent and complexity of CAD assessed by SYNTAX score and 25-hydroxyvitamin D level in patients with stable CAD. Methods In the study, 209 consecutive patients with stable CAD (age: 63.1 ± 10.0 years) and 102 healthy control subjects (age 61.3 ± 13.7 years) were included. Serum 25-hydroxyvitamin D was measured using a direct competitive chemiluminescent immunoassay and other biochemical markers were measured in all subjects. All subjects underwent coronary angiography and SS was calculated. Results The mean serum 25-hydroxyvitamin D level of CAD group was lower than the control group (P < 0,001). Multivariate regression analysis showed that serum 25-hydroxyvitamin D level was independently associated with SYNTAX score (β = −0.396, P < 0.001), hypertension (β = −0.183, P = 0.003), high sensitive C-reactive protein (β = −0.141, P = 0.014), and body mass index (β = −0.135, P = 0.023) in patient group. Conclusion 25-Hydroxyvitamin D level was associated with extent and complexity of CAD. 25-Hydroxyvitamin D may play a role in pathogenesis and severity of coronary atherosclerosis.

Systemic chemotherapy interferes in homocysteine metabolism in breast cancer patients.

Abstract: Background Hyperhomocysteinemia in breast cancer (BC) patients can be a risk factor for thromboembolic events. This study aimed to evaluate homocysteine and its cofators (folic acid and vitamin B12) concentrations and platelet count at diagnosis of BC, 3 and 6 months after the beginning of chemotherapy treatment and to correlate them with clinical data. Methods Thirty-five BC patients were included; blood samples were obtained by venipuncture. Plasmatic Hcy and cofactors concentrations were measured by competitive chemiluminescent enzyme immunoassay method. Platelet count was done using an automated analyzer. Statistical analysis was performed using the software SPSS. Results During chemotherapy, homocysteine (P = 0.032) and vitamin B12 (P < 0.001) concentrations increased, while folate and platelets decreased (P < 0.001). Among the clinical data, the menopausal status showed significant positive correlation (P = 0.022) with homocysteine concentration increase. Conclusions Evaluation of homocysteine concentrations during chemotherapy is extremely important because their levels increase during chemotherapy treatment, thus increasing the risk of thromboembolism development.

Delta Mean Neutrophil Volume (ΔMNV) is Comparable to Procalcitonin for Predicting Postsurgical Bacterial Infection

Abstract: Background The Coulter LH750 (Beckman Coulter, Brea, CA) analyzer can determine intrinsic biophysical properties of white blood cell (WBC), known as cell population data. Previous studies have shown that mean neutrophil volume (MNV) was significantly increased in postsurgical patients with bacterial infection. To further validate its potential clinical usefulness, we investigate the changes in MNV before and after surgery, called ΔMNV. We also compare the ΔMNV with procalcitonin (PCT) and C-reactive protein (CRP) in terms of diagnostic sensitivity and specificity for postsurgical bacterial infection. Methods Blood samples from 300 healthy controls, 219 cardiac surgical patients without postsurgical infection, and 31 cardiac surgical patients complicated with postsurgical bacterial infection were studied. Results There are no statistically significant differences for WBC count and neutrophil percentage prior to or after surgery between postsurgical noninfected and infected patients. However, the ΔMNV is significantly increased in postsurgical infected patients when compared with noninfected patients (P < 0.05). The receiver-operating characteristics analysis reveals the ΔMNV and PCT have largest areas under curves (0.92, 0.93 on the second day and 0.94, 0.99 on the third day postsurgery, respectively) compared to other parameters. Conclusion ΔMNV shows comparable sensitivity and specificity to PCT and superior sensitivity and specificity to WBC or CRP for predicting postsurgical bacterial infection. The potential clinical application of this parameter merits further exploration in a larger prospective study.

Issues in identifying germ tube positive yeasts by conventional methods.

Abstract: Candida speciation is vital for epidemiology and management of candidiasis. Nonmolecular conventional methods often fail to identify closely related germ tube positive yeasts from clinical specimens. The present study was conducted to identify these yeasts and to highlight issues in conventional versus molecular methods of identification. A total of 98 germ tube positive yeasts from high vaginal swabs were studied over a 12-month period. Isolates were examined with various methods including growth at 42 °C and 45 °C on Sabouraud dextrose agar (SDA), color development on CHROMagar Candida medium, chlamydospore production on corn meal agar at 25 °C, carbohydrate assimilation using ID 32C system, and polymerase chain reaction using a single pair of primers targeting the hyphal wall protein 1 (Hwp1) gene. Of all the isolates studied, 97 were molecularly confirmed as C. albicans and one isolate was identified as C. dubliniensis. No C. africana was detected in this study. The molecular method used in our study was an accurate and useful tool for discriminating C. albicans, C. dubliniensis, and C. africana. The conventional methods, however, were less accurate and riddled with many issues that will be discussed in further details.

The Diagnostic Value and Performance Evaluation of Five Serological Tests for the Detection of Treponema pallidum

Abstract: Background Syphilis is caused by the bacterium Treponema pallidum (TP). The aim of this study was to establish a clinical approach for serodiagnosis of syphilis by evaluating the performance and diagnostic value of five serological tests for the detection of TP. Methods Five tests were used to test the serum from syphilis patients and control patients, namely rapid plasma reagin (RPR) test, toluidine red unheated serum test (TRUST), TP passive particle agglutination assay (TPPA), TP-specific enzyme-linked immunosorbent assay (TP-ELISA), and TP-specific chemiluminescent immunoassay (TP-CMIA). Results The sensitivity and diagnostic efficiency of TPPA (96.25%/98.38%), TP-ELISA (100%/95.41%), and TP-CMIA (100%/94.86%) were significantly higher than that of RPR (73.13%/86.22%) and TRUST (73.75%/86.49%) (P < 0.05). The minimum detectable concentrations for the five tests were 30 mIU/ml, 20 mIU/ml, 15 mIU/ml, 150 mIU/ml, and 150 mIU/ml, respectively. According to receiver operating characteristic (ROC) curve, the optimal cut-off values for syphilis diagnosis by TP-CMIA and TP-ELISA were 2.2 and 2.0 S/CO (where S/CO = Sample/calibrator cut off), and the area under the ROC curve (AUC) were 0.998 for TP-CMIA and 0.999 for TP-ELISA. The titers/positive rates for RPR and TRUST dropped from 1:4 (100%) to 1:1 (23.3%) (both P < 0.05) after treatment. However, there were no significant differences when we compared the positive rate of syphilis patients before and after treatment by TPPA, TP-ELISA, and TP-CMIA. Conclusions Treponemal tests, such as TPPA, TP-ELISA, and TP-CMIA, are recommended for clinical routine screening of syphilis. However, nontreponemal tests, for example, RPR and TRUST, perform better in therapy response assessment. Serological test should be tailored to respective facilities and clinical demands.