Leukocyte Subgroups Research Articles

Abstract 4125093: Association of Systemic Inflammatory Index and Systemic Inflammatory Response Index with Major Adverse Cardiovascular Events in Patients with Acute Coronary Syndrome Who Had Primary Percutaneous Coronary Intervention

Introduction: Inflammation plays an important role in the pathogenesis of coronary artery disease and Acute Coronary Syndrome (ACS). Inflammatory indicators such as neutrophil count and monocyte count potentially may predict patients’ outcomes and prognosis in ACS. White blood cells count is an affordable and accessible way to assess the systemic immune response. Considering their multidirectional effect on atherosclerosis, new inflammatory biomarkers integrating various leukocyte subgroups have been proposed to calculate the systemic inflammatory response index (SIRI) and systemic inflammatory index (SII). Aim: The aim of this study was to assess the association between the systemic inflammatory response index (SIRI) and Systemic inflammatory index (SII) at admission with major adverse cardiovascular events (MACE) 30-days after Primary Percutaneous Coronary Intervention (PPCI) among patients with Acute Coronary Syndrome (ACS) at one referral hospital in Jordan. Method: A Prospective design was used to assess and follow 150 adults presented with ACS and received PPCI at one referral hospital in Jordan. Data was collected from patients and their medical records during hospital stay using a structured data collection form. The MACE was assessed 30-days after PPCI through phone calls with patients and confirmed from medical records. Results: The study included 150 patients with ACS, 82.7% (n= 124) of them were males. The mean age of patients was 57.68 (SD= 11.19) years. Types of ACS include stable angina 5.3% (n=8), unstable angina 24% (n=36), NSTEMI 28.7% (n=43), and STEMI 24% (n=36). All patients had interventional PCI with balloon and stent insertion. The mean of SIRI was 24.45 (SD=25.49) and the mean of SII was 663.30 (SD= 403.75). Rate of MACE as composite outcome 30-days after PPCI as 24.7% (n=37). The most MACE was unplanned repeat revascularization (n=14, 38% of MACE). Significant association between SIRI and SII, and MACE was found SIRI:(Χ 2 (2)> = 12.63, p = 0.001), SII:(Χ 2 (2)> = 4.23, p = 0.03) . Conclusion: The SIRI and SII are inflammatory biomarkers that should be studied as potentially predictors of MACE among patients presented with ACS.

Utilizing cell population data for diagnosis and intensive care unit requirement prediction in Fournier gangrene patients

Abstract Objectives It is vital to rapid diagnosis and to determine the intensive care unit (ICU) requirement early to reduce the mortality rate in Fournier gangrene (FG) patients. Cell population data (CPD) are the parameters obtained from complete blood count (CBC) analysis and related to the activation of different leukocyte subgroups. The study aimed to find reliable markers to diagnose and determine the ICU requirement using CPD. Methods We included 24 patients and 22 healthy controls in the study. CBC analyses were performed by using a Sysmex XN-9000 series hematology analyzer. ROC analyses and group comparisons were performed to evaluate the diagnostic accuracy and prognostic value of CPD parameters in ICU requirements. Results Statistically significant differences were observed in terms of some CPD values of lymphocytes, neutrophils, and monocytes in patients compared to healthy controls. Neutrophile-Y or reactivity index (Ne-Y or RI) (p=0.004), neutrophile-X or granularity index (Ne-X or GI) (p=0.009), monocyte-X (Mo-X) (p<0.001), and lymphocyte-WY (Ly-WY) (p<0.001) were higher in patients than controls. Ne-Y (RI) (p=0.012), Mo-X (p=0.001), Mo-Y (p=0.022), and Ne-WY (p=0.025) levels were higher in ICU patients than in non-ICU patients. Conclusions The severity of FG disease can be determined using CPD data. Ne-Y (RI) serves as a novel and reliable biomarker for determining disease severity. In addition, the neutrophile-lymphocyte ratio can be used to rule out FG, especially in combination with other well-known clinical and diagnostic parameters.

Investigation of the Associations of Novel Inflammatory Biomarkers-Systemic Inflammatory Index (SII) and Systemic Inflammatory Response Index (SIRI)-With the Severity of Coronary Artery Disease and Acute Coronary Syndrome Occurrence.

Atherosclerosis, the underlying cause of coronary artery disease (CAD), has a significant inflammatory component. White blood cell count is an affordable and accessible way to assess the systemic immune response, as it comprises many subgroups with distinct and complex functions. Considering their multidirectional effect on atherosclerosis, new biomarkers integrating various leukocyte subgroups, the Systemic Inflammatory Index (SII) and the Systemic Inflammatory Response Index (SIRI), were recently devised to describe the balance between inflammation and immune reaction. This research aimed to evaluate the relationship of the intensity of inflammation measured by these biomarkers with the severity of CAD assessed with coronary angiography and with the diagnosis of acute coronary syndrome (ACS) or stable CAD in 699 patients. SIRI, but not SII, was associated with the diagnosis, having the highest values for patients with ACS (STEMI), significantly higher than in patients with stable CAD (p < 0.01). The highest SII and SIRI values were observed in patients with three-vessel CAD. SII and SIRI require further in-depth and well-designed research to evaluate their potential in a clinical setting.

Changes in the Small Noncoding RNAome During M1 and M2 Macrophage Polarization.

Macrophages belong to a special phagocytic subgroup of human leukocytes and are one of the important cells of the human immune system. Small noncoding RNAs are a group of small RNA molecules that can be transcribed without the ability to encode proteins but could play a specific function in cells. SncRNAs mainly include microRNAs (miRNAs) and piwi-interacting RNAs (piRNAs), small nucleolar RNAs (snoRNAs), small nuclear RNAs (snRNAs) and repeat RNAs. We used high-throughput sequencing analysis and qPCR to detect the expression changes of the small noncoding RNAome during macrophage polarization. Our results showed that 84 miRNAs and 47 miRNAs with were downregulated during M1 macrophage polarization and that 11 miRNAs were upregulated and 19 miRNAs were downregulated during M2 macrophage polarization. MiR-novel-3-nature and miR-27b-5p could promote expression of TNF-α which was marker gene of M1 macrophages. The piRNA analysis results showed that 69 piRNAs were upregulated and 61 piRNAs were downregulated during M1 macrophage polarization and that 3 piRNAs were upregulated and 10 piRNAs were downregulated during M2 macrophage polarization. DQ551351 and DQ551308 could promote the mRNA expression of TNF-α and DQ551351overexpression promoted the antitumor activity of M1 macrophages. SnoRNA results showed that 62 snoRNAs were upregulated and 59 snoRNAs were downregulated during M1 macrophage polarization, whereas 6 snoRNAs were upregulated and 10 snoRNAs were downregulated during M2 macrophage polarization. Overexpression of snoRNA ENSMUST00000158683.2 could inhibit expression of TNF-α. For snRNA, we found that 12 snRNAs were upregulated and 15 snRNAs were downregulated during M1 macrophage polarization and that 2 snRNAs were upregulated during M2 macrophage polarization. ENSMUSG00000096786 could promote expression of IL-1 and iNOS and ENSMUSG00000096786 overexpression promoted the antitumor activity of M1 macrophages. Analysis of repeat RNAs showed that 7 repeat RNAs were upregulated and 9 repeat RNAs were downregulated during M1 macrophage polarization and that 2 repeat RNAs were downregulated during M2 macrophage polarization. We first reported the expression changes of piRNA, snoRNA, snRNA and repeat RNA during macrophage polarization, and preliminarily confirmed that piRNA, snoRNA and snRNA can regulate the function of macrophages.

Phenotypic and Functional Heterogeneity of Monocyte Subsets in Chronic Heart Failure Patients.

Simple SummaryA long-term condition known as chronic heart failure (CHF) is an ongoing difficulty of the heart in pumping blood enriched in oxygen and required nutrients around the body’s tissues. CHF pathogenesis is associated with various causes, and inflammation is one of the most important factors promoting the condition. In addition, monocytes, a group of cells present in the blood and infiltrating tissues, are known to participate in both pro- and anti-inflammatory processes and thus affect myocardial remodeling over time. The aim of this work was to review current studies on the function of monocyte subsets in different types of CHF with preserved and reduced left ventricle ejection fractions and to discuss the relationship of monocyte subsets to inflammatory markers. It is expected that a deeper view into CHF pathogenesis could stimulate the search for and development of individualized therapies.Chronic heart failure (CHF) results when the heart cannot consistently supply the body’s tissues with oxygen and required nutrients. CHF can be categorized as heart failure (HF) with preserved ejection fraction (HFpEF) or HF with reduced ejection fraction (HFrEF). There are different causes and mechanisms underlying HF pathogenesis; however, inflammation can be regarded as one of the factors that promotes both HFrEF and HFpEF. Monocytes, a subgroup of leukocytes, are known to be cellular mediators in response to cardiovascular injury and are closely related to inflammatory reactions. These cells are a vital component of the immune system and are the source of macrophages, which participate in cardiac tissue repair after injury. However, these monocytes are not as homogenous as thought and can present different functions under different cardiovascular disease conditions. In addition, there is still an open question regarding whether the functions of monocytes and macrophages should be regarded as causes or consequences in CHF development. Therefore, the aim of this work was to summarize current studies on the functions of various monocyte subsets in CHF with a focus on the role of a certain monocyte subset in HFpEF and HFrEF patients, as well as the subsets’ relationship to inflammatory markers.

The role of leukocytes in predicting whether laparotomy is required in patients with penetrating abdominal stab wound

BackgroundThis study aimed to determine the importance of leukocytes, leukocyte subgroups, platelets, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio in deciding whether laparotomy is required during observation in patients with penetrating abdominal stab wounds who were followed up because there was no indication for an emergency laparotomy. MethodsPatients who did not indicate an emergency laparotomy were monitored. After 48 hours from initial hospitalization, patients who did not require laparotomy were discharged nonoperatively. The total leukocytes, leukocyte subsets, platelets, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio of patients who underwent laparotomy during the follow-up compared with those who were discharged nonoperatively. The sensitivity and specificity of these laboratory values in predicting the necessity of laparotomy were calculated. ResultsIn the operated group (n = 71), leukocytes, neutrophils, monocytes, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio examined during observation were significantly higher (P < .001). Eosinophils and lymphocytes were significantly lower (P < .001) than in nonoperated (n = 476). Based on the deviation in the reference ranges of leukocyte and its subgroups, we report the sensitivity and specificity for predicting the necessity of laparotomy as 86% and 72% for leukocyte, 88% and 75% for neutrophil, 92% and 83% for neutrophil-to-lymphocyte ratio, and 72% and 77% for platelet-to-lymphocyte ratio, respectively. In receiver operating characteristic curve analysis, the cut-off value was found to be 4 for neutrophil-to-lymphocyte ratio and 125 for platelet-to-lymphocyte ratio (area under the curve/receiver operating characteristic curve of 0.929 and 0.808, respectively). ConclusionLeukocyte, leukocyte subgroups, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio may be useful to determine if a laparotomy is necessary in patients in whom the necessity of laparotomy is undetermined owing to unclear examination findings.

Leukocytes are not Reliable in Predicting Possible Diaphragmatic Injury in Patients with Penetrating Left Thoracoabdominal Stab Wounds.

The diaphragm is injured in approximately one-third of penetrating left thoracoabdominal stab wounds. Diagnostic laparoscopy or thoracoscopy is performed to reveal the diaphragmatic injury. This study investigated whether leukocytes, leukocyte subgroups, platelets, the neutrophil-to-lymphocyte ratio (NLR), and the thrombocyte-to-lymphocyte ratio (PLR) can be used to detect diaphragm injury without the need for diagnostic laparoscopy. Patients hospitalized between January 2010 and January 2020 due to penetrating left thoracoabdominal stab wounds were examined. Laparotomy was performed in patients who had indications for laparotomy, such as hemodynamic instability and peritonitis. Diagnostic laparoscopy was performed to reveal possible diaphragmatic injury in patients who did not require laparotomy after 48h of follow-up. Leukocytes, leukocyte subgroups, platelets, NLR, and PLR were measured both at admission and during follow-up, and the results were compared between patients with and without diaphragm injury during diagnostic laparoscopy. The study included 108 patients with penetrating left thoracoabdominal stab wounds that did not require laparotomy after 48h of follow-up. Of these, 102 patients were male (94.44%), and the average age was 27.68years (range 15-66years). Diaphragm injury was detected in 31 patients (28.70%) in diagnostic laparoscopy, and the diaphragm was intact in 77 patients (71.30%). In the comparison of patients with and without diaphragmatic injury, no statistically significant difference was found in terms of age, gender, platelets, leukocyte values, NLR, and PLR both at admission and during follow-up. Leukocytes, leukocyte subsets, platelets, NLR, and PLR were insufficient in the detection of asymptomatic diaphragmatic injuries caused by penetrating left thoracoabdominal stab wounds.

Electronic Immunoaffinity Assay for Differential Leukocyte Counts

Leukocytes are blood cells involved in the immune response. Leukocytes can be classified into three main groups (granulocytes, lymphocytes, and monocytes), and the measurement of the relative prevalence of each subgroup, termed differential leukocyte count, is an important clinical parameter in the diagnosis and prognosis of various health anomalies. Currently, differential leukocyte counts are obtained either manually through labor-intensive blood smears or by employing bulky hematology analyzers. Both of these measurements require a laboratory setup and hence are not amenable for decentralized and point-of-care settings. In this paper, we introduce a microfluidic assay with an electronic readout for differential leukocyte count assays. Our device classifies leukocyte subgroups based on the differential CD33 expression on cell membranes. In the device, each subgroup is differentially immunocaptured in different microfluidic chambers, while the cell capture rates are quantified by an integrated electronic sensor network. When benchmarked against a commercial hematology analyzer, our device could identify leukocyte subpopulations with differences of <6% for granulocytes and lymphocytes, and a <3% for monocytes. These results demonstrate the potential of our technique as an accurate method to perform the widely-employed hematology assay for point-of-care applications without the need for sample labeling. [2020-0197]

DİRENÇ EGZERSİZLERİ SIRASINDA SU ALIMININ HEMOGRAM ÜZERİNDEKİ AKUT ETKİSİ

The aim of this study is to determine the effects of exercises done by drinking water and not drinking water during resistance training on hemogram (leukocyte, erythrocyte, and thrombocyte). 10 male university students aged between 20 and 23 participated in the study. The participants did five different resistance exercises (3 sets 8 RM) previously determined. During the training program, they did resistance exercises by not consuming water in the first week and consuming water in the second week. (Pre-exercise and exercise days routine food and fluid intake was maintained, fluid intake restriction was applied only during exercise) The participants’ Hemogram (Erythrocyte, Leukocyte and Thrombocyte) measurements were made before and after the exercise. The obtained data were tested at p<0.05 significance level. A significant decrease in Lymph% and a significant decrease in p<0.02 in Gran% were found among exercise pre-test post-test values by taking water from sub leukocyte subgroups. A significant decrease in RBC p<0.01 and a significant decrease in HGB and HCT values of p<0.02 were observed between exercise pre-test post-test values without water intake and water intake of erythrocyte subgroups of the same resistance exercise. A significant decrease in p <0.00 was also found between the exercise pre-test post-test values of the same subgroups (RBC, HGB, HCT). A significant reduction in PDW% value of p<0.04 was observed between the exercise pre-test post-test values of the same resistance exercise without water intake and without taking water from platelet subgroups of the same resistance exercise. No statistically significant change was detected in the parameter values in the other subgroups. The exercise done without water intake; a significant difference was found in the parameters of Thrombocyte and Erythrocyte. However, no statistically significant difference was found in the parameters of Leukocyte. The exercise done with water intake; a significant difference was found in the parameters of Leukocyte and Erythrocyte. However, no statistically significant difference was found in the parameters of Thrombocyte.

The effects of preparation period exercises on the hematological parameters of the taekwondo athletes

Purpose: This study aimed at examining the effects of preparation period exercises, which were applied for eight weeks to elite level taekwondo athletes, on the hematological parameters. Material: Totally 21 athletes with international degrees from the university taekwondo team participated in this study, 12 of whom were female (Mean age: 20.3 - Mean exercise age: 8.8) and 9 of whom were male athletes (Mean age: 20.3 - Mean exercise age: 9.5). The participants attended an 8-week exercise program for 5 days a week, which included basic motoric features and technical and tactical development. 4 cc blood samples were taken from the right antecubital vein before and after the preparation period exercises. On the blood samples, WBC, GRAN, GRAN %, LYM, LYM %, MID and MID % values were examined among leukocyte sub-groups; RBC, HGB, HCT, MCV, MCH, MCHC and RDWC values were examined among the erythrocyte sub-groups, and PLT, MPV, PCT and PDW values were examined among thrombocyte sub-groups. Results: At the end of the study, we determined that there was a statistically significant increase in the MCHC values of both the male and the female taekwondo athletes. Moreover, it was determined that there were statistically significant decreases in the HCT, MCV, MCH, MPV and PCT values of the female taekwondo athletes, and in the MCV, MPV and PCT values of the male taekwondo athletes. Conclusions: As a conclusion, we can state that the 8-week exercises applied during the preparation period affected the hematological parameters of the male and female taekwondo athletes to an extent, however, this effect was limited and it did not cause an important change.

Rapid capture and labeling of cells on single domain antibodies-functionalized flow cell

Current techniques to characterize leukocyte subgroups in blood require long sample preparation times and sizable sample volumes. A simplified method for leukocyte characterization using smaller blood volumes would thus be useful in diagnostic settings. Here we describe a flow system comprised of two functionalized graphene oxide (GO) surfaces that allow the capture of distinct leukocyte populations from small volumes blood using camelid single-domain antibodyfragments (VHHs) as capture agents. We used site-specifically labeled leukocytes to detect and identify cells exposed to fungal challenge. Combining the chemical and optical properties of GO with the versatility of the VHH scaffold in the context of a flow system provides a quick and efficient method for the capture and characterization of functional leukocytes.

Neutrophil­to­lymphocyte ratio: A novel and simple prognostic marker for infective endocarditis

Infective endocarditis is a life­threatining infectious disease characterized by high morbidity and mortality. Leukocytes play a main role in infectious diseases. Neutrophils and lymphocytes are subgroup of leukocytes, and they are routinely measured as a part of automated complete blood count test. The neutrophil­to­lymphocyte ratio is an independent predictor of unfavorable clinical outcomes in infectious and cardiovascular diseases.

Ontogeny of myeloid cells.

Granulocytes, monocytes, macrophages, and dendritic cells (DCs) represent a subgroup of leukocytes, collectively called myeloid cells. During the embryonic development of mammalians, myelopoiesis occurs in a stepwise fashion that begins in the yolk sac and ends up in the bone marrow (BM). During this process, these early monocyte progenitors colonize various organs such as the brain, liver, skin, and lungs and differentiate into resident macrophages that will self-maintain throughout life. DCs are constantly replenished from BM precursors but can also arise from monocytes in inflammatory conditions. In this review, we summarize the different types of myeloid cells and discuss new insights into their early origin and development in mice and humans from fetal to adult life. We specifically focus on the function of monocytes, macrophages, and DCs at these different developmental stages and on the intrinsic and environmental influences that may drive these adaptations.

The value of C-reactive protein in symptomatic versus asymptomatic carotid artery stenosis

Inflammatory markers may help establish the diagnosis of carotid atherosclerotic disease and predict the progression and outcomes. We evaluated inflammatory markers in symptomatic and asymptomatic patients with moderate to severe carotid artery stenosis, including high-sensitivity C-reactive protein (hs-CRP), leukocyte count, leukocyte subgroups and erythrocyte sedimentation rate. There were 32 symptomatic and 16 asymptomatic patients (age ≥50 years) with moderate to severe carotid stenosis detected by color Doppler ultrasonography, and 22 individuals without atherosclerotic findings on Doppler ultrasonography. The mean hs-CRP was significantly greater in the symptomatic (p = 0.001) and asymptomatic (p < 0.001) patients than control subjects; there was no significant difference between symptomatic and asymptomatic patients in mean hs-CRP values. Compared with control subjects, asymptomatic patients had greater mean leukocyte count (p = 0.016). Compared with control subjects, symptomatic patients had greater mean neutrophil count (p = 0.012), monocyte count (p = 0.021) and erythrocyte sedimentation rate (p = 0.001). The hs-CRP was associated with moderate to severe carotid artery stenosis, but it did not differentiate symptomatic and asymptomatic patients.

Effects of acute and 4-week submaximal exercise on leukocyte and leukocyte subgroups

The aim of this study is to investigate how does an acute 4-week submaximal exercise affect both leukocyte and leukocyte subgroups. The study registered 8 healthy male subjects. The subjects were exercised on Monark 814-E and 818 model ergometric bicycles 3 times a week for 4 weeks at above threshold heart rate and 75% of Max VO 2 . Blood samples were collected three times from the subjects, before starting the exercise, immediately after finishing the last exercise and 2 hours after the last exercise. The samples were analyzed in HemaCell Counter (Skits) apparatus to quantify leukocyte, neutrophil, monocyte, eosinophil and basophil levels. Leukocyte counts obtained in both post-exercise measurements were significantly higher than those before the exercise (p < 0.05). Neutrophil percentages two hours after exercise were higher than those before and immediately after exercise (p < 0.05). Lymphocyte percentages obtained just after exercise were elevated in comparison to those both before and two hours after exercise (p < 0.05). However, lymphocyte percentages two hours after exercise were significantly lower than the pre- and post-exercise percentages (p < 0.05). Percentages of monocytes, eosinophils and basophils in both post-exercise measurements were lower relative to pre-exercise values (p<0.05). In conclusion both acute and 4-week submaximal exercise have significant effects on leukocyte and leukocyte subgroups.

A Truncating Mutation in SERPINB6 Is Associated with Autosomal-Recessive Nonsyndromic Sensorineural Hearing Loss

More than 270 million people worldwide have hearing loss that affects normal communication. Although astonishing progress has been made in the identification of more than 50 genes for deafness during the past decade, the majority of deafness genes are yet to be identified. In this study, we mapped a previously unknown autosomal-recessive nonsyndromic sensorineural hearing loss locus (DFNB91) to chromosome 6p25 in a consanguineous Turkish family. The degree of hearing loss was moderate to severe in affected individuals. We subsequently identified a nonsense mutation (p.E245X) in SERPINB6, which is located within the linkage interval for DFNB91 and encodes for an intracellular protease inhibitor. The p.E245X mutation cosegregated in the family as a completely penetrant autosomal-recessive trait and was absent in 300 Turkish controls. The mRNA expression of SERPINB6 was reduced and production of protein was absent in the peripheral leukocytes of homozygotes, suggesting that the hearing loss is due to loss of function of SERPINB6. We also demonstrated that SERPINB6 was expressed primarily in the inner ear hair cells. We propose that SERPINB6 plays an important role in the inner ear in the protection against leakage of lysosomal content during stress and that loss of this protection results in cell death and sensorineural hearing loss.

Hematologic and serum biochemical reference intervals for free-ranging common bottlenose dolphins (Tursiops truncatus) and variation in the distributions of clinicopathologic values related to geographic sampling site

To develop robust reference intervals for hematologic and serum biochemical variables by use of data derived from free-ranging bottlenose dolphins (Tursiops truncatus) and examine potential variation in distributions of clinicopathologic values related to sampling sites' geographic locations. 255 free-ranging bottlenose dolphins. Data from samples collected during multiple bottlenose dolphin capture-release projects conducted at 4 southeastern US coastal locations in 2000 through 2006 were combined to determine reference intervals for 52 clinicopathologic variables. A nonparametric bootstrap approach was applied to estimate 95th percentiles and associated 90% confidence intervals; the need for partitioning by length and sex classes was determined by testing for differences in estimated thresholds with a bootstrap method. When appropriate, quantile regression was used to determine continuous functions for 95th percentiles dependent on length. The proportion of out-of-range samples for all clinicopathologic measurements was examined for each geographic site, and multivariate ANOVA was applied to further explore variation in leukocyte subgroups. A need for partitioning by length and sex classes was indicated for many clinicopathologic variables. For each geographic site, few significant deviations from expected number of out-of-range samples were detected. Although mean leukocyte counts did not vary among sites, differences in the mean counts for leukocyte subgroups were identified. Although differences in the centrality of distributions for some variables were detected, the 95th percentiles estimated from the pooled data were robust and applicable across geographic sites. The derived reference intervals provide critical information for conducting bottlenose dolphin population health studies.

Oncofoetal fibronectin – a tumour-specific marker in detecting minimal residual disease in differentiated thyroid carcinoma

Supposedly, thyrocyte-specific transcripts such as thyroglobulin (Tg) and thyroid-stimulating hormone receptor (TSH-R) were proposed to be useful for the diagnosis of circulating tumour cells in patients suffering from differentiated thyroid carcinoma (DTC). However, several research groups reported blood-borne Tg transcripts in healthy individuals. This study determines in particular the origin of Tg mRNA in nucleated blood cells and analyses whether other tumour-associated sequences are absent in leukocytes, but widely expressed in DTC. Therefore, expression analyses for Tg, TSH-R, cytokeratin 19 (CK 19), human telomerase reverse transcriptase (hTERT) and oncofoetal fibronectin (onfFN) were carried out using cDNAs derived from (1) leukocyte fractions, (2) 18 follicular thyroid carcinomas (FTCs) and 48 papillary thyroid carcinomas (PTCs), and (3) leukocytes of two thyrocyte-depleted individuals treated for C-cell carcinoma of the thyroid. Expression of onfFN was additionally analysed by semiquantitative RT–PCR and by quantitative fluorescence-based real-time PCR. Tg and TSH-R expression was demonstrated not only in both athyroid individuals, but in all leukocyte subgroups tested, while hTERT was absent in resting CD4+ cells and only weakly expressed in the CD8+ group. CK 19 was notable in each leukocyte population except for resting CD14+, as well as for activated and resting CD19+ cells. All blood cell fractions proved negative for onfFN mRNA, whereas its presence in thyroid carcinoma was 78/98% (FTC/PTC). Threshold cycle values were calculated at: porphobilinogen deaminase (PBGD) =25.95±0.73 (FTC)/24.55±5.43 (PTC) (P=0.2878); onfFN=25.48±3.15 (FTC)/21.44±3.44 (PTC) (*P=0.0001). Finally, onfFN transcripts were detected in blood samples of six out of nine patients with known DTC metastases, demonstrating a reliable assay functionality. We propose that real-time RT–PCR of onfFN mRNA is superior to other markers in monitoring minimal residual disease in DTC with regard to both assay sensitivity and specificity.

Monoclonal antibodies identify four distinct annelid leukocyte markers

This paper describes for the first time the production and characterization of a library of monoclonal antibodies (anti-EFCC clones) raised against coelomocyte (leukocyte) markers of Eisenia fetida earthworm. Leukocyte subgroups are components of earthworm innate immunity that require a more precise characterization using immunological markers. Flow cytometry, immunocytochemistry, and immunoprecipitation analyzed and confirmed the specificity of anti-EFCC clones. Anti-EFCC mAbs revealed different leukocyte subpopulations and various staining patterns on tissues. Two functional assays (e.g. phagocytosis and encapsulation) further characterized EFCC clusters revealing a common coelomocyte marker and three subpopulation-specific markers. No crossreactivity was found on human, mouse, rat or cells from Drosophila melanogaster but immunoreactivity was detected on snail ( Planorbarius corneus) tissues. Immunohistochemical results suggest mesodermal origin of all coelomocyte subgroups that agree with classical morphological analyses.

Zellpopulationen in der Cervix uteri bei der periovulatorischen Muttermundsöffnung

Die mittzyklische Zervixöffnung ähnelt - wenn auch klinisch wesentlich geringer ausgeprägt - der Muttermundsöffnung am Geburtstermin.