Neutrophil Alkaline Phosphatase Activity Research Articles

Early Diagnosis of Neonatal Sepsis Using the Leukocyte Alkaline Phosphatase Score: A Cohort Study

Introduction: The clinical features of sepsis in neonates are subtle and non specific, requiring a high index of suspicion for early diagnosis. Blood culture is the gold standard for diagnosis, but it is time consuming. Therefore, there is a need for a costeffective and reliable screening tool. The Leukocyte Alkaline Phosphatase (LAP) activity of neutrophils is known to increase during bacterial infections in adults. Aim: To determine the activity of LAP in Neonatal Sepsis (NS) and compare the results with blood culture. Materials and Methods: This is a prospective cohort study conducted from January 2018 to June 2019 at Mandya Institute of Medical Sciences, Karnataka, India. Total of 200 neonates by their haematological profile were clinically suspected of having sepsis. A peripheral smear was prepared from a drop of blood and stained to assess the LAP activity. In each smear, a total of 100 consecutive segmented neutrophils were examined and were rated from 0 to 4 according to the red granular precipitate intensity within their cytoplasm. The possible range of 0-400. The blood culture report was obtained from the case sheet. The neonates were divided into two groups: Group 1 consisted of culture-proven sepsis cases, and Group 2 consisted of neonates with clinical suspicion of sepsis but negative blood culture results. Descriptive analysis was performed using mean and standard deviation for quantitative variables, and frequency and proportion for categorical variables. Chi-square test was conducted, and a p-value <0.05 was considered statistically significant. Results: Out of the 200 cases studied, 64 neonates showed a positive blood culture. The most common causative organism observed was Klebsiella pneumoniae, which was seen in 56 neonates. The age of the neonates ranged from newborn to 28 days old, with 115 being male. Furthermore, 116 neonates were term neonates, and 146 presented with early onset sepsis. The study revealed a wide range of LAP activity in both groups. In the culture-positive sepsis group, the LAP activity ranged from 36 to 350, while in the clinical sepsis group, it ranged from 10 to 354. Due to the broad range of LAP scores observed, it is concluded that LAP activity is not useful as a screening test. Conclusion: Findings of the present study indicate that LAP activity exhibited a wide range in both the culture-proven and clinical sepsis groups. However, due to this variability, LAP activity does not appear to be a reliable screening test for NS. While LAP activity assessment may still hold diagnostic value, further research is needed to refine its utility and explore its clinical relevance.

Clinical and laboratory characteristics of chronic neutrophilic leukemia: analyses of five cases

Objective To investigate the clinical characteristics, cell morphology, genetics, gene mutations of the patients with chronic neutrophilic leukemia (CNL). Methods Five CNL patients from the Second Hospital of Shanxi Medical University between May 2011 and May 2017 who conformed to 2016 World Health Organization (WHO) diagnostic criteria were retrospectively analyzed from clinical characteristics, laboratory features and treatment methods. Results The peripheral blood white blood cell count (WBC) of 5 CNL patients was significantly increased, and the average WBC was 81.26×109/L [(29-217)×109/L]. Morphological analysis of peripheral blood cell showed a sustained increasing number of matured neutrophilia (0.80-0.85). Neutrophil alkaline phosphatase (NAP) activity was increased (144-266). Bone marrow cell morphology typically showed granulocyte proliferation without obvious dysplasia. Gene detection showed 3 patients with CSF3R T618I mutation and 2 patients with JAK2 V617F mutation in 5 WHO-defined CNL patients. Bone marrow biopsy with reticular staining showed that marrow fibrosis (MF) degree in patients with JAK2 V617F mutation (MF≥2) was higher than that in patients with CSF3R T618I mutation(MF<2). Conclusions CNL is a rare type of chronic leukemia, and CSF3R T618I mutation is a specific diagnostic index for CNL. JAK2 V617F mutations alone may be related to myelofibrosis, which remains to be further studied. Key words: Leukemia, myeloid, chronic-phase; Cellular morphology; Genes; Receptors, colony-stimulating factor; Janus kinase 2

NAP activity in white blood cells of workers exposing to benzene

Objective To study the activity of white blood cell neutrophil alkaline phosphatase (NAP) in the workers exposing to benzene.Methods The NAP of 207 workers exposing to benzene (exposing group) and 94 healthy people (normal control group) were tested and then compared.Results The average NAP activity level was significantly higher in the exposing group than in the control group[(67.40 ± 16.12) vs.(38.69 ± 11.53),P ＜ 0.001)].There was no statistical difference in average NAP activity level either by gender (P ＞ 0.05) or working-age(P ＞ 0.05).The average NAP activity level was significantly lower in highbenzene-concentration exposing group than in low-benzene-concentration exposing group [(57.27 ± 18.78) vs.(74.26 ± 15.81),P ＜ 0.001].The average NAP activity level was higher in high-benzene-concentration exposing group than in high-benzene-concentration retiree group[(57.27 ± 18.78) vs.(49.03 ± 16.62),P ＜ 0.005].Conclusions The average NAP activity level is higher in the exposing group than in the normal controls group.Developing a screening test for NAP level may be a promising tool in protecting benzeneexposing workers from chemical hazards. Key words: Benzene; Exposing workers ; Neutrophil alkaline phosphatase (NAP)

Absent or Extremely Low Neutrophil Alkaline Phosphatase Activity Levels in Patients with Myelodysplastic Syndromes

Three patients with myelodysplastic syndrome (MDS) had absent or extremely low levels of neutrophil alkaline phosphatase (NAP) activity (arbitrarily defined as an NAP score <10). All patients showed varying degrees of hypogranulation in neutrophil morphology. The NAP activity levels transiently normalized following the administration of granulocyte colony-stimulating factor (G-CSF) in two cases. No patients experienced any severe infectious episodes. These results suggest that NAP activity is not central to the neutrophil function.

Functional and metabolic assessment of the blood cell’s response to exposure to local vibration

Physiological characteristics of blood cells in workers exposed to long-term local vibration were studied. The total cell peroxidase activity, the level of the phagocytic response, neutrophil alkaline phosphatase and acid phosphatase activity levels, and glycogen content in neutrophils and lymphocytes were measured. Changes associated with a decrease in the functional metabolic parameters, depending on the duration of contact with vibrating tools and the severity of clinical symptoms, were detected. The detected differences in changes in cell response may reflect the specific features of the mechanisms of adaptive regulation.

Numerous aggregates of “tiny” micromegakaryocytes in the bone marrow

A 70-year old male presented with leukocytosis (24.9 9 10/L) and thrombocytosis (727 9 10/L). Hb was 14.9 g/dL, neutrophils 81%, basophils 1%, eosinophils 4%, and myelocytes 1%. He had no splenomegaly. Serum LDH was 477 U/L, B12755 pg/m, and ferritin 104 ng/dL. Neutrophil alkaline phosphatase activity was normal. The marrow was moderately cellular with many aggregates of micromegakaryocytes, each aggregate comprising up to 20 ‘‘tiny’’ micromegakaryocytes (Fig. 1). Small cells were close to the size of lymphocytes. Although usual-sized micromegakaryocytes were occasionally seen, most ‘‘tiny’’ micromegakaryocytes were found to form aggregates accompanying many platelets (Fig. 2a, b) and such aggregates were scattered in the entire length of marrow film. Immature granulocytes showed hypogranulation (Fig. 3a), and occasional mature cells showed hypersegmentation (Fig. 3b, c) and cytoplasmic Dohle bodies (arrowheads Fig. 3d). Erythroid series showed mild megaloblastoid changes. These features resembled myelodysplastic syndromes (MDS). Blast cells were few. Normal marrow cytogenetic analysis and the lack of BCR-ABL in mRNA analysis excluded chronic myeloid leukemia (CML). Marrow WT1 mRNA level was 1600 copies/lg RNA. No marrow fibrosis was seen in biopsy. These findings excluded myeloproliferative neoplasm (MPN). Proliferation and marked dysplasia in megakaryocytic and myeloid lineages falls within the scope of MDS/MPN, unclassifiable, according to the current WHO system, rather than atypical BCR-ABL negative CML. The marrow findings remained essentially unchanged in repeat study although blood counts oscillated in response to the intermittent hydroxyurea (1000 mg) treatment. Y. Yoshida (&) Y. Hamakawa S. Oguma T. Katsurada The Center for Hematological Diseases, Takeda General Hospital, 28-1 Ishida-Moriminami, Fushimi-Ku, Kyoto 601-1495, Japan e-mail: yoshida@takedahp.or.jp

好中球のアルカリフォスファターゼ活性，墨粒貪食能，nitroBT還元能について

好中球のアルカリフォスファターゼ活性，墨粒貪食能，nitroBT還元能について

C/G転座，好中球アルカリフォスファターゼ低値を示す急性骨髄性白血病病型:—15例の臨床的観察—

C/G転座，好中球アルカリフォスファターゼ低値を示す急性骨髄性白血病病型:—15例の臨床的観察—

Kawasaki disease with lymphadenopathy and fever as sole initial manifestations

Initial presentation with only cervical lymphadenopathy and fever is one of the pitfalls in the diagnosis of Kawasaki disease (KD). As the number of such patients is small, their clinical features have remained uncertain. The purpose of the present study is to characterise the features of such KD patients, especially in comparison with those of patients with common onset. We conducted a retrospective review of the medical records of 136 consecutive KD patients admitted to Kobe City General Hospital from April of 2000 to March of 2006. Twenty-nine of the 136 patients initially presented with only cervical lymphadenopathy and fever and were classified into the lymphadenopathy-KD (LKD); they were compared with the remaining 107 KD patients with other presentations (other-KD). Age, days of fever to diagnosis, and duration of fever were significantly higher or longer in LKD patients, who also showed higher C-reactive protein levels and neutrophil alkaline phosphatase activity. There were no significant differences between two groups in gender, duration of hospitalization, frequency of high-dose intravenous immunoglobulin (IVIG) administration, coronary artery lesions (CALs), white blood cell or platelet counts, and levels of hemoglobin or albumin on admission. Although a delay in diagnosis and stronger inflammation were found in LKD patients, such differences did not have any significant effect on patients' outcomes as assessed by the frequency of IVIG administration and the presence of CALs.

Neutrophil alkaline phosphatase activity in respiratory viral infection

Neutrophil alkaline phosphatase (NAP) is used as a diagnostic marker in several hematological disorders. In regard to the role of NAP in infectious diseases, previous investigators have presented the hypothesis that NAP activity is useful to distinguish viral infections from bacterial infections. Because the numbers of patients enrolled in previous studies of viral infections were limited, we intended to evaluate the hypothesis by measuring NAP activity in a large number of pediatric patients with respiratory viral infections. A cytochemical analysis of NAP was performed in 160 patients with various types of respiratory infections. In patients with adenovirus or respiratory syncytial (RS) virus infection, NAP activity was significantly higher than the control value newly established at our department, while in patients with Epstein-Barr virus, measles, or influenza infection, it was comparable to the control value. On an individual basis, NAP scores (determined from NAP cytochemical activity) in 22 of 26 patients (84.6%) with adenovirus infection, and 31 of 42 patients (73.8%) with RS virus infection were found to exceed the 95% confidence upper limit of the control group. In conclusion, NAP activity is quite varied among different respiratory viral infections. When NAP activity is high in respiratory infections, adenovirus or RS virus infection, as well as bacterial infections, should be taken into consideration.

Neutrophil alkaline phosphatase activity in respiratory viral infection

<h2>Abstract</h2> Neutrophil alkaline phosphatase (NAP) is used as a diagnostic marker in several hematological disorders. In regard to the role of NAP in infectious diseases, previous investigators have presented the hypothesis that NAP activity is useful to distinguish viral infections from bacterial infections. Because the numbers of patients enrolled in previous studies of viral infections were limited, we intended to evaluate the hypothesis by measuring NAP activity in a large number of pediatric patients with respiratory viral infections. A cytochemical analysis of NAP was performed in 160 patients with various types of respiratory infections. In patients with adenovirus or respiratory syncytial (RS) virus infection, NAP activity was significantly higher than the control value newly established at our department, while in patients with Epstein-Barr virus, measles, or influenza infection, it was comparable to the control value. On an individual basis, NAP scores (determined from NAP cytochemical activity) in 22 of 26 patients (84.6%) with adenovirus infection, and 31 of 42 patients (73.8%) with RS virus infection were found to exceed the 95% confidence upper limit of the control group. In conclusion, NAP activity is quite varied among different respiratory viral infections. When NAP activity is high in respiratory infections, adenovirus or RS virus infection, as well as bacterial infections, should be taken into consideration.

Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro

We have proposed the use of the fluorimetric method with 4-methylumbelliferyl phosphate (4-MUP) specific substrate for the alkaline phosphatase determination in the neutrophil adhesion assay. We provide evidence that the endogenous neutrophil alkaline phosphatase (NAP) activity evaluation is reliable to quantify neutrophil adhesion at a wide range of cell numbers (104−106). The results obtained by fluorimetric NAP activity test correlate to the results of adherence evaluated using the MTT reduction assay. The fluorimetric NAP activity test may be applied for resting as well as activated neutrophils without the risk of the activators interferences into the test. The alkaline phosphatase survey with the use of 4-MUP substrate is recommended herein as a sensitive, repeatable, simple, and reliable method of the neutrophil adherence determination in vitro.

Neutrophil Alkaline Phosphatase Activity in Turner Syndrome

Neutrophil Alkaline Phosphatase Activity in Turner Syndrome

All-Trans-Retinoic-Acid- and Growth-Factor- Mediated Induction of Alkaline Phosphatase Activity in Freshly Isolated Chronic Myeloid Leukemia Cells

Reduced or absent neutrophil alkaline phosphatase (NAP) activity is a common feature of neutrophilic granulocytes from patients with chronic myeloid leukemia (CML). In this study we examined whether NAP activity could be restored in vitro by stimulating CML cells with different promoters such as all-trans-retinoic acid (ATRA), granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF). The results obtained indicated that ATRA and G-CSF, either alone or in combination, were effective in inducing NAP activity in CML cells, whereas GM-CSF was not. Further, NAP restoration in ATRA- and G-CSF-treated cultures was accompanied by increased morphologic differentiation of the CML clone. It might be concluded that the CML clone could be driven in vitro by ATRA and G-CSF both to achieve granulocytic maturation and to correct functional NAP-related defects.

Heat stable and urea resistant alkaline phosphatase in maternal neutrophils from normal and Down syndrome pregnancies

Activity levels of total and placental alkaline phosphatase (AP) were determined in maternal serum and neutrophils of normal and Down syndrome pregnancies. The placental iso-enzyme (PAP) was identified by its relative stability to urea and heat. Significant increase in the activity of all iso-enzymes across gestational stages was observed in maternal sera of 28 normal pregnancies. However, in the neutrophil extracts of the same blood samples no differences were detected between trimesters. Another set of experiments was aimed at finding diagnostic differences of PAP activity in maternal neutrophils of normal and trisomy 21 affected pregnancies. No differences of heat stable AP activity were found in maternal samples of 19 normal and 19 Down syndrome affected pregnancies. Urea resistant AP proportions were also similar when measured after 40 minutes of exposure (13 samples in each group). However, a marginally significant increase was observed in the mean value of the Down syndrome affected samples, after 60 minutes of urea treatment. In view of the above results we conclude that neutrophil AP activity is not as yet a useful marker for the screening of trisomy 21 fetuses. Copyright © 1999 John Wiley & Sons, Ltd.

Heat stable and urea resistant alkaline phosphatase in maternal neutrophils from normal and Down syndrome pregnancies

Activity levels of total and placental alkaline phosphatase (AP) were determined in maternal serum and neutrophils of normal and Down syndrome pregnancies. The placental iso-enzyme (PAP) was identified by its relative stability to urea and heat. Significant increase in the activity of all iso-enzymes across gestational stages was observed in maternal sera of 28 normal pregnancies. However, in the neutrophil extracts of the same blood samples no differences were detected between trimesters. Another set of experiments was aimed at finding diagnostic differences of PAP activity in maternal neutrophils of normal and trisomy 21 affected pregnancies. No differences of heat stable AP activity were found in maternal samples of 19 normal and 19 Down syndrome affected pregnancies. Urea resistant AP proportions were also similar when measured after 40 minutes of exposure (13 samples in each group). However, a marginally significant increase was observed in the mean value of the Down syndrome affected samples, after 60 minutes of urea treatment. In view of the above results we conclude that neutrophil AP activity is not as yet a useful marker for the screening of trisomy 21 fetuses.

Transition of Polycythemia Vera to Chronic Neutrophilic Leukemia

Two cases of polycythemia vera (PV) had transition to a hematological condition compatible with chronic neutrophilic leukemia (CNL) 17 and 8 years after diagnosis, respectively. One patient was treated with carboquone followed by hydroxyurea (HU) and the other with HU during PV phase. On transition, both had neutrophilia with white blood cell count above 40,000/μl, elevated neutrophil alkaline phosphatase activity, splenomegaly, normal karyotype without bcr-abl rearrangement. Busulfan was temporally effective in controlling the neutrophil count. However, one patient progressed to the so-called spent phase and the other subsequently had multiple transitions between PV and CNL. These cases may represent a form of uncommon evolution of PV and support the contention that CNL is a type of myeloproliferative disorder and that at least some CNL cases have derangement at the hematopoietic stern cell level.

Atypical neutrophil alkaline phosphatase associated with impaired neutrophil functions

We report the case of a healthy young man presenting with atypical neutrophil alkaline phosphatase (NAP) and reduced neutrophil chemotactic activity, but with no susceptibility to infection. NAP activity was low, kinetic parameters were modified and immunoreactive properties and subcellular distribution were abnormal. Neutrophil morphology was normal. A similar pattern was observed in the patient's healthy brother. The profile of the observed anomalies offers some similarity to that previously described in patients with chronic myelogenous leukaemia. However, in the present case, the NAP deficiency with impaired neutrophil function was present in two brothers with no haematological symptoms and is probably related to a non-acquired neutrophil abnormality. This observation of a primary NAP variant reinforces the hypothesis of a direct link between NAP activity and functional properties of neutrophils.

Flow cytometric measurement of neutrophil alkaline phosphatase before and during initiation of an induced Escherichia coli mastitis in cattle

In 12 healthy cows, neutrophil alkaline phosphatase (NAP) activity was measured by flow cytometer before and during an experimentally induced Escherichia coli mastitis, to study the role and increase of NAP in Gram-negative bacterial infections. Percentage of neutrophils containing alkaline phosphatase and intensity of NAP activity were measured. Preinfection percentage of neutrophils with enzyme activity varied between 64.0% and 84.4% and the intensity of enzyme activity was low in all cows. After induction of infection, percentage of neutrophils with enzyme activity showed a significant decrease on day 1 followed by an significant increase on day 3. NAP intensity increased significantly on the second and third day after infection. This increase of intensity was significantly, positively correlated with the severity of infection. From this study we may conclude that variation in susceptibility to E. coli mastitis could not be explained by preinfection NAP levels. The post-infection increase of NAP activity, that was found following an induced infection was more a result of increased enzyme intensity per neutrophil, then from an increase of percentage neutrophils with enzyme activity. Furthermore, a strong correlation was found between NAP intensity and severity of inflammation. There was evidence that the more severely diseased animals showed stronger NAP intensity increase.

Elevation in Plasma Alkaline Phosphatase Level during rhG-CSF Administration

Neutropenic patients suffering from gynecologic cancers treated with cancer chemotherapy were evaluated to investigate changes in levels of plasma alkaline phosphatase (ALP) during recombinant human granulocyte colony stimulating factor (rhG-CSF) administration, and to discern its causes. Plasma ALP, ALP isozymes, neutrophil alkaline phosphatase (NAP) activity, and morphological maturation of neutrophils were measured prior to, during, and after rhG-CSF administration. Plasma ALP values were significantly elevated (p < 0.05) during administration of rhG-CSF. ALP-3, the dominant alkaline phosphatase fraction of NAP, was the dominant isozyme that showed an increase in the plasma. Moreover, the elevation of plasma ALP-3 significantly correlated with the elevation of NAP activity (r = 0.939, p < 0.0001), and neutrophils in which NAP activity was induced were not limited to morphologically mature neutrophils. These results showed that rhG-CSF acts to stimulate neutrophils at all stages of maturation and causes an elevation of plasma ALP(ALP-3) concentrations in plasma.