Effects of paclitaxel and resveratrol on blood characteristics in rabbits.

We investigated the adverse effects of paclitaxel (PAC) on blood characteristics including the percentage of alpha naphthyl acetate esterase (ANAE)-positive peripheral blood lymphocytes, white cell count, number of neutrophil nuclear lobe and, erythrocyte diameter, and the protective effects of resveratrol (RES) on these characteristics. Male New Zealand rabbits were assigned to four groups. The control group (C) was given 40 ml normal saline, the PAC group was given 5 mg/kg PAC in 40 ml normal saline, the RES group was given 4 mg/kg RES in 40 ml normal saline, and the PAC + RES group was given 5 mg/kg PAC + 4 mg/kg RES in 40 ml normal saline. All injections were performed intravenously once/week for 8 weeks. PAC caused an increased total percentage of lymphocytes and monocytes, which was associated with neutropenia. The PAC group showed a right shift in the lobulation curve of neutrophil nuclei together with a decreased percentage of ANAE-positive lymphocytes. RES reduced the percentage of ANAE-positive lymphocytes, slightly increased the percentage of neutrophil leukocytes and reduced the total lymphocyte count. Administration of RES combined with PAC prevented, while PAC induced, neutropenia and lymphocytosis, and increased the percentage of ANAE-positive lymphocytes.

Effects of long term oral acrylamide administration on alpha naphthyl acetate esterase and acid phosphatase activities in the peripheral blood lymphocytes of rats.

Acrylamide is an important industrial chemical; it also is formed in starch-rich foodstuffs during baking, frying and roasting. Most acrylamide exposure occurs by ingestion of processed foods. We investigated possible immunotoxic effects of extended administration of low doses of acrylamide in rats. To do this, we measured alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) activities in peripheral blood lymphocytes. Male and female weanling Wistar rats were administered 2 or 5 mg acrylamide/kg/day in drinking water for 90 days. Peripheral blood was sampled at the end of the administration period. We found ANAE staining in eosinophils and T-lymphocytes, but not in monocytes, platelets, B-lymphocytes and neutrophils. ACP-ase was found in B-lymphocytes. We found a significant reduction of the ratio of ANAE:ACP-ase in lymphocytes of the experimental animals compared to controls. We found no statistically significant differences between the doses or sexes. We found that acrylamide ingested in processed foods might affect the immune system adversely by decreasing the population of mature T- and B-lymphocytes.

Protective effect of quercetin on some hematological parameters in rats exposed to cadmium.

We investigated the effects of quercetin (Q) on some hematological parameters and determined the percentage of alpha-naphthyl acetate esterase (ANAE) positive lymphocytes in rats that had been exposed to cadmium (Cd). Thirty male Wistar albino rats were divided into four groups: control (C), quercetin (Q), cadmium (Cd) and Q + Cd (CdQ). Blood samples were taken to assess erythrocytes (RBC), leukocytes (WBC), hemoglobin levels (Hb), hematocrit values (Hct), platelets (PLT), alpha-naphthyl acetate esterase (ANAE) positive lymphocytes. RBC, Hb, Hct; the number of PLT significantly decreased in the Cd group. To the contrary, these parameters were increased significantly in the CdQ group compared to the Cd group. Although we found a significant increase in total WBC count and neutrophil percentage, the number of lymphocytes decreased in the Cd group compared to the other three groups. Also, the percentage of peripheral blood ANAE positive lymphocytes decreased significantly in the Cd group (p < 0.05). Q exhibits positive effects on some hematological characteristics and the percentage of ANAE positive lymphocyte in cases of acute CD toxicity.

Cytochemical characterization of peripheral blood cell populations of two Cyprinidae, Carassius auratus and Ctenopharyngodon idellus.

Fish are the most diverse species of all vertebrate groups, and their blood cells have shown variable characteristics in terms of morphology. Cytochemical staining for enzyme activity in blood leukocytes will help assess the immune function of fish. We characterize blood cells from crucian carp (Carassius auratus) and grass carp (Ctenopharyngodon idellus) by using a Diff-Quick stain as well as different cytochemical methods. Blood specimens obtained from crucian carp and grass carp were evaluated after cytochemical staining for acid phosphatase (ACP), alkaline phosphatase (ALP), naphthol AS chloroacetate esterase (AS-DNCE), naphthyl acetate esterase (NAE), α-naphthyl butyrate esterase (NBE), peroxidase (MPO) and periodic acid-Schiff's reaction (PAS) using commercial kits. Blood cell types were evaluated based on their morphological characteristics and the presence or absence of specific chromogen. The expression pattern of enzymes was similar between the two Cyprinidae and was also broadly consistent with other fish species. However, there were some interesting differences detected between crucian carp and grass carp, including naphthol AS chloroacetate esterase activity in monocytes, peroxidase activity and location in thrombocytes. The ACP, ALP and MPO expressions of different leukocytes of the two Cyprinidae were evaluated by Image Pro Plus and were analysed for statistical significant differences. This investigation provides basic haematology and enzyme activity analyses for crucian carp and grass carp and serves as an approach to evaluating the immune response of fish.

Cytochemistry of peripheral blood leukocytes in thoroughbred foals.

We attempted to characterize the cytochemical staining patterns of leukocytes and to determine the percentages of alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACPase) positive lymphocytes in peripheral blood of thoroughbred foals at different ages. Blood samples were obtained from the jugular veins of 60 healthy thoroughbred foals, 1 day, 3 days and 1 year old. Each age group included 10 male and 10 female animals. Peroxidase (PO) activity was detected in neutrophils, eosinophils and monocytes. Lymphocytes were negative for PO staining. Periodic acid-Schiff (PAS) staining was observed in neutrophils and monocytes; eosinophils were negative. The majority of lymphocytes were negative, but a few cells showed granular PAS positivity. Monocytes were strongly positive for ANAE and ACPase, and the enzymatic reaction was common in lymphocytes. Neutrophils showed a weakly positive reaction for ANAE and ACPase. Eosinophil granules were negative or weakly positive for ANAE and usually negative for ACP, but intergranular areas were positive. Mean ANAE positive peripheral blood lymphocytes (PBL) were 67.70, 73.10 and 49.20% in females; 64.00, 70.53, and 50.60% in males 1 day, 3 days, and 1 year old, respectively. ACPase positive PBL were 27.33, 32.83, and 37.40% in females; 29.67, 31.67, and 38.40% in males 1 day, 3 days, and 1 year old, respectively. For both enzymes, the differences between mean values for the genders were not statistically significant, but significant differences were found with regard to age. We provide comparative hematological data as a guide for identifying blood cells in thoroughbred foals.

Serum chitotriosidase, a putative marker of chronically activated macrophages, increases with normal aging.

BACKGROUND: Chitotriosidase (ChT) is secreted by chronically activated macrophages in Gaucher's disease. We hypothesize that circulating levels of ChT are altered with normal aging, reflecting age-related chronic macrophage activation. Potential sources that might contribute to altered levels were assessed by measuring systemic levels of ChT are α-naphthyl acetate esterase, a macrophage lysosomal enzyme; granulocyte-macrophage colony-stimulating factor (GM-CSF), which stimulates neutrophilic granule release of ChT; interleukin-6 (IL-6); and neopterin, a macrophage activation marker. METHODS: Serum was obtained from 315 healthy participants whose age ranged from 18 to 92 years. Anthropometric measures included percent body fat and body mass index. ChT and α-naphthyl acetate esterase levels were measured by enzyme activity assays. GM-CSF, IL-6, and neopterin concentrations were measured by commercial enzyme-linked immunosorbent assays. Serum marker values were statistically analyzed using nonparametric tests. RESULTS: Six percent of the participants had undetectable ChT levels. A positive association with age was observed for ChT and IL-6, whereas a negative correlation with age was seen for α-naphthyl acetate esterase and GM-CSF. ChT values were not associated with α-naphthyl acetate esterase or GM-CSF levels. ChT was independently associated with IL-6 and neopterin levels, but statistical significance was attenuated when controlled for age. CONCLUSIONS: The data are consistent with increased serum ChT activity not arising from altered macrophage lysosomal enzyme trafficking or GM-CSF-stimulated release of neutrophil granule stores. The association of ChT with age remains significant after controlling for neopterin and IL-6 changes with age, suggesting that ChT levels reflect a macrophage state distinct from acute macrophage activation or inflammatory state.

A study of peripheral blood in hedgehogs in Turkey.

The aim of this study was to determine diameters of blood cells, differential counts of peripheral blood leukocytes, alpha-naphthyl acetate esterase (ANAE), acid phosphatase (ACP-ase) activity of some leukocyte types, and enzymatic positivity percentages of peripheral blood lymphocytes in two hedgehogs species, Hemiechinus auritus, the long-eared hedgehog, and Erinaceus concolor, the southern white-breasted hedgehog. Air-dried peripheral blood smears were stained with May-Grünwald-Giemsa stain. ANAE and ACP-ase were stained in glutaraldehyde-acetone-fixed smears. ANAE-positive lymphocytes displayed a dot-like positivity pattern characterized with 1-5 reddish brown cytoplasmic granules, whereas ACP-ase positive lymphocytes displayed a dot-like positivity pattern characterized with 1-3 pinkish cytoplasmic granules. Monocytes gave a diffuse and strong reaction while neutrophils displayed a weak positive reaction for ANAE and ACP-ase. No difference was observed in mean diameters of peripheral blood cells of these species. It was found that lymphocytes made up the majority (64.3% and 65.5%) of leukocytes, followed by neutrophils (23.9% and 23.3%), eosinophils (9.0% and 7.6%), monocytes (1.8% and 2.3%), and basophils (1.0% and 1.3%) in H. auritus and E. concolor, respectively. Mean ANAE positivity oflymphocytes was 36.6% and 51.3% and ACP-ase positivity was 32.1% and 37.5% for H. auritus and E. concolor, respectively. The ANAE positivity of lymphocytes in E. concolor was significantly (P < 0.05) higher than that of H. auritus.

Leukocyte transmigration in inflamed liver: A role for endothelial cell-selective adhesion molecule.

BACKGROUND/AIMS: This study was designed to investigate the role of endothelial cell-selective adhesion molecule (ESAM), a recently discovered receptor expressed in endothelial tight junctions and platelets, for leukocyte migration in inflamed liver. METHODS: The role of ESAM for leukocyte migration in the liver was analyzed using ESAM-deficient mice in a model of warm hepatic ischemia-reperfusion (90min/30-360min). RESULTS: As shown by immunostaining, ESAM is expressed in sinusoids as well as in venules and is not upregulated upon I/R. Emigrated leukocytes were quantified in tissue sections. Postischemic neutrophil transmigration was significantly attenuated in ESAM-/- mice after 2h of reperfusion, whereas it was completely restored after 6h. In contrast, T-cell migration did not differ between ESAM+/+ and ESAM-/- mice. Using intravital microscopy, we demonstrate that ESAM deficiency attenuates I/R-induced vascular leakage after 30min of reperfusion. The I/R-induced elevation in AST/ALT activity, the sinusoidal perfusion failure, and the number of TUNEL-positive hepatocytes were comparable between ESAM+/+ and ESAM-/- mice. CONCLUSIONS: ESAM is expressed in the postischemic liver and mediates neutrophil but not T-cell transmigration during early reperfusion. ESAM deficiency attenuates I/R-induced vascular leakage and does not affect leukocyte adherence. Despite the effect on neutrophil migration, ESAM-deficiency does not protect from I/R-induced injury.

[Cytochemical activity of cells of the lymphocytic-macrophageal system in patients with chronic acquired toxoplasmosis].

The peripheral blood lymphocytic activities of acid phosphatase (AP) and acid nonspecific esterase (ANE) (alpha-naphthylacetate esterase) were determined in 45 patients with chronic acquired toxoplasmosis on an exacerbation. There was a considerable increase in the activity of AP in the white blood cells, which being more pronounced in the monocytes. The activity of ANE was moderately enhanced in the lymphocytes. The determination of the leukocytic activities of AP and ANE may be used as additional tests in the diagnosis of an exacerbation of chronic acquired toxoplasmosis and in the evaluation of the efficiency of the treatment performed.

Hematology and enzyme histochemistry of the peripheral blood leucocytes in rock partridges (Alectoris graeca).

The purpose of this study was to determine the percentages of alpha-naphthyl acetate esterase (ANAE)-positive and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL) and the percentages of leukocytes in the rock partridge at different ages. Blood samples obtained from the vena basilica of 18 healthy rock partridges (Alectoris graeca) at 1 d, 5 wk, and 12 wk of age were used. Mean percentages of ANAE-positive PBL for the 3 age groups were 37, 29.83, and 47.83% for 1 d, 5 wk, and 12 wk of age, respectively. Heterophils also gave ANAE-positive reactions. Mean percentages of ACP-positive PBL for the 3 age groups were 70, 81, and 86.1% for 1 d, 5 wk, and 12 wk of age, respectively. Although monocytes showed a diffuse granular staining pattern, heterophils displayed a weak positive reaction for ACP. Thrombocytes showed a small granular staining pattern. This study contributes by broadening the hematological research on avian species and provides a guideline for identifying blood cells in the rock partridge.

Histologic characteristics and local cellular immunity of the gland of the third eyelid after topical ophthalmic administration of 2% cyclosporine for treatment of dogs with keratoconjunctivitis sicca.

OBJECTIVE: To evaluate the efficacy of topical administration of a 2% solution of cyclosporine (CsA) for treatment of dogs with keratoconjunctivitis sicca (KCS) and to correlate results with histopathologic characteristics and local cellular immunity of the gland of the third eyelid. ANIMALS: 24 dogs with bilateral KCS. PROCEDURE: Lacrimal secretion was measured, using Schirmer tear test (STT) strips. Leukocyte and T-lymphocyte subsets were determined in blood samples. Histopathologic changes as well as CD4+, CD8+, and alpha-naphthyl-acetate esterase-positive (ANAE+) lymphocytes were evaluated. RESULTS: Clinical signs resolved at the end of 1 month in conjunction with significantly increased STT values, compared with baseline values. Fifteen and 30 days after discontinuation of CsA treatment, a decrease was observed in STT values in both eyes; however, only values for the right eye were significantly different. There was a significant decrease in the number of lymphocytes and ANAE+ lymphocytes 15 and 30 days after discontinuation of CsA treatment, compared with baseline values. Differences were not observed in number of CD4+ lymphocytes among treatment groups. However, there was a significant decrease in number of CD8+ lymphocytes with reversal of the CD4+:CD8+ in both eyes after CsA treatment for 30 days, compared with the control group. Increased secretory activity and decreased lymphocyte infiltration were characteristic histopathologic findings. CONCLUSIONS AND CLINICAL RELEVANCE: Topical administration of a 2% solution of CsA was effective for the treatment of dogs with KCS. Strict follow-up monitoring is required after the cessation of treatment because of the possibility of recurrence of KCS.

Light and electron microscopic studies on alpha naphthyl acetate esterase activity of peripheral blood T lymphocytes in chicken.

The purpose of this study was to determine the percentage of peripheral blood T lymphocytes and the localization of Alpha Naphthyl Acetate Esterase (ANAE) enzyme at the electron microscopic level in healthy adult chickens by an alpha naphthyl acetate esterase procedure. Peripheral blood samples taken from 30 adult Leghorn breed chickens were used. The percentage of ANAE positive lymphocytes was 56.4 +/- 2.82% (SD). Pseudoeosinophil and eosinophil granulocytes showed a granular positivity. In the electron microscopic examination, ANAE positive reactions were seen in lysosomal granules found in T lymphocytes, pseudoeosinophil, and eosinophil granulocytes.

Light and electron microscopic studies on alpha naphthyl acetate esterase activity of the peripheral blood T lymphocytes in van cats.

The purpose of this study was to determine the percentage of peripheral blood T lymphocytes and the localization of ANAE enzyme at the electron microscopic level in Van cats by using an alpha-naphtyl acetate esterase procedure. Peripheral blood samples taken from 20 Van cats were used. The percentage of ANAE positive lymphocytes was 83.0%. Neutrophilic granulocytes gave a negative reaction, whereas monocytes, eosinophilic granulocytes showed a diffuse granular positivity. In the electron microscopic examination, ANAE positive reactions were seen in lysosomal granules found in lymphocytes.

Monocyte analysis in chronic myelomonocytic leukaemia.

It is suggested that monocytes in patients with chronic myelomonocytic leukaemia (CMML) or chronic myeloid leukaemia (CML) with monocytosis have morphological/functional abnormalities which cause inaccurate counting in automated analysers. In this study, monocytes in 21 normal and 14 CMML blood samples were subjected to morphological analysis and were counted by the manual reference method, three automated analysers and esterase staining. Morphological analysis showed no significant difference between control and CMML monocytes. The alpha-naphthyl acetate esterase scores, a measure of monocyte function, showed a reduction of 40% in CMML monocytes compared to controls. Counts by analysers showed that the Sysmex NE 8000 was the least accurate for CMML monocyte counts and that the Coulter STK-S and Sysmex SE 9000 gave results closer to the manually counted standards.

[Effect of acupuncture on immunologic function and histopathology of transplanted mammary cancer in mice].

This experiment mainly describes the effects of acupuncture on immunologic function and histopathology of transplanted mammary cancer in mice. The results were as follows: in acupuncture group, NK cell activity and T-lymphocyte positive rate of acid alpha-naphthyl acetate esterase (ANAE) and lymphocyte transformation rate were all increased. Compared with the control group, there was a significant difference (P < 0.01). The difference was insignificant, when compared with normal group (P > 0.05). Comparing the pathology grading of acupuncture group with control group, it showed marked difference in pathological section (P < 0.01). Adenoid structure and the degree of lymphocytic infiltration also have marked difference between acupuncture and control group (P < 0.05). Less tumour volume in acupuncture than control group were observed (P < 0.01). This indicated that acupuncture might increase the immunologic function of transplanted mammary cancer in mice and inhibit the growth of mammary cancer and enhance both differentiation level of mammary cancer cells and lymphocytic infiltration. Possibly acupuncture mightt reduce the malignancy of mammary cancer cells.

Enhancement of fibrinolytic activity of human plasma in the presence of acetone.

The fibrinolytic system was studied in normal human plasma containing increasing concentrations of acetone up to 23.4 mmol l-1. Fibrinolytic activity measured as euglobulin clot lysis time [ECLT] and amidase activities toward chromogenic peptide substrates H-D-Valyl-L-Leucyl-L-Lysine-p-nitroanilide 2 HCl [S-2251], designed for plasmin determination, H-D-Valyl-L-Phenylalanyl-L-Lysine-p-nitroanilide 2 HCl [S-2390], designed for the determination of t-PA in plasma via plasminogen activation and H-D-Prolyl-L-Phenyl-Alanyl-L-Arginine-p-nitro-anilide 2 HCl [S-2302], designed for the determination of kallikrein and activated Hageman factor, increased when 15.7 mmol l-1 concentration of acetone was reached. A parallel increase of esterolytic [substrate: naphthol-AS-acetate] activity was observed in euglobulin fractions. Crossed immunoelectrophoresis [CIE] revealed changes in fibrinogen profiles of plasma enriched with acetone as compared to native plasma. These findings suggest that acetone present in plasma in concentrations comparable to those found in some pathological states might activate fibrinolytic system.

Endotoxin-induced tissue factor in human monocytes is dependent upon protein kinase C activation.

Tissue factor (TF) is a transmembrane receptor which, in association with factors VII and VIIa, activates factor IX and X, thereby activating the coagulation protease cascades. In response to bacterial lipopolysaccharide (LPS) monocytes transcribe, synthesize and express TF on their surface. We investigated whether LPS-induced TF in human monocytes is mediated by protein kinase C (PKC) activation. The PKC agonists phorbol 12-myristate 13-acetate (PMA) and phorbol 12, 13 dibutyrate (PdBu) were both potent inducers of TF in human monocytes, whereas 4 alpha-12, 13 didecanoate (4 alpha-Pdd) had no such effect. Both LPS- and PMA-induced TF activity were inhibited, in a concentration dependent manner, by three different PKC inhibitors: H7, staurosporine and calphostin C. TF antigen determination confirmed that LPS-induced cell-surface TF protein levels decreased in parallel to TF functional activity under staurosporine treatment. Moreover, Northern blot analysis of total RNA from LPS- or PMA-stimulated monocytes showed a concentration-dependent decrease in TF mRNA levels in response to H7 and staurosporine. The decay rate of LPS-induced TF mRNA evaluated after the arrest of transcription by actinomycin D was not affected by the addition of staurosporine, suggesting that its inhibitory effect occurred at a transcriptional level. We conclude that LPS-induced production of TF and its mRNA by human monocytes are dependent on PKC activation.

[alpha-Naphthyl acetate esterase--a cytochemical marker of the T-helpers]. / al'fa-Naftilatsetatésteraza--tsitokhimicheskii marker T-khelperov.

The authors analyze the correlation between the frequency of detection of alpha-naphthylacetate esterase in peripheral blood lymphocytes and T helper/inductor ratio determined by immunological methods from CD-4 MCAB expression. The results indicate that the counts of esterase-positive lymphocytes cytochemically detected by alpha-naphthylacetate esterase activity reliably correlated with T helper counts assessed by immunoperoxidase method with Technicon H-1 flow analyzer.

Morphological and biochemical characterization of Procambarus clarki blood cells.

Morphological and cytochemical analysis of Procambarus clarki hemocytes demonstrated three cell types: hyaline, semigranular, and granular. Hyaline cells showed a higher nuclear/cytoplasmic ratio with few small electron-dense granules in the cytoplasm. Semigranular cells presented numerous round or oval eosinophilic granules (0.40-0.78 micron). Granular cell contained large eosinophilic granules (1.79-3.05 microns). Ultrastructurally, all cells showed microtubules near the borders, a poorly developed Golgi complex, and secretory-type electron-dense particles. No mitotic figures were seen. Cell monolayers showed three morphologically distinct cell types (composed of flattened and well-spread cells) depending on the presence and size of granules (hyaline, semigranular, and granular). No sex-related differences could be documented in cell features or proportions. Cytochemical studies showed that the three cell types were positive for acid phosphatase. Granular and semigranular cells were also positive for nonspecific esterase. Phenoloxidase activity was localized only in granular and semigranular hemocytes, and peroxidase activity was observed only in the granular hemocytes. These results may suggest that the semigranular and granular hemocytes participate in the prophenoloxidase system and also in phagocytic or cytotoxic function.

[Cytochemical evaluation of blast cells in acute myeloblastic leukemia after induction of their maturation in liquid culture--diagnostic usefulness]. / Ocena cytochemiczna komórek blastycznych ostrych bialaczek szpikowych indukowanych do dojrzewania w warunkach hodowli plynnej--przydatnosc diagnostyczna.

We have performed the cytochemical analysis (myeloperoxidase, ASD-chloroacetate and acid alpha-naphthylacetate esterases, Sudan black B) of blast cells from 25 acute leukemia patients, after 3, 5 and 7 days of liquid culture with conditioned medium from phytohemagglutinin stimulated leukocytes (PHA-LCM). In case of acute myeloid leukemia blast cells an increase of percentage of positive cells simultaneously with the enhancement of the cytochemical reactions was observed. This method may be useful for the precise diagnosis of poorly differentiated blasts with weak expression of cytochemical phenotype.