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The aim of the study was to investigate the effects of chitosan oligosaccharide (COS) on some hematological parameters, and define the percentage of alpha-naphthyl acetate esterase (ANAE) positive lymphocytes in rats that had been exposed to cadmium (Cd). Thirty-two (n = 32) male Wistar albino rats were randomly divided into four groups as the control (C), chitosan oligosaccharide (COS), cadmium (Cd), and Cd + COS (CdCOS) groups. Blood samples were collected to assess erythrocytes (RBC), leukocytes (WBC), hemoglobin levels (HGB), hematocrit values (HCT), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), platelets (PLT), and alpha-naphthyl acetate esterase (ANAE) positive lymphocytes. The number of WBCs significantly increased in the Cd group compared to the C, CdCOS, and COS groups (P, Cilj ovog rada bio je istražiti učinak oligosaharida hitozana (COS) na određene hematološke pokazatelje odnosno postotak limfocita pozitivnih na alfa-naftil-acetat-esterazu (ANAE) u štakora izloženih kadmiju (Cd). Mužjaci (n = 32) pasmine Wistar albino, slučajnim odabirom podijeljeni su u četiri skupine: kontrolnu (C), skupinu s oligosaharidom hitozanom (COS), skupinu s kadmijem (Cd) i skupinu s kadmijem i oligosaharidom hitozanom (CdCOS). Prikupljeni su uzorci krvi kako bi se odredili eritrociti (RBC), leukociti (WBC), hemoglobin (HGB), hematokrit (HCT), prosječni hemoglobin u eritrocitima (MCH) i njegova koncentracija (MCHC), prosječan obujam eritrocita (MCV), trombociti (PLT), te limfociti pozitivni na alfa-naftil-acetat-esterazu (ANAE). Broj leukocita znakovito je porastao u skupini Cd u usporedbi s kontrolnom skupinom te skupinama C, CdCOS, i COS (P < 0,05). Iako je broj limfocita znakovito smanjen u skupini Cd (P < 0,05), to je ublaženo primjenom COS-a u skupini CdCOS u usporedbi sa skupinom Cd (P < 0,05). Također, postotak limfocita iz periferne krvi pozitivnih na ANAE znakovito je smanjen u skupinama CdCOS i Cd (P < 0,05). COS je, u slučaju kronične izloženosti štakora kadmiju, pokazao djelomice zaštitni učinak na određene hematološke značajke, osim na postotak limfocita pozitivnih na ANAE.

We investigated the effects of melatonin (Mlt) on hematological characteristics to determine the percentage of alpha-naphthyl acetate esterase (ANAE) positive lymphocytes in rats chronically exposed to cadmium (Cd). We used four groups of male rats: control (C), melatonin (Mlt), cadmium (Cd) and Cd + Mlt (CdMlt). Blood samples were obtained to assess the erythrocytes (RBC), leukocytes (WBC), platelets (PLT), hemoglobin (HGB), hematocrit (HCT), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV) as well as alpha-naphthyl acetate esterase (ANAE) positive lymphocytes. We found that the number of WBC increased in the Cd group compared to the other three experimental groups. Low MCV and MCHC values caused by Cd were ameliorated by Mlt treatment in the CdMlt group. We found no significant change in the RBC, HGB, MCH, PLT and HCT values among the experimental groups. Mlt treatment also increased the percentage of ANAE positive lymphocytes. Mlt partially protects some hematological characteristics in cases of chronic Cd toxicity.

A 4-year, 7-month-old Holstein cow presented with anorexia. Physical examination revealed masses in the interscapular region and vagina. Blast cells were detected in the masses and peripheral blood by fine needle aspiration cytology and hematological examination. By bone marrow aspiration, blast cells constituted up to 24.2% of all nucleated cells, and 22% and 2% of non-erythroid cells stained positive for myeloperoxidase and alpha-naphthyl acetate esterase (ANAE), respectively. Pathological examination revealed the mass lesions consisted of a proliferation of tumor cells, which were positive for monocytic markers (HLA-DR and Iba-1). The cow was diagnosed with acute myelomonocytic leukemia (AMML). Even when tumor cells are ANAE-negative, AMML cannot be completely ruled out and should be considered when diagnosing cattle with leukemia/lymphoma.

WOS: 000488603500001, PubMed: 31570012, 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., Selcuk University, Scientific Research Projects Coordination UnitSelcuk University [14202004], This research is part of Sercan Kaya's Master's thesis and was supported by the Selcuk University, Scientific Research Projects Coordination Unit under Grant number 14202004.

Amaç: Lizozomal bir enzim olan alfa-naftil asetat esteraz (ANAE) enzimi pek çok türde periferik kan yaymalarında T-lenfosit, B-lenfosit ve monositlerin birbirlerinden ayırt edilmelerinde kullanılmaktadır. Bu araştırmada, Topoz olarak da bilinen Kırgızistan dağ yaklarında (Bos grinniens) periferik kan lökositlerinin ANAE enzimi pozitivitelerinin belirlenmesi amaçlandı. Gereç ve Yöntem: Bu çalışmada materyal olarak 13 adet 2–4 yaşlı sağlıklı Kırgız Dağ Yak'ından alınan periferik kan örnekleri kullanıldı. Örneklerden hazırlanan frotilerde ANAE demonstrasyonları gerçekleştirildi. Bulgular: ANAE pozitivitesi tüm monositlerde ve sadece bazı lenfositlerde gözlendi. Lenfositlerdeki pozitivite 1-4 adet kırmızı-kahverengi sitoplazmik granüllerle karakterize nokta tarzındayken monositlerin tamamı tüm sitoplâzmaları boyunca yaygın, ince taneli bir enzimatik reaksiyona sahipti. Nötrofil ve eozinofil lökositlerde ise enzimatik reaksiyon gözlenmedi. Kırgız dağ yaklarında ortalama ANAE pozitif lenfosit oranının %58,85±1,38 olduğu tespit edildi. Öneri: Kırgızistan dağ yakları hakkında yapılan çalışmalar söz konusu hayvanların yaşam koşulları ve sert mizaçları nedeniyle son derece sınırlı kalmıştır. Dolayısıyla bu hayvanlar üzerinde yapılacak olan her türlü yapısal, histolojik, enzim histokimyasal, fizyolojik, biyokimyasal ve anatomik çalışmaların yanı sıra ülkede görev yapan veteriner hekim klinisyen ya da akademisyenlerin karşı karşıya kalabilecekleri çeşitli hastalıkların olgu sunumlarının, ilerleyen yıllarda bu hayvanlar üzerinde yapılacak olan çalışmalara yön vereceği düşünülmektedir., Aim: A lysosomal enzyme, alpha naphthyl acetate esterase (ANAE), has been used to differentiate T and B lymphocytes, and monocytes in various species in peripheral blood smears. In this study, it was aimed to determine the positivity of the ANAE in peripheral blood leukocytes of Kyrgyzstan's mountain yaks, as also known Topoz,(Bos grinniens). Materials and Methods: As a material, the peripheral blood samples collected from healthy thirteen Kyrgyzstan's mountain yaks, 2–4 year old aged, were used. ANAE was demonstrated on smears prepared from blood samples. Results: ANAE-positivity was observed in all monocytes and only some lymphocytes. ANAE-positivity pattern in lymphocytes was dot like characterized with 1-4 reddish-brown cytoplasmic granules while all monocytes have a diffuse, fine granular enzymatic reaction along whole cytoplasm. No enzymatic reaction was observed in neutrophil and eosinophil leukocytes. The ANAE positive lymphocyte rate was found to be 58,85 % in Kyrgyzstan's mountain yaks. Conclusion: Studies on Kyrgyzstan mountain yaks have been extremely limited due to their living conditions and aggressive temperaments of these animals. Therefore, any kind of structural, histological, enzyme histochemical, physiological, biochemical and anatomical studies on these animals, as well as case reports of various diseases that may be faced by veterinary clinicians or academicians working in the country, will guide the studies to be carried out on these animals in future.

The aim of this study was to evaluate the effect of flumequine on the percentage of peripheral blood leukocytes of carp, alpha naphthyl acetate esterase (ANAE) activity and proliferative activity of lymphocytes stimulated with concanavalin A (Con A) and lipopolysaccharide (LPS). Flumequine was administered to 40 carp, weighing 150 ± 10 g, at a dose of 12 mg/kg, once (group I) and four times, every 2 days (group II). Among white blood cells in flumequine, treated fish (group II) observed a decrease in percentage of lymphocytes and an increase of neutrophiles. Identification of the ANAE esterase activity in fish lymphocytes of the control group showed the advantage of positive cells over the negative ones and amounted to 62.65 ± 3.22 %. After administration of flumequine in group II fish, this value decreased to 44.75 ± 3.70 %. The present study clearly demonstrated that both Con A and LPS induced lymphoid cell proliferation in vitro in group I. There was an increase in activity after stimulation of LPS and its reduction after Con A in group II. This indicates a stimulating effect of flumequine on B lymphocytes. The results of this study are not conclusive as to the positive effect of the drug on the immune system of fish and indicate the need for caution in its use. .

WOS: 000482264200008, PubMed: 30864862, 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 90days. 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., Selcuk University Scientific Research Projects Coordination Units, Konya/TURKEY [11401013], This work was supported by Selcuk University Scientific Research Projects Coordination Units, Konya/TURKEY [grant number 11401013].

WOS: 000463194200001, PubMed: 30822167, 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.

The aim of this study is to determine the functional effects of the acrylamide (AA) administrated by oral gavage on the peripheral blood lymphocytes (PBL) and the Gut Associated Lymphoid Tissue (GALT) in male Sprague-Dawley rats using alpha-naphthyl acetate esterase (ANAE) demonstration. For this purpose, two separate experiments were performed with Sprague Dawley rats. In Experiment-I rats were gavaged with 0, 30, 45 and 60 mg/kgb.w. AA for five consecutive days and in Experiment-II rats were gavaged with 0, 125, 150, and 175 mg/kg/b.w. AA for single oral dose. Animals were sacrificed 24 h after the last treatments in both experiments by servical dislocations under ether anaesthesia. Blood samples were collected from the heart in heparinized (10 UI heparin/ml(-1) of the blood) tubes before sacrification and lymphoid tissue samples from the ileal Peyer's patches (IPPs) were taken and processed for histochemical demonstration of ANAE following the sacrification. The lymphoid follicles of the IPPs of animals given 125, 150 and 175 mg/kgb.w. AA were markedly reduced in size. Germinal centres (GCs) markedly regressed in AA-treated animals compared with those of controls. ANAE-positive lymphocyte depletion of IPPs was very prominent in the high doses AA-treated animals. In the animals treated with 30, 45, and 60 mg/kg b.w. AA, the IPPs had similar histology to those of the controls. ANAE-positive peripheral blood lymphocyte levels significantly decreased in AA exposed groups in a dose dependent manner (p

Plant-esterase from wheat flour was purified via aqueous two-phase system (ATPS). The enzyme crude extract was first filtered through a polyethersulfone OmegaTM ultrafiltration membrane, followed by two-step ATPS with PEG1000, NaH2PO4 and (NH4)2SO4. Based on previous studies, the first ATPS consisting of 27.0% PEG1000 and 13.0% NaH2PO4 (w/w) was performed at room temperature and pH 5.0. The top phase was carefully separated, and 6.0% (NH4)2SO4 (w/w) was added to form the second phase system under similar conditions, to obtain the pure enzyme that partitioned in the bottom salt phase. The purification thus obtained was 10.35 fold with an enzyme yield of 72.90%. Sodium dodecylsulphate polyacrymamide gel electrophoresis (SDS-PAGE) was used to estimate the molecular weight of the target esterase to be around 68 kilo Daltons (kDa). Using a 16 mM solution of alpha naphthyl acetate as substrate, the maximum activity of the purified enzyme, alpha naphthyl acetate esterase (ANAE) was found to be 0.78 U at the optimum conditions of 40 ◦C and pH 8.0 for 15 min of incubation. The Michaelis–Menten parameters Km and Vmax of the purified enzyme were estimated using Langmuir linear plot as 22.5 mM and 4.71 U mg−1, respectively. Further purification was obtained by dialyzing the bottom ANAE rich phase in a solution of 0.04 M phosphate buffer with pH 6.5 to remove the salt, which was then concentrated using centrifugal tubes with OmegaTM membrane. The highly pure ANAE so obtained was immobilized on the multiwalled carbon nanotube for the development of inhibition based enzyme biosensors for pesticides detection.

Comparative study of isoenzymic patterns of acid phosphatase, beta-glucuronidase and alpha-naphthyl acetate esterase of normal and chronic lymphocytic leukaemia (CLL) lymphocytes was carried out by means of polyacrylamide gel electrophoresis. The isoenzymograms of acid phosphatase and beta-glucuronidase of leukaemic cells were similar to that of normal lymphocytes. However, different patterns were found in the case of alpha-naphthyl acetate esterase. Two prominent anodal bands were present in CLL lymphocytes; these bands were either absent or very weak in normal cells.

The ultrastructural localization of acid alpha-naphthylacetate esterase (ANAE) activity was studied in normal and neoplastic human monocytic and lymphocytic cells including hairy cells. Normal monocytes and malignant monocytic cells from cases of AML-M4 and -M5 (FAB-classification) displayed fundamentally the same ANAE staining pattern with reaction products localized to the external surface of the plasma membrane and to small vesicles close to this membrane, more rarely to larger intracytoplasmic vesicles containing endocytosed material or cellular debris. The enzyme activity of the neoplastic cells increased with morphological differentiation. Certain normal blood lymphocytes and T cell-derived CLL cells showed the reaction product associated with paranuclearly located clusters of vesicular structures, extending from the membranes into the adjacent cytoplasm. Gall bodies were often found in the vicinity and were invariably positive for ANAE. Now and then, plasma membrane activity was present, but never as pronounced as in monocytes. Hairy cells demonstrated a pattern of reaction very similar to that of monocytic cells, whereas B cell-derived CLL cells and lymphoblasts of T- and common type ALL were generally non-reactive.

Lymph node and peripheral blood lymphocytes in a case of Hodgkin's disease (mixed cellularity) were studied using May-Grünwald-Giemsa (MGG), acid naphthyl acetate esterase (ANAE), immunoperoxidase staining and lymphocyte surface markers, autoradiographic, and lymphocyte stimulation techniques. According to MGG staining and autoradiographic studies of lymph, node cells, small lymphocytes, intermediate lymphoid cells, and large mononuclear cells resembling in-vitro stimulated immunoblasts, Hodgkin's cells and Reed-Sternberg (RS) cells formed a morphologically continuous DNA synthetizing series. A large majority of small lymphocytes from a lymph node were ANAE positive, thus being T-lymphocytes, and formed rosetts around large mononuclear cells and RS cells. Most RS and large mononuclear cells had ANAE positive spots in the cytoplasm, thus resembling T-lymphocytes more than diffusely staining monocytes. These cells did not contain cytoplasmic immunoglobulin and were muramidase negative. Both lymph node and peripheral blood lymphocytes responded strongly to PHA. The role of T-lymphocytes in Hodgkin's disease and the origin of RS cells are are discussed on the basis of the findings.

WOS: 000347678600009 This study aimed to determine the alpha-naphthyl acetate esterase (ANAE) activity of peripheral blood leukocytes in pregnant, kitten, and adult Angora cats. In each group 9 healthy pregnant, adult, and kitten Angora cats constituted the material of the study. Optimum reaction was achieved after a 3 h incubation period at pH 5.8, by ANAE staining. Two types of reaction were observed in ANAE (+) T lymphocytes. The first reaction was a dot-like positivity pattern characterized by the presence of large granules, while the other was a granular positivity pattern characterized by the presence of 3 to 5 small granules. B lymphocytes reacted negatively in ANAE staining. The rate of ANAE (+) lymphocytes in pregnant, kitten, and adult Angora cats was 62.89 +/- 1.29%, 68.37 +/- 1.22%, and 77.71 +/- 1.63%, respectively. In conclusion, the lowest rate of ANAE-positive lymphocytes was detected in the pregnant cats, depending on the maternal immune tolerance supported by hormonal mechanisms. An increased rate of ANAE-positive lymphocytes was detected in the kittens, and the highest rate was measured in the adults, in parallel with the development of the immune system.

T. Karaca, Cemek M., Kanter M.: Lipid Peroxidation and Antioxidant Levels, and Alpha Naphthyl Acetate Esterase Activity of Peripheral Blood Lymphocytes in Mallard, Muscovy and Pekin Ducks. Acta Vet. Brno 2006, 75: 33-38. The aim of this study was to determine blood levels of malondialdehyde (MDA), reduced glutathione (GSH), ceruloplasmin and vitamin C, and the percentages of peripheral blood Tlymphocytes using the alpha-naphthyl acetate esterase (ANAE) method on Mallard (Anas platyrhynchos), Muscovy (Cairina moschata) and Pekin (Anas domestica) ducks. Blood samples were obtained from 8 adult ducks of each breed. The serum levels observed in Mallard, Muscovy and Pekin ducks respectively were 0.8, 1.07 and 1.3 nmol MDA per ml; 77.4, 66.9 and 78.7 mg GSH per 100 ml; 23.9, 26.1 and 24.1 mg ceruloplasmin per 100 ml; and 0.50, 0.52 and 0.70 mg vitamin C per 100 ml. The percentage of the ANAE (+) lymphocytes was 57.9%, 54.8% and 55.1% in Mallard, Muscovy and Pekin ducks, respectively. In this study, blood levels of lipid peroxidation and nonenzymatic antioxidants in the Mallard, Muscovy and Pekin ducks were determined. Lipid peroxidation, antioxidant, alpha-naphthyl acetate esterase, T-lymphocyte, duck

WOS: 000321750600019, This study was performed to determine the effects of pregnancy on the activities of alpha naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) of the peripheral blood lymphocytes in pregnant Merino sheep. Peripheral blood lymphocyte percentages were also estimated. Periferal blood samples were taken from animals in six different gestational stages as non-pregnant control, in the first, the second, the third, the fourth and the fifth month of pregnancy. Each group was contained 20 animals. The lowest ANAE (+) lymphocytes percentage (63.5%) was determined in the first month of pregnancy whereas the highest null lymphocytes proportion (12.75%) was detected in the same gestational period. There were statistically decreases in the proportions of peripheral blood lymphocyte (42.9%) and the ACP-ase (+) lymphocytes (43.35%) in the last gestational stages. Although the posibble hormonal changes may cause the fluctuation of peripheral blood lymphocyte proportions in all gestational periods, the most distinctive changes were observed at the begining and at the end of the pregnancy. It was concluded that the data was obtained from this study was useful for understanding of the possible mechanisms of maternal tolerance.

The purpose of our study was to determine the percentages of alpha-naphthyl acetate esterase (ANAE)-positive and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL) in both sexes of one-year-old domestic pigeons (Columba livia f. domestica). Blood samples obtained from 12 healthy domestic pigeons were used. The mean percentages of ANAE-positive PBL for females and males were 47.8% and 48.8%, respectively, whereas the mean percentages of ACP-positive PBL were 67.5% and 68.6%, respectively. The proportions of PBL were 49.3% and 48.6% in males and females, respectively.

Bu çalışmada iki farklı yer sincabı türünün perifer kan lenfosit oranları ile alfa-naftil asetat esteraz (ANAE) ve asit fosfataz (ACP) pozitif lenfosit oranlarının ışık mikroskobik düzeyde belirlenmesi amaçlanmıştır. Bu amaçla altı ergin Spermophilus taurensis (Hadim, Konya) ve dört ergin Spermophilus xanthoprymnus (Selçuklu, Konya) örneğinden usulüne uygun olarak kan alınmış ve her örnekten altı adet froti hazırlanarak havada kurutulmuştur. Frotilerden ikisi lenfosit oranının tespit edilmesi için rutin May-Grünwald Giemsa yöntemiyle boyanmıştır. Diğer frotiler fosfat tamponlu glutaraldehit-aseton solüsyonunda tespit edilmiştir. Bu frotilerden ikisine ANAE demonstrasyonu, diğer ikisine ise ACP demonstrasyonu uygulanmıştır. ANAE demonstrasyonu sonucunda her iki türde lenfositlerin çoğunluğu pozitif sonuç vermiştir. ANAE pozitif lenfositlerde iki tip granüler pozitivite tespit edilmiştir. Bu lenfositlerin çoğunda sayıları 1-5 arasında değişen kırmızı-kahverengi iri granüller (nokta tarzında pozitivite) gözlenirken, daha az orandaki lenfositlerde ise yaygın çok sayıda küçük granüller (ince granüler pozitivite) gözlenmiştir. ACP demonstrasyonu sonucunda da her iki türde lenfositlerin çoğunluğu pozitif sonuç vermiştir. ACP pozitif lenfositler sayıları 1-3 arasında değişen pembe-kırmızı küçük granüler tarzda pozitivite göstermiştir. Bunun yanı sıra her iki enzim demonstrasyonunda monositler genellikle sitoplazmalarında güçlü ve yaygın tarzda pozitivite gösterirken, nötrofillerin ise zayıf bir reaksiyon verdiği dikkat çekmiştir. S. taurensis ve S. xanthoprymnus’un lenfosit, ANAE ve ACP pozitif lenfosit oranları arasında istatistiksel olarak önemli bir fark bulunamamıştır (p>0.05). Ayrıca her bir türün ANAE ve ACP pozitif lenfosit oranları arasında da istatistiksel olarak önemli bir fark tespit edilmemiştir (p>0.05)., This study was carried out to determine peripheral blood lymphocyte (PBL), alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP) positive PBL percentages in two different species of the ground squirrel. For this purpose, smears were prepared and air dried from blood of six adult Spermophilus taurensis specimens (Hadim, Konya) and four adult Spermophilus xanthoprymnus specimens (Selçuklu, Konya). Two smears were stained with May Grünwald-Giemsa for the determination of PBL percentages. Four smears were fixed in glutaraldehyde-acetone solutions. Two of the smears were used for ANAE demonstration. The remaining two smears were used for ACP demonstration. ANAE positive PBLs showed two distinct granular positivity. While most of the ANAE positive cells had 1-5 large reddish-brown granules (dot-like granular positivity), the other positive cells had large quantity of diffusely arranged small granules (fine granular positivity). ACP positive PBLs had 1-3 pinkish-red small granules. Besides, monocytes generally gave a diffuse and strong reaction while neutrophils displayed a weak positive reaction in their cytoplasm for ANAE and ACP. The differences between PBL, ANAE and ACP positive PBL percentages of S. taurensis and S. xanthoprymnus were not statistically significant (p>0.05). The differences between ANAE and ACP positive PBL percentages of each species were also not statistically significant (p>0.05).

Bu çalışmada Konya bölgesindeki kör farelerin (Nannospalax nehringi, 2n=60 kromozomal formu) perifer kan lenfosit oranları ile alfa-naftil asetat esteraz (ANAE) ve asit fosfataz (ACP-az) pozitif lenfosit oranlarının ışık mikroskobik düzeyde belirlenmesi amaçlanmıştır. Bu amaçla beş ergin N. nehringi (2♀, Meram ve Selçuklu; 3♂, Selçuklu, Çumra ve Ilgın) örneğinden usulüne uygun olarak kan alınmış ve her örnekten altı adet froti hazırlanarak havada kurutulmuştur. Frotilerden ikisi lenfosit oranının tespit edilmesi için rutin May-Grünwald Giemsa yöntemiyle boyanmış ve lenfosit oranı %68.00±5.43 olarak belirlenmiştir. Diğer frotiler fosfat tamponlu gluteraldehit-asetonda (-10°C) 3 dakika tespit edilmiştir. Bu frotilerden ikisine ANAE demonstrasyonu, son ikisine de ACP-az demonstrasyonu uygulanmıştır. ANAE demonstrasyonu sonucunda lenfositlerin çoğunluğu pozitif sonuç verdiği halde, daha az orandaki lenfositlerin (%22.29±5.79) negatif sonuç verdiği gözlenmiştir. ANAE pozitif lenfositlerde iki tip granüler pozitivite tespit edilmiştir. Bu lenfositlerin çoğunda (%73.45±6.87) sayıları 1-5 arasında değişen kırmızı-kahverengi iri granüller (nokta tarzında pozitivite) gözlenirken, daha az orandaki lenfositlerde (%4.26±2.36) ise diffüz çok sayıda küçük granüller (ince granüler pozitivite) gözlenmiştir. Bunun yanı sıra monositler genellikle sitoplazmalarında güçlü ve yaygın tarzda pozitivite gösterirken, nötrofiller ise zayıf bir reaksiyon vermiştir. ACP-az demonstrasyonu sonucunda ise lenfositler çoğunlukla (%83.40±5.90) negatif reaksiyon verirken, daha az orandaki lenfositler (%16.60±5.90) sayıları 1-3 arasında değişen pembe-kırmızı granüler tarzda pozitivite göstermiştir., The aim of this study was to determine peripheral blood lymphocyte (PBL), alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) positive PBL percentages of mole rats (Nannospalax nehringi (2n=60 chromosomal form); 2♀, Meram and Selçuklu; 3♂, Selçuklu, Çumra and Ilgın) in Konya region. For this purpose, six smears were prepared and air dried from each animal. Two of them were stained with May Grünwald-Giemsa for the determination of PBL percentages. The PBL percentage was 68.00±5.43%. Four smears were fixed in glutaraldehydeacetone solutions (pH 4.8) at -10°C for 3 minutes. Two of the smears were used for ANAE demonstration. The remaining two smears were used for ACP-ase demonstration. ANAE staining revealed that large majority of the PBLs were positive, whereas smaller proportion (22.29±5.79%) were negative. Positive PBLs showed two distinct granular positivity. While most of the ANAE positive cells (73.45±6.87%) had 1-5 large reddish-brown granules (dot-like granular positivity), 4.26±2.36% of positive cells had large quantity of diffusely arranged small granules (fine granular positivity). Besides, monocytes generally gave a diffuse and strong reaction while neutrophils displayed a weak positive reaction in their cytoplasms for ANAE. ACP-ase staining revealed that large majority of the PBLs (83.40±5.90%) were negative, whereas smaller proportion (16.60±5.90%) were positive. ACP-ase positive cells had 1-3 large pinkish-red granules.

WOS: 000275982900010, The aims of this study were to determine the percentages of A-Naphthyl Acetate Esterase (ANAE) positive and Acid phosphatase (Acp)-postive Peripheral Blood Lymphocytes (PBL) and the proportion of peripheral blood lymphocytes in the Arabian race horses. Blood samples obtained from the Vena jugularis of 40 healthy Arabian race horses at four years old were analyzed. The animals were selected randomly from found as 46.20%. In conclusion, the enzyme histochemical techniques are simple, inexpensive and well performed for laboratory conditions. Also, these techniques may help early diagnosis of different diseases. Further studies about enzyme histochemistry in Arabian race horses shouled be performed.

Analysis and diagnostic interpretation of leukemic cell histochemistries are relatively subjective processes that reflects a number of factors including observer experience, methodological idiosyncrasies and limitations, and a theoretical understanding of the technologies involved. In particular, although the application of alpha-naphthyl acetate esterase (ANAE) cytochemistry for the recognition and delineation of monocytic1histiocytic components is generally straightforward, a significant proportion of cases (particularly acute myeloid leukemias) display cytochemical staining patterns that are difficult to interpret. The incorporation of sodium fluoride has been widely used to help resolve these difficulties even though its mode of inhibition is poorly understood and its influence on different esterase species inconsistent. An alternative inhibitor, phenylmethylsulphonyl fluoride (PMSF) was evaluated in this study for its potential usefulness in leukemic cell ANAE histochemical analysis. The res...

In order to study the differentiation level of two chicken monocytic cell lines, IN24 and LSCC-NP1 cells, alpha-naphthyl acetate esterase (ANAE) was examined by ultrastructural cytochemical technique. Morphologically and functionally, IN24 cells showed immature character compared to LSCC-NP1 cells. In IN24 and LSCC-NP1 cells, ANAE activity was localized on the external face of the plasma membrane and coated vesicles, but not in lysosomes and phagocytic vacuoles. ANAE reactivity showed a dense linear reaction product on the entire cell surface in IN24 cells and granular reaction product in LSCC-NP1 cells. On treatment with interferon-gamma (IFN-gamma), ANAE reactivity of IN24 cells became similar to that of LSCC-NP1 cells. IN24 cells treated with lipopolysaccharide (LPS) had less amounts of ANAE surface product, and reactivity was partially dispersed on the cell surface.

We examined gazelle peripheral blood leucocytes using the alpha-Naphthyl acetate esterase (ANAE) staining technique (pH 5.8). Our purpose was to determine the percentage of ANAE positive lymphocytes. The proportion of ANAE positive T-lymphocytes was 72%. T-lymphocytes showed an ANAE positive reaction, but eosinophilic granulocytes and monocytes also showed a positive reaction. By contrast, no reaction was detected in B-lymphocytes, neutrophil granulocytes or platelets. The reaction observed in T-lymphocytes was a red-brown coloration, usually 1-2 granules, but enough granules to fill the cytoplasm were detected rarely. As a result of ANAE enzyme staining, we concluded that the staining technique can be used as a cytochemical marker for gazelle T-lymphocytes.

WOS: 000253855000009, PubMed: 18079450, 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.

Cytochemical analysis of leukemic blasts from 46 patients with acute myeloblastic M2 leukemia (according to the FAB classification) was performed before and after cytostatic therapy, and compared with findings obtained in 20 age- and sex-matched control subjects. Cytochemical findings for myeloperoxidase (MPO), Sudan black B, acid phosphatase and alpha-naphthyl-acetate esterase (ANAE) were related to the achievement of the first complete remission (CR), i.e. data were compared after the patients had been divided into CR and non-CR groups. The analysis clearly showed that a high proportion of myeloperoxidase- and, to a lesser extent, Sudan black B-positive blasts before treatment may have constituted a significantly unfavourable prognostic factor.

The average percentage of acid alpha naphthyl acetate esterase reacting lymphocytes (APARL) was enumerated in the peripheral blood of chickens challenged with Marek's disease after vaccination with either turkey herpesvirus (HVT), inactivated Marek's disease virus (IMDV) or a mixture of the two (bivalent vaccine). A gradual increase in APARL value was noticed in the vaccinated chickens from day 7 to 70 after challenge with a virulent Marek's disease virus. The increase was consistent and significantly higher in bivalent (HVT plus IMDV) than in HVT-vaccinated chickens while the slight increase noticed in IMDV vaccinated-challenged birds was inconsistent.

The histochemical demonstration of nonspecific acid α-naphthyl acetate esterase (ANAE) activity was evaluated as a T lymphocyte marker primarily with the sheep erythrocyte (E) assay. A distinctive staining pattern characterized T lymphocytes which could be readily distinguished from monocyte staining. The percentage of E + and ANAE + lymphocytes was nearly always comparable in the peripheral blood and lymphoid tissue from normal and selected patients, including those with acute and chronic lymphocytic leukemia. Divergences were noted in certain other tissues including spleen and thymus. Certain mitogen-stimulated cells lost their ANAE activity while retaining their ability to form e n rosettes. Atypical and variable staining patterns were observed in established lymphoid cell lines. The histochemical demonstration of ANAE is simple and reproducible; preparations may be counterstained for cytomorphologic detail and mounted as a permanent record. Certain disadvantages are discussed. The method represents a practical alternative to E rosette assays. It is particularly well suited for certain routine laboratories.

A non-specific acid alpha-naphthyl-acetate esterase activity was investigated in human lymphoid cell populations from tonsils, blood, thymus and in different leukaemias. Four patterns have emerged. (i) The vast majority of T lymphocytes (peripheral blood, tonsils) showed a localized, intense reaction product (‘T-like’). (ii) A thymocyte subpopulation expressed faint, localized enzyme activity (‘Thy-like’) but the majority of thymocytes showed no esterase activity. Some cells from acute lymphoblastic leukaemias with thymocyte surface characteristics had a ‘Thy-like’appearance. (iii) Most B lymphocytes and cells from chronic lymphocytic leukaemias were esterase negative. No activity was seen in mitogen activated peripheral blood T and B lymphoblasts from peripheral blood and in leukaemic blasts from the common form of childhood acute ‘lymphoblastic’leukaemia. (iv) Myeloid cells, including peripheral blood monocytes, and blast cells from acute myeloblastic and chronic granulocytic leukaemias showed an intense, diffuse reaction product (M-like). It is concluded that the modified esterase technique is a convenient marker for small mature human T lymphocytes in tissue sections and smears. It may also be helpful in the differential diagnosis of acute lymphoblastic leukaemia with T characteristics, the common form of acute ‘lymphoblastic’leukaemia and acute myeloblastic leukaemia.

Peripheral blood mononuclear cells of 21 patients undergoing allogeneic bone marrow transplantation (BMT) were monitored post-BMT for immunologic markers (E rosettes OKT3, OKT8, DR, and BT5/9, a monoclonal antibody which stains helper T cells), cytochemical markers (acid phosphatase [AP], beta-glucuronidase [BGLU], and acid alpha naphthyl acetate esterase [ANAE] ), and morphology. The cytochemical T score, was obtained from typical AP, BGLU, and ANAE reactivity. The same was done for the cytochemical non-T score and macrophage (Mo) score. All patients received cyclosporine A (CyA) for graft-v-host disease (GvHD) prophylaxis. In univariate analysis there was no significant correlation between the proportion of E rosettes, OKT3-, OKT8-, DR-, and BT5/9-positive cells, and GvHD. The first three showed instead a positive correlation with time from transplant: E rosettes (P = .02), OKT3 (P = .01), and OKT8 (P = .003). In contrast, a significant negative correlation was found in univariate analysis, between the cytochemical T score and GvHD (P = .0001), and a positive correlation between non-T score and GvHD (P = .0008), as well as between the Mo score and GvHD (P = .03). There was no influence of time from transplant on the T (P = .8), non-T (P = .8), or Mo score (P = .4). In multivariate analysis comparing E rosettes, OKT3, T score, non-T score, GvHD, and time from BMT, the only variable associated with GvHD was the T score (P less than .05). These results suggest that T cell activation during GvHD is associated with a loss of hydrolase expression in T cells, but does not imply relevant modifications of immunologic surface markers. In addition, lysosomal enzymes appear early (before day 10) after transplantation, indicating that T cells at this stage are well differentiated.

Acid alpha-naphthyl acetate esterase (ANAE) activity is characteristic of resting human T lymphocytes. The expression of the ANAE marker by activated human T and B lymphocytes (blasts) and by corresponding 'secondary' lymphocytes has been investigated. Human blood lymphocytes were stimulated by selective T-cell (phytohemagglutinin (PHA) and concanavalin A (Con A)) or B-cell (Staphylococcus aureus strain Cowan 1) mitogens or in the mixed lymphocyte culture (MLC), and the percentage of blasts expressing the marker was quantiated. Whereas 95% of Con-A-activated blasts expressed the marker, approximately 25%-30% of MLC-activated blasts and only 10%-25% of PHA-activated blasts were ANAE-positive. After reversion to secondary lymphocytes, the PHA- and MLC-activated cells regained the ANAE activity, and more than 90% of the blast-derived secondary T lymphocytes were ANAE-positive. Only 2%-8% of blast cells activated by Staphylococcus aureus strain Cowan 1 were ANAE-positive. We therefore conclude that ANAE is not a reliable marker for T cells when activated cells (blasts) are considered.

The shape of alpha-naphthyl acetate esterase (ANAE)-positive cells (other than T lymphocytes) has been measured in 40 lymph nodes. The specimens comprised 15 high-grade lymphomas, 15 low-grade lymphomas and 10 reactive lymph nodes. The parameter used for the measurement of shape was form factor (FF), which is readily calculated by the Reichert-Jung (Kontron) MOP-AMO3 user-controlled image analyzer. Perfectly round cells have an FF value of 1.0, whereas the FF of irregularly-shaped cells diverges from unity. It has been demonstrated that the ANAE-positive cells in high-grade lymphomas have mean values for FF of 0.8-0.9, whereas in low-grade lymphomas and reactive nodes the mean value is 0.4-0.5. Thus, high-grade lymphomas contain many more rounded ANAE-positive macrophage type cells than do low-grade lymphomas and reactive nodes. In the latter two types of specimen there is an excess of branching and spindle-shaped ANAE-positive cells.

Nonspecific acid alpha naphthyl acetate esterase (ANAE) activity has been demonstrated in bovine lymphocytes and monocytes-macrophages. The lymphocytes exhibited a solitary reddish brown globular reaction adjacent to the cell membrane (T-pattern) whereas the monocytes-macrophages revealed a diffuse reddish brown cytoplasmic reaction (M-pattern) product. Cy tocen tri fuge smears of lymphocytes forming rosettes with sheep erythrocytes also revealed T-pattern ANAE reaction in the majority of the cells. Similar T- and M-patterns of ANAE activity were observed in cryostat sections of biopsies collected from normal bovine lymph nodes and horn cancer tissue sections from bullocks.

Acid α-naphthyl acetate esterase (ANAE) activity was investigated to be one of porcine T lymphocyte markers by modified fixation procedures in normal porcine lymphocytes prepared from spleen, blood, mesenteric lymph nodes, thymus and Peyer's patch. Smears were fixed for 2 to 3 seconds with buffered formalin acetate (pH 6.6). The majority of ANAE positive cells had the red-brown spot. The cells diffusely stained with densely red color were found to be monocytes. Other stained cells had stained granules. There were no differences between the ratios of ANAE positive cells with stained spots or granules and those of E-rosette positive cells in lymphocyte preparations from spleen, blood, mesenteric lymph nodes and Peyer's patch. However, the ANAE positive cells in thymus were stained faintly and the ratio of ANAE positive cells was by far lower than that of E-rosette positive cells. In Con A or PHA-P stimulated lymphocytes, ANAE staining density was more faint than that of non-stimulated cells and the percentages of ANAE positive cells were lower than those of E-rosette positive cells. These results indicated that ANAE activity could be one of T cell markers for porcine mature lymphocytes (spot or granular pattern) of spleen, blood, mesenteric lymph nodes and Peyer's Patch and one of markers for porcine monocytes (diffuse pattern).

Diagnostic features (cytochemistry, immunophenotyping and serum biochemistry) were examined in 51 cases of acute monocytic leukaemia (AMoL). Peroxidase, Sudan black B and alpha naphthyl acetate esterase (ANAE) cytochemical reactions were unrelated to morphological (FAB groups M5a and M5b) or immunological subtype. ANAE cytochemistry, however, indicated that AMoL cases could be subdivided into those with typical (M-type) reactions and those with insignificant staining or monocytic ANAE isoenzymes (defined by IEF). All cases were phenotypically CD13/CD33 positive and, with one exception, had greater than 30% HLA-DR positive cells. Membrane CD14 expression was insignificant or variable in 33% of M5a cases in contrast to 23/24 M5b cases which showed high proportions of CD14-staining cells with at least two monoclonal antibodies. Serum lysozyme, LDH and beta-2 microglobulin (beta 2m) were increased in 88%, 68% and 81% of cases respectively but, with the exception of statistically higher lysozyme levels in CD14+ cases, were unrelated to the morphological, cytochemical or immunological diagnostic subgroups. Clinical and diagnostic features were also examined as possible prognostic indicators. The morphological, cytochemical and immunological subgroups of AMoL were not found to be of prognostic relevance but age (P = 0.004), renal failure (P = 0.005) and serum beta 2m levels (P = 0.002) were related to patient survival. Moreover, renal failure and serum beta 2m remained significant (P = 0.012 respectively) when age was taken into account and were shown to be independent prognostic variables.

A strong activity of acid alpha-naphthyl acetate esterase (ANAE) in T lymphocytes and macrophages within mouse lymphoid organs is demonstrable even after paraffin-embedding, if a rapid embedding procedure with acetone as dehydrating agent is used. The lymphoid tissue structure is better preserved in paraffin sections procured after this rapid embedding procedure than in frozen sections commonly used for histochemical demonstration of ANAE. In the paraffin sections the pattern of the ANAE reaction allows a clear delineation of the T and B areas in the lymphoid organs, and the intracellular location of the focal ANAE activity in T lymphocytes is readily discernible.

The numbers of alpha-naphthyl acetate esterase (ANAE)-positive cells (other than T cells) have been counted in 32 specimens of Hodgkin's disease and two specimens of histiocytic lymphoma. The different Rye subtypes of Hodgkin's disease contain varying numbers of enzyme-positive cells, ranging from 1.8-16% in the lymphocyte-predominant form to 39.8-47% in lymphocyte-depleted Hodgkin's disease. The percentage of enzyme-positive cells in the mixed cellularity variety was from only 6.5 to 14.6%. In the two specimens of apparently genuine histiocytic lymphoma, the enzyme-positive cells constituted 95.2 and 97.5% respectively of all cells. Thus, the numbers of macrophages and macrophage-like cells in true histiocytic lymphoma are much greater than in Hodgkin's disease.

Alpha-naphthyl acetate esterase (ANAE) activity was demonstrated in ovine lymphocytes harvested from blood on Ficoll-metrizoate gradients. The enzyme's specificity for T lymphocytes, identified by immunofluorescent staining of T cell-specific antigens, was assessed. Correlation analysis of the results obtained using unfractionated lymphocytes from 12 sheep showed no correlation between ANAE activity and the expression of T cell antigens ( r = 0.22). When lymphocytes from 4 sheep were fractionated on nylon wool columns a mean of only 43.2% of the cells in the non-adherent population were ANAE-positive whereas 94.7% of these cells were identified as T lymphocytes. Blood lymphocytes from 5 animals were separated into 3 fractions using Percoll discontinuous density gradients. No significant relationship was seen between ANAE activity and T cells in Fractions 1 and 3 ( r = 0.41 and 0.21). Fraction 2 cells, however, did show a significant positive relationship ( r = 0.91) between these two features but the biological significance of this relationship is unknown. It was concluded that ANAE activity is not a specific marker for ovine T lymphocytes.

The distribution of mononuclear cell subsets has been studied in human adenoids, tonsils and peripheral blood (PB) by evaluating the presence of surface immunoglobulins, E-rosette formation, receptors for IgG Fc and for complement, alpha-naphthyl acetate esterase (ANAE) cytochemistry, reactivity with peanut lectin (PNA) and with monoclonal antibodies (McAb) (OK panel). Adenoids and tonsils, compared to PB, contain (1) fewer macrophages and T cells but more B cells; (2) higher proportions of ANAE negative, complement receptors and Ia-like antigens bearing T lymphocytes; (3) higher percentages of cells reacting with the McAbs OKT9 and OKT10 ("immature" lymphoid cells). In both adenoids and tonsils, clusters, formed by a central heavily ANAE stained interdigitating cell surrounded by lymphocytes with a sickle-shaped ANAE reaction, were found. Analogous clusters have been previously described in mice and human thymus. Two major hypotheses could be put forward: (1) adenoids and tonsils contain "immature" lymphoid cells undergoing education process, or (2) the above organs contain lymphocytes activated by a constant exposure to bacterial antigens or mitogens.