Positive Alkaline Phosphatase Reaction Research Articles

Proposal of Helicobacter canicola sp. nov., previously identified as Helicobacter cinaedi, isolated from canines

During the course of our taxonomic investigation of Helicobacter cinaedi, it was realized that the strains isolated from dogs, which have been identified as H. cinaedi, showed different biochemical traits than did the isolates obtained from humans. None of the three dog isolates could reduce nitrate to nitrite, whereas all of the human H. cinaedi isolates could do so. The dog isolates showed a strong positive alkaline phosphatase reaction and could grow at 42°C, however the human isolates showed negative to very weak responses to those tests. The GyrA protein based phylogenetic analysis showed that the three isolates from dogs formed a slightly distinct cluster from the human isolate cluster. Phylogenetic analysis of the 16S rRNA, 23S rRNA, gyrB, and hsp60 gene sequences further confirmed that the dog isolates differed from the human H. cinaedi isolate cluster. The whole-genome in silico DNA similarities of each isolate based on their full genome sequences revealed that the isolates from dogs shared more than 94.9% ANIb (average nucleotide identity based on BLAST), while 94.0% ANIb were found between the isolates from dogs and the humans, including the H. cinaedi type strain ATCC BAA-847T (=CCUG 18818T). From these data, we propose a new species, ‘H. canicola’ sp. nov., for the isolates from dogs. The type strain is PAGU 1410T (CCUG 33887T=LMG 29580T).

Valores hematológicos e identificação morfo-citoquímica de células sangüíneas de capivaras (Hydrochoerus hydrochoeris) parasitadas por carrapatos e capivaras livres de infestação

Blood samples were collected from thirty seven adult parasited capybaras and twenty one free of parasitism aiming the characterization of different circulating cell types and complete hemogram. After a morphological analysis, the blood cells were identified as heterophils, eosinophils, basophils, lymphocytes, monocytes and platelets. The PAS reaction revealed glycogen in heterophils and platelets cytoplasm. A positive reaction was found only in heterophils round granules and in all cytoplasmic granules of eosinophils for the demonstration of myeloperoxidase activity and lipids. The bromofenol blue method revealed strong positivity in heterophils, eosinophils, basophils granules and red blood cells. A positive reaction for alkaline phosphatase was observed in eosinophils and heterophils. The blood parameters studied showed statistic differences (p 6 /µL and 2,81±0,62 x 10 6 /µL) , haematocrit (48.14±4.75% and 41,22±6.21%) and heterophils (39,38±11,02% and 16,14±7,13%) /eosinophils (8,57±3,78% and 21,92±10,24%) in differential count. In this case the ticks involved in the parasitism of these capybaras were identified as Amblyomma cajenense and Amblyomma cooperi . These data showed that the highly infested capybaras presented some anemia and a remarkable eosinophilia.

Extraskeletal osteosarcomas: a clinicopathologic study of 25 cases.

A follow-up investigation of 25 cases of extraskeletal osteosarcomas diagnosed at the Center for Bone and Soft Tissue Tumors, Aarhus University Hospital, Denmark, in the period from 1970-1995 was undertaken. The immunohistochemical profile of these tumors was evaluated using a panel of 10 antibodies, and the value of alkaline phosphatase staining in differential diagnostic situations also was considered. The study revealed that this tumor is high-grade malignant and affects adults (median age, 67 years; range, 35-82 years) at diagnosis. The thigh (52%) was the most common tumor location. Seven tumors were superficial, whereas the remaining 18 were intramuscular. Two patients with superficial tumors previously received radiation to the area. Local recurrences developed in 9 (36%) patients and distant metastases developed in the lungs in 15 (60%) patients as the most common site. Median survival time was 24 months, and the cause-specific survival rate at 5 years was less than 25%. Thirteen (52%) intramuscularly located extraskeletal osteosarcomas were of the fibroblastic subtype, often with sparse amounts of osteoid. They could be separated from malignant fibrous histiocytoma on the basis of a strongly positive alkaline phosphatase reaction. Immunohistochemistry did not reveal characteristic features because positivity for vimentin, occasional positivity for desmin, actin, S-100, epithelial membrane antigen, cytokeratin, and p-53 may be observed in many other pleomorphic sarcomas. Various histopathologic factors, such as tumor size, tumor depth, histopathologic subtype, malignancy grade (IIIA versus IIIB), MIB-1, and p53 reactivity were analyzed in relation to clinical course. Only MIB proliferation was correlated to prognosis, with significantly longer survival in patients with tumors with MIB-1 values less than 24%. Our study has shown extraskeletal osteosarcoma to behave in a highly aggressive fashion. Alkaline phosphatase staining compared with immunohistochemistry proved to be superior in the differentiation from other pleomorphic sarcomas.

Mineralization in serially passaged human alveolar bone cells.

Well-characterized human bone cell cultures have been regarded as a useful tool to study bone control mechanisms and also to analyse bone/biomaterials interactions. In the present study, human alveolar bone cells were cultured in alpha-minimal essential medium (alpha-MEM) containing 10% foetal bovine serum (FBS), 50 microg/ml ascorbic acid, 10 mM sodium beta-glycerophosphate and either in the presence or in the absence of 10 nM dexamethasone (Dexa). Cultures were characterized concerning cell viability/proliferation, alkaline phosphatase (ALP), acid phosphatase (ACP) and tartraric acid resistant phosphatase (TRAP) activities, and formation of mineralized areas. Cell proliferation increased gradually for approximately 20 days. In the presence of Dexa, cells formed isolated or interconnected multilayered clusters that increased with culture time. Histochemical assays revealed strong positive reactions for ALP and calcium and phosphates deposits, mainly in relation t! o ce lls associated with the clusters. High levels of ALP activity (biochemical determination) were observed. Cells cultured in the absence of Dexa showed significantly lower ALP activity and no calcium and phosphates deposits were present. Serially passaged cells kept the proliferation rate constant but a decrease in ALP activity was observed either in the presence or in the absence of Dexa. The ability to form mineralized areas (cultures fed with Dexa) also decreased on serial subculture.

Ultracytochemical characteristics of cultured goat brain microvascular endothelial cells [corrected

This ultrastructural study was undertaken to determine the localization of cytochemically demonstrable blood-brain barrier (BBB)-associated enzymatic activities and of some nonenzymatic constituents in goat [corrected] brain microvascular endothelial cells (ECs) growing in vitro. Positive reactions for alkaline phosphatase (AP), 5'-nucleotidase (5'N), transport ATPase (Na+,K(+)-ATPase), and adenosine diphosphatase (ADPase) were present on both apical and basolateral plasma membranes (PMs) of the ECs. The reaction for calcium-dependent ATPase (Ca(2+)-ATPase) was less intense and was restricted to basolateral PM and associated plasmalemmal pits. These cells also revealed an abundance of anionic sites labeled with cationic colloidal gold (CCG) and Ricinus communis agglutinin 120 (RCA)-binding sites, specific for beta-D-galactosyl residues, on the apical PM. The labeling of the apical PM with Ulex europaeus agglutinin (UEA)-gold complex, specific for alpha-L-fucosyl residues, was negligible. When compared with results of cytochemical examination of the ECs of goat [corrected] brain capillary in vivo, these observations indicate that although cells cultivated in vitro retain at confluence the enzymatic activities typical for BBB-type ECS, they lose their characteristic (polar) localization. This loss is interpreted as a reflection of lost functional polarity of the microvascular endothelium in vitro resulting from deprivation of the normal influence of the components of brain parenchyma.

Calcification capacity of dental papilla mesenchymal cells transplanted in the isogenic mouse spleen

The capacity of the dental pulp to form calcified tissue was examined in papilla cells dissociated from first molar tooth germs of the neonatal mouse and isografted in the spleen for up to 7 days. To obtain papilla cell populations without odontoblasts, pulpal mesenchyme was isolated mechanically from the enamel organ after 0.1% trypsin treatment and rolled on a membrane filter. On day 3 after transplantation, the grafted papilla cells had changed into large, spindle-shaped cells, and initial calcification with needle-like crystals began in association with the collagenous matrix surrounding those cells. On day 7 after transplantation, the spindle cells transformed into odontoblast-like cells containing well-developed secretory organelles, and irregular, but nontubular, calcified tissues were commonly observed surrounding the extracellular collagenous matrix. The calcified tissue matrix with cellular inclusions displayed a structure similar to that of osteodentin. During this period, an intense positive reaction for alkaline phosphatase (ALPase) activity was demonstrated along the cell membranes of the odontoblast-like cells aligned at the periphery of forming calcified tissue. Enzymatic activity could not be detected on the cells incorporated completely into osteodentin-like matrix. The present results show that the papilla cell population transplanted into the spleen formed osteodentin-like material, thus demonstrating the capacity of papilla cells to produce calcified tissue.

Enzyme profiles of oral spirochetes in RapID-ANA system.

Enzyme profiles of oral Treponema species were determined by using RapID-ANA (Innovative Diagnostic System, Atlanta, Ga.), a 4-h test system which detects 18 enzymatic reactions, including aminopeptidases and glycosidases. Seventy-two clinical isolates of Treponema denticola, four reference strains of T. denticola (ATCC 35404, ATCC 35405, ATCC 35520, and ATCC 33521), one strain of T. vincentii (ATCC 35580), and two strains of T. socranskii subspecies (T. socranskii subsp. buccale ATCC 35534 and T. socranskii subsp. socranskii ATCC 35536) were used in this study. All T. denticola strains produced indole and a variety of aminopeptidases and glycosidases. These organisms could be differentiated into two groups on the basis of tetrazolium reductase and serine, phenylalanine, and glycine aminopeptidase activities. T. vincentii produced N-acetylglucosaminidase and arginine aminopeptidase, which facilitated the differentiation of this organism from T. socranskii subspecies and the T. denticola group. T. socranskii subspecies gave positive reactions for alkaline phosphatase only. These findings suggest that the RapID-ANA system is useful for enzymatic characterization and differentiation of oral spirochetes.

Identification of ‘Haemophilus somnus’ by rapid tests for pre-formed enzymes

In rapid tests for pre‐formed enzymes, all strains of ‘Haemophilus somnus’ examined gave positive reactions for alkaline phosphatase, cytochrome oxidase, β‐glucosidase, β‐glucuronidase and hippurate hydrolase but were negative for α‐mannosidase, β‐galactosidase and β‐xylosidase, tributyrin esterase, cystine aminopeptidase and γ‐glutamyl aminopeptidase. The pattern of results differed from those given by Actinobacillus, Haemophilus, ‘Histophilus’, Pasteurella and Taylorella species.

Ultrastructural analysis of the midgut of Drosophila auraria larvae - Distribution of alkaline phosphatase, acid phosphatase, leucine aminopeptidase, and glycogen.

The posterior midgut of Drosophila auraria larvae, and more specifically its anterior half, must be considered an important region for the digestion and the uptake of nutrients. The cells, and the region of the central lumen they define, are unique in that they give positive reaction for alkaline phosphatase; they also give the strongest histochemical reaction for leucine aminopeptidase. There are histochemical indications that the middle midgut, and especially its posterior region, can be a differentiated area of sugar storage.

Description and interpretation of morphological changes in canine kidneys after preservation with hypothermic solutions and subsequent reperfusion

The morphological appearance of canine kidneys preserved with hypothermic solutions of three different types (Rheomacrodex, Collins 3 and Sacks-II) was studied in acute autologous reperfusion models. While at the end of preservation the renal structure still seemed intact (with the exception of the Rheomacrodex model), reperfusion was associated with the manifestation of hitherto latent structural lesions. This is confirmed by light-microscopic features (hydropic swelling and destruction of the tubular epithelium), enzyme histochemical studies (positive alkaline phosphatase reaction of the Bowman space) and topo-optical reactions (loss of anisotropy of the cytomembranes in the presence of fairly well preserved birefringence of the basement membranes).

唇弁反転法における移植部の治癒過程に関する実験的研究 : 神経組織学的ならびに Alkaline Phosphatase 活性の変化について

Lip switch operation is a grafting method with the full thickness pedicle flap of one lip rotated to repair a defect of the other lip. This pedicle flap contains all tissues of the lip skin, orbicularis oris muscle, and oral mucosa. It is therefore considered that the method would reveal a slightly different healing process from the skin graft performed in the field of Plastic Surgery. Nerve-histological studies reported of the healing process of the flap are few. Consequently, there are unknown facts about the change of the nerve distribution in the grafted flap. Lip switch operations were therefore performed experimentally on the lip of dogs and the healing process was examined nerve-histologically. Furthermore, it was considered that an observation of the changes in alkaline phosphatase activity might clarify a phase of the healing process of the grafted tissue. The changes of enzyme therefore were observed histochemically. The results were as follows. 1. The epithelial layer of the grafted flap began to proliferate by the 3rd day after transplantation, but the mucous epithelial layer of the top of the flaps were degenerating from the 1st day to the 7th day after transplantation. 2. In the subcutaneus and submucous tissue layers of the flap, there were much inflammatory cell infiltration, edema, and vasodilatation at the first stage after transplantation. On the 10th day, they almost disappeared, but the dilated vessels were still seen to the 25th day after transplantation. 3. The muscle fibers in the grafted flap were degenerating to the 10th day after transplantation, but on the 120th day, they came to resemble the normal tissue. 4. Around the 5th day after transplantation, the neoformation of muscle fibers took place by continuous regeneration from muscle fibers of the grafted flap and adjacent tissues at the muscle layer of the junction, and more was seen at the distal (labial angular side) junction. 5. The nerve fibers in the flaps chiefly from the central part to the top degenerated after rotation operation. The nerve fibers in the pedicle part of the flaps degenerated after cutting off the pedicle on the 14th day. 6. By the 5th day after transplantation, the regenerating nerve fibers were seen budding out from the end and the lateral of the nerve fibers at the margin of the muscle layer of the adjacent tissues. On the 15th day, they came to enter the flaps, and on the 20th day, they could be observed in every part of the flaps. 7. Every regenerating nerve fiber in the grafted flap was connected with Schwann cell band. But at the junction, there were also excessive regenerating nerve fibers that had no connection with Schwann cell band, and most of them were degenerating on the 30th day. On the 60th day, the nerve fibers of the junction decreased in number. 8. Around the 45th day after transplantation, the formation of nerve endings was observed in the regenerating fibers. But on the 120th day, the number of endings was less than that of the normal tissue. 9. On the 120th day after transplantation, the condition and maturity of the nerve distribution in the grafted flaps were almost similar to the normal tissue, but in the pedicle part of the flaps, they were still less progressed. 10. In the epidermis of the grafted flap and the adjacent tissues, the basal cell layer showed a slight positive alkaline phosphatase reaction from the 3rd day to the 10th day, while the epithelium was proliferating. 11. In the muscle layer of the flap and the adjacent tissues, the capillaries showing a positive alkaline phosphatase activity increased in number remarkably from the 5th day to the 10th day after transplantation, and from the 6th day to the 11th day after cutting off the pedicle. 12. The changes in alkaline phosphatase activity of the junction were similar to those of the incised wound healing process, that is, the regenerating epithelium showed a positive reaction from the

Alkaline phosphatase and tartrate resistant acid phosphatase activity in cells of prolymphocytic leukemia.

In a typical case of prolymphocytic leukemia, blood smears and lymph node imprints have been investigated cytologically and cytochemically. It could be shown that many leukemic cells in both blood smears and lymph node imprints contained tartrate resistant acid phosphatase activity. Furthermore, the lymph node imprints disclosed many cells with a positive alkaline phosphatase reaction. Such a reaction hitherto has not been described in malignant cells of lymphoproliferative diseases. The cytochemical results underline that prolymphocytic leukemia indeed is a separate entity which can be differentiated from hairy cell leukemia and chronic lymphatic leukemia not only morphologically but also cytochemically. In addition, the case shows that leukemic blood cells are not inevitably identical with those occurring in organ infiltrates.

Ultrahistochemistry of matrix vesicles in elastic cartilage

Matrix vesicles in the elastic cartilage of epiglottis were negative for acid phosphatase, alkaline phosphatase, and ATPase. This is in agreement with the very rare occurrence of mineralization of elastic cartilage. Only the lysosomes of the chondrocytes showed a positive reaction for acid phosphatase, and a positive reaction for alkaline phosphatase and ATPase was found in relation to the cells of the perichondrium.

Isolation and metabolism of granular pneumocytes from rat lungs.

A population of rat lung cells enriched in granular pneumonocytes (type II cells) was prepared by modification of the method of Kikkawa (Lab. Invest.30, 76, 1974). Lungs were minced, mechanically agitated and incubated with a fluorochemical emulsion and 1% trypsin. The isolated cells were centrifuged in a discontinuous Ficoll gradient. Granular pneumonocytes were identified by the presence of cytoplasmic inclusions on Papanicolaou and acid phosphatase staining and a strongly positive alkaline phosphatase reaction. The yield at the interface between 1.058 and 1.100 density Ficoll was (4.0 ± 1.5) × 106 (mean ± SD) cells per g of lung tissue consisting of 72.7 ± 9.0% granular pneumonocytes and 20.3 ± 8.4% alveolar macrophages. O2 uptake by the cells at room temperature was 46.9 ± 5.8 nmol/h per 106 cells (mean ± SE; n=5). Respiration was inhibited by oligomycin and subsequently stimulated by an uncoupler of oxidative phosphorylation. During incubation with [U-14C] glucose, the cells produced14CO2, lactate and pyruvate at rates of 17.0 ± 2.9, 17.1 ± 1.6 and 8.7 ± 0.8 nmol/h/106 cells, respectively. These results show that a cell population enriched in granular pneumonocytes can be isolated by lung trypsinization. The cells in this preparation show respiration that is coupled to oxidative phosphorylation and intact glycolytic pathways.

Zur Darstellung der Vasa vasorum in der Tunica media der Arterienwand

A strongly positive alkaline phosphatase reaction in endothelial cells of capillaries and some precapil laries and a considerable activity of α-naphthylacetate esterase in endothelial cells of postcapillary vessels allow a complete demonstration of vasa vasorum in arteries of the muscular type.This method gives results also in such parts of arteries or pieces of their wall which do not allow a demonstration of their vasa vasorum by means of injection methods. In the internal spermatic artery of the bull a dense rope-ladder shaped capillary network is demonstrable within the outer parts of the tunica media with the alkaline phosphatase and the α-naphthylacetate esterase reactions. The two methods also may be combined. Most of the vasa vasorum arise directly from the lumen of the artery. They set through the whole of the arterial wall and spread over the vessel with retrograde branches. A uniformely thick inner portion of the arterial wall has no capillaries.

Cytological studies of the ductuli efferentes of the opossum.

AbstractDuctuli efferentes in four examined genera of family Didelphidae are organized into highly convoluted tubules, located within a conical body adjacent to the vascular plexus supplying the testis. They have yellowish‐green color in the adult Didelphis virginiana and grey pink color in other genera. The ductuli efferentes in all examined animals are lined by very low columnar cells, covered by microvilli or booth cilia and microvilli.Histochemical analysis reveals several types of cytoplasmic granules in the genera examined. A single sperical body about 1.5 μ in diameter is found in epithelial cells of the four eyed opossum (genus Philander); this body stains positively for RNA and non‐histone protein, and appears granular under the electron microscope, without a limiting membrane. Abundant cytoplasmic bodies in the Virginia opossum show strong reactions for SH groups; these granules may be responsible for the green color of the ductuli in this species. Ultrastructural studies show that the morphology of membrane limited granules in the ductuli of the examined genera is characteristic for each genus. Abundant pinocytotic vacuoles in the apices of these cells and a strongly positive alkaline phosphatase reaction suggest a marked absorptive activity of epithelial cells.

The presence of a common embryonic blastema for ovarian and testicular parenchymal (follicular, interstitial and tubular) cells in cattle, Bos taurus

The early embryonic gonadal development in the cattle is characterized by the appearance of an alkaline phosphatase positive blastema. Its derivatives in gonads of both sexes, follicular cells in the female and interstitial cells in the male, also show positive alkaline phosphatase reaction. Primordial germ cells are equally alkaline phosphatase positive, but loose this activity when they later transform to oogonia and oocytes, or to spermatogonia respectively. Using the enzyme activity as label to trace these constituents in the developmental steps of the bovine gonads, the following results were obtained.

Histochemical Studies on the Hypothalamo-hypophysial Neurosecretory System. III. Vitamin C, Phosphatase, Peroxydase and Zinc

Histochemical investigation was made on the hypothalamo-hypophyseal system of dog as to the vitamin C, phosphatase, peroxydase and zinc.1. Nerve cells of neurosecretory nuclei (supraoptic and paraventricular nuclei) were found to contain a large amount of vitamin C.2. Capillary in the hypothalamo-hypophyseal system shows a definitely positive alkaline phosphatase reaction, but this was found to be negative in the neurosecretory cells. In case yeast nucleic acid was used as substrate, some of the pituicytes showed positive reaction.3. Nerve cells of neurosecretory nuclei and some of the pituicytes were found to have markedly great amount of acid phosphase.4. No peroxydase was observed in neurosecretory cells.5. Zinc reaction was found to be highly positive in the pituitary intermediate lobe cells.

Alkaline phosphatase in the rabbit mammary gland.

STRONG positive reactions for alkaline phosphatase have been reported in the basket or ‘myo-epithelial’ cells of the mammary gland of some pregnant and lactating mammals1,2. Progressive transfer of this activity from the basket cells to the adjacent capillary endothelium later in pregnancy has also been described1.

'Kernechtrot' or nuclear fast red in the histochemical detection of calcareous corpuscles in Taenia saginata.

Taenia saginata, like many other cestodes1, is known to contain numerous calcareous corpuscles or calcium balls distributed throughout the parenchyma. These corpuscles, in addition to their calcium content, consist of an organic base, containing2 deoxyribo- and ribo-nucleic acids, simple proteins, glycogen and a hyaluronic acid - type of polysaccharide. With Gomori's modified technique these corpuscles also gave a positive reaction for alkaline phosphatase. The presence of calcium in these balls was indicated by von Kossa's silver test for calcium3.