Localization Of Acid Phosphatase Research Articles

HISTOCHEMICAL LOCALIZATION OF ACID AND ALKALINE PHOSPHATASES IN MONIEZIA EXPANSA (RUDOLPHI, 1805)

Cestodes or true tapeworms are important group of the Platyhelminthes, endoparasitic in the alimentary tract and associated ducts of various vertebrates and therefore, of great interest to the medical and veterinary profession.Moniezia expansa, the tapeworm under study is the common parasite of oldest domesticated animal sheep. It inhabits the small intestine especially the lower portion and absorb the nutrition of host from the intestine and deprive it of many important nutrients and thus deteriorates the quality of meat, wool, skin of host which leads to ecomical losses as well.Phosphatases are enzymes hydrolysing esters of phosphoric acid and play an important role in many metabolic processes. The histochemical localization of acid and alkaline phosphatase has been studied in Moniezia expansa as an effort to study some of its biological aspects which would help in its control and chemotherapy.Acid phosphatase was found to be localized in tegument, subtegumental cells, parenchymal muscles, interproglottidal gland tegument, Mehlis glands, testes, uterine wall and eggs and was not observed in parenchyma, interproglottidal gland cells, excretory vessels, ovaries and vitellaria. Alkaline phosphatase activity was observed in tegument, subtegumental cells, parenchymal muscles, parenchyma, interproglottidal glands, excretory vessels, membranes enclosing ovaries, vitellaria, Mehlis gland, testes, uterine wall and eggs.The presence of acid phosphatase in testes, mehlis' glands, uterine wall and eggs as well as alkaline phosphatase in eggs in Moniezia expansa has been reported for the first time.Both the acid and alkanine phosphatases showed the variations in their activity in the tegument along the strobila with highest in mature region. This gives the idea of high metabolic activity in the mature region of the cestode. These phosphatases play important role in metabolism of the parasite and seems to be involved in transportation, digestion, distribution of nutrients and metabolites and absorption of metabolic end products.

Toxicopathological impact of sub-lethal concentrations of lead nitrate on the gill of the catfish Heteropneustes fossilis

In recent studies, fish are heavily used as biomarkers of aquatic pollution, and heavy metals are among the main contributors to water pollution. In the present study, we investigated histopathological changes along with alterations in localization and activity of enzymes alkaline phosphatase (ALP), acid phosphatase (ACP), catalase (CAT), peroxidase (PER) and Na+/K+-ATPase in the gill tissues of Indian stinging catfish Heteropneustes fossilis exposed to two different concentrations (0.4 and 4 mg/L) of lead nitrate for 15 days. Histopathological examination of gill tissues revealed hypertrophy and swelling of epithelial cells, the fusion of epithelium of gill filaments and secondary lamellae, and alteration of secondary lamellae structure. Biochemical assays and histochemical localization show a pronounced effect on enzyme alkaline phosphatase activity and acid phosphatase in the gills of both groups of treated groups. In contrast, a significant decrease was noticed in the enzymatic response including catalase and peroxidase activity. Being a vital organ gill reflects the fish's physiological condition and the severity of the contamination in the surrounding environment. Gill is also the prime organ of osmoregulation in teleosts. Decreased activity of Na+/K+-ATPase suggests lead as a potent inhibitor of Na+/K+-ATPase that causes sodium hyperregulation. Alteration in the activity of metabolic enzymes reflects the level of tissue damage and metabolic disruption. At the same time, the increased activity of antioxidant enzymes states the condition of oxidative stress. Haematological parameters also altered with the lead nitrate exposure, reflecting metal toxicity and immune response against it. Meanwhile, this study also provides a potential use of H. fossilis as a biomarker for aquatic pollution.

Rice ACID PHOSPHATASE 1 regulates Pi stress adaptation by maintaining intracellular Pi homeostasis.

The concentration and homeostasis of intracellular phosphate (Pi) are crucial for sustaining cell metabolism and growth. During short-term Pi starvation, intracellular Pi is maintained relatively constant at the expense of vacuolar Pi. After the vacuolar stored Pi is exhausted, the plant cells induce the synthesis of intracellular acid phosphatase (APase) to recycle Pi from expendable organic phosphate (Po). In this study, the expression, enzymatic activity and subcellular localization of ACID PHOSPHATASE 1 (OsACP1) were determined. OsACP1 expression is specifically induced in almost all cell types of leaves and roots under Pi stress conditions. OsACP1 encodes an acid phosphatase with broad Po substrates and localizes in the endoplasmic reticulum (ER) and Golgi apparatus (GA). The phylogenic analysis demonstrates that OsACP1 has a similar structure with human acid phosphatase PHOSPHO1. Overexpression or mutation of OsACP1 affected Po degradation and utilization, which further influenced plant growth and productivity under both Pi-sufficient and Pi-deficient conditions. Moreover, overexpression of OsACP1 significantly affected intracellular Pi homeostasis and Pi starvation signalling. We concluded that OsACP1 is an active acid phosphatase that regulates rice growth under Pi stress conditions by recycling Pi from Po in the ER and GA.

Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells

BackgroundTartrate–resistant acid phosphatase (TRAP/ ACP5) belongs to the binuclear metallophosphatase family and is present in two isoforms. The primary translation product is an uncleaved TRAP 5a isoform with low phosphatase activity. TRAP 5a can be post-translationally processed to a cleaved TRAP 5b isoform with high phosphatase activity by e.g. cysteine proteinases, such as Cathepsin K (CtsK). The relevance of the phosphatase activity of TRAP 5b has been demonstrated for proliferation, migration and invasion of cancer cells. TRAP-overexpressing MDA-MB-231 breast cancer cells displayed higher levels of TRAP 5a and efficient processing of TRAP 5a to TRAP 5b protein, but no changes in levels of CtsK when compared to mock-transfected cells. In TRAP-overexpressing cells colocalization of TRAP 5a and proCtsK was augmented, providing a plausible mechanism for generation of TRAP 5b. CtsK expression has been associated with cancer progression and has been pharmacologically targeted in several clinical studies.ResultsIn the current study, CtsK inhibition with MK-0822/Odanacatib did not abrogate the formation of TRAP 5b, but reversibly increased the intracellular levels of a N-terminal fragment of TRAP 5b and reduced secretion of TRAP 5a reversibly. However, MK-0822 treatment neither altered intracellular TRAP activity nor TRAP-dependent cell migration, suggesting involvement of additional proteases in proteolytic processing of TRAP 5a. Notwithstanding, CtsK was shown to be colocalized with TRAP and to be involved in the regulation of secretion of TRAP 5a in a breast cancer cell line, while it still was not essential for processing of TRAP 5a to TRAP 5b isoform.ConclusionIn cancer cells multiple proteases are involved in cleaving TRAP 5a to high-activity phosphatase TRAP 5b. However, CtsK-inhibiting treatment was able to reduce secretion TRAP 5a from TRAP-overexpressing cancer cells.

Technique of histochemical staining on the activity of acid phosphatase for identification of larvae of the first stage of nematodes of Dirofilaria immitis, D. repens, Acanthocheilonema dracunculoides and A. reconditum species from the blood of dogs

The technique of staining on the activity of acid phosphatase has been modified, which makes it possible to differentiate between larvae of the first stage of nematodes of four species: Dirofilaria immitis, D. repens, Acanthocheilonema dracunculoides, A. reconditum from the blood of dogs. The method is based on the localization of acid phosphatase in different parts of the body in different types of microfilariae. For visualization, the substrate hydrolysis reaction, which catalyzes acid phosphatase, and the formation of an insoluble stained precipitate in these areas is used. The list of equipment and reagents is attached, a brief description of the process of preparation of clinical material, coloring and criteria of differentiation of species by color is given.

The effects of diversified phosphorus nutrition on the growth of oat (Avena sativa L.) and acid phosphatase activity

We studied the effect of differential phosphorus (P) supply on the development of oat seedlings (<em>Avena sativa</em> L. ‘Arab’) as well as localization and activity of acid phosphatases in tissues and root exudates. Plants were grown for 1–3 weeks on nutrient media with inorganic phosphate (+P, control), reduced Pi (0.1 P), phytic acid (PA) as organic P source, and without P addition (−P), in standard conditions or in a split-root culture system. Phosphate starvation reduced shoot growth but increased root elongation and root/shoot ratio, whereas 0.1 P and PA oat plants had similar growth parameters to +P plants. The growth on −P medium significantly decreased Pi content in all tissues, but only a slight Pi decrease was observed in plants grown on 0.1 P and PA media or various split-root system conditions. Pi starvation led to an increase in acid phosphatase (APase) activity in root exudates when compared to +P, 0.1 P, and PA plant samples. APase activity was especially intensive in root cross sections in rhizodermis and around/in vascular tissues of −P plants. For plants grown on 0.1 P medium and on phytic acid, APase activity did not change when compared to the control. Three major isoforms of APases were detected in plant tissues (similar in all studied conditions, with a higher activity of one isoform under Pi deficit). Generally, lowered Pi content (0.1 P) was not stressful to oat plants for up to 3 weeks of culture. Oat plants grew equally well on nutrient media with Pi and on media with phytate, although phytate was considered not available for other plants. The oat plants activated mainly extracellular APases, but also intracellular enzymes, rather via nonlocal signals, to acquire Pi from external/internal sources under Pi deficiency.

Mechanisms of oat (Avena sativa L.) acclimation to phosphate deficiency.

BackgroundDeficiency of available forms of phosphorus is common in most soils and causes reduction of crop plants growth and yield. Recently, model plants responses to phosphate (Pi) deficiency have been intensively studied. However, acclimation mechanisms of cereals like oat (Avena sativa L.), to low Pi stress remains not fully understood. Oat plants have been usually cultured on poor soils, with a low nutrient content, but their responses to such conditions are not well known, therefore the main goal of the study was to investigate the mechanisms that enable oat plants to grow under low Pi conditions.MethodsFour oat cultivars (A. sativa, cv. Arab, Krezus, Rajtar and Szakal) were grown for three weeks in a nutrient media with various P sources: inorganic—KH2PO4 (control), organic—phytate (PA) and with no phosphate (−P). The effects of Pi deficiency on the level of P, oat growth parameters, intensity of photosynthesis, plant productivity, root exudation ability, localization, activity and isoforms of acid phosphatases, enzymes involved in Pi mobilization, were estimated. In addition, the effect of mycorrhization on plant growth was also observed.ResultsAll studied oat cultivars grown on Pi-deficient media had significantly decreased Pi content in the tissues. Pi deficiency caused inhibition of shoot growth, but generally it did not affect root elongation; root diameter was decreased, root/shoot ratios increased, whereas PA plants showed a similar growth to control. Photosynthesis rate and productivity parameters decreased under low Pi nutrition, however, sugar content generally increased. Studied oat cultivars did not respond to low Pi via increased exudation of carboxylates from the roots, as pH changes in the growth media were not observed. Pi starvation significantly increased the activity of extracellular and intracellular acid phosphatases (APases) in comparison to the control plants. Three major APase isoforms were detected in oat tissues and the isoform pattern was similar in all studied conditions, usually with a higher level of one of the isoforms under Pi starvation. Generally no significant effects of mycorrhizal colonization on growth of oat cultivars were observed.DiscussionWe postulated that acid phosphatases played the most important role in oat cultivars acclimation to Pi deficiency, especially extracellular enzymes involved in Pi acquisition from soil organic P esters. These APases are mainly located in the epidermis of young roots, and may be released to the rhizosphere. On the other hand, intracellular APases could be involved in fast Pi remobilization from internal sources. Our study showed that oat, in contrast to other plants, can use phytates as the sole source of P. The studied oat cultivars demonstrated similar acclimation mechanisms to Pi deficiency, however, depending on stress level, they can use different pools of acid phosphatases.

Acid Phosphatase and Vitamin A Induced Abnormal Tail Regeneration in Frog Tadpoles: an Immunohistochemical Study

Application of vitamin A palmitate or retinoic acid causes abnormal tail regeneration and sometimes leads to formation of ectopic hind limbs along with pelvic girdles at the cut end of tail. Such a phenomenon referred to as homeotic transformation, has been reported in different anuran species. Acid phosphatase, a lysosomal marker enzyme is known to be associated with regeneration in amphibians. However, role of this enzyme during homeotic transformation of tail of an anuran tadpole into limbs is not explored. Thus, aim of the present study is immunohistochemical localization of acid phosphatase in non-amputated, normally regenerated and vitamin A treated abnormally regenerated tails (a prerequisite for ectopic organ development) of tadpoles of the Indian tree frog Polypedates maculatus. In the non-amputated and normally regenerated tails, acid phosphatase is majorly restricted to the epidermis and muscle patches although in the normally regenerated tails notochordal sheath and spinal cord also stain for acid phosphatase. In vitamin A treated tails, acid phosphatase is mostly localized in the epidermis, notochord precursor cells and undifferentiated cells of the mesenchyme. Notochordal cells and notochordal sheath also show positive staining. Since, acid phosphatase is majorly expressed by tissue forming precursor cells, this enzyme is suggested to be involved in tissue remodelling processes.

Acid and alkaline phosphatase localization in the digestive tract mucosa of the Hemisorubim platyrhynchos

This cytochemical study investigated the acid and alkaline phosphatase of the digestive tract of Hemisorubim platyrhynchos. Acid phosphatase was detected in the lining epithelium throughout the digestive tract, whereas alkaline phosphatase was only observed in the intestine. In the esophagus, an acid phosphatase reaction occurred in the apical cytoplasm of the epithelial cells and was related to epithelial protection and freeing of superficial cells for sloughing. Similar results were also observed in epithelial cells of gastric epithelium. In the gastric glands, acid phosphatase occurred in lysosomes of the oxynticopeptic cells acting in the macromolecule degradation for use as an energy source, whereas in the vesiculotubular system, its presence could be related to secretion processes. Furthermore, acid phosphatase in the intestine occurred in microvilli and lysosomes of the enterocytes and was correlated to absorption and intracellular digestion. However, no difference was reported among the regions of the intestine. However, alkaline phosphatase reaction revealed a large number of reaction dots in the anterior intestine, with the number decreasing toward the posterior intestine. This enzyme has been related to several functions, highlighting its role in the nutrient absorption primarily in the anterior intestine but also being essential in pH regulation because this is a carnivorous species with many gastric glands with secretions that could damage the intestine.

Cytochemical Localization of ATPase and Acid Phosphatase in Minor Veins of the Leaf of Vicia faba During Different Developmental Stages

Cytochemical Localization of ATPase and Acid Phosphatase in Minor Veins of the Leaf of <i>Vicia faba</i> During Different Developmental Stages

The R740S mutation in the V‐ATPase a3 subunit results in osteoclast apoptosis and defective early‐stage autophagy

Vacuolar-type H(+)-ATPases (V-ATPases) are located in lysosomes and at the ruffled border in osteoclasts. We showed previously that the R740S mutation is dominant negative for V-ATPase activity, uncouples proton transport from ATP hydrolysis and causes osteopetrosis in heterozygous mice (+/R740S). Here we show mice homozygous for R740S (R740S/R740S) have more severe osteopetrosis and die by postnatal day 14. Although R740S/R740S osteoclasts express wild-type levels of a3, it is mislocalized. Acridine orange staining of R740S/R740S osteoclasts grown on a Corning resorptive surface reveals no resorption and no acidification of intracellular compartments. Whereas osteoblast and osteocyte apoptosis is normal, R740S/R740S osteoclasts exhibit increased apoptosis compared with wild-type osteoclasts. Localization of the enzyme tartrate-resistant acid phosphatase (TRAP) is also aberrant. Transmission electron microscopy reveals that R740S/R740S osteoclasts do not polarize, lack ruffled borders, and contain fewer autophagosomes. Consistent with an early stage defect in autophagy, expression of LC3II is reduced and expression of p62 is increased in R740S/R740S compared to wild-type osteoclasts. These results indicate the importance of intracellular acidification for the early stages of autophagy as well as for osteoclast survival, maturation, and polarization with appropriate cytoplasmic distribution of key osteoclast enzymes such as TRAP.

慢性中耳炎における耳小骨の酸フォスタァーゼ活性 特に真珠腫, および非真珠腫における骨破壊との関係

慢性中耳炎における耳小骨の酸フォスタァーゼ活性 特に真珠腫, および非真珠腫における骨破壊との関係

Histochemical Distribution of Digestive Enzymes in the Intestine of the Common Two‐Banded SeaBream, Diplodus vulgaris, Geoffroy St‐Hilaire 1817

The histochemical localization of non-specific esterase, alkaline and acid phosphatase as well as aminopeptidase in the intestine of the free-living common two-banded sea bream (Diplodus vulgaris) was investigated. Fish were caught near the town of Zadar (Adriatic Sea, Croatia). Samples of pyloric caeca and three parts of the intestine proper (anterior, middle and posterior) were used for the description of non-specific esterase, alkaline and acid phosphatase as well as aminopeptidase. Non-specific esterase activity was found in the cytoplasm of enterocytes in pyloric caeca and in all investigated intestinal segments. The activity was stronger in the anterior and posterior part of the intestine than in the pyloric caeca and middle segment of the intestine. Intestinal alkaline phosphatase was detected in brush border of enterocytes of all investigated intestinal segments. Enzymatic activity gradually decreased in a posterior direction. Acid phosphatase activity was observed as a fine granular reaction product in the supranuclear region of enterocytes. This activity was almost equal in pyloric caeca as well as in the anterior intestinal segment, while it was stronger in the middle and posterior intestinal segment. Aminopeptidase was present along the intestinal epithelium brush border in all investigated parts of the digestive tube. The intensity of aminopeptidase increased posteriorly. The possible role of investigated enzymes in intracellular digestion and transport is discussed.

Fluorescence spectroscopy measures yeast PAH1-encoded phosphatidate phosphatase interaction with liposome membranes

Phosphatidate (PA) phosphatase, the enzyme that catalyzes the penultimate step in triacylglycerol synthesis, is a cytosolic enzyme that must associate with the membrane where its substrate PA resides. Fluorescence spectroscopy was used to measure the interaction of yeast PAH1-encoded PA phosphatase with model liposome membranes. PA phosphatase contains five tryptophan residues and exhibited inherit fluorescence that increased upon interaction with phosphatidylcholine liposomes. The interaction was enhanced by inclusion of other phospholipids and especially the substrate PA. Interaction was dependent on both the concentration of phosphatidylcholine-PA liposomes as well as the surface concentration of PA in liposomes. Mg(2+) ions, which were required for catalysis, did not affect PA phosphatase interaction with phosphatidylcholine-PA liposomes. PA phosphatase was a substrate for protein kinase A, protein kinase C, and casein kinase II, and these phosphorylations decreased PA phosphatase interaction with phosphatidylcholine-PA liposome membranes.

Relationship of glucose and oleate metabolism to cardiac function in lipin-1 deficient (fld) mice

Lipin-1 is the major phosphatidate phosphatase (PAP) in the heart and a transcriptional coactivator that regulates fatty acid (FA) oxidation in the liver. As the control of FA metabolism is essential for maintaining cardiac function, we investigated whether lipin-1 deficiency affects cardiac metabolism and performance. Cardiac PAP activity in lipin-1 deficient [fatty liver dystrophy (fld)] mice was decreased by >80% compared with controls. Surprisingly, oleate oxidation and incorporation in triacylglycerol (TG), as well as glucose oxidation, were not significantly different in perfused working fld hearts. Despite this, [³H]oleate accumulation in phosphatidate and phosphatidylinositol was increased in fld hearts, reflecting the decreased PAP activity. Phosphatidate accumulation was linked to increased cardiac mammalian target of rapamycin complex 1 (mTORC1) signaling and endoplasmic reticulum (ER) stress. Transthoracic echocardiography showed decreased cardiac function in fld mice; however, cardiac dysfunction was not observed in ex vivo perfused working fld hearts. This showed that changes in systemic factors due to the global absence of lipin-1 could contribute to the decreased cardiac function in vivo. Collectively, this study shows that fld hearts exhibit unchanged oleate esterification, as well as oleate and glucose oxidation, despite the absence of lipin-1. However, lipin-1 deficiency increases the accumulation of newly synthesized phosphatidate and induces aberrant cell signaling.

Nuclear Envelope Phosphatase 1-Regulatory Subunit 1 (Formerly TMEM188) Is the Metazoan Spo7p Ortholog and Functions in the Lipin Activation Pathway

Lipin-1 catalyzes the formation of diacylglycerol from phosphatidic acid. Lipin-1 mutations cause lipodystrophy in mice and acute myopathy in humans. It is heavily phosphorylated, and the yeast ortholog Pah1p becomes membrane-associated and active upon dephosphorylation by the Nem1p-Spo7p membrane complex. A mammalian ortholog of Nem1p is the C-terminal domain nuclear envelope phosphatase 1 (CTDNEP1, formerly "dullard"), but its Spo7p-like partner is unknown, and the need for its existence is debated. Here, we identify the metazoan ortholog of Spo7p, TMEM188, renamed nuclear envelope phosphatase 1-regulatory subunit 1 (NEP1-R1). CTDNEP1 and NEP1-R1 together complement a nem1Δspo7Δ strain to block endoplasmic reticulum proliferation and restore triacylglycerol levels and lipid droplet number. The two human orthologs are in a complex in cells, and the amount of CTDNEP1 is increased in the presence of NEP1-R1. In the Caenorhabditis elegans embryo, expression of nematode CTDNEP1 and NEP1-R1, as well as lipin-1, is required for normal nuclear membrane breakdown after zygote formation. The expression pattern of NEP1-R1 and CTDNEP1 in human and mouse tissues closely mirrors that of lipin-1. CTDNEP1 can dephosphorylate lipins-1a, -1b, and -2 in human cells only in the presence of NEP1-R1. The nuclear fraction of lipin-1b is increased when CTDNEP1 and NEP1-R1 are co-expressed. Therefore, NEP1-R1 is functionally conserved from yeast to humans and functions in the lipin activation pathway.

Autophagy in Tobacco BY-2 Cells Cultured under Sucrose Starvation Conditions: Isolation of the Autolysosome and its Characterization

Tobacco culture cells carry out a large-scale degradation of intracellular proteins in order to survive under sucrose starvation conditions. We have previously suggested that this bulk degradation of cellular proteins is performed by autophagy, where autolysosomes formed de novo act as the major lytic compartments. The digestion process in autolysosomes can be retarded by addition of the cysteine protease inhibitor E-64c to the culture medium, resulting in the accumulation of autolysosomes. In the present study, we have investigated several properties of autolysosomes in tobacco cells. Electron microscopy showed that the autolysosomes contain osmiophilic particles, some of which resemble partially degraded mitochondria. It also revealed the presence of two kinds of autolysosome precursor structures; one resembled the isolation membrane and the other the autophagosome of mammalian cells. Immunofluorescence microscopy showed that autolysosomes contain acid phosphatase, in accordance with cytochemical enzyme analyses by light and electron microscopy in a previous study. Autolysosomes isolated by cell fractionation on Percoll gradients showed the localization of acid phosphatase, vacuolar H(+)-ATPase and cysteine protease. These results show that starvation-induced autophagy in tobacco cells follows a macroautophagic-type response similar to that described for other eukaryotes. However, our results indicate that, although the plant vacuole is often described as being equivalent to the lysosome of the animal cell, a new low pH lytic compartment-the autolysosome-also contributes to proteolytic degradation when tobacco cells are subjected to sucrose deprivation.

Evasion of IFN-γ Signaling by Francisella novicida Is Dependent upon Francisella Outer Membrane Protein C

Background Francisella tularensis is a Gram-negative facultative intracellular bacterium and the causative agent of the lethal disease tularemia. An outer membrane protein (FTT0918) of F. tularensis subsp. tularensis has been identified as a virulence factor. We generated a F. novicida (F. tularensis subsp. novicida) FTN_0444 (homolog of FTT0918) fopC mutant to study the virulence-associated mechanism(s) of FTT0918.Methods and FindingsThe ΔfopC strain phenotype was characterized using immunological and biochemical assays. Attenuated virulence via the pulmonary route in wildtype C57BL/6 and BALB/c mice, as well as in knockout (KO) mice, including MHC I, MHC II, and µmT (B cell deficient), but not in IFN-γ or IFN-γR KO mice was observed. Primary bone marrow derived macrophages (BMDM) prepared from C57BL/6 mice treated with rIFN-γ exhibited greater inhibition of intracellular ΔfopC than wildtype U112 strain replication; whereas, IFN-γR KO macrophages showed no IFN-γ-dependent inhibition of ΔfopC replication. Moreover, phosphorylation of STAT1 was downregulated by the wildtype strain, but not the fopC mutant, in rIFN-γ treated macrophages. Addition of NG-monomethyl-L-arginine, an NOS inhibitor, led to an increase of ΔfopC replication to that seen in the BMDM unstimulated with rIFN-γ. Enzymatic screening of ΔfopC revealed aberrant acid phosphatase activity and localization. Furthermore, a greater abundance of different proteins in the culture supernatants of ΔfopC than that in the wildtype U112 strain was observed.Conclusions F. novicida FopC protein facilitates evasion of IFN-γ-mediated immune defense(s) by down-regulation of STAT1 phosphorylation and nitric oxide production, thereby promoting virulence. Additionally, the FopC protein also may play a role in maintaining outer membrane stability (integrity) facilitating the activity and localization of acid phosphatases and other F. novicida cell components.

Ultrastructure and cytochemical localization of acid phosphatase of laticifers in Euphorbia kansui Liou

Acid phosphatase (AcPase) activities are involved in the degeneration process of cytoplasm in plants. In this study, acid phosphatase was detected by the method of lead nitrate and cytochemical electron microscopy during the development of nonarticulated laticifers in Euphorbia kansui Liou. The most important feature in the differentiation of the laticifers in E. kansui is that the development of small vacuoles arises from endoplasmic reticulum (ER). The mature laticifers possess a thin layer of electron-dense peripheral cytoplasm in which the organelle cannot be distinguished and a large central vacuole filled with latex particles. AcPase cytochemistry studies show AcPase reaction products congregated into heaps are distributed along the tonoplast of central vacuole and around the mitochondria and plastids. Some small vacuoles which develop at later developmental stages of laticifers contain AcPase reaction products. As a result, the central vacuole is formed by cellular autophagy and fusion of small vacuoles which apparently arises from ER.

Combined apoptosis and autophagy, the process that eliminates the oocytes of atretic follicles in immature rats

We studied the alterations of dying oocytes in 1-28 days old rats using TUNEL method, immunolocalizations of active caspase 3, lamp1, localization of acid phosphatase, and DAPI staining. All procedures were performed in adjacent sections of each oocyte. In most dying oocytes exist simultaneously features of apoptosis as active caspase 3 and DNA breaks, and a large increase of lamp1 and acid phosphatase characteristic of autophagy. Large clumps of compact chromatin and membrane blebbing were absent. Electron microscope observations demonstrated the presence of small clear vesicles and autophagolysosomes. All these features indicate that a large number of oocytes are eliminated by a process sharing features of apoptosis and autophagy. In dying oocytes of new born rats the markers of apoptosis predominate over those of autophagy. However, fragmentation and apoptotic bodies were not found. These features suggest that in different cytophysiological conditions the processes of cell death may be differently modulated.