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Synthesis and antimicrobial activity of pyridines bearing thiazoline and thiazolidinone moieties.

Two series of new pyridines bearing thiazoline (3a--n) and thiazolidinone (5a--e) moieties were prepared via the cyclization of the corresponding substituted pyridyl thiourea (2a--g) with an appropriately substituted phenacyl bromide or chloroacetic acid, respectively. The antimicrobial activity was determined for representative compounds and most of them showed moderate activity against Gram-positive bacteria.

Anti-Bacterial Agents↗

A facile preparation of 1-(6-hydroxyindol-1-yl)-2,2-dimethylpropan-1-one.

An effective synthesis of 1-(6-hydroxyindol-1-yl)-2,2-dimethylpropan-1-one (4) was developed starting from 1H-indole (2). The key step involved suitable utilization of 4-(1-pyrrolidino)pyridine for the removal of the chloroacetyl moiety from chloroacetic acid 1-(2,2-dimethylpropionyl)-1H-indol-6-yl ester (3); a possible mechanism is, also, presented. Compound 4 might lead to selectively substituted derivatives, either on the phenolic-OH or the indolyl-NH, with putative biological interest. In this respect, we found that the core structure of 1H-indol-6-ol (1) possesses a degree of aldose reductase inhibitory potential, at a concentration of 100 microM.

Aldehyde Reductase↗

Histamine synthesis by cells of the macrophage lineage in bone marrow of mice.

An injection of Escherichia coli lipopolysaccharide (LPS) increased the activity of histidine decarboxylase (HDC) in bone marrow (BM) cells of C3H/HeN mice much more than in C3H/HeJ mice, which are resistant to various effects of LPS. In WBB6/F1 (W/Wv) mice, which are genetically deficient in mast cells, HDC activity increased more than in C3H/HeN mice. Cultured BM cells of W/Wv mice spontaneously synthesized histamine in a HDC-dependent way. LPS caused a slight increase in HDC-associated histamine synthesis by these cells. Treatment of the BM cells with murine recombinant granulocyte-macrophage colony stimulating factor (mrGM-CSF) increased the histamine synthesis. In addition, treatment with mrGM-CSF made the cells respond to LPS by a dose-dependent increase in HDC activity and histamine synthesis. Most dish-adherent BM cells that had been treated with both mrGM-CSF and LPS for 48 h were stained for nonspecific esterase and not for chloroacetate esterase, and had twice as much HDC activity as the nonadherent cells had. Immunocytochemical analysis of the BM cells of W/Wv mice treated with both mrGM-CSF and LPS showed that HDC was in the cytoplasm of cells having Mac-1, a macrophage-differentiation antigen. These results suggest that cells of the macrophage lineage in the BM of mice synthesize histamine.

Animals↗

[Morphological characteristics of human mast cells in normal and pathological thyroid glands. Functional aspect of human mast cells in comparison with rat mast cells].

We examined the distribution and population density of human mast cells in thyroid glands. The results were compared with those of Sprague-Dawley (SD) rats because they thyroid function of SD rats is known to be under the control of bioactive amines discharged from mast cells. Normal thyroid tissues were obtained either form autopsy or from a normal portion of the tissue distant from nodular lesions. Thyroid tissues were surgically removed from cases of Graves' disease and other tumorous lesions such as follicular adenoma, follicular carcinoma, papillary carcinoma and medullary carcinoma. The tissues were fixed with buffered formaldehyde or Carnoy fluid and embedded in paraffin. Mast cells were stained with toluidine blue and naphthol ASD chloroacetate esterase (esterase). Immunoperoxidase reactions to antihuman tryptase and chymase monoclonal antibodies were then observed. The mast cells were also observed by electron microscopy. The histamine content of the thyroid tissues was estimated by the high-performance liquid chromatography method. The mast cells in SD rat thyroid glands were scattered in perifollicular connective tissues which were comprised of capillaries, fibroblasts, nerve fibers and occasional fine deposits of collagen fibrils. Their cytoplasmic granules appeared to be distinct, electron dense and amorphous. In contrast, the mast cells in normal human thyroid glands were scattered exclusively over relatively thick interstitial spaces like the interlobular and subcapsular connective tissues. These mesenchymal tissues were composed of bundles of collagen fibrils, fibroblasts, histiocytes and thin cytoplasmic processes of unknown origin. In pathologic thyroid tissues, the mast cells were distributed in a similar pattern over the connective tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

1,1-Dichloroethylene hepatotoxicity: proposed mechanism of action and distribution and binding of 14C radioactivity following inhalation exposure in rats.

1,1-Dichloroethylene is reported to produce renal tumors in male mice. It is an hepatotoxin in fasted rats after inhalation. We found that trichloropropane epoxide, an inhibitor of epoxide hydrase, enhances hepatic injury as measured by serum sorbitol dehydrogenase elevation. A significant elevation of hepatic citric acid concentration was seen in fasted but not fed rats. We hypothesized that mitochondrial injury was associated with inhibition of the tricarboxylic acid cycle and postulated that monochloroacetic acid was a toxic metabolite of 1,1-DCE. Fluoroacetic acid and chloroacetic acid were similar in their ability to inhibit oxygen uptake when pyruvic and malic acids were substrates in isolated mitochondria supplemented with adenosine diphosphate. In experiments where 1,1-DCE metabolism was estimated, no difference between the rate of uptake in a 2-hr period was detected between fed and fasted animals. Urinary output of radioactivity at 26 hr for fed and fasted rats was similar. Water-soluble (i.e. TCA-soluble) 1,1-DCE metabolites were found in tissues of fasted rats in excess of that seen in fed rats. The kidney had the largest concentration of total metabolites. Tissue-bound, or TCA-insoluble, radioactivity was associated with the mitrochondrial and microsomal fraction of fasted rats in excess of that seen in fed rats. The disappearance of TCA-insoluble radioactivity from the mitochondrial and microsomal fractions was comparable in rate between fed and fasted rats respectively. These results suggest that 1,1-DCE is metabolized quite rapidly in the organism to TCA-soluble components which are excreted by the kidneys. Metabolites of 1,1-DCE may enter the metabolic pool, since a reasonably short turnover of (14)C-labeled, bound material was observed. The metabolite of 1,1-DCE appears to inhibit the mitochondria so that citric acid accumulates. This may occur by a process of lethal synthesis.

Acetates↗

Occurrence of by-products of strong oxidants reacting with drinking water contaminants--scope of the problem.

This paper describes results of a detailed literature review of the organic and inorganic by-products that have been identified as being formed in aqueous solution with four of the strong oxidizing/disinfecting agents commonly employed in drinking water treatment. These agents are: chlorine, chlorine dioxide, chloramine, and ozone. Significant findings include the production of similar nonchlorinated organic oxidation products from chlorine, chlorine dioxide, and ozone. In addition, all three chlorinous oxidants/disinfectants can produce chlorinated by-products under certain conditions. The presence of chloronitrile compounds in drinking waters is indicated to arise from reactions of chlorine or chloramine to amine/amide functions in amino acids or proteinaceous materials, followed by dehydrohalogenation. These nitriles could hydrolyze to produce the corresponding chloroacetic acids. It is concluded that to minimize the presence of oxidation by-products in drinking waters, the concentrations of oxidizable organic/inorganic impurities should be lowered before any oxidizing agent is added.

Chemical Phenomena↗

NaF induces early differentiation of murine bone marrow cells along the granulocytic pathway but not the monocytic or preosteoclastic pathway in vitro.

The stimulatory effects of sodium fluoride (NaF) on bone formation have been explained solely by its activation of osteoblasts. However, whether and how NaF acts on the osteoclast lineage is poorly understood. We previously found that NaF differentiates HL-60 cells to granulocytic cells. To further test this action, we have employed here primary cultures of progenitor cells derived from murine bone marrow. NaF at subtoxic concentrations (<0.5 mM) significantly up-regulated activities of several intracellular enzymes (lactate dehydrogenase, beta-glucuronidase, acid phosphatase), cellular reduction of nitroblue tetrazolium, and nitric oxide (NO) production; which are all accepted as general differentiation markers. NaF (<0.5 mM) also up-regulated granulocyte-specific markers (chloroacetate esterase, cell surface antigens [Mac-1, Gr-1]) but not any of the monocyte-specific markers (nonspecific esterase, cell surface antigens [F4/80, MOMA-2]). Although other general differentiation markers (phagocytosis, adhesion, appearance, nuclear:cytoplasmic ratio) were not appreciably influenced by NaF, essentially in support of our previous data from HL-60 cells, the present findings suggest that NaF induces early differentiation of bone marrow hemopoietic progenitor cells along the granulocytic pathway but not the monocytic pathway that is linked to osteoclast formation. Therefore, in addition to its potent stimulatory effects on osteoblastic bone formation, NaF applied to patients with osteoporosis could be expected to indirectly reduce osteoclastic bone resorption.

Acid Phosphatase↗

Ultrastructure and cytochemical characteristics of leukocyte infected with Hepatozoon canis.

We have observed the ultrastructure of a canine leukocyte infected with Hepatozoon canis by transmission electron microscope. The infected leukocytes contained many small vesicles and a few typical granules in the cytoplasm. They also contained a few rough surfaced endoplasmic reticula, mitochondria and a segmented nucleus with condensed chromatin. Cytochemical analysis indicated that the infected leukocytes with H. canis contained little alkaline phosphatase, peroxidase, naphthol AS-D chloroacetate esterase or alpha naphthol acetate esterase. These results suggested that the infected leukocytes were neutrophils or monocytes which lost their typical characteristics.

Alkaline Phosphatase↗

Myeloid sarcoma involving the gynecologic tract: a report of 11 cases and review of the literature.

Myeloid sarcoma can involve any anatomic site, but involvement of the gynecologic tract is uncommon. We describe 11 women, 17 to 60 years old, with myeloid sarcoma involving the gynecologic tract, including 5 patients in whom myeloid sarcoma presented as an isolated mass. The uterus was the most frequently involved anatomic site, in 8 patients (5 corpus, 3 cervix). Each neoplasm diffusely infiltrated normal structures, and, cytologically 7 tumors were immature, 3 were differentiated, and 1 was blastic. In 9 cases assessed, immunohistochemical stains showed that all neoplasms were positive for myeloperoxidase and lysozyme; CD117 was positive in 7 of 8 cases, and cytochemical staining for naphthol AS-D chloroacetate was positive in all 6 neoplasms analyzed. Following chemotherapy, complete remission and long-term survival were achieved in a subset of patients, as was particularly true for 2 patients (cases 8 and 10), with complete remission 12.5 and 31 years after diagnosis, respectively.

Adolescent↗

Myeloid sarcoma involving the testis.

Myeloid sarcoma is a neoplasm of immature granulocytes, monocytes, or both involving any extramedullary site. Myeloid sarcoma involving the testis, however, is uncommon and very rarely occurs as an isolated mass. We describe 4 patients with myeloid sarcoma involving the testis, including 2 patients in whom the neoplasm was isolated to the testis (1 unilateral and 1 bilateral). Histologically, 4 neoplasms were poorly differentiated and 1 was blastic. Each neoplasm was shown to be of myeloid lineage, and negative for T- and B-cell specific antigens using immunohistochemical methods. One case was also positive for chloroacetate esterase. In the literature, most cases of myeloid sarcoma involving the testis represent relapse or the initial presentation of acute myeloid leukemia. Including the 2 cases we report here, only 7 cases of myeloid sarcoma isolated to the testis have been reported.

Adult↗

Immunohistochemical and histochemical stains for differentiating canine cutaneous round cell tumors.

Immunohistochemical and histochemical stains are useful adjunct techniques in the diagnosis of canine cutaneous round cell tumors, which can appear histologically similar. We applied a panel of monoclonal antibodies (recognizing tryptase, chymase, serotonin for mast cells; CD1a, CD18, MHC class II for histiocytes; CD3 for T lymphocytes; CD79a for B lymphocytes and plasma cells) and one histochemical stain (naphthol AS-D chloroacetate for chymase activity) to formalin-fixed, paraffin-embedded sections of canine cutaneous mast cell tumors, histiocytomas, lymphosarcomas, plasmacytomas, and unidentified round cell tumors. Of 21 tumors with a histologic diagnosis of mast cell tumor, 7/7 (100%) grade I, 6/7 (85.7%) grade II, and 3/7 (42.9%) grade III tumors were diagnosed as mast cell tumors based on positive staining for tryptase antigen and chymase activity. Mast cells were positive for both tryptase antigen and chymase activity, indicating equal efficacy of tryptase immunohistochemistry and chymase histochemistry. Chymase was detected immunohistochemically in both tumor and nontumor cells, while serotonin was not detected in most mast cell tumors, and thus, neither was useful in the diagnosis of mast cell tumors. Immunohistochemistry to detect CD18 and MHC class II was equally effective in staining histiocytomas, although lymphosarcoma must be ruled out through the use of CD3 and CD79a immunohistochemistry. Immunohistochemistry using three different monoclonal antibodies to human CD1a showed no cross-reactivity in canine histiocytomas and was not useful. A final diagnosis was obtained for 4/5 (80%) of the unidentified tumors, indicating the usefulness of multiple stains in poorly differentiated round cell tumors.

Animals↗

Cyclosporin A-induced changes of the thymic microenvironment. A review of morphological studies.

Cyclosporin A is an immunosuppressive drug, which disrupts the activation of peripheral T-lymphocyte pool and blocks the maturation of thymocytes within the thymus. Normally, thymic nonlymphoid cells provide the optimal inductive microenvironment for development of T-lymphocytes. After application of cyclosporin A the complex alterations of the thymic microenvironment occur, affecting all types of nonlymphoid cells. All subsets of thymic epithelial cells are thoroughly changed. The subcapsular epithelial cells show the prominent enlargement of cytokeratin contents. In electron microscopy, however, these cells present the morpho-functional aspect of resting cells. The epithelial cells in deeper cortex become enlarged and stockier, whereby their cell processes appear more ramified and thicker. Thus, the cytoreticulum they create seems much denser. These cells strongly express MHC antigens. Their subcellular organization is suggestive of increased synthetic and secretory activity. The number of medullary epithelial cells is decreased. The cells with the most mature phenotype are the most prominently depleted and the ones with phenotypically and morphologically immature appearance predominate. The number of Hassall's bodies is also decreased. The number of cortical macrophages does not increase. However, these cells become enlarged showing the prominent changes in enzyme capacity, histochemical features and ultrastructural organization. Thus, they become similar to macrophages located in the cortico-medullary zone of the normal rat thymus. Cortical macrophages increase the activity of hydrolytic enzymes, acid phosphatase and nonspecific esterase, develop the strong activity of chloroacetate esterase, the strong activity of respiratory enzyme succinic dehydrogenase and begin to show the marked presence of prostaglandin synthase. Moreover, the cytoplasmic inclusions, which are aldehyde fuchsin- and PAS-positive and show sudanophilia, appear within cortical macrophages. In electron microscopy these cells show an abundant cytoplasm a very active appearance and the variety of vacuolar cytoplasmic inclusions. The mitoses of neighboring thymocytes are often seen. The number of interdigitating cells is decreased due to reduced size of thymic medulla, but these cells do not show the substantial phenotype changes. The description and classification of all types of nonlymphoid cells, which constitute the normal thymic microenvironment, is also presented. The functional significance and possible mechanisms of CSA-induced changes of the thymic microenvironment are discussed.

Animals↗

Site-directed mutagenesis of the 2-haloalkanoic acid dehalogenase I gene from Pseudomonas sp. strain CBS3 and its effect on catalytic activity.

Two 2-haloalkanoic acid dehalogenases from Pseudomonas sp. strain CBS3 catalyse hydrolytic dehalogenation of chloroacetate and 2-chloropropionate. We used site-directed mutagenesis to introduce specific changes in the dehalogenase I encoding gene (dehCI). Substitution of Asp-10 by Ala-10 resulted in complete loss of dehalogenating activity although expression of the 2-haloacid dehalogenase I was not affected in the mutant as shown by western blot analysis, and although comparison of the mutated enzyme with the wild type enzyme indicated that extensive rearrangements in the three-dimensional structure of the enzyme had not occurred. From these data we suggest that Asp-10 of 2-haloacid dehalogenases I from Pseudomonas sp. strain CBS3 may be the nucleophilic residue in the active-site of this enzyme essential for halide release.

Amino Acid Sequence↗

Protonation of excited state pyrene-1-carboxylate by phosphate and organic acids in aqueous solution studied by fluorescence spectroscopy.

Pyrene-1-carboxylic acid has a pK of 4.0 in the ground state and 8.1 in the singlet electronic excited state. In the pH range of physiological interest (pH approximately 5-8), the ground state compound is largely ionized as pyrene-1-carboxylate, but protonation of the excited state molecule occurs when a proton donor reacts with the carboxylate during the excited state lifetime of the fluorophore. Both forms of the pyrene derivatives are fluorescent, and in this work the protonation reaction was measured by monitoring steady-state and time-resolved fluorescence. The rate of protonation of pyrene-COO(-) by acetic, chloroacetic, lactic, and cacodylic acids is a function of DeltapK, as predicted by Marcus theory. The rate of proton transfer from these acids saturates at high concentration, as expected for the existence of an encounter complex. Trihydrogen-phosphate is a much better proton donor than dihydrogen- and monohydrogen-phosphate, as can be seen by the pH dependence. The proton-donating ability of phosphate does not saturate at high concentrations, but increases with increasing phosphate concentration. We suggest that enhanced rate of proton transfer at high phosphate concentrations may be due to the dual proton donating and accepting nature of phosphate, in analogy to the Grotthuss mechanism for proton transfer in water. It is suggested that in molecular structures containing multiple phosphates, such as membrane surfaces and DNA, proton transfer rates will be enhanced by this mechanism.

Acids↗

Myeloid sarcoma occurring in the maxillary gingiva: a case without leukemic manifestations.

Myeloid sarcoma (MS) is a localized extramedullary mass of immature granulocytic cells that usually occurs in patients with acute myeloid leukemia (AML) or myeloproliferative disorders. It may rarely precede peripheral blood or bone marrow involvement, presenting a diagnostic challenge. Although MS may be found in any location, an intraoral occurrence is rare. In this report we describe a rare case of a patient with nonleukemic MS of the maxillary gingiva. The histologic specimen was first interpreted as non-Hodgkin's lymphoma. The correct diagnosis was reached after extensive immunohistologic studies. The malignant cells were myeloperoxidase positive, lysozyme positive, CD45+, CD68+, CD3-, CD10-, CD19-, CD20-, CD30-, CD34-, CD56-, CD79a-, S100-, and chloroacetate esterase negative. Induction therapy with FLAND (fludarabine, Ara-C, mitoxantrone, and dexamethasone) was started, but the patient did not achieve a remission. Some weeks later, the patient presented pleural effusion and paralysis of the seventh cranial nerve on the left side. She died a few days later. The present case indicates the importance of a correct initial diagnosis for adequate therapy, which is often delayed because of a high misdiagnosis rate. If the MS is treated without intensive chemotherapy for AML as soon as possible, the prognosis will be poor.

Antineoplastic Combined Chemotherapy Protocols↗

15N-labeled glycine synthesis.

This work describes a method for 15N-isotope-labeled glycine synthesis, as well as details about a recovery line for nitrogen residues. To that effect, amination of alpha-haloacids was performed, using carboxylic chloroacetic acid and labeled aqueous ammonia (15NH3). Special care was taken to avoid possible 15NH3 losses, since its production cost is high. In that respect, although the purchase cost of the 13N-labeled compound (radioactive) is lower, the stable tracer produced constitutes an important tool for N cycling studies in living organisms, also minimizing labor and environmental hazards, as well as time limitation problems in field studies. The tests were carried out with three replications, and variable 15NH3aq volumes in the reaction were used (50, 100, and 150 mL), in order to calibrate the best operational condition; glycine masses obtained were 1.7, 2, and 3.2 g, respectively. With the development of a system for 15NH3 recovery, it was possible to recover 71, 83, and 87% of the ammonia initially used in the synthesis. With the required adaptations, the same system was used to recover methanol, and 75% of the methanol initially used in the amino acid purification process were recovered.

Journal Article↗

Auer rods in mature granulocytes and monocytes.

In a 73-year-old man with acute myelomonocytic leukemia (M4 on FAB classification) Auer rods were demonstrated in mature granulocytes and monocytes in the blood as well as the bone marrow. Cytochemically, a significant number of leukemic blasts were positive for both alpha-naphthyl-acetate esterase and naphthol-AS-D-chloroacetate esterase. Auer rods in mature granulocytes were associated with immature cytoplasm and not infrequently with pseudo-Pelger anomaly. The significance of these observations is discussed.

Aged↗

Myelogenous leukemia in a bearded dragon (Acanthodraco vitticeps).

A 3-yr-old bearded dragon (Acanthodraco vitticeps) presented with lethargy, a swollen right elbow joint, inability to move its rear limbs normally, and marked leukocytosis. The majority of leukocytes were an abnormal mononuclear lymphoid-type cell with a high nuclear to cytoplasmic ratio, a slightly blue cytoplasm, nuclei with coarsely granular chromatin, and some nuclear clefts. Acute leukemia of lymphoid or myeloid origin was tentatively diagnosed. The abnormal mononuclear leukocyte cell population stained positively for the myeloid cytochemical stains: peroxidase, chloroacetate esterase, and L1-calprotectin. The abnormal cell population of the peripheral blood did not stain with the lymphoid cytochemical stains: alpha-naphthyl butyrate esterase, CD3, and CD79a.

Animals↗