Adenosine inhibits IL-12 and TNF-[alpha] production via adenosine A2a receptor-dependent and independent mechanisms.

Interleukin 12 (IL-12) is a crucial cytokine in the regulation of T helper 1 vs. T helper 2 immune responses. In the present study, we investigated the effect of the endogenous purine nucleoside adenosine on the production of IL-12. In mouse macrophages, adenosine suppressed IL-12 production. Although the order of potency of adenosine receptor agonists suggested the involvement of A2a receptors, data obtained with A2a receptor-deficient mice showed that the adenosine suppression of IL-12 and even TNF-alpha production is only partly mediated by A2a receptor ligation. Studies with adenosine receptor antagonists or the adenosine uptake blocker dipyridamole showed that adenosine released endogenously also decreases IL-12. Although adenosine increases IL-10 production, the inhibition of IL-12 production is independent of the increased IL-10. The mechanism of action of adenosine was not associated with alterations of the activation of the p38 and p42/p44 mitogen-activated protein kinases or the phosphorylation of the c-Jun terminal kinase. Adenosine failed to affect steady-state levels of either IL-12 p35 or p40 mRNA, but augmented IL-10 mRNA levels. In summary, adenosine inhibits IL-12 production via various adenosine receptors. These results support the notion that adenosine-based therapies might be useful in certain autoimmune and/or inflammatory diseases.     

Fine-needle aspiration of mucin-producing thyroid tumors

Objective: Mucin-producing thyroid tumors are extremely rare. Nonetheless, we have encountered three such cases in fine-needle aspiration. We report the cytologic and histologic findings and review the literature. Study Design: Cytologic features were studied on direct smears using Romanovsky stain to detect background substance and Papanicolaou stain to analyze nuclear and cytoplasmic features. The cytologic features were correlated with histology. Mucin was demonstrated by mucicarmine, Alcian Blue/PAS, and Alcian Blue (pH 2.5). Results: The cytologic features related to mucin include: (1) thick luminal mucin globules and signet ring cells aspirated from an 83-year-old woman with a 3-cm signet ring cell follicular adenoma, (2) abundant fluffy mucin containing signet ring cells in cohesive fragments aspirated from a 75-year-old man with the bilateral signet ring cell follicular variant of papillary carcinoma, which is the first case in the English literature, and (3) abundant thin mucoid mucin aspirated from the lymph node of an 86-year-old woman with a 5-cm mucinous poorly differentiated thyroid carcinoma. Conclusion: The occurrence of mucin in thyroid fine-needle aspiration does not necessarily indicate metastasis, and the presence of mucin in cervical lymph nodes does not exclude the thyroid gland as a possible primary.     

Macromolecular synthesis during embryogenesis of Habrotrocha rosa Donner I. Replication of DNA

DNA synthesis was inhibited during embryogenesis of Habrotrocha rosa with mitomycin C and hydroxyurea. Inhibition of DNA replication in early stages of embryogenesis, at the beginning of organogenesis, just after cavitation of the stomodeum, resulted in a complete inhibition of further development. After this stage of embryogenesis development was insensitive to inhibition of DNA replication.     

Germ-line cysts are formed during oogenesis in Erpobdella octoculata (Annelida,Clitellata, Erpobdellidae)

Abstract

Erpobdella octoculata (Clitellata, Hirudinea, Erpobdellidae) has paired ovarian sacs, each containing several rod-shaped structures termed ovarian bodies. Oogenesis takes place within the ovarian bodies. We show that in the apical part of the bodies the germ-line cells form syncytial cysts of cells interconnected by stable intercellular bridges. Germ-line cyst architecture is broadly similar to that of other clitellate annelids; that is, each germ cell has only one intercellular bridge connecting it to the anuclear cytoplasmic mass, the cytophore. Unlike germ-line cysts described in other leech species, the cytophore in cysts of E. octoculata is poorly developed, taking the form of thin cytoplasmic strands. Oogenesis in E. octoculata is meroistic because the germ cells forming the cysts (cystocytes) have diverse fates, i.e., nurse cells and oocytes appear. One large ramified cell (apical cell) occurs within the apical part of the ovarian body. We compare the ultrastructure of the apical cell found in E. octoculata with that of apical cells described recently in some hirudiniform leeches. The germ-line cysts as well as the oocytes are enveloped by somatic follicular cells. As in other leeches, the follicular cells surrounding the growing oocytes have cytoplasm perforated by intracellular canals. In view of the many similarities between E. octoculata ovarian bodies and the ovary cords described in glossiphoniids and especially in hirudiniform leeches, we suggest that the ovarian bodies found in E. octoculata are in fact modified ovary cords.     

Simplified heavy metal staining techniques demonstrated with Fast Plant leaf tissue

Fast Plant(Brassica rapa,Cruciferae)leaf tissuefixed in glutaraldehyde-acrolein and post-fixed in os-mium,was examined for response to several easily-prepared heavy metal stains.Lead and uranium,separately and in combination,gave typical resultsacross the spectrum of cell organellets.As a single stainfollowing osmium,bismuth produced images seeminglyequivalent to lead and uranium.Phosphotungstic acidproduced very good membrane delineation but produceda washed-out background image similar to that from leadstaining.Carbohydrate compounds were especiallyresponsive to ruthenium;the cytoplasm and the matrixof all organelles were also stained very well.Theprocedures were no more demanding than traditionalstaining methods and may be easily used in research andteaching.Fast Plant materials are a reliable,quick andeasy source of living material.     

HIV protease inhibitor induces fatty acid and sterol biosynthesis in liver and adipose tissues due to the accumulation of activated sterol regulatory element-binding proteins in the nucleus

The mechanism by which human immunodeficiency virus (HIV) protease inhibitor therapy adversely induces lipodystrophy and hyperlipidemia has not been defined. This study explored the mechanism associated with the adverse effects of the prototype protease inhibitor ritonavir in mice. Ritonavir treatment increased plasma triglyceride and cholesterol levels through increased fatty acid and cholesterol biosynthesis in adipose and liver. Ritonavir treatment also resulted in hepatic steatosis and hepatomegaly. These abnormalities, which were especially pronounced after feeding a Western type high fat diet, were due to ritonavir-induced accumulation of the activated forms of sterol regulatory binding protein (SREBP)-1 and -2 in the nucleus of liver and adipose, resulting in elevated expression of lipid metabolism genes. Interestingly, protease inhibitor treatment did not alter SREBP mRNA levels in these tissues. Thus, the adverse lipid abnormalities associated with protease inhibitor therapy are caused by the constitutive induction of lipid biosynthesis in liver and adipose tissues due to the accumulation of activated SREBP in the nucleus.     

The plastidic Arabidopsis protoporphyrinogen IX oxidase gene, with or without the transit sequence, confers resistance to the diphenyl ether herbicide in rice

Agrobacterium‐mediated gene transformation was used to introduce plastidic protoporphyrinogen IX oxidase (Protox) genes from Arabidopsis, with and without the transit sequence, into the rice genome. They were placed under the control of the constitutive and ubiquitous maize ubiquitin promoter, and their abilities to confer resistance to the diphenyl ether‐type herbicide, oxyfluorfen were compared. The integration and expression of the transgene in the T₁ generation was examined by Southern, northern and western blot analyses. Surprisingly, as judged by an in vivo seed germination assay and an in vitro cellular leakage assay, both lines were similarly resistant to oxyfluorfen. The tolerance to cellular damage (lipid peroxidation and electrolyte leakage) was higher in transgenic plants than in wild‐type plants. In transgenic plants, the degree of herbicide resistance varied directly with the absolute amount of Protox protein expression. Both the intact protein and the protein with the transit sequence deleted were accumulated in plastids.     

Adenovirus-mediated human pancreatic lipase gene transfer to rat bile: gene therapy of fat malabsorption

This study explored the potential of using the gene therapy approach, based on adenovirus-mediated expression of pancreatic lipase in the hepatobiliary tract, to increase lipid digestion in the intestinal lumen and promote lipid absorption through the gastrointestinal tract. Recombinant adenovirus containing the human pancreatic lipase cDNA (AdPL) was shown to transduce and mediate pancreatic lipase biosynthesis in rat IEC-6 epithelial cells in vitro. Retrograde infusion of recombinant adenovirus (3 x 10(8) plaque-forming units) containing the bacterial LacZ gene (AdLacZ) into the bile duct of rats resulted in positive X-gal reaction products in the periportal liver cells 7 days after AdLacZ infusion. A high level of human pancreatic lipase was detected in bile after retrograde bile duct infusion of rats with AdPL but not in the bile of animals infused with AdLacZ. Triglyceride hydrolytic activity in the bile of AdPL-infused rats was equivalent to that present in pancreatic juice. In contrast, serum obtained from these animals did not contain any detectable pancreatic lipase activity. These results suggest that ectopic expression of pancreatic enzymes in the hepatobiliary tract may be an alternative therapeutic strategy for treating fat malabsorption due to pancreatic insufficiency.