Reversibility of morphine effects on phagocytosis by murine macrophages.

Proneness of addicts to infections may be partially due to opiate effects on immune cells. We find that acute morphine inhibits phagocytosis in murine peritoneal macrophages in vitro with apparent desensitization at high concentrations, whereas chronic exposure results in a state akin to tolerance/dependence where macrophages seem to require morphine to phagocytize at a control level. However, both putative desensitization and tolerance/dependence are reversible, since drug re-addition several hours after withdrawal results in inhibition, as in acute exposure. This shows that opiate effects on immune cells are variably related to the experimental context in which they are administered, which is of relevance for understanding their potential role in immunosuppression.

[Obstructive syndrome caused by trichobezoars: historical disease or disease still current? Description of a case in adolescence]. / Sindrome ostruttiva da tricobezoari: malattia storica o patologia ancora attuale? Descrizione di un caso in adolescente.

Trichobezoars are masses of entangled material, found in the stomach and intestines, composed of hair ingested by the patient. When the mass grows, symptoms of intestinal occlusion can appear. Trichobezoars in pediatrics are usually found in adolescent females presenting personality disorders and trichophagia. This work describes a case of trichobezoar diagnosed in a 13 year-old girl, wearing a brace for serious scoliosis but absolutely normal from the emotional and psychical point of view, with normal scholastic and familiar situation. Already hospitalized three months before for pneumonia from mycoplasma, the girl comes to our observation for the appearance of vomiting and constipation. The clinical examination reveal an epigastric mass as big as an orange. General conditions and hematochemical tests are good. Lab tests are performed (abdomen echography and upper abdomen MNR) but is the oesophago-gastroscopy which allows the diagnosis. A big trichobezoar is then surgically removed and the gastroenteric symptoms completely disappear.

Partially oxygenated sickled cells: sickle-shaped red cells found in circulating blood of patients with sickle cell disease.

A previously uncharacterized type of sickled cell was found in venous blood of patients with sickle cell disease when blood was collected without exposure to air and fixed immediately with 1% glutaraldehyde solution equilibrated with 5% oxygen. These cells were either elongated, resembling irreversibly sickled cells (ISCs), or nonelongated, with a raisin-like shape. Both types assumed a normal discoidal shape upon full oxygenation. Since these cells exist only under partially oxygenated conditions, they are described as partially oxygenated sickled cells (POSCs). POSCs are morphologically distinct from partially deoxygenated sickled cells formed during deoxygenation by having rounded edges, while the latter have sharp edges. Transmission electron microscopy of POSCs revealed various amounts of misaligned Hb S polymers. Investigations in vitro demonstrated the formation of POSC-like cells by partial oxygenation of deoxygenated cells. Since POSCs contain intracellular fibers and sickle readily upon deoxygenation, they may have clinical and pathological significance.

Monokine-mediated increase in human immunodeficiency virus type 1 expression in chronically infected promonocyte- and T-cell-derived lines.

The ACH-2 cell clone derived from a human T-cell line and chronically infected with human immunodeficiency virus 1 (HIV-1) and the U1 cell clone derived from a human promonocyte cell line and also chronically infected with HIV-1 produce HIV-1 in a response to stimulation with monokine-enriched supernatants prepared from highly purified populations of peripheral blood-derived human monocytes. Monokine-mediated expression of HIV-1 in these cell lines resulted in augmented virus production reflected by increases in reverse transcriptase (RT) activity, production of p24 antigen, and synthesis of major viral proteins. Examination of the cells by electron microscopy revealed numerous HIV-1 virions in the cells treated with the supernatants. This stimulation of virus production by monokine-enriched supernatants resulted in approximately 100-fold increases in RT activity and p24 antigen expression in comparison with those in untreated U1 and ACH-2 cells. Absorption of monokine-enriched supernatants with rabbit anti-tumor necrosis factor alpha antibody removed most, but not all, of the induced HIV-1 RT activity and p24 antigen expression in U1 and ACH-2 cell lines, suggesting that tumor necrosis factor alpha in the monokine-enriched supernatants is a major factor in the induction of HIV-1 expression in these cells.

A Paneth cell analogue in Xenopus small intestine expresses antimicrobial peptide genes: conservation of an intestinal host-defense system.

Antimicrobial peptides are a widespread component of host defense. We characterized the tissue distribution and cellular localization of expression of the magainin family of antimicrobial peptide genes in Xenopus laevis. Two genes from this family, magainin and PGLa, are expressed at high levels in the skin and throughout the gastrointestinal tract. Magainin and PGLa mRNAs are synthesized in the granular multinucleated cell (GMC) of the gastric mucosa, a cell shown previously to contain magainin and PGLa peptides by immunohistochemical methods. In addition, we have localized magainin and PGLa mRNAs to distinct cells of Xenopus small intestine. Further characterization of this large, granule-filled cell by electron microscopy demonstrates features in common with the Paneth cell of mammalian small intestine, previously identified as a site of expression of antimicrobial peptide genes of the defensin family in mouse and human. Our identification of granule-laden, eosinophilic intestinal cells in Xenopus as a site of magainin and PGLa antimicrobial peptide gene expression suggests that these cells are functional analogues of mammalian Paneth cells and further supports a conserved role of antimicrobial peptides in host defense of the vertebrate small intestine.

Expression of magainin antimicrobial peptide genes in the developing granular glands of Xenopus skin and induction by thyroid hormone.

The granular gland of amphibian skin is an epithelial derivative that first appears during metamorphosis. The granular glands of Xenopus laevis are an abundant source of magainins, a family of peptides with antimicrobial activity in vitro. We demonstrate the developmental expression and thyroid hormone induction of magainin and PGLa (peptide with aminoterminal glycine and carboxyterminal leucinamide), the two most abundant members of the magainin peptide family. Magainin and PGLa mRNA levels increase dramatically over the 2-day period between stage 58 and stage 60 and remain high during the remainder of metamorphosis. We used in situ hybridization to localize magainin and PGLa mRNAs to the developing granular gland. Magainin and PGLa peptides are also first detected during metamorphosis, as demonstrated by extraction of active peptides from stage 60 tadpoles and immunogold histochemical localization of magainin and PGLa to the granules of the skin granular gland. We demonstrate the premature induction of magainin and PGLa expression upon administration of exogenous thyroid hormone to stage 54 tadpoles. Further investigation of the developmental expression and induction of the magainin peptides will provide insight into their possible biological functions during Xenopus metamorphosis.

A novel peptide-producing cell in Xenopus: multinucleated gastric mucosal cell strikingly similar to the granular gland of the skin.

We have characterized a novel peptide-containing cell within the gastric mucosa of Xenopus laevis. The cell is a spherical, multinucleated syncytial structure containing a cytoplasmic space filled with dense rice-shaped granules, and is strikingly similar in morphology to the well-studied granular gland of the amphibian skin. Immunohistochemical and immunogold methods were used to demonstrate that several peptides previously isolated from the granular glands of the skin, including the antimicrobial peptides magainin and PGLa (a peptide with amino-terminal glycine and carboxy-terminal leucinamide), are also stored in granules present in these enteric cells. These data demonstrate that this enteric peptide-producing cell is strikingly similar both morphologically and biochemically to the granular gland, previously considered a highly specialized structure of the amphibian integument. This novel gastric mucosal cell, which we have designated a "granular multinucleated cell," is distinct in its morphology and its diversity of stored peptide products from other well-characterized peptide-containing cells in the vertebrate gastrointestinal tract.

Antimicrobial peptides in the stomach of Xenopus laevis.

Antimicrobial peptides are widely distributed in nature and appear to play a role in the host defense of plants and animals. In this study we report the existence of antimicrobial peptides in the stomach of the vertebrate Xenopus laevis, an animal previously shown to store high concentrations of antimicrobial peptides in its skin. Antimicrobial activity was detected in extracts of X. laevis stomach tissue and nine antimicrobial peptides were then purified. A novel 24-amino acid peptide, designated PGQ, was isolated from these extracts, and has the following amino acid sequence: GVLSNVIGYLKKLGTGALNAVLKQ. PGQ is relatively basic and has the potential to form an amphipathic alpha-helix. The other peptides isolated are members of the magainin family of antimicrobial peptides, and include magainins I and II, PGLa, xenopsin precursor fragment, and four caerulein precursor fragments. None of these peptides had been previously identified in tissues other than the skin. The purification of the peptides from stomach extracts and subsequent protein sequence analysis reveals that the peptides have undergone the same processing as their dermal counterparts, and that they are stored in their processed forms. Northern blot analysis indicates that the magainin family of peptides are synthesized in the stomach, and immunohistochemical studies demonstrate that magainin is stored in a novel granular multinucleated cell in the gastric mucosa of Xenopus. This study demonstrates that the magainin family of antimicrobial peptides is found in the gastrointestinal system of X. laevis and offers an opportunity to further define the physiological role of these defense peptides.

Preservation of brush border transport systems for proline and alpha-methyl-D glucoside from rat, dog, and human kidney.

The effects of freezing renal tissue from rat, dog, and man on the time course of uptake of proline and alpha-methyl-D-glucoside by subsequently isolated brush border membrane vesicles was examined and compared with uptake patterns by membranes isolated from tissue that had never been frozen. The overshoot phenomenon was used as the critical criterion for viability of the transport systems. Membranes isolated from frozen rat and dog kidney possessed intact transport systems for proline and alpha-methyl-D-glucoside capable of producing normal or even enhanced overshoot patterns. Freezing human kidney prior to membrane isolation resulted in severe impairment of the vesicle transport capabilities. Freezing a crude membrane suspension, however, allowed the subsequent purification of only partially intact systems.