Structural and functional definition of the human chitinase chitin-binding domain.

Mammalian chitinase, a chitinolytic enzyme expressed by macrophages, has been detected in atherosclerotic plaques and is elevated in blood and tissues of guinea pigs infected with Aspergillus. Its normal physiological function is unknown. To understand how the enzyme interacts with its substrate, we have characterized the chitin-binding domain. The C-terminal 49 amino acids make up the minimal sequence required for chitin binding activity. The absence of this domain does not affect the ability of the enzyme to hydrolyze the soluble substrate, triacetylchitotriose, but abolishes hydrolysis of insoluble chitin. Within the minimal chitin-binding domain are six cysteines; mutation of any one of these to serine results in complete loss of chitin binding activity. Analysis of purified recombinant chitin-binding domain revealed the presence of three disulfide linkages. The recombinant domain binds specifically to chitin but does not bind chitosan, cellulose, xylan, beta-1, 3-glucan, beta-1,3-1,4-glucan, or mannan. Fluorescently tagged chitin-binding domain was used to demonstrate chitin-specific binding to Saccharomyces cerevisiae, Candida albicans, Mucor rouxii, and Neurospora crassa. These experiments define structural features of the minimal domain of human chitinase required for both specifically binding to and hydrolyzing insoluble chitin and demonstrate relevant binding within the context of the fungal cell wall.

Profile of human macrophage transcripts: insights into macrophage biology and identification of novel chemokines.

High throughput partial sequencing of randomly selected cDNA clones has proven to be a powerful tool for examining the relative abundance of mRNAs and for the identification of novel gene products. Because of the important role played by macrophages in immune and inflammatory responses, we sequenced over 3000 randomly selected cDNA clones from a human macrophage library. These sequences represent a molecular inventory of mRNAs from macrophages and provide a catalog of highly expressed transcripts. Two of the most abundant clones encode recently identified CC chemokines. Macrophage-derived chemokine (MDC) plays a complex role in immunoregulation and is a potent chemoattractant for dendritic cells, T cells, and natural killer cells. The chemokine receptor CCR4 binds MDC with high affinity and also responds by calcium flux and chemotaxis. CCR4 has been shown to be expressed by Th2 type T cells. Recent studies also implicate MDC as a major component of the host defense against human immunodeficiency virus.

Thrombosis of the proximal greater saphenous vein: ultrasonographic diagnosis and clinical significance.

Eleven of 985 (1.1%) patients referred for leg ultrasonographic examinations to evaluate for deep venous thrombosis over a 3 year period were found to have isolated proximal greater saphenous vein thrombosis. Retrospective review of their treatments and sequelae showed that six patients (55%) initially treated with anticoagulant medication or high vein ligation had no further sequelae. Of four patients (36%) not initially treated, two subsequently developed pulmonary embolus, one had clot propagation into the common femoral vein, and one developed noncontiguous deep venous thrombosis. The findings of this review would suggest aggressive treatment for isolated upper greater saphenous vein thrombosis.

Diagnostic associations of p53 immunostaining in fine needle aspiration cytology of the breast.

Duplicate cytospin preparations were made from 46 symptomatic breast fine needle aspirates. One of each pair was assigned to benign or malignant categories by one experienced observer as part of the "triple approach" patient assessment. The other was immunostained with DO7, a monoclonal antibody to recombinant p53 protein, and rated by another observer as positive or negative for nuclear staining, unaware of the cytodiagnosis. Positive controls included carcinomas known to have mutant p53, while negative controls were of the reagent substitution type. Of the 26 aspirates with a benign cytodiagnosis (verified by the triple approach), 23 were p53 protein-negative and three positive. Of the 20 with a malignant cytodiagnosis (histologically confirmed), six were p53 protein-negative and 14 positive (exact P < 0.0001). As a diagnostic test this would give 70% sensitivity and 88% specificity.

Partial subclavian steal.

The phenomenon of subclavian steal syndrome has recently been found to have a subset group that has been described as partial or "pre-steal" in physiology. Our case of a Marine Corps pilot with this partial subclavian steal physiology is reviewed, in addition to the maneuvers we performed to temporarily convert him to complete steal physiology. A discussion of the symptomatology and prognosis of this diagnosis is included, with emphasis on military service.

From the archives of the AFIP. Malignant germ cell tumors of the ovary: radiologic-pathologic correlation.

Malignant germ cell tumors are the most common malignant ovarian neoplasms in girls and young women. The most well known of these neoplasms are dysgerminoma, immature teratoma, endodermal sinus tumor, and a mixture of two or more of these elements. All malignant germ cell tumors, especially immature teratoma, can be associated with mature (benign) teratoma. These tumors are generally large and have a complex but predominantly solid appearance on cross-sectional images. Elevated serum alpha-fetoprotein and human chorionic gonadotropin levels can help establish the diagnosis. Cystic areas of calcifications suggestive of mature teratoma do not exclude a co-existing malignant neoplasm.