Do patients with antiphospholipid syndrome have autoantibodies to beta 2-glycoprotein I?

High positive anticardiolipin antibody tests have been associated with recurrent thrombosis and pregnancy loss. Although these antibodies were believed to bind negatively charged phospholipids, recent reports have suggested that a serum protein, beta 2-glycoprotein I (beta 2-GPI), may be the true antigen for these antibodies. To resolve this issue, we compared binding of 75 anticardiolipin-positive and 71 anticardiolipin-negative serum samples from patients with rheumatic diseases to beta 2-GPI by using an enzyme-linked immunosorbent assay (ELISA). Serum samples from 30 healthy blood donors and 10 laboratory personnel were used as normal controls. We found no difference in binding between the three groups of serum samples. In addition, when binding to beta 2-GPI coated plates was compared with binding to ELISA plates without beta 2-GPI (blank), no difference was observed. Finally, binding of anticardiolipin-positive serum samples to plates coated with cardiolipin-beta 2-GPI mixture varied directly with the cardiolipin concentrations. Based on these findings, we conclude that anticardiolipin-positive serum samples do not bind beta 2-GPI.     

Study on structural gene expression in human insulinoma

A human insulinoma cDNA library was constructed in the expression plasmid vector pUEX1. The clone pUEX1Ins12 was selected by means of hybridization with an insulin probe. It codes for full size amino acid sequence preproinsulin. The bacterial strain pUEX3Ins8 producing proinsulin as beta-galactosidase fusion protein was obtained for the use of recombinant protein as an antigen in an ELISA to detect serum antibodies in subjects with IDDM. Recombinant clones containing the middle, N- and C-terminal domains of the GAD65, the major autoantigen in IDDM, were constructed in pVEX1. These clones may become important tools to study the nature of GAD autoreactivity in IDDM. The clone pHICEO.9 was selected from the human insulinoma cDNA library by immunoscreening with total human insulinoma protein antibodies. This clone expresses the C-terminal fragment of human cholesterol esterase/lipase containing its antigenic determinant and can be used for blood lipase determination. Four clones containing cDNA inserts (0.47-1.42 kb) without any significant homologies to the known sequences in the Gene Bank were obtained by means of statistic selection.     

Lung capillary albumin leak in oxygen toxicity. A quantitative immunocytochemical study

Uterine leiomyomas or fibroids are common among women of reproductive age, but their biology is poorly understood. The PTH-related protein (PTHrP) has been identified in a number of sites throughout the reproductive tract. We, therefore, examined whether fibroids express PTHrP mRNA and compared their level of expression with that in normal myometrium. Total RNA prepared from fibroid tissue and corresponding normal myometrium from seven patients was examined by RNase protection analysis. In all cases, fibroid and myometrial tissue expressed PTHrP, and in six of seven cases, PTHrP expression was higher in fibroids than in normal myometrium. Cultured fibroid cells from four patients also expressed higher levels of PTHrP mRNA than corresponding cultured normal myometrial cells. Tissue extracts from eight patients and conditioned medium from cultured cells from nine patients were examined for PTHrP immunoreactivity using a two-site immunoradiometric assay. In tissue extracts and conditioned medium, the mean PTHrP concentration was significantly higher in fibroids than normal myometrium. Immunohistochemical staining of fibroid and myometrial tissue was positive for PTHrP. Finally, PTHrP-(1-34) induced a dose-dependent increase in cAMP in fibroid and myometrial cells in vitro. These findings suggest that PTHrP may have an autocrine/paracrine function in regulating myometrial physiology and may play a role in regulating fibroid growth or differentiation.     

pH-dependent microspheres from modified soybean protein hydrolysate

Soybean hydrolysate is a hydrophilic mixture of amino acids and low molecular peptides which are soluble over the whole pH range. Chemical modification of soybean hydrolysate with aromatic acyl chlorides resulted in a product that yielded pH-dependent microspheres. Investigation into the physiochemical properties of the microsphere forming material indicated that acylation had alerted the hydrophobic/hydrophilic ratio as evidenced by an increased column retention time on reverse phase HPLC. This was further confirmed by analysis of the amino acid composition of the modified material. The data indicated that the hydrophobic/hydrophilic ratio and low molecular were critical factors in the formation of this supramolecular complex. An estimation based on sedimentation rate revealed an average molecular weight of these microspheres as 10(7)-10(8) Daltons. Light scattering experiments indicated that the microspheres have discrete chambers in the interior. Included among the properties of the microspheres that have been determined are the pH range of the phase transition, size distribution and zeta-potential. The physiochemical characteristics of the microspheres prepared from modified soybean protein are similar to the microsphere forming material produced by thermal condensation of amino acids. Formation of microspheres in solution containing either porcine insulin or salmon calcitonin resulted in the encapsulation of nearly 60% of the dissolved proteins. Oral gavage of encapsulated porcine insulin or salmon calcitonin into the stomach of rats resulted in significant titers of either protein in the systematic circulation.     

Therapy after skeletal fixation in the hand and wrist

Skeletal fixation can be very beneficial to the recovery of wrist and hand function after a displaced fracture because it allows mobilization of soft tissues before the completion of fracture healing. The benefits of skeletal fixation can be greatly diminished, however, if excessive force causes the fixation to fail before fracture healing has occurred, infection occurs around the implant, or the patient develops reflex sympathetic dystrophy. Those complications, as well as others, are often caused by inappropriate or inadequate hand therapy. This article discusses the techniques needed to avoid many such complications while providing the best possible functional result for every patient.