Lessons from TGF-beta transgenic mice

Aberrant expression of TGF-beta and/or receptor/signaling function is present in a wide variety of disease processes. Overexpression of TGF-beta isoforms in transgenic mice using tissue-specific promoters has provided model systems to study the effects of increased activity of TGF-beta in the intact organism. We will review the pertinent features of some of these models, and discuss new insights provided by these studies into regulation and role of TGF-beta in health and disease.     

Electron beam computed tomography (EBCT)--funding protocol. EBCT Funding Protocol Working Group

BACKGROUND: Pressure measuring systems using fluid-filled catheters can result in the recording of distorted pressure waveforms. It results in phase delay, overestimation of systolic and, to a lesser extent, of diastolic pressure. We designed and evaluated a method to correct this pressure waveform distortion using an appropriate transfer equation obtained from the dynamic response of the fluid-filled catheter. This transfer equation is based on the principle that a fluid-filled catheter recording system is considered as an underdamped dynamic system fully characterized by its natural frequency (omega n) and damping ratio (zeta). METHODS: Pressure waveforms, simultaneously recorded in vitro or in vivo by a fluid-filled catheter (Pc) and a micromanometer-tipped catheter (Pref), were used to validate the method. Dynamic response of the catheter used was obtained from a fastflush test. The corrected signal (Ppred) was obtained using omega n, zeta and the following transfer equation: d2Pc/dt2 + 2 omega n zeta dPc/dt + omega n 2Pc = C Ppred (t) After correction of Pc, Ppred was compared, using a linear regression, with Pref taken as reference. RESULTS: Our results showed that Ppred was fitted to Pref with excellent coefficient correlation (0.99). The mean error and the standard error of estimate were respectively -1.16 mmHg and 1.4 mmHg. CONCLUSION: This new method can convert the distorted pressure waveforms transmitted by any fluid-filled catheters into high-fidelity signals. It suppresses the phase delay and the over-estimation of systolic pressure induced by fluid-filled catheters.     

Bending and fracture toughness of woven self-reinforced composite poly(methyl methacrylate)

Loosening remains an impediment to the long-term success of total hip replacements despite numerous improvements in the materials used. In cemented prostheses, fatigue and fracture of bone cement have been implicated in the failure of these devices. A new material, self-reinforced composite poly(methyl methacrylate). (SRC-PMMA), has been developed. SRC-PMMA is formed by a novel processing method that will be described. The composite consists of high strength, highly oriented PMMA fibers embedded in a matrix of PMMA. Using a woven form of SRC-PMMA, an in vitro physical and mechanical evaluation was performed to assess the feasibility of its use in an orthopedic prosthesis. Three different weaves of SRC-PMMA were evaluated in bending and fracture toughness in air, after immersion for 30 days in 37 degrees C saline, and after gamma irradiation followed by immersion. Bending modulus and strength were decreased by gamma irradiation followed by saline immersion. The effect of saline immersion alone on bending strength and modulus was negligible. Saline immersion and gamma irradiation followed by saline immersion was shown to have little or no effect on the fracture toughness of woven SRC-PMMA. Differences in the fracture processes of the different weaves were found and can be related to the differing orientation of fibers to the fracture toughness pre-crack. Optimally incorporated SRC-PMMA absorbs the same amount of water as bone cement. Comparison to previous and current work with bone cement controls shows that SRC-PMMA is a material equal to or better than bone cement in all tests performed. It deserves further consideration as a candidate biomaterial.     

An experimental study of the anti-ulcer properties of an extract of poplar bark

Marked antiulcerative properties of a Populus tremula bark extract were demonstrated in experiments on mice and rats with stress-, reserpine-induced, and acetylsalicylic ulcers. The gastroprotective activity of the extract was expressed in decrease in the number of animals with mucosal ulcers and in marked decrease in the number of ulcers. It exceeded similar parameters of alanton, plantaglucide, and Hippopheae oil.     

Zyme, a novel and potentially amyloidogenic enzyme cDNA isolated from Alzheimer's disease brain

The deposition of the beta amyloid peptide in neuritic plaques and cerebral blood vessels is a hallmark of Alzheimer's disease (AD) pathology. The major component of the amyloid deposit is a 4.2-kDa polypeptide termed amyloid beta-protein of 39-43 residues, which is derived from processing of a larger amyloid precursor protein (APP). It is hypothesized that a chymotrypsin-like enzyme is involved in the processing of APP. We have discovered a new serine protease from the AD brain by polymerase chain reaction amplification of DNA sequences representing active site homologous regions of chymotrypsin-like enzymes. A cDNA clone was identified as one out of one million that encodes Zyme, a serine protease. Messenger RNA encoding Zyme can be detected in some mammalian species but not in mice, rats, or hamster. Zyme is expressed predominantly in brain, kidney, and salivary gland. Zyme mRNA cannot be detected in fetal brain but is seen in adult brain. The Zyme gene maps to chromosome 19q13.3, a region which shows genetic linkage with late onset familial Alzheimer's disease. When Zyme cDNA is co-expressed with the APP cDNA in 293 (human embryonic kidney) cells, amyloidogenic fragments are detected using C-terminal antibody to APP. These co-transfected cells release an abundance of truncated amyloid beta-protein peptide and shows a reduction of residues 17-42 of Abeta (P3) peptide. Zyme is immunolocalized to perivascular cells in monkey cortex and the AD brain. In addition, Zyme is localized to microglial cells in our AD brain sample. The amyloidogenic potential and localization in brain may indicate a role for this protease in amyloid precursor processing and AD.     

Pseudotyping of murine leukemia virus with the envelope glycoproteins of HIV generates a retroviral vector with specificity of infection for CD4-expressing cells

CD4-expressing T cells in lymphoid organs are infected by the primary strains of HIV and represent one of the main sources of virus replication. Gene therapy strategies are being developed that allow the transfer of exogenous genes into CD4(+) T lymphocytes whose expression might prevent viral infection or replication. Insights into the mechanisms that govern virus entry into the target cells can be exploited for this purpose. Major determinants of the tropism of infection are the CD4 molecules on the surface of the target cells and the viral envelope glycoproteins at the viral surface. The best characterized and most widely used gene transfer vectors are derived from Moloney murine leukemia virus (MuLV). To generate MuLV-based retroviral gene transfer vector particles with specificity of infection for CD4-expressing cells, we attempted to produce viral pseudotypes, consisting of MuLV capsid particles and the surface (SU) and transmembrane (TM) envelope glycoproteins gp120-SU and gp41-TM of HIV type 1 (HIV-1). Full-length HIV-1 envelope glycoproteins were expressed in the MuLV env-negative packaging cell line TELCeB6. Formation of infectious pseudotype particles was not observed. However, using a truncated variant of the transmembrane protein, lacking sequences of the carboxyl-terminal cytoplasmic domain, pseudotyped retroviruses were generated. Removal of the carboxyl-terminal domain of the transmembrane envelope protein of HIV-1 was therefore absolutely required for the generation of the viral pseudotypes. The virus was shown to infect CD4-expressing cell lines, and infection was prevented by antisera specific for gp120-SU. This retroviral vector should prove useful for the study of HIV infection events mediated by HIV-1 envelope glycoproteins, and for the targeting of CD4(+) cells during gene therapy of AIDS.     

Comparison of right ventricular outflow tract and apical lead permanent pacing on cardiac output

Interactions of gastrointestinal prokinetic benzamides with 5-hydroxytryptamine (5-HT)3 and 5-HT4 receptors and the relation to their effects on gastrointestinal propulsion were investigated. Renzapride and zacopride potently inhibited 5-HT3-receptor-mediated contractions in the guinea pig colon, whereas RS67506 (1-(4-amino-5-chloro-2-methoxyphenyl)-3-[1-(2-methyl sulphonylamino)ethyl-4-piperidinyl]-1-propanone hydrochloride), a selective 5-HT4-receptor agonist, showed no inhibition. RS67506, renzapride and zacopride all exerted 5-HT4 receptor-mediated relaxation in the carbachol-precontracted rat oesophagus. In mice, RS67506 shortened the whole gut transit time, whereas renzapride and zacopride were reported to prolong it. Gastrointestinal prokinetic benzamides, which are selective for 5-HT4-receptor agonistic over 5-HT3-receptor antagonistic action, may be useful in treating gastrointestinal disorders associated with impaired lower intestinal propulsion such as constipation.     

Inhibition of steroidogenesis in rat adrenal cells by 18-ethynyldeoxycorticosterone: evidence for an alternative pathway of aldosterone biosynthesis

The effect of the mechanism-based inhibitor 18-ethynyldeoxycorticosterone (18-E-DOC) on the late steps of the aldosterone biosynthetic pathway was examined in freshly isolated cells of the zona glomerulosa (ZG) and fasciculata (ZF) from rat adrenal glands. ZG synthesis of aldosterone was inhibited by 18-E-DOC in a time- and concentration-dependent manner with a Ki of approximately 0.05 microM. The maximal degree of inhibition of ZG production of aldosterone and 18-hydroxycorticosterone (18-OH-B) was approximately 80%. ZF cells, perhaps surprisingly, were found to secrete 18-OH-B at levels approximately one-third to one-fourth those of ZG cells and the Ki of 18-E-DOC inhibition of 18-OH-B secretion was approximately 10 microM for ZF cells, 200-fold higher than for ZG cells. The inhibitor had no effect on the secretion of corticosterone by either ZG or ZF, and the secretion of 18-hydroxydeoxycorticosterone (18-OH-DOC) by both the ZG and ZF was inhibited only to a minor degree. 18-E-DOC inhibited the biosynthesis of aldosterone by ZG cells incubated with 10 microM added DOC or 18-OH-DOC by approximately 75%, similar to the degree of inhibition of aldosterone biosynthesis from endogenous substrate, whereas ZF biosynthesis of 18-OH-B from either substrate was inhibited by less than 40%. ZF cells do not express aldosterone synthase, the only enzyme known to convert 18-OH-DOC into 18-OH-B. Incubation of MA-10 cells stably transfected with the cDNA of the rat aldosterone synthase with 18-E-DOC resulted in a complete inhibition of the conversion of DOC to aldosterone with a Ki of approximately 0.02 microM. In addition, transfected cells expressing 11beta-hydroxylase convert DOC to 18-OH-B in very small quantities only and cannot convert 18-OH-DOC to 18-OH-B. These data suggest that neither 11beta-hydroxylase nor aldosterone synthase are responsible for the biosynthesis of 18-OH-B by ZF cells from DOC or 18-OH-DOC, that 20% of aldosterone synthesis appears not to be attributable to the actions of aldosterone synthase and that an unknown CYP11B enzyme is also involved in the biosynthesis of 18-OH-B.