In situ staining for poly(ADP-ribose) polymerase activity using an NAD analogue

Poly(ADP-ribose) polymerase (PARP) is a highly abundant nuclear enzyme which metabolizes NAD, in response to DNA strand breakage, to produce chains of poly(ADP-ribose) attached to nuclear proteins. PARP activation has been implicated in ischemia/reperfusion injury, but its biological significance is not fully understood. We have modified an existing in situ method for detection of PARP activity by using an NAD analogue in which adenine is modified by an "etheno" (vinyl) bridge. Etheno-NAD serves as a PARP substrate in an initial enzymatic reaction; a specific antibody to ethenoadenosine is then used in an immunohistochemical reaction to detect the production of modified poly(ADP-ribose). The method produces strong and specific labeling of nuclei in which PARP has been activated, i.e., those in which DNA strand breaks have been produced, and the results can be analyzed by microscopy, flow cytometry, or colorimetry. The method is applicable to cultured cells in several formats and to frozen tissue sections. The particular characteristics of the new method may assist in future in situ studies of PARP activation.     

Measurement of cyclooxygenase inhibition in vivo: a study of two non-steroidal anti-inflammatory drugs in sheep

The anti-inflammatory effects of the non-steroidal anti-inflammatory drugs phenylbutazone (PBZ) and flunixin meglumine (FM) and the relationship between the effects and drug concentration in vivo were studied using a subcutaneous tissue-cage model in sheep. Intracaveal injection of carrageenan induced prostaglandin (PG) E2 production in tissue-cage exudate (maximal concentration, 101 nM) with significant increases in white blood cell (WBC) numbers, skin temperature over the inflamed cage and exudate leukotriene B4 (LTB4) concentration (P < 0.05). Intravenous PBZ, 4.4 mg kg-1 produced mild inhibition of exudate PGE2 generation (10%), but greater inhibition of serum TXB2 (75.3%). The IC50 for TXB2 was 36.0 microM. Phenylbutazone did not alter effects on skin temperature, WBC numbers or exudate LTB4 concentrations. Intravenous FM, 1.1 mg kg-1, significantly inhibited carrageenan-induced exudate PGE2 formation (Emax, 100%, IC50, < 0.4 nM) and serum TXB2 generation (Emax, 100%, IC50, 17 nM) for up to 32 h. Flunixin meglumine significantly inhibited the rise in skin temperature but had a limited effect on exudate WBC. Phenylbutazone and FM have distinct effects on carrageenan-induced cyclooxygenase (COX-2) and platelet COX (COX-1). Flunixin meglumine was a more potent COX inhibitor than PBZ and was more selective for the inducible form of COX in vivo.     

Femoral hypoplasia-unusual facies syndrome with bifid hallux, absent tibia, and macrophallus: a report of a Bedouin baby

We report a case where Lactobacillus rhamnosus was isolated from pericardial effusion and blood in a child following a bone marrow transplant for aplastic anaemia. A resume of cases in which this organism has been implicated as a pathogen is also presented.     

Comparative pharmacokinetics of paracetamol (acetaminophen) and its sulphate and glucuronide metabolites in desert camels and goats

Functional and aesthetic nasal surgery has been undergoing a process of fine-tuning. The surgical approaches lean toward greater conservation-particularly aesthetic- and functional selectivity. This has been made possible by improved diagnostic methods and pre-operative programming techniques such as computerized morphometry, computerized axial tomography, rhinosinus endoscopy, rhinomanometry, acoustic rhinometry and electromyography. Another important point in functional and aesthetic nasal surgery is that a) it can be divided into different techniques for each anatomo-functional sector considered and b) these can be used together in various combinations. Experience and the literature have underscored some issues which are most pertinent to the modern methodological viewpoint. The first large-scale controversy involves a comparison of open and closed techniques for access. The advantages and disadvantages of both techniques are described, although the disadvantages can be reduced to a minimum if the right indication is chosen. In the authors' opinion, the open technique should be reserved for general revision surgery, particularly in conjunction with difficult tips, labiopalatoschisis reconstruction, septal perforation and saddle nose. The second basic point involves how to deal with the perichondrium-periostium-mucosal flaps. The two main techniques for nasal skeletizing in rhinoseptoplasty include the extra-mucosal approach which is conservative and the trans-mucosal approach which is apparently more traumatic. Curiously, however, it is the latter technique which leads to fewer medium-long term complications. The nasal tip is the crux for the neophyte to rhinoseptoplasty. Through knowledge of the indications and the risks of the various techniques makes it possible to predict the result before-hand, although with certain margin of error. Indeed, this is one of the regions of the nose where errors reap the greatest damage. In this anatomical site, the use of conservative techniques is, therefore, strongly suggested. Finally, not only has research into the field of functional nasal surgery been unable to find concrete applications in humans (for obvious ethical reasons), it has likewise been unable to provide an answer to the question most rhinosurgeons pose: why do some patients still complain of difficulty breathing, even after successful surgery? It is our conviction that the problem is due to the fact that research into the endonasal receptors has not progressed.     

Androgen-repressed phenotype in human prostate cancer

An androgen-repressed human prostate cancer cell line, ARCaP, was established and characterized. This cell line was derived from the ascites fluid of a patient with advanced metastatic disease. In contrast to the behavior of androgen-dependent LNCaP and its androgen-independent C4-2 subline, androgen and estrogen suppress the growth of ARCaP cells in a dose-dependent manner in vivo and in vitro. ARCaP is tumorigenic and highly metastatic. It metastasizes to the lymph node, lung, pancreas, liver, kidney, and bone, and forms ascites fluid in athymic hosts. ARCaP cells express low levels of androgen receptor mRNA and prostate-specific antigen mRNA and protein. Immunohistochemical staining shows that ARCaP cells stain intensely for epidermal growth factor receptor, c-erb B2/neu, and c-erb B3. Staining is negative for chromogranin A and positive for bombesin, serotonin, neuron-specific enolase, and the c-met protooncogene (a hepatic growth factor/scatter factor receptor). ARCaP cells also secrete high levels of gelatinase A and B and some stromelysin, which suggests that this cell line may contain markers representing invasive adenocarcinoma with selective neuronendocrine phenotypes. Along with its repression of growth, androgen is also found to repress the expression of prostate-specific antigen in ARCaP cells as detected by a prostate-specific antigen promoter-beta-galactosidase reporter assay. Our results suggest that the androgen-repressed state may be central to prostate cancer progression and that advanced prostate cancer can progress from an androgen-independent to an androgen-repressed state.