Retroperitoneal haemorrhage caused by a renal angiomyolipoma.

Renal angiomyolipoma (AML) is a benign renal tumour and is nowadays considered a relatively common lesion. When an AML increases in size or becomes symptomatic, embolisation via the renal artery should then be considered, because rupture is an important complication and interventional therapies are required to stop bleeding. We present a 21 year old woman who was seen at the emergency department following a low velocity trauma. After a period of 9 weeks, clinical examination and radiological examination revealed a haemorrhage from a renal AML, which was treated by selective embolisation. A discussion of the relevant literature is also presented.

Acidic fibroblast growth factor overexpression partially protects 3T3 fibroblasts from apoptosis induced by synthetic retinoid CD437.

Retinoids are proapoptotic compounds with therapeutic potential for treating cancer. We evaluated the apoptotic effect of the novel retinoid CD437, and particularly its relationship to Akt and acidic fibroblast growth factor (aFGF). We hypothesized that the novel synthetic retinoid CD437 would exert its apoptotic effect by reducing the activity of Akt. We further hypothesized that aFGF would protect against CD437 apoptosis by preserving the activity of Akt. Initially we demonstrated that CD437 produces apoptotic cell death in NIH 3T3 fibroblasts, and that this effect is attenuated in fibroblasts transfected to express aFGF. Next we assessed Akt activity and showed that phospho-Akt is significantly reduced in 3T3 cells exposed to CD437. We showed that this effect is less pronounced in aFGF transfected 3T3 cells. Furthermore, we observed that the addition of exogenous aFGF to 3T3 cells significantly increases Akt phosphorylation. These findings tend to confirm our hypothesis that reduction in Akt activation is a mechanism involved in the apoptotic effect of the retinoid CD437, and that preservation of Akt phosphorylation occurs in response to aFGF and appears to explain the partially protective effect of aFGF for 3T3 cells vis a vis CD437.

A hypereosinophilic syndrome with retinal arteritis and tuberculosis.

A 35-year-old man was initially seen with a decrease in visual acuity, renal insufficiency, and elevation of the eosinophil count in the blood. The ocular syndrome was caused by extensive arterial occlusions of the retina. The subsequent apparition of cardiac, pulmonary, and neurologic signs fulfilled the criteria for the diagnosis of hypereosinophilic syndrome (HES). Most symptoms, including ocular, were temporarily but notably improved by hydroxyurea. The patient died after two years. An autopsy showed an endomyocardial fibrosis and disclosed destruction of the left kidney by an active tuberculosis. A pathogenic relationship between the infectious disease and the HES is envisaged.

Synthetic retinoid CD437 induces apoptosis of esophageal squamous HET-1A cells through the caspase-3-dependent pathway.

6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (AHPN/CD437) is a novel retinoid that has shown to be a potent inducer of apoptosis in cancer cells. We investigated the apoptosis-inducing activity of CD437 and its mechanism of action in the human esophageal squamous epithelial cell line (HET-1A). CD437 decreased HET-1A cell viability in a time and dose dependent manner. CD437 was found to induce DNA fragmentation. Apoptosis was accompanied by markedly increased activity of caspase-3 as well expression of caspase-3 and -8 and APRP fragmentation. The CD437-mediated activation of caspase-3 was inhibited by Z-DEVD-FMK These data indicate that CD437-induced apoptosis in HET-1A cells may be mediated through the caspase-3 dependent pathway. In addition, apoptosis was also accompanied by increased expression of Bad protein and down-regulation of Bcl-2 expression. Phosphorylation of Jun occurred following CD437 exposure indicating that Jun kinase (JNK) activity was strongly induced by CD437. Understanding these apoptotic pathways of esophageal epithelial cells may allow therapeutic induction of cell death in malignant orpremalignant lesions of the esophagus by agents such as CD437.

Evidence of androgen receptor expression in squamous and adenocarcinoma of the esophagus.

Androgen receptors (AR) are known to stimulate cellular proliferation in certain tumors. We have assessed the androgen receptor status of esophageal carcinoma in surgically resected specimens as well as in established human esophageal carcinoma cells lines. In these initial studies we sought to characterize the frequency of expression of androgen receptors in squamous versus adenocarcinoma and in male versus female patients, and to assess the possible influence of AR expression on survivaL We analyzed androgen receptor expression utilizing immunohistochemistry in adenocarcinoma and squamous cell carcinoma of the esophagus in surgical specimens from 25 patients treated at Johns Hopkins Bayview Medical Center. Tumors in 7 of 21 males (33%) and 1 of 4 females (25%) showed positive androgen receptor staining with the monoclonal body antibody AR 441. There was no suggestion of a difference in expression of AR between males and females. Five of 11 adenocarcinomas (45%) and 3 of 14 squamous carcinomas were positive. Survival was similar in AR+ and AR- patients. Studies with established tissue culture cell lines showed AR expression by RT-PCR, with stronger expression of AR in adenocarcinoma lines than in squamous carcinoma lines. The presence of AR in human esophageal cancer is an impetus for further studies to assess anti-androgen therapy for treatment and or prevention of these tumors.

Left atrial and right ventricular cardiac myxoma. A case report.

A case is presented with a tumour in the left atrium as well as in the right ventricle. During the initial investigation of the atrial myxoma, the ventricular tumour was overlooked and a second operation was necessary. Once the diagnosis of myxoma is made, a second synchronous tumour should always be carefully sought.

In Situ Localization of Barley Yellow Dwarf Virus-PAV 17-kDa Protein and Nucleic Acids in Oats.

ABSTRACT Barley yellow dwarf virus strain PAV (BYDV-PAV) RNA and the 17-kDa protein were localized in BYDV-PAV-infected oat cells using in situ hybridization and in situ immunolocalization assays, respectively. The in situ hybridization assay showed labeling of filamentous material in the nucleus, cytoplasm, and virus-induced vesicles with both sense and antisense nucleic acid probes, suggesting that the filamentous material found in BYDV-PAV-infected cells contains viral RNA. BYDV-PAV negative-strand RNA was detected before virus particles were observed, which indicates that RNA replication is initiated before synthesis of viral coat protein in the cytoplasm. The 17-kDa protein was associated with filamentous material in the cytoplasm, nucleus, and virus-induced vesicles. The labeling densities observed using antibodies against the 17-kDa protein were similar in the nucleus and cytoplasm. No labeling of the 17-kDa protein was observed in plasmodesmata, but filaments in the nuclear pores occasionally were labeled. Since BYDV-PAV RNA and 17-kDa protein colocalized within infected cells, it is possible that single-stranded viral RNA is always associated with the 17-kDa protein in vivo. The 17-kDa protein may be required for viral nucleic acid filaments to traverse the nuclear membrane or other membrane systems.

The activity of powdery-mildew haustoria after feeding the host cells with different sugars, as measured with a potentiometric cyanine dye.

The biotrophic parasite Erysiphe graminis f. sp. hordei produces haustoria within the cells of its host Hordeum vulgare. To determine the physiological activity of these haustoria, the electric potential across the membranes in the mitochondria of the haustorium was studied. The membrane potential was estimated with the fluorescent potentiometric cyanine dye 3,3'-dibutyloxacarbocyanine iodide. The addition of depolarizing agents (carbonylcyanide m-chlorophenylhydrazone, 2,4-dinitrophenol or KCN) to infected cells resulted in an increase of fluorescence after the addition of low concentrations or a decrease of fluorescence after the addition of higher concentrations. When the infected host cell was fed with increasing concentrations of D-glucose (25, 50, 75 mM), corresponding decreases of fluorescence were measured immediately in the mitochondria of the fungal haustoria. Sucrose induced a similar reduction of fluorescence about 20 min late. D-Galactose and D-fructose induced a somewhat smaller reduction of fluorescence, L-glucose and D-glucitol had no effect. The results indicate that haustoria take up glucose from the host cells immediately. Sucrose, D-galactose and D-fructose seem to require time to be metabolized before their products reach the fungal haustorium or mitochondria.

Protective immunity against herpes simplex virus generated by DNA vaccination compared to natural infection.

To evaluate the utility of plasmid DNA vaccination against disease caused by herpes simplex virus (HSV), we compared the strength of protection against lethal challenge following natural virus infection with that following vaccination with a plasmid encoding HSV glycoprotein gD (gD-DNA). We further determined the cellular basis of each type of protection using lymphocyte deficient knockout mice. Establishment of immunity to HSV using live virus immunization required CD8+ T cells and B cells, but not CD4+ or gamma/delta+ T cells, and was related to specific antibody levels; surprisingly, CD4 knockout mice had large quantities of IgG anti-HSV serum antibodies. Establishment of immunity to HSV using gD-DNA immunization approached the strength of that generated following sublethal infection, but was dependent on alpha/beta+ CD4+ T cells, CD8+ T cells, B cells, and even partially on gamma/delta+ T cells, and not strictly correlated with antibody levels.

Antibody response and protective capacity of plasmid vaccines expressing three different herpes simplex virus glycoproteins.

Plasmid expression vectors were constructed that contained the genes encoding herpes simplex virus 1 (HSV-1) glycoproteins C (gC), D (gD), and E (gE). Mice receiving two intramuscular injections of expression plasmid (50 microg) produced a specific HSV-1 antibody response. Mice receiving the gD plasmid were protected against a lethal intraperitoneal challenge of HSV-1 (5 x 10(4) pfu) but not against more demanding challenge doses. Protection with gC or gE plasmid vaccination could be demonstrated only if the inoculating dose of DNA was increased to 250 microg. In contrast, all mice immunized with vaccinia recombinants expressing either gC or gE survived HSV-1 challenge. Analysis of the HSV-1 antibody isotype produced by plasmid immunization revealed a response dominated by IgG2a. Combination delivery of all three glycoprotein expression plasmids provided better protection against lethal challenge, but mice receiving the combination were still not able to withstand increased challenge doses of virus.

Success and limitations of a naked plasmid transfection protocol for keratinocyte growth factor-1 to enhance cutaneous wound healing.

Our group and others have previously reported enhancement of cutaneous wound healing following the transfection of tissue with plasmid vectors expressing the DNA for growth factors. In these experiments, growth factor treated animals were usually compared to animals treated with control plasmid vector. To achieve consistent transfection, high DNA plasmid load and repeated penetrations of the wound by needle or gene gun were required. In the current experiments, we assessed the effect of the plasmid load and repeated tissue penetrations on wound healing of excisional wounds in diabetic C57 mice. Animals received 5 mm excisional wounds, and were assigned to the following groups, no treatment, phosphate buffered saline solution injections, and plasmid vector injection with and without the keratinocyte growth factor-1 gene. Intradermal injections of 100 microg plasmid were given adjacent to the wounds at days 1-5, 7 and 11. At day 9, wound closure was more advanced in keratinocyte growth factor-1 treated animals compared to those treated with control plasmid. But a detrimental effect of the DNA plasmid injection was evident from a comparison of the DNA control group versus the non-injected group. Therefore, the challenge for developing an effective system for the enhancement of wound healing lies in improving transfection efficiency.

Iliac artery stenoses after percutaneous transluminal angioplasty: follow-up with duplex ultrasonography.

PURPOSE: To assess iliac artery stenosis before and up to 1 year after percutaneous transluminal angioplasty (PTA) with duplex ultrasound (DUS) to determine the incidence of residual and recurrent stenoses and correlate these findings to clinical outcome. PATIENTS AND METHODS: Sixty-one patients with 70 iliac artery segments treated with PTA were examined. The peak systolic velocity (PSV) ratio (PSV ratio = PSV in stenosis divided by PSV proximal or distal to stenosis) was determined by DUS before PTA and 1 day, 3 months and 1 year after PTA. Three categories of results were identified by using PSV ratios at the site of the treated stenosis 1 day and 1 year after PTA (good result, residual stenosis, and recurrent stenosis). The DUS-determined anatomic result was correlated with the clinical outcome at 1 year. Clinical outcome was classified according to Society for Vascular Surgery/International Society for Cardiovascular Surgery (SVS/ISCVS) criteria. RESULTS: Good results with DUS (PSV ratio 1 day and 1 year after PTA > or = to 2.5) were found in 45 of 70 segments (64.3%), residual stenoses (PSV ratio > or .5 1 day after PTA) in 15 of 70 segments (21.4%), and recurrent stenosis (PSV ratio 1 day after PTA < 2.5 and 1 year after PTA > or = 2.5) in 10 of 70 segments (14.3%). PSV ratios of residual stenoses decreased significantly between 1 day and 1 year after PTA because some residual stenoses improved hemodynamically in time. Clinical results were significantly better in patients with a good result compared with other patients. However, the clinical outcome of patients with residual stenoses was not significantly different from the patients with good DUS results. CONCLUSION: Some residual stenoses improved sonographically after PTA. Clinical results at 1 year are highly variable within different groups. Clinical outcome of patients with residual stenoses did not differ from patients with good DUS results, whereas clinical outcome in patients with recurrent stenoses was worse than in the other groups.

Characterization of an aFGF gene expression vector with therapeutic potential.

BACKGROUND: Topical application of growth factors to wounds has proven to be suboptimal in achieving epithelial growth and accelerating healing. We propose transfection of fibroblasts with a gene for acidic fibroblast growth factor (aFGF) which will allow continuous, local delivery of the growth factor to wounds, ulcerative lesions, or healing tissues. METHODS: We utilized a pMEXneo vector containing the human aFGF gene with a secretory signal sequence from the hst/KS3 gene to obtain continuous secretion of therapeutic doses of aFGF. NIH 3T3 fibroblasts were transfected using a liposomal transfection reagent and grown in selective media. RESULTS: Dot blot hybridization with labeled complementary DNA probes revealed the presence of plasmid DNA in transfected but not wild type fibroblasts. Intracellular concentrations of aFGF remained low in transfected cells; however, the media contained high levels (32 +/- 7 nM) of aFGF as measured by ELISA. Concentrations of aFGF capable of stimulating cell proliferation were maintained for several weeks. CONCLUSIONS: The aFGF cDNA was transcribed and translated into a functional polypeptide that is secreted from NIH 3T3 cells at physiologically significant concentrations. Stable transfection with a eukaryotic vector which induces secretion of aFGF at levels promoting cell growth holds promise for clinical application in wounds or healing tissue. Transfection could be achieved by topical or endoscopic injection of this type of vector.

Evaluation of femoropopliteal arteries with duplex ultrasound after angioplasty. Can we predict results at one year?

OBJECTIVES: To determine if Duplex ultrasound (DUS) 1 day after percutaneous transluminal angioplasty (PTA) is prognostic for haemodynamic and clinical results at 1 year. DESIGN: Prospective study. PATIENTS AND METHODS: Thirty-four femoropopliteal artery segments were treated with PTA. The peak systolic velocity ratio (PSV ratio = PSV in stenosis: PSV in normal segment) was determined with DUS before PTA, 1 day after PTA and 1 year after PTA. Clinical results were assessed with the SVS/ISCVS (Society for Vascular Surgery/International Society for CardioVascular Surgery) results classification. RESULTS: A 1 year, clinical benefit from PTA was seen in 16 of 25 patients (64%) and haemodynamic improvement in 20 of 34 treated segments (59%). With DUS three residual stenoses were found 1 day after PTA; all occluded within 1 year. Segments with good DUS results after PTA showed haemodynamic deterioration in 30%. Clinical improvement was seen in most patients with DUS improvement, whereas no change or deterioration was found in patients with both good and poor DUS results at 1 year. CONCLUSIONS: Residual stenosis on DUS 1 day after PTA is prognostic for failure within 1 year. However, good DUS results after PTA cannot predict haemodynamic success. Haemodynamic success at 1 year does not imply clinical success.