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The vast amount of solid waste produced each year is one of the greatest problems associated with greenhouse horticulture in some European countries. In particular, the disposal of used growing media arising from the soil-less cultivation of vegetables in mineral wool creates serious difficulties. The nonbiodegradability of these mainly inorganic substrates causes environmental concern and has prompted the search for alternative growing media such as cocos derivatives, perlite and resin foam (Fytocell®). Organic substrates in combination with biodegradable material such as plastic, rope and clippings have the advantage that re-use or recycling of the waste is easier, cheaper and more environmentally friendly than for mineral wool. However, the differing physical and chemical characteristics of the alternative substrates may affect yield significantly. Substrates based respectively on peat and peat with cocos derivatives were tested against a mineral wool control for the production of tomato in three consecutive years. Both organic substrates were placed in biodegradable plastic bags. Greenhouse experiments demonstrated that plants grown in the pure peat substrate rooted more easily than plants grown in the peat-cocos substrate or mineral wool, and that they developed less blossom-end rot in both peat substrates than in mineral wool. Due to the buffering capacity of the organic substrates, the electrical conductivity of the draining water appeared to be more stable during cultivation. The total yield of tomato fruits was similar for all substrates, and no differences between substrates could be observed in the quality of the fruits produced. On the other hand, flavour tests demonstrated that plants grown on peat substrate produced more tasty fruits under certain conditions. The results of this study show that organic growbags are promising and competitive alternatives to mineral wool. [ABSTRACT FROM AUTHOR]

Blepharoplasty is a frequent request in consultations of plastic surgery. Patients are often presenting with a progressive swelling of the eyelids. For functional or aesthetic reasons, we commonly perform a superior and/or inferior blepharoplasty to correct this problem. We present the case of a 72-year-old woman who consulted us with a prominent unattractive swelling of both lower eyelids. Because of the atypical and suspicious presentation, supplementary examinations were conducted, and the fat was also sent for analysis. A diagnosis of unilateral orbital lymphoma was obtained. The patient was treated as a primarily localized lymphoma with a positive outcome. When correctly diagnosed and treated, orbital lymphomas can have a very good prognosis.

Background/aims: Acute hemorrhage of the upper gastrointestinal tract occurs at a rate of 50 to 100 per 100,000 annually in the Western adult population. With the increased use of therapeutic endoscopy, the role of surgery is decreasing; surgical intervention is now only used in cases of failure of endoscopic hemostasis. The goal of this study is to determine whether there are predictive factors associated with high-risk post-operative mortality., Methodology: This retrospective study included 30 patients treated from March 1996 to September 2008 at Brugmann Hospital. These patients presented with upper gastrointestinal non-variceal hemorrhage that was treated first endoscopically then surgically for recurrent hemorrhage. Multiple risk factors (variable and fixed) and parameters were evaluated to determine their influence on mortality., Results: Of 30 patients, 10 (33%) developed recurrent hemorrhage following surgical treatment. A total of 8 (26.6%) deaths occurred of which 4 were related to hemorrhage. Three deaths occurred after the first intervention and 5 occurred after a second intervention. Logistic regression analysis revealed that the total number of blood units transfused and the presence of at least one surgical reintervention both significantly increased mortality rate (p = 0.0426 and p = 0.0068). Other parameters were not significant. However, there is a lack of power due to the small sample size., Conclusion: For recurrent massive upper gastrointestinal hemorrhage following endoscopic treatment and necessitating more than 19 blood transfusions, early surgical intervention is recommended and surgical reintervention should be avoided. If reintervention is necessary, radical surgery is recommended. However, the small number of patients treated over a 12-year period limits the results of this study, and these results may represent simple coincidences.

Bilateral humeral artery aneurysm is a very rare condition. Simultaneous bilateral complication of these aneurysms makes it even more unusual. We present a case of a 44-year-old man with bilateral digital ischaemia as a complication of a bilateral humeral artery aneurysm. Anamnestic, clinical and per-operative findings led to several hypotheses. A venous bypass graft was done to exclude both aneurysms and to arrest the embolisation into the digital arteries.

Congenital high airway obstruction syndrome (CHAOS) is a rare prenatal diagnosis consisting of a typical fetal triad of large hyperechogenic lungs, flattened or inverted diaphragms and ascites. Most cases are sporadic with unknown incidence. Before attempts of fetoscopic fetal salvage or ex utero intrapartum treatment (EXIT) are considered, additional malformations must be carefully excluded as CHAOS may be part of various monogenic conditions or chromosomal disorders. We report an unique family with autosomal dominant inheritance of CHAOS and variable expression in the affected father and two affected children. It is concluded that minor expression in one of the parents may be an important indicator for genetic counseling in CHAOS and management of future pregnancies.

The presence of L-type Ca2+ channels at the frog neuromuscular junction (nmj) was studied by monitoring changes in intracellular Ca2+ evoked in presynaptic terminals and perisynaptic Schwann cells (PSCs) and by studying the distribution of Ca2+ channels using a monoclonal antibody directed against the alpha 2/delta subunit of L channels. L-type Ca2+ channel agonist and antagonist had no effect on resting level of fluorescence and nerve-evoked Ca2+ responses in presynaptic terminals. However, depolarization of PSCs induced by KCl (25 mM) produced entry of Ca2+, which was prevented by L-type Ca2+ channel blockers, in (+)R Bay K 8644 of nimodipine. Labeling of Ca2+ channels revealed an intracellular epitope with an irregular and spotty distribution along the endplate. Similar results were obtained with a fluorescent phenylalkylamine [(-)DM-BODIPY-PAA], a blocker of L-type Ca2+ channels. Ca2+ channel labeling remained in absence of nerve terminals but was absent after mechanical removal of nerve terminals and PSCs. Most Ca2+ channel spots were distributed in between bands of cholinergic receptors labeled with alpha-bungarotoxin-TRITC. Cross sections of motor endplates revealed that labeling of Ca2+ channels was found only at the level of the synaptic cleft and not all around the PSCs. We conclude that L-type Ca2+ channels are located in perisynaptic glial cells in an appropriate location to sense depolarization induced by neurotransmitters and thus may support possible roles of glial cells on synaptic function.

The autoradiographic distribution of Ca(2+)-sensitive high- and low-affinity binding sites for the N-type calcium channel ligand, [125I]omega-conotoxin GVIA, has been examined in rat brain. Both classes of sites were rather evenly distributed, exhibiting their highest density in cell body rich layers, such as the pyramidal layer of Ammon's horn and the granular layer of the dentate gyrus. In cerebellum, the granular layer was more densely labelled than the molecular layer at low ligand concentration, and this pattern was reversed at high concentration. The presence of 30 mM Ca2+ in the incubation medium revealed the existence of a third class of sites, insensitive to Ca2+, comparably distributed at both concentrations, and apparently absent from cell body rich layers.

The surface distribution of the alpha 2/delta subunit of the 1,4-dihydropyridine receptor and its topographical relationship with the neural cell adhesion molecule (N-CAM) were investigated during early myogenesis in vitro, by double immunocytochemical labeling with the monoclonal antibody 3007 and an anti-N-CAM polyclonal antiserum. The monoclonal antibody 3007 has been previously shown to immunoprecipitate dihydropyridine receptor from skeletal muscle T-tubules. In further immunoprecipitation experiments on such preparations and muscle cell cultures, it was demonstrated here that the monoclonal antibody 3007 exclusively recognizes the alpha 2/delta subunit of the 1,4-dihydropyridine receptor. In rabbit muscle cell cultures, the labeling for both alpha 2/delta and N-CAM was first detected on myoblasts, in the form of spots on the membrane and perinuclear patches. Spots of various sizes organized in aggregates were then found on the membrane of myotubes. At fusion (T0), aggregates of N-CAM spots alone were found at the junction between fusing cells. At T6 and later stages, all alpha 2/delta aggregates present on myotubes co-localized with N-CAM, while less than 3% of N-CAM aggregates did not co-localize with alpha 2/delta. A uniform N-CAM staining also made its appearance. At T12, when myotubes showed prominent contractility, alpha 2/delta-N-CAM aggregates diminished in size. Dispersed alpha 2/delta spots of a small regular size spread over the whole surface of the myotubes and alignments of these spots became visible. Corresponding N-CAM spots were now occasionally seen, and uniform N-CAM staining was prominent. These results show that alpha 2/delta and N-CAM are co-localized and that their distributions undergo concomitant changes during early myogenesis until the T-tubule network starts to be organized. This suggest that these two proteins might jointly participate in morphogenetic events preceding the formation of T-tubules.

The Purkinje cell protein 2 (Pcp-2) is expressed in cerebellar Purkinje cells and retinal bipolar neurons. To illuminate how Pcp-2 expression is restricted to only two neuronal types and to derive tools to express heterologous genes in these neuronal subpopulations, genomic sequences of the mouse Pcp-2 gene have been cloned and flanking sequences have been evaluated as a source of neuron-specific regulatory elements. An upstream region with homology to other genes expressed in neurons was identified and a hybrid gene containing this sequence was constructed by ligating 0.4 kb of upstream and 0.3 kb of downstream Pcp-2-flanking DNA to lacZ. Transgenic mice bearing this construct exhibited beta-galactosidase in a wide array of neuron types, suggesting that this sequence may play an important role in specifying neuronal expression. Addition of a further 3.1 kb of Pcp-2 upstream sequences restricted expression of beta-galactosidase to a small number of neuron types and most notably to Purkinje cells within parasagitally oriented cerebellar compartments. The presence of elements lying within the 3.1-kb upstream region and acting to specifically restrict Pcp-2 expression is therefore suggested. Moreover, as beta-galactosidase was not expressed in the bipolar cells of these transgenic mice, retinal expression of the endogenous Pcp-2 gene must involve elements in addition to those conferring expression within Purkinje cells.

GTP binding proteins have been proposed to play a role in excitation--contraction coupling. In a precedent study [Toutant et al., (1988), Biochem. J., 405-409], we determined that Bordetella pertussis toxin is able to catalyse ADP-ribosylation of two substrates in the detergent soluble fraction of total muscle extracts. Purified fractions of transverse tubule membranes (T-tubule membranes), a key element of the excitation--contraction coupling, were shown to exhibit a major ADP-ribosylated substrate at 40 kd and an immunoreactivity with antisera raised against purified bovine brain Go alpha or G beta. In the present study, we have investigated the cellular distribution of G protein subunits in comparison with that of the voltage-dependent Ca2+ channels by immunofluorescence on transverse and longitudinal sections of fast and slow muscles. With affinity-purified antibodies against G beta subunits, a fluorescent labelling underlined the myofibrils and sarcolemma, whereas a strong immunoreaction in a dotted pattern evoked the presence of the subunit in repetitive triadic structures. With anti-Go alpha antibodies, the immunofluorescence was more clearly focussed on a dotted pattern and the co-location with the voltage-dependent Ca2+ channel immunoreactivity indicates that both proteins were located in very close subcellular structures. Immunoblot analysis and PTX ADP-ribosylation of the purified light sarcoplasmic reticulum (LSR), heavy sarcoplasmic reticulum (HSR) and T-tubule subcellular fractions indicate the discrete presence of G proteins in LSR, an unambiguous labelling of the HSR fraction, while T-tubule membranes clearly appear very rich in a Go-like protein, confirming the observed preferential immunocytochemical distribution of G protein subunits.(ABSTRACT TRUNCATED AT 250 WORDS)

We compared the maturation pathway of normal and psoriatic epidermis using three different markers: (1) Involucrin, which is normally detected in the stratum granulosum in normal skin, was detected in all but the basal layer of involved psoriatic skin; (2) an antigen, recognized by the murine monoclonal antibody psi 3, was present in all but the basal layer of involved psoriatic skin but was absent from uninvolved and normal skin; (3) fibronectin, which normally localizes in the dermis and the epidermal-dermal junction, was also detected intra- and extracellularly in the psoriatic epidermis. These results indicate that the alterations in keratinocyte maturation found in psoriasis do not arise from a truncation of the normal maturation pathway but rather reflect the onset of an abnormal pathway of differentiation characterized by the expression of psi 3 antigen and fibronectin and the premature appearance of involucrin.

Monoclonal hybridoma cell lines secreting antibodies against the (+)-PN 200-110 and the (-)-demethoxyverapamil binding components of the voltage-dependent calcium channel from rabbit transverse-tubule membranes have been isolated. The specificity of these monoclonal antibodies was established by their ability to coimmunoprecipitate (+)-[3H]PN 200-110 and (-)-[3H]demethoxyverapamil receptors. Monoclonal antibodies described in this work cross-reacted with rat, mouse, chicken, and frog skeletal muscle Ca2+ channels but not with crayfish muscle Ca2+ channels. Cross-reactivity was also detected with membranes prepared from rabbit heart, brain, and intestinal smooth muscle. These antibodies were used in immunoprecipitation experiments with 125I-labeled detergent [3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) and digitonin] solubilized membranes. They revealed a single immunoprecipitating component of molecular weight (Mr) 170,000 in nonreducing conditions. After disulfide bridge reduction the CHAPS-solubilized (+)-PN 200-110-(-)-demethoxyverapamil binding component gave rise to a large peptide of Mr 140,000 and to smaller polypeptides of Mr 30,000 and 26,000 whereas the digitonin-solubilized receptor appeared with subunits at Mr 170,000, 140,000, 30,000, and 26,000. All these results taken together are interpreted as showing that both the 1,4-dihydropyridine and the phenylalkylamine receptors are part of a single polypeptide chain of Mr 170,000.

The serum of a patient suffering from myeloma was found to decorate microtubules and mitotic spindles of cultured cells. Immunoblots performed after one- and two-dimensional electrophoresis showed a reaction with a certain subset of beta-tubulin isoforms, but not with beta'- and alpha-tubulins. The tubulin subset contained both ubiquitous (beta-3) and neurospecific (beta-4,5,6) isoforms. An IgM lambda and an IgA kappa myeloma protein were found in this serum. Immunoblots performed with specific anti-isotype second antibodies showed that the tubulin subset could be evidenced using anti-mu, alpha, lambda, and kappa-specific antisera. Moreover, the tubulin subset was also evidenced using an anti-gamma second antibody. These results, which do not exclude a participation of the myeloma proteins in the anti-tubulin reactivity, indicate, however, that the antibody response was polyclonal. The same restricted specificity of all classes of anti-tubulin antibodies of this serum favours the hypothesis that the immune response of the patient was directed against an antigen sharing epitopes with tubulin rather than with tubulin itself.

Partially purified fractions of dihydropyridine and phenylalkylamine receptors associated with voltage-dependent calcium channels in rabbit skeletal muscle were found to contain two glycopeptides of similar molecular weight. A peptide of approximately 165 kDa was photoaffinity labelled with an arylazido-phenylalkylamine Ca channel inhibitor and also was phosphorylated with cAMP-dependent protein kinase. Another peptide of 170 kDa could be distinguished from the 165 kDa peptide by peptide mapping and differences in electrophoretic mobility. The results suggest that the 165 kDa peptide contains the sites responsible for regulation of calcium channel activity by calcium channel inhibitors as well as by neurotransmitters that regulate its activity in a cAMP-dependent manner.

The dihydropyridine-sensitive voltage-dependent Ca2+ channel from cardiac tissue was purified 900-fold using DEAE-Sephadex A-25, concanavalin A-Sepharose, and wheat germ agglutinin-Sepharose. The purified preparation was highly enriched in a peptide of 140,000 daltons when electrophoresed on sodium dodecyl sulfate gels in the presence of 2-mercaptoethanol, or 170,000 when electrophoresed in the presence of iodoacetamide. Polyclonal antibodies raised against the purified subunits of the rabbit skeletal muscle Ca2+ channel recognized the 170-kDa protein in preparations electrophoresed under nonreducing conditions, and the large peptide of 140 kDa and smaller peptides of 29-32 kDa in preparations analyzed under reducing conditions. Monoclonal antibodies, which were raised against the native Ca2+ channel from skeletal muscle, immunoprecipitated [3H]PN 200-110 binding activity from solubilized cardiac membranes and immunoprecipitated 125I-labeled peptides (from the purified cardiac Ca2+ channel preparation) which migrated as a single species of 170 kDa under nonreducing conditions, or as 140, 32, and 29 kDa under reducing conditions. The results show that the purified cardiac Ca2+ channel, like that previously purified from skeletal muscle, consists of a major component of 170 kDa which is comprised of a 140-kDa peptide linked by disulfide bonds to smaller peptides of 32-29 kDa. Peptide maps of the 140-kDa peptide purified from cardiac and skeletal muscle preparations were strikingly similar, suggesting a high degree of homology in their primary sequence.