The simultaneous appearance of a nasal natural killer-cell lymphoma and acute myelogenous leukemia.

INTRODUCTION AND AIM: Sinonasal malignant neoplasms are uncommon, with an annual incidence of less than 1/100,000. About 80% of these are squamous cell carcinoma. Adenocarcinoma and adenoid cystic carcinoma are next in frequency. Lymphoma of the nasal cavity, paranasal sinuses and nasopharynx are rare, constituting less than 5% of all extranodal lymphomas. CASE REPORT: A 47-year-old man was referred to our hospital because of severe headache and progressive facial pain. He also complained of right-sided visual acuity. He had a manifest exopthalmia with disturbed eye movements. Nasoscopy showed a large mass with atypical appearance. CT and MRI showed a bilateral ethmoid mass invading the frontal sinuses, the right orbit, the lamina cribrosa and the right frontal cerebral region, and growing posteriorly through the choana. The first biopsies were inconclusive, showing only necrotic cells and purulent inflammation with epithelial elements. A larger biopsy demonstrated a high-grade malignant tumour with necrosis. The differential diagnosis of undifferentiated sinonasal carcinoma, undifferentiated neuro-endocrine tumour or T-cell lymphoma was suggested. In the meantime our patient developed high fever and sudden-onset pancytopenia. Bone marrow punction showed 65% blasts, leading to the diagnosis of AML type M2. He was immediately referred for chemotherapy, but died in intensive care before his first session. The biopsy of the sinonasal mass was diagnosed surprisingly as a natural killer cell lymphoma stage IVB. CONCLUSIONS: Natural killer cell lymphoma is rare in Europe. The simultaneous appearance of a NK-cell lymphoma and acute myelogenous leukemia has, as far as we know, never been described in the English literature before.

Pott's puffy tumor: CT and MRI findings.

Pott's puffy tumor, a subperiosteal abscess of the frontal bone with cranial osteomyelitis, is a rare complication of frontal sinusitis. In this report, we describe the radiological findings of a 24-year-old man, presenting with swelling of the right upper eyelid. Correct and early diagnosis of this infrequent, but potentially life-threatening condition is of utmost importance.

The feather touch rasp, a powered instrument for hump reduction.

INTRODUCTION AND AIM: Standard rhinoplasty procedure involves bony profile alignment with an osteotome, followed by profile refinement with a manual rasp. The entire bony hump may sometimes be addressed with a rasp. Manual rasps and osteotomes, however, can be traumatic instruments, resulting in tissue oedema and bruising. The feather touch rasp is one of the powered instruments developed in recent years to improve precision and technical ease while preventing tissue trauma. The powered rasp has been frequently used at our department in the last eighteen months for hump reductions. MATERIAL AND METHODS: Retrospective evaluation of 72 rhinoplasty procedures performed in 68 patients between January 2004 and June 2005. A bony hump reduction was necessary in 52 of the 68 patients. 60% of the patients were male. The mean age was 30 years. The open rhinoplasty technique was used in 65% of the patients. All humps were addressed with the feather touch rasp only. Patients were seen 10 days, 4 weeks and three months after surgery. RESULTS: In one patient the nasal dorsum remained too high. We found one asymmetric nasal dorsum. Another patient had a low nasofrontal angle, creating a false impression of a remaining hump. In two patients, bony irregularities appeared a few months after the rhinoplasty procedure. Overall, most patients were satisfied with the results of the hump reduction. CONCLUSIONS: The feather touch rasp makes safe and gradual bony hump reduction possible, with fragments being aspirated and the overlying skin being well protected.

Signaling through a novel domain of gp130 mediates cell proliferation and activation of Hck and Erk kinases.

Interleukin-6 (IL-6) induces the activation of the Src family kinase Hck, which is associated with the IL-6 receptor beta-chain, gp130. Here we describe the identification of an "acidic" domain comprising amino acids 771 to 811 of gp130 as a binding region for Hck, which mediates proliferative signaling. The deletion of this region of gp130 (i.e., in deletion mutant d771-811) resulted in a significant reduction of Hck kinase activity and cell proliferation upon stimulation of gp130 compared to wild-type gp130. In addition, d771-811 disrupted the growth factor-stimulated activation of Erk and the dephosphorylation of Pyk2. Based on these findings, we propose a novel, acidic domain of gp130, which is responsible for the activation of Hck, Erk, and Pyk2 and signals cell proliferation upon growth factor stimulation.

Signal transduction of interleukin-6 involves tyrosine phosphorylation of multiple cytosolic proteins and activation of Src-family kinases Fyn, Hck, and Lyn in multiple myeloma cell lines.

Binding of interleukin-6 to its receptor (IL-6R) induces the association of the IL-6R alpha chain (IL-6Ralpha) with a 130-kDa transmembrane glycoprotein, gp130. This event activates tyrosine kinases of the Janus kinase (JAK) family and transduces signals to the cytosol or nucleus. To further characterize the biochemical mechanisms by which IL-6 promotes cell proliferation, we investigated the effects of IL-6 on the growth and transmembrane signaling of several lymphoid cell lines. In the IL-6-dependent cell line B-9, IL-6 induced a rapid, transient, and concentration-dependent tyrosine phosphorylation of several cytosolic proteins as detected by antiphosphotyrosine immunoblots. The molecular weight of major bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 44, 65, 70, 80, 137, 148, 184, and 190 kDa, respectively. Similar effects of IL-6 on tyrosine phosphorylation were observed in the human multiple myeloma cell line LP-1. Because JAKs were unlikely to mediate all the biological effects of IL-6, we investigated whether members of the Src family of tyrosine kinases were also activated in B-9 or LP-1 cells. IL-6 induced the activation and tyrosine phosphorylation of p59Fyn, p56/59Hck, and p56Lyn. Coprecipitation experiments with anti-Hck, anti-Lyn, anti-Fyn, and anti-gp130 antibodies revealed a physical association with gp130 of p56/59Hck and p56Lyn, but not p59Fyn, in LP-1 cells. Together, these results show for the first time that several Src kinases may become activated by IL-6 (p59Fyn, p56/59Hck, and p56Lyn) and associate with gp130 (p56/59Hck and p56Lyn).

Familiarity interferes with filial imprinting.

The present study was performed to investigate whether and how pre-exposure to an object affects subsequent filial imprinting to that object. In Experiment 1 junglefowl chicks (Gallus gallus spadiceus) were first exposed to either a red object alone (control group), or a red and a yellow object simultaneously (experimental group; phase 1). Subsequently, all chicks were exposed to the yellow object in the presence of a black and blue one (phase 2). At the end of phase 1, most experimental chicks had developed a preference for the red object over the yellow one. At the end of phase 2, preferences of experimental chicks were shifted away from the yellow object towards the novel black and blue object, relative to preferences of control chicks. This shows that pre-exposure may interfere with imprinting. Experiment 2 revealed that when control chicks were tested with the yellow object at the end of phase 1, filial responses were as strong as in experimental chicks. This shows that the yellow object had not acquired control over filial behaviour during phase 1, and also that the relatively impaired imprinting on that object in phase 2 was not due to reduced generalization from the red object. One possible explanation why pre-exposure may interfere with imprinting is that familiarity alters the level of attention attracted by an object, a mechanism suggested to underlie 'latent inhibition' in conditioning.

Parallel genome analysis by two-dimensional DNA typing.

By two-dimensional (2-D) DNA typing a restriction enzyme digest of genomic DNA can be resolved on the basis of both size and base-pair sequence and subsequently analysed by repeat probe hybridization to reveal sequence variants at multiple genomic sites in parallel. The system has been partly automated and allows for large-scale comparative analysis of complex genomes in a cost-effective manner.

[Orbital involvement of sinusitis]. / Orbitale verwikkelingen van een sinusitis.

Sinusitis can spread to the orbital tissues. 'Preseptal' soft-tissue bacterial infections are relatively common during childhood and with appropriate antimicrobial therapy they usually resolve quickly. 'Orbital' soft-tissue infections, in contrast, are relatively rare and frequently cause serious morbidity. Four cases are presented. Symptomatology and pathogenesis are reviewed. Diagnostic procedures, especially CT-scan, are discussed and recommendations are given for medical and surgical management.

An investigation into blocking of filial imprinting in the chick during exposure to a compound stimulus.

The occurrence of "blocking" was investigated in jungle fowl chicks (Gallus gallus spadiceus B.) in an imprinting situation. In Experiment 1, chicks were simultaneously exposed to two stationary coloured cylinders, either two red cylinders (Group RR), a yellow and a red cylinder (YR), or two yellow cylinders (YY). After six days of exposure, the cylinders were removed from the cages and replaced by a yellow and a blue cylinder (i.e. YB) for each chick. This second phase of the experiment lasted for seven days. When the blue cylinder was presented alone during tests at different stages in Phase 2, the RR birds spent significantly more time with this cylinder and emitted fewer shrill calls than the chicks in the YR and YY groups. In Experiment 2, RR and YY birds were reared as in Experiment 1, except that in the second phase of the experiment they were exposed to a blue cylinder only. In this experiment the development of an attachment to the novel blue cylinder proceeded similarly in the RR and YY birds. In Experiment 3, it was found that chicks that were reared with a yellow and a red cylinder preferred the latter stimulus. Thus, although in the first phase of Experiment 1 the RR birds had been exposed to a more attractive stimulus, in tests during the second phase they spent more time with a novel stimulus (B) than the YY birds. These results are consistent with the suggestion that imprinting to a novel stimulus is "blocked" to some extent when that stimulus is presented in compound with another stimulus to which the animal has previously been exposed.

The virB operon of Agrobacterium tumefaciens pTiC58 encodes 11 open reading frames.

Agrobacterium tumefaciens genetically transforms plant cells by transferring a copy of its T-DNA to the plant where it is integrated and stably maintained. In the presence of wounded plant cells this process is activated and mediated by the products of the vir genes which are grouped into six distinct loci. The largest is the virB locus spanning 9.5 kb. Transposon mutagenesis studies have shown that virB gene products are required for virulence but their functions remain largely unknown. To provide information relevant to understanding the function of VirB polypeptides, the nucleotide sequence of the virB operon from a nopaline plasmid, pTiC58, is presented here. Eleven open reading frames (ORFs) are predicted from this sequence. The predicted sizes of 10 of the 11 VirB polypeptides are verified by specific expression in Escherichia coli. Only the product of the smallest ORF potentially encoding a 5.8 kDa polypeptide has not been detected. The initiation of translation of five virB ORFs occurs at codons that overlap the termination codons of the ORF immediately upstream; thus, translational coupling may be an important mechanism for efficient translation of the large virB polycistronic mRNA. Based on hydropathy plot analysis nine of the virB ORFs encode proteins that may interact with membranes; these data support the earlier hypothesis that virB gene products may form a membrane pore or channel to mediate exit of the T-DNA copy (T-strands) from Agrobacterium into the plant cell. A comparison of the two published octopine virB sequences with the nopaline sequence presented here is made.

Activation of the T-DNA transfer process in Agrobacterium results in the generation of a T-strand-protein complex: Tight association of VirD2 with the 5' ends of T-strands.

The T-DNA transfer process of Agrobacterium is activated following induction of expression of the Ti plasmid virulence (vir) genes. The virD1 and virD2 gene products are required for the production of nicks at the T-DNA borders and for the generation of free linear single-stranded copies of the T-DNA region, T-strands. T-strands are complexed with proteins in vir-induced bacteria, since T-strands partition to the aqueous/phenol interface in non-Pronasetreated total cell extracts. To determine whether the proteins are tightly associated with T-strands, DNA-protein complexes were purified away from bulk proteins by adsorption to glass beads. A 58-kDa protein was specifically released from vir-induced DNA-protein complexes after treatment with S1 nuclease to digest single-stranded DNA. The 58-kDa protein was identified as VirD2 by using VirD2-specific antibodies. The tight association of VirD2 with T-strands was shown directly by using VirD2-specific antibody to isolate T-strands. The 5' side of the border nick sites on the Ti plasmid was also shown to be tightly associated with protein. The data suggest that after VirD1/VirD2-dependent nicking at the T-DNA borders, the VirD2 protein remains bound to the 5' end of the nick, and the VirD2 protein continues to bind tightly to this 5' end during unwinding (or displacement) of the T-strand from the Ti plasmid T-DNA region. The tight binding of VirD2 to T-strands suggests that this protein has additional functions in T-strand generation and potentially in the later steps of T-DNA transfer.

Expression of Agrobacterium nopaline-specific VirD1, VirD2, and VirC1 proteins and their requirement for T-strand production in E. coli.

Induction of Ti plasmid virulence (vir) genes during early stages of the genetic transformation of plant cells by Agrobacterium tumefaciens results in several molecular events that are involved in generating a transferable T-DNA copy. These events include site-specific nicking at the T-DNA borders and synthesis of free, unipolar, linear, single-stranded copies of the T-DNA (T-strands). Here E. coli was used as a heterologous cell to assay the requirements for T-strand synthesis. Cells of E. coli harbored two compatible plasmids, one containing coding sequences overlapping the virC and virD regions of the nopaline Ti plasmid, and a second plasmid containing a T-DNA region. The amount of vir proteins produced was varied by placing their expression under the control of either native Agrobacterium, tac, or T7 promoters. The data show that VirD1 and VirD2 proteins are absolutely essential for T-strand production in E. coli, and the relative amounts of these polypeptides produced correlate with the amounts of T-strand observed. When VirD1 and VirD2 products are limiting, the VirC1 protein increases T-strand production. The yield of T-strands also varies as a function of the plasmid vector used to clone the T-DNA region substrate; the same T-DNA cloned into pLAFR1 produces more T-strands than that cloned into the higher copy number plasmid pACYC184. In summary, VirD1 and VirD2 proteins are the minimal requirements for T-strand production; however, other factors such as VirC1, the relative concentration of VirD1, VirD2, and the T-DNA substrate, and possibly additional functions (e.g., those specified by pLAFR1) influence the efficiency of T-strand production. Additional results regarding the requirements for expression of VirD1 and VirD2 polypeptides are presented.

Single-Stranded DNA Binding Protein Encoded by the virE Locus of Agrobacterium tumefaciens.

The transfer process of T (transfer)-DNA of Agrobacterium tumefaciens is activated after the induction of the expression of the Ti plasmid virulence (vir) loci by plant signal molecules such as acetosyringone. The vir gene products then act to generate a free transferable single-stranded copy of the T-DNA, designated the T-strand. Although some vir proteins are responsible for the synthesis of the T-strand, others may mediate T-strand transfer to plant cells as part of a DNA-protein complex. Here, a novel 69-kilodalton vir-specific single-stranded DNA binding protein is identified in Agrobacterium harboring a nopaline-type Ti plasmid. This protein binds single-stranded but not double-stranded DNA regardless of nucleotide sequence composition. The molecular size of the vir-specific single-stranded DNA binding protein and its relative abundance in acetosyringone-induced Agrobacterium suggested that it might be the product of the virE locus; molecular cloning and expression of the virE region in Escherichia coli confirmed this prediction.

Genetic manipulations in Rhizobium meliloti utilizing two new transposon Tn5 derivatives.

Two derivatives of the prokaryotic transposon Tn5 were constructed in vitro. In Tn5-233, the central area of Tn5, which carries resistance to kanamycin/neomycin, bleomycin and streptomycin, is replaced by a fragment carrying resistance to the aminocyclitol antibiotics gentamycin/kanamycin and streptomycin/spectinomycin. In Tn5-235, the Escherichia coli beta-galactosidase gene is inserted within the streptomycin resistance gene of Tn5, and constitutively expressed from a Tn5 promoter. Both constructs transpose with about the same frequency as Tn5 in Escherichia coli and Rhizobium meliloti. When a Tn5-derivative is introduced into an R. meliloti strain which already contains a different Tn5-derivative, in situ transposon replacement is obtained at high frequency, presumably by a pair of crossovers between the IS50 sequences at the ends of the incoming and resident transposons. In this way we converted a previously isolated recA::Tn5 mutant into the corresponding recA::Tn5-233 strain, which can now be used as a genetic background in the study of complementation of other Tn5-induced mutations. We also replaced the drug markers of several Tn5-induced exo mutants, which we were then able to map relative to each other by transduction with phage phi M12. In a strain carrying Tn5-235 located near Tn5-233, we were able to isolate deletions of the intervening markers, presumably resulting from general recombination between the two transposons, by screening for loss of the Lac+ phenotype. Unlike Tn5 itself, resident Tn5-233 does not appear to suppress transposition of another incoming Tn5-derivative.

Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes.

Using physical and genetic data, we have demonstrated that Rhizobium meliloti SU47 has a symbiotic megaplasmid, pRmeSU47b, in addition to the previously described nod-nif megaplasmid pRmeSU47a. This plasmid includes four loci involved in exopolysaccharide (exo) synthesis as well as two loci involved in thiamine biosynthesis. Mutations at the exo loci have previously been shown to result in the formation of nodules which lack infection threads (Inf-) and fail to fix nitrogen (Fix-). Thus, both megaplasmids contain genes involved in the formation of nitrogen-fixing root nodules. Mutations at two other exo loci were not located on either megaplasmid. To mobilize the megaplasmids, the oriT of plasmid RK2 was inserted into them. On alfalfa, Agrobacterium tumefaciens strains containing pRmeSU47a induced marked root hair curling with no infection threads and Fix- nodules, as reported by others. This plant phenotype was not observed to change with A. tumefaciens strains containing both pRmeSU47a and pRmeSU47b megaplasmids, and strains containing pRmeSU47b alone failed to curl root hairs or form nodules.

Pattern ERG and glaucomatous visual field defects.

In the past five years numerous reports have suggested that ganglion cell function can be tested by means of a specialized form of electroretinography, the so-called pattern electroretinogram (PERG). Because of the important potentials of a ganglion cell test for clinical use this technique has been applied by several investigators to patients with (presumed) ganglion cell dysfunction, especially glaucoma. On grounds of principle we had reason to question whether the reported positive results should be attributed to ganglion cell dysfunction or to other factors such as optical disturbances. We investigated in this study the PERG as a function of visual field loss in glaucoma patients with careful control of optical factors. We did not find changes in PERG as a function of field loss. So either field loss is not related to the mass behaviour of ganglion cells, or ganglion cells are not the prime basis of the PERG. We believe the latter to be true.

Pigment organization and energy transfer in the green photosynthetic bacterium Chloroflexus aurantiacus : I. The cytoplasmic membrane.

We have studied the pigment arrangement in purified cytoplasmic membranes of the thermophilic green bacterium Chloroflexus aurantiacus. The membranes contain 30-35 antenna bacteriochlorophyll a molecules per reaction center; these are organized in the B808-866 light-harvesting complex, together with carotenoids in a 2:1 molar ratio. Measurements of linear dichroism in a pressed polyacrylamide gel permitted the accurate determination of the orientation of the optical transition dipole moments with respect to the membrane plane. Combination of linear dichroism and low temperature fluorescence polarization data shows that the Qy transitions of the BChl 866 molecules all lie almost perfectly parallel to the membrane plane, but have no preferred orientation within the plane. The BChl 808 Qy transitions make an average angle of about 44° with this plane. This demonstrates that there are clear structural differences between the B808-866 complex of C. aurantiacus and the B800-850 complex of purple bacteria. Excitation energy transfer from carotenoid to BChl a proceeds with about 40% efficiency, while the efficiency of energy transfer from BChl 808 to BChl 866 approaches 100%. From the minimal energy transfer rate between the two spectral forms of BChl a, obtained by analysis of low temperature fluorescence emission spectra, a maximal distance between BChl 808 and BChl 866 of 23 Å was derived.