Retrospective 25-year follow-up of treatment outcomes in Angle Class III patients : Success versus failure.

OBJECTIVES: Despite recommendations for early treatment of hereditary Angle Class III syndrome, late pubertal growth may cause a relapse requiring surgical intervention. This study was performed to identify predictors of successful Class III treatment. MATERIALS AND METHODS: Thirty-eight Class III patients treated with a chincup were retrospectively analyzed. Data were collected from the data archive, cephalograms, and casts, including pretreatment (T0) and posttreatment (T1) data, as well as long-term follow-up data collected approximately 25 years after treatment (T2). Each patient was assigned to a success or a failure group. Data were analyzed based on time (T0, T1, T2), deviations from normal (Class I), and prognathism types (true mandibular prognathism, maxillary retrognathism, combined pro- and retrognathism). RESULTS: Compared to Class I normal values, the data obtained in both groups yielded 11 significant parameters. The success group showed values closer to normal at all times (T0, T1, T2) and vertical parameters decreased from T0 to T2. The failure group showed higher values for vertical and horizontal mandibular growth, as well as dentally more protrusion of the lower anterior teeth and more negative overjet at all times. In adittion, total gonial and upper gonial angle were higher at T0 and T1. A prognostic score-yet to be evaluated in clinical practice-was developed from the results. The failure group showed greater amounts of horizontal development during the years between T1 and T2. Treatment of true mandibular prognathism achieved better outcomes in female patients. Cases of maxillary retrognathism were treated very successfully without gender difference. Failure was clearly more prevalent, again without gender difference, among the patients with combined mandibular prognathism and maxillary retrognathism. Crossbite situations were observed in 44% of cases at T0. Even though this finding had been resolved by T1, it relapsed in 16% of the cases by T2. CONCLUSION: The failure rate increased in cases of combined mandibular prognathism and maxillary retrognathism. Precisely in these combined Class III situations, it should be useful to apply the diagnostic and prognostic parameters identified in the present study and to provide the patients with specific information about the increased risk of failure.

Variety of angiotensin receptors in 3T3-L1 preadipose cells and differentiated adipocytes.

A local paracrine acting angiotensin (ANG) system of preadipocytes and mature adipocytes is involved in metabolic effects and tissue differentiation. The present study reports on the investigation of binding affinities for various angiotensin receptors including their relevance in 3T3-L1 adipocytes and preadipocytes and 3T3-442A preadipocytes. Competitive binding studies using both 125I-ANG II and its more stable analogue 125I-SARILE for investigating AT1/AT2 binding sites in 3T3-L1 preadipocytes reveal a biphasic competition curve with KDs at a low and high nanomolar range. By using the AT2 receptor selective ligand 125I-CGP4112A the presence of high affinity AT2 binding sites in preadipocytes was observed. High nonspecific binding and a low receptor number is characteristic for all these experiments. An AT4 binding site (binding site for ANG IV) exists in 3T3-L1 and F442A preadipocytes and adipocytes with a high nanomolar KD. This low binding affinity was confirmed by a biological assay, the IRAP assay (=insulin regulated aminopeptidase assay). IRAP is associated with the AT4 receptor, which is a binding site at the luminal part of membrane bound IRAP. The curves for competition binding and for inhibition of IRAP activity are superimposable with respect to angiotensin IV. In conclusion, AT1 and AT2 binding sites are present in preadipocytes. AT2 receptor binding affinities are shown in preadipocytes for the first time. The description of a non-AT1/AT2 binding site with low affinity remains speculative albeit of high interest because antidiabetic and obesity related effects of angiotensin peptides and sartanes as antagonists are observed at these high concentrations. Local concentrations of ANG II and their degradation products may be extremely high. The low amounts of AT1 and AT2 binding sites emphasize the relevance of other binding sites in adipose tissue development and metabolic effects. The AT4 binding site seems to be one of the predominant receptors in adipose cells. Other degraded, but still bioactive peptides like ANG III, IV and ANG(1-7), activating receptors not influenced by ANG II, could be of importance.

Appearance of the bona fide spiral tubule of ORF virus is dependent on an intact 10-kilodalton viral protein.

Parapoxviruses can be morphologically distinguished from other poxviruses in conventional negative staining electron microscopy (EM) by their ovoid appearance and the spiral tubule surrounding the virion's surface. However, this technique may introduce artifacts. We have examined Orf virus (ORFV; the prototype species of the Parapoxvirus genus) by cryoelectron microscopy (cryo-EM) and cryo-negative staining EM. From these studies we suggest that the shape and unique spiral tubule are authentic features of the parapoxviruses. We also constructed an ORFV mutant deleted of a gene encoding a 10-kDa protein, which is an orthologue of the vaccinia virus (VACV) 14-kDa fusion protein, and investigated its ultrastructure. This mutant virus multiplied slowly in permissive cells and produced infectious but morphologically aberrant particles. Mutant virions lacked the spiral tubule but displayed short disorganized tubules similar to those observed on the surface of VACV. In addition, thin extensions or loop-like structures were appended to the ORFV mutant particles. We suggest that these appended structures arise from a failure of the mutant virus particles to properly seal and that the sealing activity is dependent on the 10-kDa protein.

Constant versus dissipating forces in orthodontics: the effect on initial tooth movement and root resorption.

The aim of this clinical and confocal laser scanning microscopic study was to compare the effects of two frequently used archwires on tooth movement and root resorption. A total of 84 premolars in 27 individuals (10 boys, 17 girls, with a mean age of 12.5 years) was moved buccally with an experimental fixed orthodontic appliance. In a split mouth experimental design the premolar on one side was activated with a stainless steel wire with a buccal offset of 1 mm, which was reactivated every four weeks and the contralateral premolar was moved with a superelastic wire with a force plateau of 0.8-1 N. This wire had an initial activation of 4.5 mm and was not reactivated during the 12-week experimental period. At the end of the experimental period the teeth were extracted. Six premolars were used as control teeth and were extracted before the experiment started. Tooth displacement was studied three-dimensionally on dental casts with a co-ordinate measuring machine. The depth, perimeter, area, and volume of the resorption lacunae was measured using three-dimensional digital images made with a confocal laser scanning microscope (CLSM). On these images the resorbed portions of the root surface were 'reconstructed' mathematically. The results show that the teeth activated with the superelastic wire moved significantly more than the teeth with the steel wire during the experimental period. The depth of the resorption lacunae did not differ significantly between the groups; however, perimeter, area, and volume of the resorption lacunae on the teeth of the 'superelastic group' were 140 per cent greater than on the teeth of the 'steel group'. It may be concluded that a greater amount of tooth movement occurred with superelastic wires, offering a force level of 0.8-1 N compared with stainless steel wires, with initially higher but rapidly declining forces in an experimental set up for a period of 12 weeks. The amount of root resorption was significantly larger in the superelastic group.

Localizing infection with a technetium-99m-labeled peptide: initial results.

In vitro-labeled leukocyte imaging is useful for the detection of infection, but an in vivo labeling method is preferable. This study sought to evaluate the safety and efficacy of a leukocyte-avid peptide for the detection of infection, to determine the effects of peptide dose on performance and to compare the peptide with in vitro-labeled leukocytes. A 23-amino acid peptide, P483, containing the platelet factor-4 heparin-binding sequence, was labeled with 99mTc and complexed with heparin (P483H). Thirty patients were injected with 29 microg (n = 11), 145 microg (n = 10) or 290 microg (n = 9) of labeled peptide, and imaged 15 min and 90-120 min later. Early and late images were interpreted individually and jointly. Twenty patients underwent (111)In-labeled leukocyte scintigraphy. Fourteen patients had infection: osteomyelitis (n = 7), vascular graft (n = 2), abscess (n = 2), joint replacement (n = 1), surgical wound (n = 1) and pneumonia (n = 1). There were 10 adverse events in six patients; all were mild and resolved spontaneously, and without any intervention. The sensitivity, specificity and accuracy were the same for both early and late imaging: 0.86, 0.81 and 0.83, respectively. Interpreting early and late images together did not improve the results. No relationship between peptide dose and study accuracy was found. In patients undergoing both examinations, the accuracies of the peptide and in vitro-labeled leukocyte imaging were identical: 0.80. In summary, 99mTc-P483H safely, rapidly and accurately detected focal infection, was comparable with in vitro-labeled leukocyte imaging and therefore merits further investigation.

Monoclonal antibodies directed against conserved epitopes on the nucleocapsid protein and the major envelope glycoprotein of equine arteritis virus.

We recently developed a highly effective immunization procedure for the generation of monoclonal antibodies (MAbs) directed against the porcine reproductive and respiratory syndrome virus (E. Weiland, M. Wieczorek-Krohmer, D. Kohl, K. K. Conzelmann, and F. Weiland, Vet. Microbiol. 66:171-186, 1999). The same method was used to produce a panel of 16 MAbs specific for the equine arteritis virus (EAV). Ten MAbs were directed against the EAV nucleocapsid (N) protein, and five MAbs recognized the major viral envelope glycoprotein (G(L)). Two of the EAV G(L)-specific MAbs and one antibody of unknown specificity neutralized virus infectivity. A comparison of the reactivities of the MAbs with 1 U.S. and 22 newly obtained European field isolates of EAV demonstrated that all N-specific MAbs, the three nonneutralizing anti-G(L) MAbs, and the weakest neutralizing MAb (MAb E7/d15-c9) recognized conserved epitopes. In contrast, the two MAbs with the highest neutralization titers bound to 17 of 23 (MAb E6/A3) and 10 of 23 (MAb E7/d15-c1) of the field isolates. Ten of the virus isolates reacted with only one of these two MAbs, indicating that they recognized different epitopes. The G(L)-specific MAbs and the strongly neutralizing MAb of unknown specificity (MAb E6/A3) were used for the selection of neutralization-resistant (NR) virus variants. The observation that the E6/A3-specific NR virus variants were neutralized by MAb E7/d15-c1 and that MAb E6/A3 blocked the infectivity of the E7/d15-c1-specific NR escape mutant confirmed that these antibodies reacted with distinct antigenic sites. Immunoelectron microscopy revealed for the first time that the antigenic determinants recognized by the anti-G(L) MAbs were localized on the virion surface. Surprisingly, although the immunofluorescence signal obtained with the neutralizing antibodies was relatively weak, they mediated binding of about three times as much gold granules to the viral envelope than the nonneutralizing anti-G(L) MAbs.

Identification and phylogenetic comparison of Salem virus, a novel paramyxovirus of horses.

A virus that could not be identified as a previously known equine virus was isolated from the mononuclear cells of a horse. Electron microscopy revealed enveloped virions with nucleocapsid structures characteristic of viruses in the Paramyxoviridae family. The virus failed to hemabsorb chicken or guinea pig red blood cells and lacked neuraminidase activity. Two viral genes were isolated from a cDNA expression library. Multiple sequence alignments of one gene indicated an average identity of 45% as compared to Morbillivirus N protein sequences. A weaker relationship was found with Tupaia paramyxovirus (TPMV) and Hendra virus (HeV) N proteins. In the second gene, multiple open reading frames (ORFs) were identified, corresponding to the arrangement of the P, V, and C ORFs in the Morbillivirus and Respirovirus viruses. Short stretches in the C-terminal regions of the P and C proteins showed limited homologies to viruses in the Morbillivirus genus but no obvious relationship to viruses in other genera. The V ORF translation product contained a highly conserved, cysteine-rich domain that is common to most viruses in the Paramyxovirinae subfamily. Sequencing of P gene cDNA clones confirmed the use of a cotranscriptional editing mechanism for the regulation of P/V expression. Based on the location of its origin it has been named Salem virus (SalV).

Polyhistidine-tagged hepatitis B core particles as carriers of HIV-1/gp120 epitopes of different HIV-1 subtypes.

The hepatitis B core antigen is a widely accepted carrier particle to enhance the immunogenicity of foreign epitopes. From electron cryomicroscopy, the immunodominant region between amino acid positions 79 to 81 is known to protrude from the surface of the shells. It can be replaced by heterologous sequences without interfering with the particle-forming capacity in many cases. Here we have introduced various V3 sequences of the envelope protein of different subtypes (A, B, O) of HIV-1/gp120 in order to enhance their immunogenicity and broaden the immune response against the virus. To improve purification efficiency and solubility of the E. coli-expressed hybrids, six histidine residues were fused to amino acid 156. An adjustable purification scheme was utilised including denaturation, Ni(2+)-NTA affinity chromatography and particle renaturation under high salt conditions, resulting in highly pure antigen preparations. The hybrids reacted specifically with sera of HIV-1-infected patients. They further induced an autologous, subtype-specific anti-HIV-1 antibody response superior to that of Keyhole limpet-haemocyanine-coupled peptides.

Intranuclear inclusions in cells infected with Newcastle disease virus.

Cells infected by Newcastle Disease Virus were observed to contain both intracytoplasmic and intranuclear inclusion bodies. Ultrastructurally, they consisted of twisted strands of about 18-20 nm diameter resembling nucleocapsids. The presence of these inclusions was detected irrespective of host cell or pathogenicity of the virus. In immunofluorescence and immunogold labelling experiments, these structures were tagged by an anti-P protein monoclonal antibody. In summary, we show that intracytoplasmic and intranuclear inclusion bodies, hitherto used as a taxonomic characteristic for the genus Morbillivirus of the Paramyxoviridae, also occur in a member of the genus Rubulavirus.

Localization of pestiviral envelope proteins E(rns) and E2 at the cell surface and on isolated particles.

The glycoproteins E(rns) of classical swine fever virus (CSFV) and E(rns) and E2 of bovine viral diarrhoea virus (BVDV) are shown to be located at the surface of infected cells by the use of indirect immunofluorescence and by cytofluorometric analysis. The positive immunostaining of the cell surface was further analysed by immunogold electron microscopy and it could be shown that only extracellular virions were labelled. Gold granules were not seen at the cellular plasma membrane. In contrast to BVDV E2, the CSFV E2 of virions sticking to the plasma membrane was not accessible to the respective monoclonal antibodies. However, CSFV particles isolated from culture supernatant were able to bind both monoclonal anti-E(rns) and anti-E2 antibodies. For CSFV and BVDV, binding of anti-E(rns) antibodies to the virions was more pronounced than that of anti-E2. This finding was unexpected since E2 is considered to be the immunodominant glycoprotein.

Monoclonal antibodies to the GP5 of porcine reproductive and respiratory syndrome virus are more effective in virus neutralization than monoclonal antibodies to the GP4.

The arterivirus porcine reproductive and respiratory syndrome virus (PRRSV) contains six structural proteins the roles of which are not completely understood. In a preceding study, immunization with the dutch isolate I10 of PRRSV had led to the development of MAbs against four structural proteins [Wieczorek-Krohmer, M., 1994. Herstellung und Charakterisierung von monoklonalen Antikörpern gegen das Virus des Porzinen Reproduktiven und Respiratorischen Syndroms (PRRSV). Inaugural-Dissertation, Ludwig-Maximilians-Universität, München] here finally identified by reaction with individual plasmid-expressed PRRSV proteins as products of ORFs 3 (GP3), 4 (GP4), 5 (GP5) and 7 (N). Surprisingly, the MAbs against GP5 revealed the presence of two antigenically distinct virus populations in the isolate I10, the population PRRSV-'PPV', isolated from plaques and the PRRSV-'EPV', gained by end point dilution. MAbs against GP3, GP4 and N reacted with both I10 populations as well as with natural PRRSV isolates. However, the anti-GP5 MAbs exclusively recognized PRRSV-'PPV'. In this study immunization of mice with both separated I10 populations confirmed that solely PRRSV-'PPV' possesses the property to induce an immune response ultimately leading to the establishment of MAbs against GP5. Whereas the 15 anti-GP5 MAbs (derived from four independent fusions) reacted exclusively with PRRSV-'PPV' of the isolate I10, anti-GP4 MAbs detected their target antigen on various isolates of European origin and were able to neutralize them. As indicated by competition assays and selection of neutralization-resistant virus mutants, all GP5 MAbs are directed against a single antigenic site on the ORF 5 protein. Both groups of neutralizing antibodies bound to the surface of purified virions demonstrating that the recognized epitopes represent surface structures of the virion envelope. However, anti-GP5 MAbs mediated the binding of more gold granules than anti-GP4 MAbs. Comparison of the neutralizing effect of anti-GP4 and anti-GP5 MAbs revealed the anti-GP5 MAbs as the more efficient antibodies. For the complete neutralization of about 100 ID50 of PRRSV-'PPV' anti-GP5 culture supernatant was effective up to a dilution of 1:1280 whereas the most effective anti-GP4 antibodies exhibited a comparable effect only up to 1:64. These results indicate that PRRSV GP5 in principle is a major target for neutralizing antibodies, as is found for other arteriviruses, but that in nature 'ORF 5 escape mutants' may develop as easily as in vitro.

Optimizing images of acute deep-vein thrombosis using technetium-99m-apcitide.

OBJECTIVE: The purpose of this paper is to introduce the nuclear medicine technologist to a new radiopharmaceutical, 99mTc-apcitide, for imaging acute venous thrombosis. After reading this paper, the technologist should be able to: (a) describe patient preparation for imaging with 99mTc-apcitide; (b) state the amount of 99mTc-apcitide that is administered to patients for imaging acute venous thrombosis; (c) explain patient positioning for optimal image acquisition; and (d) discuss gamma camera acquisition parameters and their importance in obtaining high-quality images. Clinical cases illustrate both the whole-body distribution and diagnostic value of 99mTc-apcitide in detecting acute deep-vein thrombosis.

Matrix protein of rabies virus is responsible for the assembly and budding of bullet-shaped particles and interacts with the transmembrane spike glycoprotein G.

To elucidate the functions of rhabdovirus matrix (M) protein, we determined the localization of M in rabies virus (RV) and analyzed the properties of an M-deficient RV mutant. We provide evidence that M completely covers the ribonucleoprotein (RNP) coil and keeps it in a condensed form. As determined by cosedimentation experiments, not only the M-RNP complex but also M alone was found to interact specifically with the glycoprotein G. In contrast, an interaction of G with the nucleoprotein N or M-less RNP was not observed. In the absence of M, infectious particles were mainly cell associated and the yield of cell-free infectious virus was reduced by as much as 500,000-fold, demonstrating the crucial role of M in virus budding. Supernatants from cells infected with the M-deficient RV did not contain the typical bullet-shaped rhabdovirus particles but instead contained long, rod-shaped virions, demonstrating severe impairment of the virus formation process. Complementation with M protein expressed from plasmids rescued rhabdovirus formation. These results demonstrate the pivotal role of M protein in condensing and targeting the RNP to the plasma membrane as well as in incorporation of G protein into budding virions.

[Characterization of viral hemorrhagic septicemia (VHS) virus isolates in trout]. / Charakterisierung von Isolaten des Virus der Viralen Hämorrhagischen Septikämie (VHS) der Forellen.

Virus diseases of fish can seriously impair the economy of aquacultur. Control and prevention of fish diseases in the European Union (EU) are focussed on the viral haemorrhagic septicaemia (VHS) and the infectious haematopoeitic necrosis (IHN). The diagnosis of VHS and IHN is performed in the Federal Republic of Germany (FRG) on the basis of the legislation of the EU. Since 1994 we received an increasing number of VHS virus (VHSV) isolates which did not react with a commercially available anti-VHSV monoclonal antibody (MAb) in the indirect immuno fluorescence test. With our own MAb ID8, however, as well as with additional diagnostic methods these virus isolates could be identified. These isolates of rainbow trouts were designated as VHSV type "Wi". Electron microscopically all stages of rhabdovirus maturation could be detected. Morphologically the isolates were undistinguishable from other rhabdoviruses. By immuno electron microscopy using the MAb ID8 rhabdovirus nucleocapsid structures were demonstrated. The virulence of the new VHSV type Wi was not different from that of a VHSV isolate with conventional reaction patterns as well as of a VHSV laboratory strain.

Upper extremity radionuclide bone imaging: the wrist and hand.

Bone scintigraphy of the hands and wrists represents an important adjunct imaging technique that complements plain film radiographic examination. The use of the three-phase bone scan provides clinical information not only regarding osseous uptake but the blood flow and extravascular distribution of the radiotracer as well. Scintigraphic evaluation of the hands and wrists is employed in acute and chronic conditions. In the event of an equivocal or negative plain film, the bone scan can identify occult fractures. Of particular concern is the identification of scaphoid fractures due to the higher incidence of osteonecrosis. Work related injuries represent a significant health issue. The bone scan can be a part of the algorithm for evaluating chronic pain syndromes including reflex sympathetic dystrophy. The complimentary roles of bone scanning and imaging with gallium-67 citrate or radiolabeled leukocytes has proven useful in the evaluation of acute or chronic osteomyelitis. In addition, the diphosphonates are useful in identifying solitary and multiple primary bone tumors. In the case of primary bone tumor, thallium-201 can be used to evaluate response to therapy. Although uncommon in the hand and wrist, the bone scan can identify metastatic tumors or tumor-related conditions such as hypertrophic osteoarthropathy. Finally, bone scintigraphy may be useful in identifying location and extent in a variety of conditions such as fibrous dysplasia, histiocytosis X, and Paget's disease.

A CXCR4/CD4 pseudotype rhabdovirus that selectively infects HIV-1 envelope protein-expressing cells.

We show that a cellular virus receptor functions in the envelope of a virus, allowing selective infection of cells displaying the receptor ligand. A G-deficient rabies virus (RV) pseudotyped with CD4- and CXCR4-derived proteins selectively infected cells expressing HIV-1 envelope protein. Envelope protein or CD4 antibodies blocked virus entry. Pseudotype virus formation was most efficient with chimeric receptor proteins possessing the cytoplasmic tail of the RV G spike protein (CXCR4-RV and CD4-RV). While CXCR4-RV was incorporated when expressed alone, CD4-RV incorporation required CXCR4-RV as a carrier protein, indicating a mechanism by which oligomeric surface proteins are sorted into the RV envelope. Viral vectors bearing virus receptors in their envelope may be useful reagents for targeting virus-infected cells in vivo.

Secular trends in malocclusion in Austrian men.

A comparison between occlusal deviations in the permanent dentition in the skulls of 94 19th century and 157 present-day Austrian males was made by means of the PAR Index. It was found that the contemporary dentitions showed significantly higher malocclusion scores than the 19th century sample (weighted PAR Index 11.79 and 6.62, respectively). The results show that secular changes in malocclusion have occurred during the last 100 years.

Initial effects of treatment of Class II malocclusion with the Herren activator, activator-headgear combination, and Jasper Jumper.

The initial effects of treatment of Class II, Division 1 malocclusion with an activator, according to Herren (27 patients), with an activator-headgear combination (20 patients), or with the Jasper Jumper appliance (25 patients) were studied on lateral cephalograms from before and after 6 to 8 months of treatment. The patients' ages ranged from 9 to 12 years. At the end of the period of observation, the correction in overjet and molar relationship was more complete in the patients with the Jasper Jumper than in the patients with the activator. Whereas all the patients with the Jasper Jumper showed neutral occlusion, this was the case in only 20 of the 47 patients with the activator. The correction of the distal occlusion occurred through a combination of skeletal and dentoalveolar adaptations. Skeletal changes accounted for 42%, 35%, and 48% of the overjet correction by the Herren-type activator, the headgear-activator, and the Jasper Jumper, respectively. The correction of the molar relationship occurred to 55%, 46%, and 38% by skeletal changes in the respective groups. Dentoalveolar compensation (distal movement of the upper molars, mesial movement of the lower molars) appeared to be inversely related to skeletal adaptation. The patients with the Jasper Jumper showed a marked intrusion of the lower incisors with a consequent reduction in overbite.