Panoramic radiographic examination of edentulous jaws.

Many dentists routinely make panoramic radiographs of edentulous patients who request replacement dentures. A study was conducted in patients who attended a Jordanian dental school seeking a new set of dentures. Patients who had at least one set of complete dentures and were treated elsewhere, were included in this study. Any patient whose clinical findings suggested a need for further radiographic study was excluded. Of 286 patients (572 edentulous arches), only three were found to have impacted molars that required extraction and altered the treatment plan. The use of radiography in dentistry generates a large expense and there are risks of radiation hazards. The routine use of radiographs for patients who wear complete dentures and need replacement dentures should be discouraged. A thorough history must be obtained and a careful clinical examination must be performed for the diagnosis and treatment planning.

GP3 is a structural component of the PRRSV type II (US) virion.

Glycoprotein 3 (GP3) is a highly glycosylated PRRSV envelope protein which has been reported as being present in the virions of PRRSV type I, while missing in the type II PRRSV (US) virions. We herein present evidence that GP3 is indeed incorporated in the virus particles of a North American strain of PRRSV (FL12), at a density that is consistent with the minor structural role assigned to GP3 in members of the Arterivirus genus. Two 15aa peptides corresponding to two different immunodominant linear epitopes of GP3 derived from the North American strain of PRRSV (FL12) were used as antigen to generate a rabbit monospecific antiserum to this protein. The specificity of this anti-GP3 antiserum was confirmed by radioimmunoprecipitation (RIP) assay using BHK-21 cells transfected with GP3 expressing plasmid, MARC-145 cells infected with FL12 PRRSV, as well as by confocal microscopy on PRRSV-infected MARC-145 cells. To test if GP3 is a structural component of the virion, (35)S-labelled PRRSV virions were pelleted through a 30% sucrose cushion, followed by a second round of purification on a sucrose gradient (20-60%). Virions were detected in specific gradient fractions by radioactive counts and further confirmed by viral infectivity assay in MARC 145 cells. The GP3 was detected in gradient fractions containing purified virions by RIP using anti-GP3 antiserum. Predictably, the GP3 was less abundant in purified virions than other major structural envelope proteins such as GP5 and M. Further evidence of the presence of GP3 at the level of PRRSV FL12 envelope was obtained by immunogold staining of purified virions from the supernatant of infected cells with anti-GP3 antiserum. Taken together, these results indicate that GP3 is a minor structural component of the PRRSV type II (FL12 strain) virion, as had been previously described for PRRSV type I.

A quick method for boxing complete denture elastomeric impressions.

Rubber base impressions must be poured immediately for maximum accuracy. A method of boxing is presented that uses a previously formed thermoplastic box for pouring the complete denture rubber base impressions in a few minutes.

A boxing procedure for corrected cast impressions.

This new method for boxing a corrected cast impression for a distal extension removable partial denture improves the procedure. The framework with its impression is seated on the remainder of the cast after anatomic ridges have been removed and luted with sticky wax, and the assembly is inserted into a previously formed thermoplastic box and poured to make a new corrected cast.

A one-appointment impression and centric relation record technique for compromised complete denture patients.

This article describes a two-in-one modified custom tray and record block system that is recommended for compromised elderly patients. Custom trays, which are made on primary casts and formed from a patient's functionally corrected old dentures, are used to make final impressions and centric jaw relation records in one clinical appointment. The clinical visits are reduced without compromising the quality of denture construction.

Establishing the posterior palatal seal during the final impression stage.

A procedure for adding a posterior palatal seal at the final impression stage with green stick modeling compound is described. The location and degree of tissue displacement of the posterior palatal seal area are controlled by the dentist. This procedure is suggested to be more accurate than the arbitrary scraping of the master cast.

Making a custom tray for elastomeric impression materials without a primary cast.

The custom tray is recommended for impressions in which it is desirable to establish precise borders, such as Kennedy class I maxillary partial dentures, where an accurate postpalatal seal is a requirement. The peripheries, including the postpalatal seal, can be established with a high degree of accuracy with a custom tray. A procedure is described that does not require a primary impression or a primary cast to make a custom tray for elastomeric impression materials.

A new procedure for separating the edentulous distal extension portion from the master cast when an altered cast is made.

Making an altered cast requires separating the edentulous portion from the rest of the master cast. A plaster saw, which is commonly used for this purpose, requires considerable time and effort. A simplified and efficient procedure is presented for separating the edentulous portion from the rest of the master cast without the use of a plaster saw.

Duplicating an existing complete denture to make a replica.

Sometimes it is desirable to make a few changes when making new complete dentures for a patient who has been wearing old dentures for a long time and is satisfied with them. Replica dentures can be made for these patients, and success with the new dentures can be assured. A procedure is presented for duplication of the old dentures to make replicas.

A procedure for reorienting a cast on a surveyor.

The tilt of a cast must be recorded so that it can be easily transmitted to the dental technician, who should then be able to transfer the original tilt from the master cast directly to any necessary duplicate casts. A procedure is presented that can easily record and transfer the tilt with the help of an impression made with a custom-made U-shaped universal tray.

Simplified clinical remount for complete dentures.

Occlusal errors in new dentures can result from clinical errors and processing changes. Corrections must be made the dentures in the patient's mouth or by making new jaw relation records and correcting the occlusion on an articulator. The latter method is far more accurate because the dentures are seated on rigid bases, errors are more easily seen, and soft tissues and saliva do not interfere with selective grinding. This article describes a simple and quick clinical remount procedure using putty material.

The in vitro-synthesized RNA from a cDNA clone of hepatitis E virus is infectious.

Hepatitis E virus (HEV) is an important etiological agent of epidemic and sporadic hepatitis, which is endemic to the Indian subcontinent and prevalent in most of the developing parts of the world. The infection is often associated with acute liver failure and high mortality, particularly in pregnant women. In order to develop methods of intervention, it is essential to understand the biology of the virus. This is particularly important as no reliable in vitro culture system is available. We have constructed a cDNA clone encompassing the complete HEV genome from independently characterized subgenomic fragments of an Indian epidemic isolate. Transfection studies were carried out with HepG2 cells using in vitro-transcribed RNA from this full-length HEV cDNA clone. The presence of negative-sense RNA, indicative of viral replication, was demonstrated in the transfected cells by strand-specific reverse transcription-PCR and slot blot hybridization. The viral proteins pORF2 and pORF3 and processed components of the pORF1 polyprotein (putative methyltransferase, helicase, and RNA-dependent RNA polymerase) were identified in the transfected cells by metabolic pulse-labeling with [(35)S]methionine-cysteine, followed by immunoprecipitation with respective antibodies. The expression of viral proteins in the transfected cells was also demonstrated by immunofluorescence microscopy. Viral replication was detected in the transfected cells up to 33 days posttransfection (six passages). The culture supernatant from the transfected cells was able to produce HEV infection in a rhesus monkey (Macaca mulatta) following intravenous injection, indicating the generation of viable HEV particles following transfection of cells with in vitro-synthesized genomic RNA. This transient cell culture model using in vitro-transcribed RNA should facilitate our understanding of HEV biology.

Protracted viremia during acute sporadic hepatitis E virus infection.

BACKGROUND/AIMS: Hepatitis E virus (HEV) is associated with epidemic and sporadic hepatitis in developing countries. The disease is largely self-limited with no long-term sequelae. The source of HEV for maintenance of the disease in an endemic area is unknown. This study investigated the occurrence and duration of viremia in patients with acute sporadic HEV infection. METHODS: In 26 of 37 patients with sporadic acute non-A, non-B viral hepatitis, HEV infection was diagnosed based on positivity for immunoglobulin M anti-HEV and/or presence of viremia as shown by reverse-transcription polymerase chain reaction. In 4 patients, fecal samples were analyzed for presence of virus using polymerase chain reaction. Multiple samples were studied at varying times in 20 patients. RESULTS: Viremia was detected in 19 of 26 patients. Two patients had viremia in the absence of immunoglobulin M anti-HEV. Four patients had protracted viremia of 45-112 days' duration. One patient showed fecal virus shedding up to the 52nd day of illness. CONCLUSIONS: Protracted viremia and prolonged fecal shedding of HEV were shown in a small group of patients. These patients may serve as temporary virus carriers responsible for continuous contamination of the sewage water.

An Indian strain of hepatitis E virus (HEV): cloning, sequence, and expression of structural region and antibody responses in sera from individuals from an area of high-level HEV endemicity.

Hepatitis E virus (HEV) is responsible for a majority of sporadic and epidemic viral hepatitides in India and other developing countries. Even though the genomes of four geographically distinct strains of HEV have been cloned and sequenced, the Indian strain of HEV remains largely uncharacterized. We have cloned and sequenced about 2.2 kb of the HEV genome constituting the structural region from an Indian strain of HEV. The nucleotide and amino acid sequences show a high degree of conservation with sequences from other HEV strains. Open reading frames (ORF) 2 and 3 were expressed in Escherichia coli as N-terminal hexahistidine epitope fusions. The purified proteins were then used in an immunoblot assay to evaluate the antibody status in sera from individuals from an area of high-level HEV endemicity. The anti-ORF2 antibodies were found to be nonspecific and could not be correlated to clinical disease. The immunoglobulin M anti-ORF3 was found to be specific for the presence of acute disease. The implications of these findings in HEV diagnosis and vaccine development are discussed.

Hepatitis E infection in children: study of an outbreak.

BACKGROUND: Hepatitis E virus (HEV) is responsible for most of the hepatitis epidemics in the developing world and it frequently affects young adults. Therefore, common perception is that it does not affect children. METHODS: A group of 20 school children (13 years old) were possibly exposed to hepatitis E virus infection during a 2 day trekking trip. Epidemiological and clinical information was correlated to the presence of the hepatitis E virus genome and antibodies to HEV structural and non-structural proteins found in the blood of the children, using polymerase chain reaction and line immunoassay techniques. RESULTS: Ten children developed icteric hepatitis, seven prodrome-like illness without jaundice while three remained asymptomatic. Immunoglobulin M (IgM) antibodies to open reading frame (ORF)2 protein (pORF2) were detected in all 19 children tested, whereas 11 and 10 of the children were positive for IgM antibodies against ORF1 (pORF1) and ORF3 (pORF3) proteins, respectively. The rate of HEV infection was found to be 85%. Viraemia was observed in 11 children and was present in four of the seven anicteric patients (55%) compared with six of the nine (66%) icteric patients. One child without any symptom also had viraemia. CONCLUSIONS: The data obtained indicate a high susceptibility of children for HEV infection and a frequently prolonged viraemia in those infected.

Cloning, sequencing, and expression of the hepatitis E virus (HEV) nonstructural open reading frame 1 (ORF1).

Hepatitis E virus (HEV) causes enterically transmitted epidemic and sporadic viral hepatitis affecting millions of people in the developing world. Different geographical isolates of HEV show a high degree of homology at the nucleotide and amino acid levels. The approximately 7.2 kb RNA genome has three open reading frames of which ORF1 is predicted to code for the viral nonstructural polyprotein. The expression, processing and properties of the nonstructural ORF1 polyprotein have not been reported so far. In this study, the complete HEV ORF1 was reconstructed from overlapping fragments amplified by polymerase chain reaction (PCR) of total RNA isolated from the bile fluid of a rhesus monkey experimentally infected with HEV isolate from an epidemic. The complete assembled ORF1 was sequenced using HEV specific primers. The ORF1 polyprotein was expressed in E. coli, in a cell free translation system and in HepG2 cells, and was characterized by western blotting and immunoprecipitation using acute phase patient serum as well as polyclonal antibodies raised against defined parts of the ORF1 polyprotein. The nonstructural polyprotein of HEV was expressed as a 186 kDa protein. No processing was observed into discrete units, either in-vitro based on a kinetic analysis, or in HepG2 cells based on immunoprecipitation.

Acute viral hepatitis types E, A, and B singly and in combination in acute liver failure in children in north India.

The aetiological agents responsible for, and the outcome of, acute liver failure were investigated prospectively in 44 children (29 males, 15 females) attending a tertiary health care facility in India. The children were between the ages of 2 months and 13 years. Studies for viral infections and other etiologies could be carried out in 40 patients. Specific aetiological labels were possible in 35 (87.5%) patients. Thirty (75%) had evidence of acute viral hepatitis. Acute hepatitis E virus (HEV) infection was found in a total of 18 children, with hepatitis A (HAV) in 16, hepatitis B in 5, and C in 1. Seven had isolated infection with hepatitis E, five with A, and four with B. Nine had both E and A infection. Superinfection of HEV was observed in a child with Indian childhood cirrhosis (ICC). Acute HEV infection was confirmed by immunoblot assay in all the patients and in eight of these, HEV-RNA was also detected in the serum. HAV was involved in 37.5% of cases with isolated infection in 10% (4 of 40). The aetiological factors associated with acute liver failure, apart from HAV and HEV, were other hepatotropic viruses (22.5%), Wilson's disease (5%), ICC (5%), and hepatotoxic drugs (7.5%). In five patients, no serological evidence of acute viral hepatitis could be found, neither did the metabolic screen yield any result. It was observed that enterically transmitted hepatitis viruses (HAV and HEV) were associated with 60% of acute hepatic failure in children. Mixed infection of HAV and HEV formed the single largest aetiological subgroup. In developing countries, where hepatitis A and E infections are endemic, severe complications can arise in the case of mixed infection. This may contribute to most of the mortality from acute liver failure during childhood.