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Biomedical subjects

M E Johansson

Publications and source records attributed to M E Johansson.

At least 19 recordsLinked to original sources

The glycosylation of rat intestinal Muc2 mucin varies between rat strains and the small and large intestine. A study of O-linked oligosaccharides by a mass spectrometric approach.

The large glycosylated domains obtained from the rat intestinal mucin Muc2 were isolated from the large and small intestine of the inbred rat strains GOT-W and GOT-BW. The expression of the rat Muc2 in the large intestine was confirmed immunochemically and by Northern blotting. Released oligosaccharides were structurally characterized by gas chromatography-mass spectrometry (neutral and sialylated species) or by tandem mass spectrometry (sulfated species), and a total of 63 structures was assigned. The large intestinal oligosaccharides were found to be identical between the strains, while the small intestinal glycosylation differed. Until now, detailed structural analysis of oligosaccharides isolated from a single mucin core or mucin domain with different origin have not been performed, and the information of different mucin glycoforms has been limited to immunochemistry. Blood group A-determinants (GalNAcalpha1-3(Fucalpha1-2)Galbeta1-, and structures related to the blood group Sda/Cad-related epitope NeuAc/NeuGcalpha1-3(GalNAcbeta1-4)Galbeta1-, were found in GOT-BW small intestine, and also in both large intestines. Blood group H-determinants and NeuAc/NeuGcalpha1-3Galbeta1- were found in all samples. Core 1 (Galbeta1-3GalNAcalpha1-), core 2 (Galbeta1-3(GlcNAcbeta1-6)GalNAcalpha1-), core 3 (GlcNAcbeta1-3GalNAcalpha1-), and core 4 (GlcNAcbeta1-3(GlcNAcbeta1-6)GalNAcalpha1- were also found in all the samples. The large intestine were enriched in sulfated oligosaccharides and the small intestine contained higher amounts of sialylated species. Sulfation were found exclusively on C-6 of GlcNAc.

ABO Blood-Group System↗

Molecular characterization of the large heavily glycosylated domain glycopeptide from the rat small intestinal Muc2 mucin.

The largest high-glycosylated domain, glycopeptide A, of the "insoluble' mucin complex of the rat small intestine has earlier been purified and characterized (Carlstedt et al., 1993, J Biol Chem 268: 18771-81). A rabbit antiserum raised against deglycosylated glycopeptide A was used to clone part of a mucin showing homology to the human MUC2 mucin (Hansson et al., 1994, Biochem Biophys Res Commun 198. 181-90). This serum specifically stained goblet cells (paranuclear) in the mouse small intestine. The size of the coding sequence of glycopeptide A was estimated by using reversed transcriptase PCR of mRNA from an inbred rat strain (GOT-W) using primers in the unique central and C-terminal parts of the proposed rat Muc2 sequences. The PCR and Southern blot of the PCR products showed a fragment of about 5.5 kb corresponding to about 1700 amino acids when the known Cys-rich sequences used for the primers were subtracted. This is slightly larger than the size estimated earlier by biochemical studies. The mRNA encoding the rat Muc2 was slightly smaller than the mRNA encoding the human MUC2 in a colorectal cell line. Although the size of glycopeptide A estimated from biochemical results and by PCR is not identical, the results obtained here further support that the "insoluble' mucin of the rat small intestine is encoded by the Muc2 gene. Most of the oligosaccharides in glycopeptide A were either neutral (40%) or sialylated (40%). The remaining ones were sulfated with the sulfate group attached to C-6 of N-acetylglucosamine linked to C-6 of the N-acetylgalactosaminitol as revealed by tandem mass spectrometry of the perdeuteroacetylated oligosaccharides. Eighteen oligosaccharides were found of which fourteen were characterized and found to be mostly novel. Our findings thus expand the current knowledge of the core peptide of the rat intestinal goblet cell mucin and provide a relatively complete picture of the glycosylation of a defined mucin domain.

Animals↗

Adenoviruses isolated in the Stockholm area during 1987-1992: restriction endonuclease analysis and molecular epidemiology.

Adenoviruses recovered from the northern Stockholm area during 1987-1992 have been subjected to restriction endonuclease analysis. Adenoviruses of all subgenera (A-F) were represented and a considerable degree of serotype variation was seen, e.g. the rarely encountered subgenus A viruses were frequently isolated in the present study. Of 16 subgenus A isolates, Ad31 predominated with 12 strains which were equally distributed into the DNA-variants D2 and D7. Analysis of 38 Ad3 isolates revealed four DNA-variants: D1, D3, D10, and "Sto1". The ten Ad7 isolates belonged all to the DNA-variant D5 of Ad7. Of 27 Ad1 strains, 11 belonged to D10, followed by the DNA-variants D4 and D7 with four strains each. Among Ad2 isolates, D2 or D2-like strains prevailed (23/28). Of six Ad5 strains, three belonged to the DNA-variant D2. The most notable subgenus D event was a nosocomial outbreak of keratokonjunctivitis due ot Ad19a. In addition, a collection of heterogenous subgenus D strains was detected, most of which untypable by RE-analysis. Among the six Ad4 isolates of subgenus E, both genomic clusters (p and a, respectively) of Ad4 were recognized. Concerning the clinical important subgenus F adenoviruses, only two strains of Ad40 were detected as compared to 12 strains of Ad41, all of which ascribed to the DNA-variant D12 of Ad41.

Adenovirus Infections, Human↗

Characterization of two divergent adenovirus 31 strains.

Two divergent strains of adenovirus type 31 were analyzed by neutralization test and restriction endonuclease (RE) patterns in an effort to find the basis for their genetic variability. One strain, isolated from the throat of a child in Maryland during an upper respiratory illness in 1968, was partially neutralized by Ad 31 antisera (to 16-fold lower than homologous titer) while its own antiserum fully neutralized prototype Ad 31 virus, but shared only 9% of comigrating RE fragments with Ad 31 prototype (vs. 30% with Ad 18 prototype); however, PCR tests specific for the inverted terminal repeat (ITR) sequence of Ads 12 and 18 were negative. The other strain, recovered from a stool sample from an infant with diarrhea in Georgia in 1979, was inhibited by Ad 31 antiserum to within 4-fold homologous titer, but shared only 15% of comigrating fragments with Ad 31 prototype (vs. 91% with Ad 18 prototype); ITR-specific PCR tests with this virus were positive for Ad 12/Ad 18. These data suggest that both strains are from separate evolutionary lines of Ad 31 unrelated to all other isolates studied to date by RE analysis, and that the partial neutralization by prototype Ad 31 antisera might represent small mutations in the hexon gene.

Adenovirus Infections, Human↗

Molecular epidemiology of adenoviruses isolated in Iceland during 1988-1990.

All human adenoviruses isolated in Iceland during 1988-1990 have been subjected to restriction endonuclease analysis. Of 55 isolates altogether, subgenus C (Ad1, Ad2, and Ad5) predominated with 42 isolates followed by subgenus B (Ad3 and Ad7) with 12. Analysis of the 9 Ad1 isolates revealed 6 DNA-variants. Among these the established DNA-variants D4, D7, and D10 were recognized. The remaining 3 DNA-variants were primarily found in Iceland. Among the 22 Ad2 isolates, 7 DNA-variants could be distinguished. D2 predominated with 15 isolates whereas the prototype was isolated only once. The novel 5 DNA-variants of Ad2 were all closely related to D2. Analysis of the 11 Ad5 isolates revealed 6 DNA-variants, 2 of which (D2 and D5) were already established. Ice2 and D3 were the most common occurring DNA-variants of Ad5. Ad5 showed the highest degree of genomic variability within subgenus C, both in terms of the low degree of pair-wise comigration of restriction fragments and the number of principal variants of RE-patterns. Analysis of the 9 Ad3 isolates revealed 3 DNA-variants: D3, D10, and Ice1 (a novel DNA-variant that resembles D10). The DNA-variants D3 and D10 were each represented by 4 isolates. The three Ad7 isolates belonged all to the DNA-variant D5.

Adenovirus Infections, Human↗

Restriction endonuclease patterns of adenovirus type 12 and 18.

Restriction endonuclease analysis using 10 restriction enzymes was performed on six and three wild isolates of adenovirus (Ad) type 12 and 18, respectively. Among the Ad12 strains, five DNA variants could be identified. The degree of pairwise comigration of restriction fragments suggests a high degree of genomic diversity within Ad12. The wild isolates of Ad18, on the other hand, displayed a low degree of genetic variability and comprised one DNA variant closely related to the prototype strain. The BglII, BstEII, and HindIII restriction endonuclease patterns of Ad18 were inconsistent with those originally presented. Identical RE-patterns among Ad18 prototype strains (DC) obtained from four different sources, including directly from the American Type Culture Collection, verify that the genuine Ad18 prototype was used in the present study. Using the revised restriction patterns of BglII, BstEII, and HindIII, a proper identification of Ad18 will be facilitated.

Adenovirus Infections, Human↗

Intrauterine fetal death due to echovirus 11.

In a case of intrauterine fetal death in the 29th week of gestation, echovirus 11 could be isolated from the umbilical cord of the fetus. The mother had no apparent signs of infection but serological evidence of current echovirus 11 infection. Enterovirus PCR performed on paraffin-embedded specimens of various tissues (myocardium, lung, liver and placenta) from the fetus yielded positive results in all cases. These findings, together with supporting serological and epidemiological findings--e.g. proven echovirus 11 infection 3 weeks before in the 18-month-old son of the woman--constituted strong evidence that echovirus 11 infection was responsible for the fetal death.

Adult↗

Genome analysis of adenovirus type 31 strains from immunocompromised and immunocompetent patients.

Adenovirus type 31 (Ad31) was isolated from 15 immunocompromised patients in 12 of whom seroconversion was also recorded. Ad31 infection has a substantial clinical relevance since 8 of 10 with lower respiratory tract infection and 4 of 4 with hepatitis died. Therefore, Ad31 isolates from immunocompetent and immunodeficient hosts were compared by restriction endonuclease analysis. Nine genome types were identified among the 79 Ad31 isolates. Pairwise comparison of comigrating restriction fragments indicated that the genome types could be divided into three genomic clusters. Several Ad31 genome types were isolated from immunocompromised patients, but no highly virulent genome type could be found. A genome type was identified in a child with severe combined immunodeficiency who originally was infected with another genome type. This observation is suggested to have evolutionary implications.

Adenovirus Infections, Human↗

Characterization of two genome types of adenovirus type 31 isolated in Stockholm during 1987.

The authors isolated during 1987 seven adenovirus type 31 (Ad31) within a 9-month period. The isolates were obtained from urine, throat, and feces, implying a systemic spread of the infection. Most patients displayed gastrointestinal symptoms, but some had respiratory symptoms and fever. All of the strains differed by restriction endonuclease analysis from the prototype strain (1315) by an additional Bgl II restriction site. Ad31 isolates 1-6 could be divided into two groups by the enzymes Bam HI, Msp I, and Xho I. Each enzyme gave rise to the same group distribution: isolates 1-3 and 4-6, respectively. Digestion with Bst EII, Hind III, Kpn I, and Sma I resulted in identical patterns for isolates 1-6. Isolate 7, however, demonstrated a DNA deletion of approximately 0.8 kbp, but it was otherwise identical to isolates 4-6. In conclusion, two separate genome types of Ad31 were isolated, one of which included a DNA deletion mutant. The increased isolation rate may reflect an epidemiological situation, as the same isolation procedure had been used both before and after this period.

Adenovirus Infections, Human↗

Investigation of the film formation of magnesium stearate by applying a flow-through dissolution technique.

The film formation of magnesium stearate on the surface of acetylsalicylic acid was investigated by applying a flow-through dissolution technique. The effect of mixing time, lubricant surface area, and the addition of colloidal silica was studied. The film formation increased by increasing mixing time. The final level reached was independent of the specific surface area of the lubricants, but granular magnesium stearate gave a lower surface coverage than the powdered lubricants. The lubricating effect was independent of the mixing time and specific surface area of the lubricants. Colloidal silica was found to interact primarily with the free fraction of magnesium stearate.

Aspirin↗

Enzyme-linked immunosorbent assay for detection of enteric adenovirus 41.

An enzyme-linked immunosorbent assay (ELISA) for direct detection of enteric adenovirus 41 (Ad41) in stool specimens was developed and compared with an Ad40-specific ELISA described previously [Johansson et al, 1980]. Rabbit antiserum to Ad41 was obtained by immunization with purified virions. To eliminate genus-specific reactivity the serum was passed through an immunosorbent column containing soluble adenovirus components of members of subgenera A to E. The anti-Ad41 serum still displayed high reactivity against Ad40 and had to be immunoabsorbed with soluble virus components of Ad40 to be rendered type-specific. The absorbed antiserum was used in an indirect ELISA and proved to be specific for Ad41. No heterotypic reactivity against Ad40 or Ad1 through Ad35 was found. The Ad41-specific ELISA proved to be of equal sensitivity to electron microscopy. The type-specific ELISAs for Ad40 and Ad41 were evaluated by testing 76 stool specimens containing enteric adenoviruses originating from England and Scandinavia. All specimens could be typed--41 (54%) as Ad40 and 35 (46%) as Ad41. These results were confirmed by DNA restriction site analysis. The type-specific ELISA proved to be a specific, sensitive, and a rapid technique for detection of Ad41 and allowed clear-cut discrimination from Ad40 in clinical specimens.

Adenoviruses, Human↗

Preformulation studies in a drug development program for tablet formulations.

Recently developed techniques were applied in a preformulation program to select a compound and a suitable salt for use in a one-month toxicological test. The program showed that valuable information can be obtained prior to the choice of a compound as a candidate for a solid-dosage form. Information about the physical and mechanical properties of the chosen compound can be obtained with a limited amount of substance, i.e., surface color and hygroscopicity (1-2 g), dissolution rate and solubility (1-2 g), powder properties (1-2 g), and compaction properties (5-7 g).

Chemical Phenomena↗

Influence of the granulation technique and starting material properties on the lubricating effect of granular magnesium stearate.

Magnesium stearate has been granulated in four ways to produce lubricant granulations with different properties. The lubricating properties, as well as the tablet properties with the granulated lubricant, were evaluated on tablets prepared from a mixture of dicalcium phosphate, corn starch and microcrystalline cellulose. The lubricating effect of the magnesium stearate granulations showed a similar pattern regardless of the granulation technique used except for a granulation with providone. Increasing the particle size of the magnesium stearate granulation increased the amount of lubricant required to obtain lubrication similar to powdered magnesium stearate. Variations in the specific surface area of the starting materials could be masked by using them in granular form.

Chemistry, Pharmaceutical↗

Importance of enteric adenoviruses 40 and 41 in acute gastroenteritis in infants and young children.

In a prospective 1-year study of acute infantile gastroenteritis, adenoviruses were detected in the stools or by seroconversions, or both, in 56 of 416 (13.5%) ill children. By use of DNA restriction enzyme analysis, enzyme immunoassay, and culture techniques, 33 of 56 (59%) adenovirus specimens were identified as enteric adenoviruses 40 and 41 (Ad40 and Ad41). They were found as the sole recognizable cause of diarrhea in 30 of 416 (7.2%) ill children and in 0 of 200 controls. Three additional ill children had enteric adenoviruses as a part of a dual infection. Evidence for established adenoviruses (Ad1 through Ad39) in gastroenteritis was found in 15 of 416 (3.6%) ill children but also in 3 of 200 (1.5%) controls. Eight adenovirus specimens remained untyped. Seroconversions were demonstrated in 17 of 18 (94%) paired serum samples from patients shedding enteric adenoviruses. The predominant symptom of infections with enteric adenoviruses was diarrhea, with a mean duration of 8.6 days (Ad40) and 12.2 days (Ad41). One-third of the children with Ad41 infections had prolonged symptoms (greater than or equal to 14 days). The frequency of respiratory symptoms was low (21%). The established adenoviruses presented a different clinical picture, characterized by diarrhea of shorter duration, higher fever, and significantly increased occurrence of respiratory symptoms (79%). In conclusion, enteric adenoviruses appear to be an important cause of acute infantile gastroenteritis, second only to rotaviruses in this study.

Acute Disease↗

Direct identification of enteric adenovirus, a candidate new serotype, associated with infantile gastroenteritis.

Two enzyme-linked immunosorbent assay techniques for detection of adenovirus in stools were developed. The first, which is group-specific, detects the 35 established adenovirus types and, in addition, enteric adenoviruses associated with infantile gastroenteritis. The second technique, which is type-specific, selectively detects enteric adenovirus. The efficiency of these techniques was assayed on nine coded stool specimens from Glasgow children. Eight of nine was classified as adenovirus by the group-speicific enzyme-linked immunosorbent assay. The six enteric adenovirus specimens were antigenically distinct from each of the 35 established adenovirus types but not from each other. They are suggested to represent a new adenovirus serotype which appear to be associated with gastroenteritis without clear-cut respiratory symptoms.

Adenoviruses, Human↗