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Biomedical subjects

K Poulsen

Publications and source records attributed to K Poulsen.

At least 19 recordsLinked to original sources

Activation of the Notch signaling pathway in response to pulp capping of rat molars.

Notch signaling is an evolutionarily conserved pathway that controls the developmental choices made by individual cells. Cells communicate via Notch receptors and their ligands, which direct decisions on the fate of stem cells according to the states of their neighbors. In this study we explored Notch signaling after the pulp capping of adult first upper rat molars. The wound was capped with calcium hydroxide. In situ hybridization revealed an increased expression of Notch signaling genes on day 1, which showed a tendency to decrease on day 3. Notch1 increased in the subodontoblast zone and close to the lesion limited to a few cells. Notch2 increased in pulp stroma surrounded by coronal odontoblasts. Notch1 and, especially, Notch3 expression increased, corresponding to perivascular cell groups. A low increase of ligand expression was observed near the injury with Delta1 expression along the dentin wall and Jagged1 in the stroma. Expression of the downstream target, Hes1, was observed along the lesion and adjacent dentin walls. Hes5 expression was not observed. The results indicate that Notch signaling is activated in response to injury and associated with the differentiation of pulp cells into perivascular cells and odontoblasts. The findings are consistent with the concept that the Notch pathway controls stem cell fate during pulp regeneration.

Animals↗

The highly leukotoxic JP2 clone of Actinobacillus actinomycetemcomitans and progression of periodontal attachment loss.

The JP2 clone of Actinobacillus actinomycetemcomitans has been implicated in the etiology of periodontitis in adolescents. The aim of this two-year longitudinal study was to describe clinical attachment loss (CAL) progression and to assess its association with baseline occurrence of the JP2 and non-JP2 types of A. actinomycetemcomitans. Clinical re-examination of 121 adolescents in Morocco was performed. Progression of CAL > or = 1 mm, > or = 2 mm, > or = 3 mm, and > or = 4 mm on at least one site was found in 58%, 48%, 22%, and 6% of the subjects, respectively. Subjects who, at baseline, harbored the JP2 clone had a significantly higher progression of CAL than did subjects harboring non-JP2 types of A. actinomycetemcomitans. Subjects harboring non-JP2 types displayed a marginally higher CAL progression than did subjects who were culture-negative for A. actinomycetemcomitans.

Adolescent↗

Angiotensin converting enzyme in bovine ovarian follicular fluid and its relationship with oestradiol and progesterone.

CONTENT: The purpose of the present study was to identify angiotensin converting enzyme (ACE) in bovine ovarian follicular fluid and to relate the ACE activity to the phase of the oestrous cycle, pregnancy, and the follicular fluid concentrations of oestradiol and progesterone. The ACE activity was similar to that found in bovine serum and was completely inhibited by the specific ACE inhibitor captopril. The 50% inhibitory concentration (IC50) was 1.4 x 10(-8) mol/l (range 0.8 x 10(-8) to 5.0 x 10(-8) mol/l; n=6), which is similar to that found in bovine and human serum. The ACE activity did not differ in the pre-ovulatory and luteal phase, pregnancy or cystic follicles. It correlated with the follicular fluid concentration of progesterone in cycling cows (rho=0.476; p < 0.005; n=36), but did not correlate with the diameter of the follicles, the follicular fluid concentration of oestradiol or the ratio between the oestradiol and progesterone concentrations. The demonstration of ACE in bovine ovarian follicular fluid provides further evidence for the presence of a local renin-angiotensin system in the bovine ovary.

Angiotensin-Converting Enzyme Inhibitors↗

Angiotensin-converting enzyme activity in the bovine uteroplacental unit changes in relation to the cycle and pregnancy.

The expression of the angiotensin-forming enzymes, renin and angiotensin-converting enzyme (ACE), were examined in the bovine uteroplacental unit. The ACE activity was determined in cell membrane fractions, and ACE and renin were localized by autoradiography and immunohistochemistry, respectively. In the myometrium, the ACE activity was significantly higher in dioestrous than in oestrous. ACE activity correlated negatively with the day of gestation in the endometrium and myometrium but positively in the placentome and allantoamniotic membrane. Autoradiography showed, that ACE was localized in vascular endothelial cells in all compartments. ACE was also expressed in the endometrial stroma and uterine glands, most pronounced in the outer part of the basal zone. In the intercotyledonary membrane and the placentome, the mesenchymal cells located near the trophoblast cells expressed ACE. Solitary macrophage- or monocyte-like cells showing intense renin immunoreactivity were found in the uterus, while the uterine and the glandular epithelial cells displayed inconsistent reactivity. No renin was observed in the placentomes or in the fetal membranes. The findings demonstrate a regulated expression of angiotensin-forming enzymes throughout the bovine uteroplacental unit. Whether this local renin-angiotensin system contributes to the highly regulated morphological and functional changes throughout the oestrous cycle and gestation remains to be established.

Angiotensin-Converting Enzyme Inhibitors↗

Localization of the renin-angiotensin system in the bovine ovary: cyclic variation of the angiotensin II receptor expression.

Angiotensin (Ang) II may modulate reproductive function in the bovine ovary. Therefore, expression and localization of a local ovarian renin-angiotensin system (RAS) were investigated by elucidating the influence of the estrus cycle, pregnancy, and the presence of follicular cysts. Receptor analysis and autoradiography were used to characterize and localize Ang II receptors. Cyclic variations in the density of ovarian Ang II receptors were found with a higher value in estrus than in diestrus. The density in ovaries with follicular cysts was in the same order of magnitude as in estrus. The Ang II receptor type 2 (AT(2)) dominated in all three groups. Autoradiography showed that the majority of antral follicles and follicular cysts had intense AT(2) receptor binding in the theca externa. Binding was less intense in the theca interna, whereas there was no binding in the granulosa layer. In the corpora lutea, the AT(2) receptor was dominant in the capsule and in connective tissue infoldings, whereas no binding was observed in the luteal tissue. The type 1 Ang II receptor (AT(1)) was dominant in the stroma and showed no cyclic changes. Angiotensin-converting enzyme (ACE) activity was detected in all aspirated follicular fluids and homogenates of ovarian tissue. Autoradiography showed that most of the ACE was localized on endothelial cells. Renin immunoreactivity was found in granulosa and thecal cells of antral follicles and in luteal cells. Furthermore, solitary cells in the stroma, presumably macrophages, displayed intense staining. Our finding of cyclic changes support the concept of an active and regulated RAS in the bovine ovary.

Animals↗

Asphyxial homicide in two Scandinavian capitals.

In the Oslo and Copenhagen capital areas, 94 asphyxial homicides were committed in the 10-year period 1985-1994, accounting for 22% of all homicides in that period. Sixty-nine (73%) of the asphyxia victims were female. The most common method of asphyxiation was manual strangulation. Seventeen (18%) of the victims were below the age of 10, accounting for 59% of all homicides in that age group. Whereas 38% of the female victims were killed by their spouse, this was the case for only one male victim. The motive was not known in a great proportion of cases. Fifty-seven percent of the victims had been subjected to additional violence, and in this respect there was no difference between the sexes. In 12 of the cases the offender was female; in 9 such cases the victim was her offspring. More than half of the victims had no blood alcohol. When disregarding the victims less than 10 years of age, 33% of the male and 49% of the female victims had no blood alcohol. The crime scene was the victim's domicile among 72% of female and 52% of male victims. Forty-two percent of the female and 11% of the male victims above the age of 10 years were married or cohabitant.

Adolescent↗

Epidemic population structure of Pseudomonas aeruginosa: evidence for a clone that is pathogenic to the eye and that has a distinct combination of virulence factors.

The genetic structure of a population of Pseudomonas aeruginosa, isolated from patients with keratitis, endophthalmitis, and contact lens-associated red eye, contact lens storage cases, urine, ear, blood, lungs, wounds, feces, and the environment was determined by multilocus enzyme electrophoresis. The presence and characteristics of virulence factors were determined by restriction fragment length polymorphism analysis with DNA probes for lasA, lasB, aprA, exoS, exoT, exoU, and ctx and by zymography of staphylolysin, elastase, and alkaline protease. These analyses revealed an epidemic population structure of P. aeruginosa, characterized by frequent recombination in which a particular successful clone may increase, predominate for a time, and then disappear as a result of recombination. Epidemic clones were found among isolates from patients with keratitis. They were characterized by high activity of a hitherto-unrecognized size variant of elastase, high alkaline protease activity, and possession of the exoU gene encoding the cytotoxic exoenzyme U. These virulence determinants are not exclusive traits in strains causing keratitis, as strains with other properties may cause keratitis in the presence of predisposing conditions. There were no uniform patterns of characteristics of isolates from other types of infection; however, all strains from urinary tract infections possessed the exoS gene, all strains from environment and feces and the major part of keratitis and wound isolates exhibited high elastase and alkaline protease activity, and all strains from feces showed high staphylolysin activity, indicating that these virulence factors may be important in the pathogenesis of these infectious diseases.

ADP Ribose Transferases↗

Evidence of recombination in Porphyromonas gingivalis and random distribution of putative virulence markers.

The association of Porphyromonas gingivalis to periodontal disease is not clearly understood. Similar proportions of P. gingivalis may be cultivated from both inactive and actively degrading periodontal pockets. Differences in virulence among strains of P. gingivalis exist, but the molecular reason for this remains unknown. We examined the population structure of P. gingivalis to obtain a framework in which to study pathogenicity in relation to evolution. Phylogenetic trees derived from the sequencing of fragments of four housekeeping genes, ahp, thy, rmlB, and infB, in 57 strains were completely different with no correlation between clustering of strains in the four dendrograms. Combining the various alleles of the four gene fragments sequenced resulted in 41 different sequence types. The index of association, I(A), based on a single representative of each sequence type was 0.143 +/- 0.202, indicating a population at linkage equilibrium. Inclusion of all isolates for the calculation of I(A) resulted in a value of 0.206 +/- 0.171. This suggests an epidemic population structure supported by the finding of genetically identical strains in different parts of the world. We observed a random distribution of two virulence-associated mobile genetic elements, the ragB locus and the insertion sequence IS1598, among 132 strains tested. In conclusion, P. gingivalis has a nonclonal population structure characterized by frequent recombination. Our study suggests that particular genotypes, possibly with increased pathogenic potential, may spread successfully in the human population.

Bacterial Proteins↗

Early-onset periodontitis in Morocco is associated with the highly leukotoxic clone of Actinobacillus actinomycetemcomitans.

A particular clone (JP2) of Actinobacillus actinomycetemcomitans with increased leukotoxin production has been isolated from individuals with early-onset periodontitis (EOP). The aim of this study was to determine the frequency of carriers of this clone and its association with EOP in Moroccan schoolchildren. Of 217 plaque samples, 131 (60.4%) were culture-positive for A. actinomycetemcomitans. A total of 19 of these isolates had a 530-bp deletion in the leukotoxin promoter region characteristic of the JP2 clone. A strong association between the presence of A. actinomycetemcomitans with the 530-bp deletion and EOP was found (adjusted OR = 29.4; 95% Cl = 8.3 - 104.4; p < 0.0005), while no association could be demonstrated between the presence of A. actinomycetemcomitans without the deletion and EOP (adjusted OR = 1.3; 95% CI = 0.5 -2.9; p = 0.750). The study demonstrates that the endemic presence, in a human population, of the highly leukotoxic JP2 clone may result in an unusually high prevalence of EOP.

Adolescent↗

Distinct pathways of mannan-binding lectin (MBL)- and C1-complex autoactivation revealed by reconstitution of MBL with recombinant MBL-associated serine protease-2.

Mannan-binding lectin (MBL) plays a pivotal role in innate immunity by activating complement after binding carbohydrate moieties on pathogenic bacteria and viruses. Structural similarities shared by MBL and C1 complexes and by the MBL- and C1q-associated serine proteases, MBL-associated serine protease (MASP)-1 and MASP-2, and C1r and C1s, respectively, have led to the expectation that the pathways of complement activation by MBL and C1 complexes are likely to be very similar. We have expressed rMASP-2 and show that, whereas C1 complex autoactivation proceeds via a two-step mechanism requiring proteolytic activation of both C1r and C1s, reconstitution with MASP-2 alone is sufficient for complement activation by MBL. The results suggest that the catalytic activities of MASP-2 split between the two proteases of the C1 complex during the course of vertebrate complement evolution.

Carrier Proteins↗

Homicide by sharp force in two Scandinavian capitals.

In the Oslo and Copenhagen capital areas, 141 homicides by sharp force were committed in the ten-year period 1985-1994. This method accounted for 33% of the homicides in this period. Thirty-five percent of the victims were female, and most of the victims were between 20 and 50 years of age. The majority of the male victims were killed by an acquaintance, the females by their spouse. Sixty-five percent of the male and 37% of the female victims had alcohol in their blood. The majority of the female victims had lesions in 3-4 anatomical regions, while the males most often had lesions in only one anatomical region. Seventy-nine percent of the females and 36% of the males had self-defence injuries in the upper extremities. In 21 cases (15%) the offender was a woman, 19 of their victims being male; the weapon in these cases was most often a kitchen knife. Seventy-eight percent of the females and 49% of the males were killed in their own home. The most common circumstance was family row when the victim was female, while a fight was the most common circumstance when the victim was male. Three offenders committed suicide after having committed homicide(s) (seven victims; three offenders).

Adolescent↗

Abiotrophia species in early dental plaque.

The most characteristic traits that distinguish Abiotrophia strains from other streptococci are nutrient requirements, satellitism and pyrrolidonyl arylamidase activity. The presence of Abiotrophia strains was studied in early dental plaque formed on sucrose-containing and sucrose-free (glucose-containing) diets. The isolates were from 0- to 3-day dental plaque formed on the buccal surface of a lower premolar in six subjects. Identification of Abiotrophia strains was based on their pyrrolidonyl arylamidase activity. They were among the predominant cultivable microorganisms isolated from diluted suspensions of early dental plaque formed on both diets. Using biochemical tests without testing for pyrrolidonyl arylamidase activity, the Abiotrophia strains would be biochemically identified as Streptococcus mitis or unidentified streptococci, and many isolates first classified as unidentified streptococci were Abiotrophia strains. Generally, identifications using partial 16S rRNA sequences confirmed the identifications obtained biochemically. However, Abiotrophia elegans and Gemella strains were biochemically identified as Abiotrophia adiacens. Abiotrophia strains produce hydrogen sulfide, which may influence the metabolism and ecology of dental plaque and act as a virulence factor in periodontal disease. They are not able to grow on tryptic soy agar. In the present study, some S. mitis strains were not able to grow on mitis salivarius agar, and some Abiotrophia strains were able to grow on this medium. These observations indicate that the use of these media for estimation of total viable counts and number of streptococci introduces systematic error in studies of microorganisms in dental plaque.

Aminopeptidases↗

Resident aerobic microbiota of the adult human nasal cavity.

Recent evidence strongly suggests that the microbiota of the nasal cavity plays a crucial role in determining the reaction patterns of the mucosal and systemic immune system. However, little is known about the normal microbiota of the nasal cavity. The purpose of this study was to determine the microbiota in different parts of the nasal cavity and to develop and evaluate methods for this purpose. Samples were collected from 10 healthy adults by nasal washes and by swabbing of the mucosa through a sterile introduction device. Both methods gave results that were quantitatively and qualitatively reproducible, and revealed significant differences in the density of the nasal microbiota between individuals. The study revealed absence of gram-negative bacteria that are regular members of the commensal microbiota of the pharynx. Likewise, viridans type streptococci were sparsely represented. The nasal microbiota was dominated by species of the genera Corynebacterium, Aureobacterium, Rhodococcus, and Staphylococcus, including S. epidermis, S. capitis, S. hominis, S. haemolyticus, S. lugdunensis and S. warneri. These studies show that the microbiota of the nasal cavity of adults is strikingly different from that of the pharynx, and that the nasal cavity is a primary habitat for several species of diphtheroids recognized as opportunistic pathogens. Under special circumstances, single species, including IgA1 protease-producing bacteria, may become predominant in a restricted area of the nasal mucosa.

Actinomycetales↗

The Prevotella intermedia group organisms in young children and their mothers as related to maternal periodontal status.

Currently, the Prevotella intermedia group includes three biochemically and phylogenetically related species: Prevotella intermedia, Prevotella nigrescens, and the newly described Prevotella pallens. The two first-named species are mentioned with varying emphasis in connection with periodontal diseases, while such a connection of P. pallens is not known. Mothers serve as a plausible source of bacteria to their children, and conceivably, a mother with periodontitis as a recurrent reservoir of periodontally infecting organisms. In the present study, 23 mothers and their young children were examined for the presence of the P. intermedia group organisms in relation to maternal periodontal status (I: periodontal health, II: initial periodontitis, and III: advanced periodontitis). Species differentiation was based on established biochemical methods, electrophoretic mobility patterns, SDS-PAGE, and DNA hybridization. P. intermedia was not recovered from children but nearly exclusively from mothers in group III, thus confirming its association with periodontitis. P. nigrescens and P. pallens were frequently found in mothers and children. To determine bacterial transmission between a mother and her child, 72 isolates from 13 mother-child pairs were analyzed by arbitrarily primed PCR (AP-PCR). Similar AP-PCR types of P. nigrescens and/or P. pallens were recovered from 3/4 pairs in group I, 2/5 pairs in group II, and none in group III. Our results indicate that different species within the P. intermedia group have a different colonization pattern in childhood and that the periodontal status reflects qualitatively their presence in maternal saliva. Intra-familial transmission of P. nigrescens and P. pallens can occur in early childhood, however similar AP-PCR types were most obvious within periodontally healthy mother-child pairs.

Adult↗

Homicide by firearms in two Scandinavian capitals.

In the Oslo and Copenhagen areas, 83 homicides by firearms were registered in the 10-year period 1985 to 1994, accounting for 19.3% of all homicides in that period. The majority of the victims were between 20 and 50 years old, and 58% were male, 42% female. The yearly number of firearm homicides varied between 4 and 15, with neither an increase nor decrease throughout the period. Most of the victims had no detectable blood alcohol at autopsy. Female victims were typically shot in their own domicile with a shotgun by their spouse, mostly because of jealousy or in a family argument, whereas male victims were shot on different locations, predominantly with a handgun, for many different reasons. Most victims were shot in the head, and few had entrance wounds in more than one anatomic region.

Adolescent↗

Two allelic forms of the aureolysin gene (aur) within Staphylococcus aureus.

Proteinases of Staphylococcus aureus are emerging as potential virulence factors which may be involved in the pathogenecity of staphylococcal diseases. We describe here the structure of the gene encoding the metalloproteinase referred to as aureolysin. This gene occurs in two allelic forms and is strongly conserved among S. aureus strains, implying the possibility that the proteinase may have important housekeeping functions.

Alleles↗

Conserved residues and their role in the structure, function, and stability of acyl-coenzyme A binding protein.

In the family of acyl-coenzyme A binding proteins, a subset of 26 sequence sites are identical in all eukaryotes and conserved throughout evolution of the eukaryotic kingdoms. In the context of the bovine protein, the importance of these 26 sequence positions for structure, function, stability, and folding has been analyzed using single-site mutations. A total of 28 mutant proteins were analyzed which covered 17 conserved sequence positions and three nonconserved positions. As a first step, the influence of the mutations on the protein folding reaction has been probed, revealing a folding nucleus of eight hydrophobic residues formed between the N- and C-terminal helices [Kragelund, B. B., et al. (1999) Nat. Struct. Biol. (In press)]. To fully analyze the role of the conserved residues, the function and the stability have been measured for the same set of mutant proteins. Effects on function were measured by the extent of binding of the ligand dodecanoyl-CoA using isothermal titration calorimetry, and effects on protein stability were measured with chemical denaturation followed by intrinsic tryptophan and tyrosine fluorescence. The sequence sites that have been conserved for direct functional purposes have been identified. These are Phe5, Tyr28, Tyr31, Lys32, Lys54, and Tyr73. Binding site residues are mainly polar or charged residues, and together, four of these contribute approximately 8 kcal mol-1 of the total free energy of binding of 11 kcal mol-1. The sequence sites conserved for stability of the structure have likewise been identified and are Phe5, Ala9, Val12, Leu15, Leu25, Tyr28, Lys32, Gln33, Tyr73, Val77, and Leu80. Essentially, all of the conserved residues that maintain the stability are hydrophobic residues at the interface of the helices. Only one conserved polar residue, Gln33, is involved in stability. The results indicate that conservation of residues in homologous proteins may result from a summed optimization of an effective folding reaction, a stable native protein, and a fully active binding site. This is important in protein design strategies, where optimization of one of these parameters, typically function or stability, may influence any of the others markedly.

Acyl Coenzyme A↗