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Biomedical subjects

H E Schroeder

Publications and source records attributed to H E Schroeder.

At least 19 recordsLinked to original sources

Bean alpha-amylase inhibitor 1 in transgenic peas (Pisum sativum) provides complete protection from pea weevil (Bruchus pisorum) under field conditions.

Two alpha-amylase inhibitors, called alphaAI-1 and alphaAI-2, that share 78% amino acid sequence identity and have a differential specificity toward mammalian and insect alpha-amylases are present in different accessions of the common bean (Phaseolus vulgaris). Using greenhouse-grown transgenic peas (Pisum sativum), we have shown previously that expression of alphaAI-1 in pea seeds can provide complete protection against the pea weevil (Bruchus pisorum). Here, we report that alphaAI-1 also protects peas from the weevil under field conditions. The high degree of protection is explained by our finding that alphaAI-1 inhibits pea bruchid alpha-amylase by 80% over a broad pH range (pH 4.5-6.5). alphaAI-2, on the other hand, is a much less effective inhibitor of pea bruchid alpha-amylase, inhibiting the enzyme by only 40%, and only in the pH 4.0-4.5 range. Nevertheless, this inhibitor was still partially effective in protecting field-grown transgenic peas against pea weevils. The primary effect of alphaAI-2 appeared to be a delay in the maturation of the larvae. This contrasts with the effect of alphaAI-1, which results in larval mortality at the first or second instar. These results are discussed in relationship to the use of amylase inhibitors with different specificities to bring about protection of crops from their insect pests or to decrease insect pest populations below the economic injury level.

Animals↗

Mutations of the human hepatic lipase gene in patients with combined hypertriglyceridemia/hyperalphalipoproteinemia and in patients with familial combined hyperlipidemia.

Hepatic lipase is an enzyme which hydrolyzes triglycerides from plasma lipoproteins and thus takes part in the metabolism of intermediate density lipoproteins and high-density lipoproteins. The search described here concentrated on mutations of the HL gene in 129 patients with combined hypertriglyceridemia/hyperalphalipoproteinemia and in 184 members of 19 families with familial combined hyperlipidemia. Controls were 100 subjects with favorable lipid values (age 46-51 years). Mutation screening and analysis were performed by temperature-gradient gel electrophoresis, allele-specific restriction genotyping, and sequencing. Six different missense mutations and four different silent mutations were found in the HL gene. The alleles Phe-267 and Gln-343 were detected only once in the patient group with hypertriglyceridemia and hyperalphalipoproteinemia and were not detected in the control group. The allele Met-383 was rare in both patients and controls. We found 9.3% of the patients and only 3.0% of controls to be carrying the Val-73-Met missense mutation. The allele Phe-334 was found in 5.43% of patients and in 2.0% of controls. The difference between the frequencies of these alleles was significant between male patients and male controls (Met-73 P=0.044; Phe-334 P=0.047). Also, the summarized odds ratio of 3.28 (95% confidence interval 1.23-8.73) demonstrates that mutation carriers are significantly more prevalent in the patients. Fifteen carriers of the Met-73 allele were found in six families of the familial combined hyperlipidemia group. Furthermore, six carriers of the Phe-334 allele were found in three families of the same group. In comparison to the controls the summarized odds ratio of 2.45 (95% confidence interval 0.89-6.71) barely missed the level of significance. The linkage between genotype and phenotype was incomplete. These results show an association of the missense mutations Val-73-Met and Leu-334-Phe as susceptibility alleles for combined forms of hyperlipidemia.

Adolescent↗

Expression of the insecticidal bean alpha-amylase inhibitor transgene has minimal detrimental effect on the nutritional value of peas fed to rats at 30% of the diet.

The effect of expression of bean alpha-amylase inhibitor (alpha-AI) transgene on the nutritional value of peas has been evaluated by pair-feeding rats diets containing transgenic or parent peas at 300 and 650 g/kg, respectively, and at 150 g protein/kg diet, supplemented with essential amino acids to target requirements. The results were also compared with the effects of diets containing lactalbumin with or without 0.9 or 2.0 mg bean alpha-AI, levels equivalent to those in transgenic pea diets. When 300 and 650 g peas/kg diet were fed, the daily intake of alpha-AI was 11.5 or 26.3 mg alpha-AI, respectively. At the 300 g/kg level, the nutritional value of the transgenic and parent line peas was not significantly different. The weight gain and tissue weights of rats fed either of the two pea diets were not significantly different from each other or from those of rats given the lactalbumin diet even when this was supplemented with 0.9 g alpha-AI/kg. The digestibilities of protein and dry matter of the pea diets were slightly but significantly lower than those of the lactalbumin diet, probably due to the presence of naturally occurring antinutrients in peas. The nutritional value of diets containing peas at the higher (650 g) inclusion level was less than that of the lactalbumin diet. However, the differences between transgenic and parent pea lines were small, possibly because neither the purified recombinant alpha-AI nor that in transgenic peas inhibited starch digestion in the rat small intestine in vivo to the same extent as did bean alpha-AI. This was the case even though both forms of alpha-AI equally inhibited alpha-amylase in vitro. Thus, this short-term study indicated that transgenic peas expressing bean alpha-AI gene could be used in rat diets at 300 g/kg level without major harmful effects on their growth, metabolism and health, raising the possibility that transgenic peas may also be used at this level in the diet of farm animals.

Animals↗

Impact of concentrations of glycated hemoglobin, alpha-tocopherol, copper, and manganese on oxidation of low-density lipoproteins in patients with type I diabetes, type II diabetes and control subjects.

The late organ complications in diabetic patients are associated with enhanced oxidation of low-density lipoproteins (LDL). The role of vitamin and trace metal concentrations in this process is not clear. Therefore, we compared the oxidative susceptibility and alpha-tocopherol concentration of LDL with the levels of glycated hemoglobin (HbA1c), copper and manganese. Sixty-three diabetic patients (23 female and 40 male; 53 of type II, 10 of type I) and 35 control subjects (17 female and 18 male) were investigated. The in vitro-formation of conjugated dienes in purified LDL preparations in the presence of copper was followed as absorbance at 234 nm. LDL exhibited a shorter lagtime (44.5 +/- 10.1 vs. 67.8 +/- 16.0 and 50.1 +/- 14.3 vs. 68.8 +/- 14.6 min) for type I and type II diabetic patients vs. sex and age-matched controls, P < 0.001. For all subjects together the lagtime was inversely correlated to HbA1c (r = -0.230, P = 0.023) and positively correlated to LDL alpha-tocopherol/LDL (mol/mol). This ratio was lower in diabetic patients (P < 0.01 for type II) than in control subjects. The copper and manganese plasma levels were not different between diabetic and nondiabetic groups. However, parameters of LDL oxidizability (amount and rate of oxidation) were positively correlated with both copper and manganese concentrations. We conclude that in diabetes the resistance of LDL against oxidation is diminished in relation to the quality of glucose control.

Adult↗

Cementogenesis reviewed: a comparison between human premolars and rodent molars.

BACKGROUND: Cementum continues to be the least-known mineralized tissue. Although recent advances in the field of molecular biology have contributed to an understanding of the involvement of molecular factors in cementum formation during development and regeneration, cementogenesis on a cell biological basis is still poorly understood. Virtually nothing is known about cementoblast origin, differentiation, and the cell dynamics during normal development, repair, and regeneration. This review describes the recent findings of cementogenesis on roots of human premolars and opposes them to those of teeth from other mammals, particularly the rodent molar. METHODS: Using light and electron microscopy, light microscopic radioautography, and various measurements, a comprehensive insight into the development and repair of cementum during and after root formation and tooth eruption has been achieved for human premolars. RESULTS: Cementum is a highly responsive mineralized tissue. This biological activity is necessary for root integrity and for bringing and maintaining the tooth in its proper position. With regard to cementum formation and periodontal fiber attachment, considerable species-particularities exist that are mainly based on differences in growth rates and tooth sizes. Since root development and initial cementogenesis last on the average 5-7 years in human premolars, cementum formation in these teeth is characterized by along-lasting phase of prefunctional development, with occurs independent of principal periodontal fiber attachment to the root and which may take 5 years or more. The first molar of the rat, however, is in functional occlusion 3 1/2 weeks after the onset of root formation. Since initial cementum formation and periodontal fiber attachment to the root occur almost at the same time in this tooth, the distinction between cells associated with one or the other process is very difficult to achieve, and cementogenesis cannot be described independent of periodontal fiber attachment to the root. Therefore, the determination of cementoblast origin in the rodent molar may be intricate. CONCLUSIONS: Taking into account these species differences, the current description on the origin and differentiation of cementoblasts is inconsistent and the description of cementogenesis is still incomplete. This review calls into question the currently held concept of cementogenesis and offers a possible alternative.

Animals↗

Tooth eruption: theories and facts.

The mechanisms of tooth eruption (i.e., the answer to the question of how and why teeth erupt) has been a matter of long historical debate. This review focuses on human and other mammalian teeth with a time- and spacewise limited period of eruption and analyzes recent observations and experimental data on dogs, rats, primates, and humans in a framework of basic biological parameters to formulate a guiding theory of tooth eruption. Acknowledging basic parameters (i.e., that teeth move in three-dimensional space, erupt with varying speed, and arrive at a functional position that in inheritable) eliminates a number of previously held theories and favors those that accommodate basic parameters, such as alveolar bone remodeling in association with root elongation, with possible correction factors in the form of cementum apposition and periodontal ligament formation. We have critically analyzed, summarized, and integrated recent findings associated with preeruptive movements of developing teeth, the intraosseous stage of premolar eruption in dogs, molar eruption in rodents, and premolar and molar eruption in primates. The variable speeds of eruption are particularly important. We conclude with basic principles of tooth eruption--that is, the type of signals generated by the dental follicle proper, the conditions under which teeth are moved and the clinical understanding to be derived from this knowledge.

Animals↗

Types and incidence of human periapical lesions obtained with extracted teeth.

OBJECTIVES: To determine (1) the frequency of the incidence of abscess, granuloma, and radicular cyst among human periapical lesions obtained with extracted teeth; and (2) whether periapical cysts occur in two categories when histologically analyzed in relation to the root canals. STUDY DESIGN: A total of 256 lesions were analyzed. The specimens were decalcified and embedded in plastic. Serial sections or step-serial sections were prepared, and the sections were evaluated on the basis of predefined histopathologic criteria. RESULTS: The 256 specimens consisted of 35% periapical abscess, 50% granuloma, and 15% cysts. The latter occurred in two categories, the apical true cysts and the apical pocket cysts. CONCLUSIONS: These results show (1) the low incidence of radicular cysts among periapical lesions as against the widely held view that almost half of all periapical lesions are cysts; and (2) the occurrence of two classes of radicular cysts. We are of opinion that the pocket cysts may heal after root canal therapy but the true cysts are less likely to be resolved by conventional root canal treatment.

Decalcification Technique↗

Number and size spectra of non-myelinated axons of human premolars.

The aim of this study was to determine the number and size of apical non-myelinated (C) axons of healthy human premolars. The material was derived from a large collection of specimens prepared for a previous quantitative investigation on the myelinated (A) axons of human premolars. A total of 16 teeth (six maxillary first and five each of mandibular first and second premolars), removed from adolescents for orthodontic reasons, were used. Root discs of about 0.6 mm thickness were prepared at about 2 mm cervical to the root apex and processed for light and electron microscopy. The number of non-myelinated axons was determined by taking a total census of such fibres that could be identified and reconstructed by standardized composite electron micrographs from each root disc. The measurement of axons was done on a statistically representative sample of axons (n = 1810) using an electronic image processing unit. The 16 teeth had an average of 2000 +/- 1023 non-myelinated axons at the juxta-apical level (range 534-3912). The average diameter of the non-myelinated axons was found to be 0.5 +/- 0.4 microns (range 0.05-2.4 microns).

Axons↗

How repair cementum becomes attached to the resorbed roots of human permanent teeth.

Human premolars extracted for orthodontic reasons were fixed with a mixture of formaldehyde and glutaraldehyde and processed for light and transmission electron microscopy. About 15% of all available teeth showed signs of superficial root resorption. Quantitatively, the resorption lacunae were more frequent at the apical third of the root, when compared to the mid-root and the cervical levels. Qualitatively, the resorption areas appeared as shallow irregular concavities. The degree of resorption and/or repair varied considerably both within and between sites, ranging from arrested resorption to advanced repair. An about 1- to 2-microns-thick seam of exposed collagen fibrils of the residual dentinal matrix lined the resorbed root surfaces in the arrested stage of resorption. A particular class of mononuclear cells repopulated these surfaces and commenced to attach newly produced collagenous matrix fibrils to the exposed dentinal matrix. Increasing fibril formation and bundling resulted in a collagenous fiber fringe oriented perpendicular to the bottom of Howship's lacunae. Following the implantation and establishment of this initial fiber fringe, the junctional zone between the two matrices and further portions of the repair matrix became progressively obscured by a fine granular and electron-dense material, indicating the advancing front of mineralization. In addition, the junctional zone later attained a high degree of electron density and basophilia. Subsequently formed matrix derived from cementoblasts was structurally heterogeneous and resembled cellular intrinsic fiber cementum (CIFC). It is suggested that the initially produced matrix along resorbed root surfaces closely resembles that seen during initial formation of acellular extrinsic fiber cementum (AEFC), whereas further apposition results in a tissue with the characteristics of CIFC.

Adolescent↗

[Attrition, dentin apposition and protein synthesis in the pulp and desmodont of young and old rats].

Occlusal attrition and responsive adaptations in the pulp-dentinal unit are still problematic. In this study, up to 2 years-old rats were used to measure the amount of occlusal attrition, of dentin apposition and of the incorporation of 3H-proline into the pulpal and periodontal ligament tissues. Two to four SIV-rats each, 45, 365 and 730 days of age, were once injected with 3H-proline 2 hours or 5 days prior to exitus. After cardiac perfusion, the molar blocks of both jaws were decalcified in EDTA and divided into bucco-oral and mesio-distal slices. The latter were embedded in Epon. Semithin sections served to measure the amount of attrition and of tertiary dentin apposition, as well as for autoradiographic labelling and determination of the rate of dentin apposition and proline incorporation in pulpal and periodontal ligament tissues. The results show that (1) the molar cusps lost up to 40% of their initial height during the two years of life, (2) occlusal dentin apposition in the intercuspal region led to an increase in dentinal thickness by 50 to 70%, while in the pulp horn region dentinal apposition occurred early in life and later did not keep phase with attrition, and (3) protein synthesis in pulp and periodontal ligament essentially did not change within the two years of life. These findings are related to that of other authors and discussed in connection with dentinal innervation.

Aging↗

Attempts to label matrix synthesis of human root cementum in vitro.

The present study describes the dynamic process of both acellular extrinsic (AEFC) and acellular/cellular intrinsic fiber cementum (AIFC/CIFC) matrix production on growing human teeth. Selected erupting maxillary and mandibular premolars with roots grown to about 70%-95% of their final length were placed in organ culture immediately following extraction. Twelve teeth for short-time labeling were pulse-incubated for 15 min in medium containing 3H-proline and chased for various times in order to follow the migration and secretion of the tracer. Eight teeth for long-time incubation were labeled continuously for 5 h before being chased for 1-8 days in order to label cementum matrix accumulation. After decalcification in ethylene diaminetetraacetic acid (EDTA), their roots were subdivided into about 20 slices each. Epon-embedded sections were prepared for light- and electron-microscopic as well as autoradiographic examination. During CIFC-formation, cementoblasts revealed high intracytoplasmic silver grain concentrations within the first hour after 3H-proline administration. The release of the tracer occurred between 60 to 120 min after administration. After 2 h, cementoblasts and the cementum matrix appeared to be labeled about equally. After 5 h, most of the labeled proteins appeared to be localized in the cementoid. Silver grains increased in number over the cementum matrix from 5-24 h. Very high intracellular grain concentrations within very large cementoblasts corresponded to regions of rapid cementum formation. Tracer-halos around entrapped cells lend support to a multipolar mode of matrix production during CIFC-initiation. The fate of the tracer during the development of early AEFC-matrix was less clear. However, fibroblasts revealed dense intracytoplasmic grain accumulations within the first hour after 3H-proline administration. Thereafter, the tracer localization was vague. This indistinct grain localization reflected the particular mode of AEFC-matrix production characterized by addition of new fibril segments to pre-existing fibers of a collagenous fringe.

Adolescent↗

[Age-related changes in the pulp chamber and its wall in human canine teeth].

A total of 21 extracted, vital human canines (i.e., 8 maxillary and 13 mandibular) of patients between 40 and 78 years of age has been used to study, by means of X-rays and scanning electron microscopy, the age-related restriction of the pulp chamber and of the root canal, and the structure of its inorganic dentinal walls. This was done at the mesial and distal as well as the vestibular and oral aspects of the pulp chamber walls. In addition, the density of the openings of the dentinal tubules was, under standardized conditions, determined at the coronal part of the pulp chamber, in the middle of the root canal and apically on each of the four aspects of the pulp chamber, i.e. vestibularly, mesially, orally and distally. The results demonstrated that (1) pulp chamber and root canal of both maxillary and mandibular canines, with increasing age of the patients, does not become very much restricted in the vestibulo-oral direction and only very little or patchwise in the mesio-distal direction, (2) fibrodentin that causes such restriction, is deposited in greater amounts only after 60 to 70 years of age, and (3) the density of the tubules is very variable, rather similar in the coronal crown chamber and the middle of the root canal, both mesio-distally and vestibulo-orally, and decreases sharply towards the root apex. These findings are discussed mainly with respect to the age-related changes in the pulp chamber and root canals of human central and lateral incisors.

Adult↗

Human cellular mixed stratified cementum: a tissue with alternating layers of acellular extrinsic- and cellular intrinsic fiber cementum.

Following the first description of the formal genesis of acellular extrinsic- and cellular intrinsic fiber cementum on growing human teeth, the present paper introduces cellular mixed stratified cementum as a particular tissue. Based on a large and diverse material (human premolars and molars) that was examined by various methods, such as microradiography, light microscopy using transmitted and polarized light, and scanning- and transmission-electron microscopy, the composition and structure of this type of cementum is described. Cellular mixed stratified cementum covers the apex, the apical third of the roots or more, and the furcational aspects of human teeth. With all methods employed, it was shown that this tissue consists in principle of acellular extrinsic- and cellular intrinsic fiber cementum, i.e. two major types of cementum that alternate or are in irregular sequences deposited upon one another. The dynamics of this layering and the fact, that, temporarily, root surfaces may remain unsupported by Sharpey's fibers as a consequence of this layering, are discussed.

Adolescent↗