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Biomedical subjects

E Reich

Publications and source records attributed to E Reich.

At least 55 records · Page 3Linked to original sources

A pilot study of confocal laser scanning microscopy for the assessment of undisturbed dental plaque vitality and topography.

Confocal microscopy and vital fluorescence techniques were combined for the first time to investigate ex vivo human dental plaque. The vital fluorescence technique used discriminates vital from dead cells, while confocal laser scanning microscopy allows the optical sectioning of undisturbed biofilms leaving the samples intact during analysis. The concomitant use of both methods made an examination of the three-dimensional architecture of dental plaque possible. The topography of plaque biofilms that were allowed to accumulate in situ on glass and enamel was recorded. The distribution of plaque microflora vitality as well as its accumulation varied according to plaque age. A plaque thickness of up to 8, 35 and 45 microm was estimated ex vivo on enamel after 1, 2 and 3 days, respectively. Young and sparse plaque biofilms consisted mainly of dead material. Vital bacteria were observed on top of this dead layers.

Dental Plaque↗

Distribution of reducible 4-hydroxynonenal adduct immunoreactivity in Alzheimer disease is associated with APOE genotype.

Two major risk factors for late-onset familial and sporadic Alzheimer disease (AD), a leading cause of dementia worldwide, are increasing age and inheritance of the epsilon4 allele of the apolipoprotein E gene (APOE4). Several isoform-specific effects of apoE have been proposed; however, the mechanisms by which apoE isoforms influence the pathogenesis of AD are unknown. Also associated with AD is increased lipid peroxidation in the regions of the brain most damaged by disease. 4-hydroxynonenal (HNE), the most potent neurotoxic product of lipid peroxidation, is thought to be deleterious to cells through reactions with protein nucleophiles. We tested the hypothesis that accumulation of the most common forms of HNE-protein adducts, borohydride-reducible adducts, is associated with AD and examined whether there was a relationship to APOE. Our results demonstrated that reducible HNE adducts were increased in the hippocampus, entorhinal cortex, and temporal cortex of patients with AD. Furthermore, our data showed that the pattern of reducible HNE adduct accumulation was related to APOE genotype; AD patients homozygous for APOE4 had pyramidal neuron cytoplasmic accumulation of reducible HNE adducts, while AD APOE3 homozygotes had both pyramidal neuron and astrocyte accumulation of reducible HNE adducts. This is in contrast to our previous observations that a distinct HNE protein adduct, the pyrrole adduct, accumulates on neurofibrillary tangles in AD patients. We conclude that APOE genotype influences the cellular distribution of increased reducible HNE adduct accumulation in AD.

Aged↗

Acro-renal-ocular syndrome: expansion of the phenotype.

We report the sixth described family with acro-renal-ocular syndrome in a boy and more mildy in his mother. Severe upper limb deficiency, dysplastic kidneys, and strabismus are noted in this child in addition to developmental delay, dysplastic corpus callosum, and incomplete myelination. Developmental central nervous system (CNS) malformations have not been described in this syndrome previously and may represent an expansion of the phenotype.

Abnormalities, Multiple↗

Regeneration of oxytalan fibres in different types of periodontal defects: a histological study in monkeys.

The aim of the present study was to investigate in monkeys the regrowth of oxytalan fibres in different types of acute and chronic periodontal defects following regenerative periodontal treatment. One-wall intrabony and mandibular furcation III-defects were produced surgically in 3 monkeys (Macaca fascicularis). After a 6-wk dental plaque accumulation period the defects were exposed using a full thickness flap procedure. The granulation tissue was removed and the root surfaces were scaled and planed. Additionally, fenestration-type defects were produced at the vestibular surfaces of the maxillary and mandibular canines by surgically removing the vestibular bone plates and the root cementum. Subsequently, the defects were treated with guided tissue regeneration (GTR), enamel matrix proteins (EMP), combination of EMP and GTR or with coronally repositioned flaps. The postoperative care included tooth cleaning once a week during the experiment. After 5 months the animals were sacrificed and and the block sections were embedded in paraffin. Eight microns histological sections were cut and stained with the oxone-aldehyde-fuchsin-Halmi method. The results revealed that in all specimens where a regenerated periodontal ligament could be observed newly formed oxytalan fibers were present. They had a mainly apico-occlusal orientation and were localized closer to the cementum than to the alveolar bone. The regenerated oxytalan fibers had a similar morphological appearance than those observed in the original periodontal ligament regardless of the treatment modality by which regeneration was accomplished. Their presence was related to that of newly formed cementum suggesting a strong relationship between these 2 tissues. The neoformation of oxytalan fibres can thus be observed in some types of periodontal defects where the cementum and the periodontal ligament have been regenerated.

Animals↗

High-dose multimodality therapy with autologous stem-cell support for stage IIIB breast carcinoma.

PURPOSE: Women with locally unresectable and inflammatory breast carcinoma (IBC) have an approximately 30% 5-year disease-free survival (DFS) rate with conventional multimodality therapy. A short but dose-intensive multimodality phase II trial was designed in an attempt to improve outcome in stage IIIB disease. Mastectomy was performed after high-dose therapy to evaluate pathologic response to treatment. METHODS: Women with newly diagnosed disease received four 2-week cycles of doxorubicin 90 mg/m2 with granulocyte colony-stimulating factor (G-CSF), followed by cyclophosphamide 6,000 mg/m2, thiotepa 500 mg/m2, and carboplatin 800 mg/m2 (CTCb) with marrow and peripheral-blood progenitor cell (PBPC) support. Local therapy consisted of mastectomy and radiotherapy. Tamoxifen (5 years) was begun if the patient was estrogen receptor-positive (ER+). RESULTS: Fifty women (46 stage IIIB [91% IBC], four stage IIIA) entered the study and 47 are assessable. Ten had mastectomy before any systemic therapy (seven with pathologic IBC, three with residual tumor after mastectomy). Eighty percent received full-dose doxorubicin with 60% on schedule. Clinical response rates to induction were 15% complete response (CR), 5% very good partial response (VGPR), 59% partial response (PR), and 21% minor response (MR)/stable disease (SD). Mastectomy after CTCb in 37 patients showed a 14% pathologic CR rate, 29% microscopic foci in breast and/or axilla, and 57% gross tumor. Fifteen (32%) patients have relapsed (median, 17 months post-CTCb). The 30-month DFS is estimated at 64%. For those in pathologic CR, with microscopic, or with gross disease remaining after CTCb, the 30-month DFS is estimated at 100%, 70%, and 38%, respectively. Those with zero, one to three, or > or = four positive nodes at axillary dissection had a median DFS of 31, 18, and 13 months, respectively. CONCLUSION: This short but dose-intensive multimodality approach for stage IIIB breast carcinoma is feasible with encouraging results to date.

Adult↗

Structure and function of microplasminogen: reconstitution of microplasminogen and microplasmin from isolated fragments.

We describe limited chemical proteolysis of microplasminogen/microplasmin (mPlg/mPlm) and their reconstitution from isolated fragments. A V141-->M141 substitution in methionineless human mPlg/mPlm allowed the protein(s) to be cleaved in CNBr/formic acid. The resulting two fragments (141 and 118 residues, respectively), each internally disulfide bonded, were separated by preparative non-reducing gradient SDS-PAGE, and could then be mixed to reconstitute the characteristic mPlg/mPlm, including their activation by urokinase (uPA) and streptokinase (SK), and inhibition by macromolecular inhibitors. The isolated larger, N-terminal fragment, which contains the mPlg activation site in a normal disulfide configuration, was not cleaved by uPA in the absence of its smaller C-terminal companion, showing that the linear amino acid sequence is not by itself sufficient to confer substrate character, even when its conformation is constrained by the disulfide structure.

Amino Acid Sequence↗

Photodynamic effects of sulfonated aluminum chlorophthalocyanine in human urinary bladder carcinoma cells in vitro.

OBJECTIVES: Topically applied photosensitizers are particularly suited for use in the photodynamic therapy of urinary bladder tumors. The objective of the present study was to investigate the photodynamic effects of sulfonated aluminum chlorophthalocyanine on human bladder carcinoma cells. METHODS: The photodynamic efficiency and the morphologic changes induced by sulfonated aluminum chlorophthalocyanine on human bladder carcinoma cells and normal bladder wall cells were investigated in vitro using light microscopy and in order to determine the subcellular target organelles using electron microscopy. RESULTS: Examination of tumor cells with light microscopy after previous photodynamic therapy initially showed distension of the cells and, depending on the duration, a significant blistering of the cell membrane, leading, in some cases, to rupture of the cells. Electron microscopy revealed destruction of the mitochondria. No changes in other cell organelles or structures were observed. CONCLUSIONS: Sulfonated aluminum chlorophthalocyanine exhibits good photodynamic activity in bladder tumor cells with relative tumor selectivity. The primary cellular target of photodynamic therapy seems to be the mitochondria.

Aluminum↗

Ion beam slope cutting in dentistry: a catalog of applications.

It is very difficult to prepare structures of heterogeneous materials, material interfaces, or sensitive biologic samples. The usual grinding, cutting, or fracturing methods are mostly material destructive because of resulting shear strengths. Ion beam slope cutting allows microcuts to be accurately placed into subsurface areas. The material reduction is based on the principles of ion sputtering, whereby atoms are removed in the area of ion incidence. The speed of removal depends on the sputtered material, amounting to several micrometers per hour. Approximately 40 biologic dental specimens of various origin were examined. Depending on the incidence angle of the ion beam, the typical structures of ion etching can be reduced or intensified. Both variants are used to obtain a precise judgment of morphologic details or an exact topography of failures.

Dental Materials↗

[Fluorides in group prophylaxis].

In most of the industrialised countries there has been a decline in the incidence of dental caries during the past few years largely due to the use of fluorides. For many years now, fluorides have taken root both in private homes and on a professional basis. Side effects caused by them are mostly due to chronic overdosage producing whitish stains in the dental enamel, but in Germany such stains are rare and by no means severe. Application of fluorides within the overall framework of group prophylaxis can be practised in Germany only with the written consent of parents or guardians. Various systems are being used in this country, either brushing with highly concentrated fluoride jelly or applying fluoride lacquer. Considering the present prevalence of dental caries in Germany, it can be expected that group prophylaxis with fluorides would further reduce the incidence. Realisation of correspondingly more intensive prevention programmes must be planned on the spot. Dentists should supervise and instruct trained dental staff in the application of fluorides in group prophylaxis.

Adolescent↗

Aminolevulinic acid for photodynamic therapy of bladder carcinoma cells.

A new concept in photosensitizing tumor cells is photosensitizer synthesis in situ. Aminolevulinic acid (ALA) is a precursor of protoporphyrin IX (PP IX), a potent photosensitizer. The goal of our study was to examine dark toxicity, phototoxic potential, metabolism of ALA and morphological alterations in Waf bladder cancer cells. Dark toxicity of Waf cells was observed after incubation with ALA, beginning at a concentration of 15 mM. Photodynamic treatment with ALA at concentrations of 1, 5 and 10 mM showed a drug- and light-dose-dependent cell survival rate in comparison to a control group. Two incubation times of 3.5 and 5.5 h were compared for cell survival. After a longer incubation time of 5.5 h, cell survival was decreased in all experiments; this is consistent with our extraction data where higher fluorescence was found after 5.5 than after 3.5 h. The results show that ALA-induced photosensitization has a high potential for photodynamic therapy (PDT) of superficial bladder carcinoma.

Aminolevulinic Acid↗

Double dose-intensive chemotherapy with autologous stem-cell support for metastatic breast cancer: no improvement in progression-free survival by the sequence of high-dose melphalan followed by cyclophosphamide, thiotepa, and carboplatin.

PURPOSE: Twenty-one percent of responding metastatic breast cancer patients remain progression-free a median 50 months following one intensification cycle of cyclophosphamide (6,000 mg/m2), thiotepa (500 mg/ m2), and carboplatin (800 mg/m2) (CTCb) with autologous bone marrow transplantation (ABMT). This trial studied whether the sequence of high-dose melphalan followed by CTCb resulted in improved disease response and duration. METHODS: Women with at least partial responses (PRS) to induction received melphalan (140 or 180 mg/ m2) with peripheral-blood progenitor cell (PBPC) and granulocyte colony-stimulating factor (G-CSF) support. They were monitored as outpatients. After recovery, patients were hospitalized for CTCb with marrow, PBPC, and G-CSF support. RESULTS: Data on 67 women, at a median of 25 months from CTCb, were examined. After melphalan, 49 (73%) required admission for fever (89%), mucositis (35%), or infection (15%) (median stay, 8 days). All received CTCb. For the first 33 patients, the median days from start of melphalan to CTCb was 24. After liver toxicity (one death from venoocclusive disease [VOD]) developed in 11 patients during CTCb, the interval between intensifications was increased to 35 days without incident. Twenty-three patients (34%) are progression-free a median of 16 months post-CTCb. The median progression-free survival (PFS) and survival times for the whole group are estimated at 11 and 20 months, respectively. CONCLUSION: Treatment with this sequence of high-dose melphalan followed by CTCb has not resulted in superior PFS to date, when compared with single-intensification CTCb. This report discusses factors related to patient selection, the role of induced drug resistance, and the schedule of administration of alkylating agenting that may adversely influence outcome.

Adolescent↗

Prenatal diagnostic testing for familial dysautonomia using linked genetic markers.

Familial dysautonomia (FD), a recessively inherited disease, has been mapped to chromosome 9q31. Highly polymorphic dinucleotide repeat markers flanking the genetic locus and at the same genetic location have been identified. We describe the prenatal diagnosis of FD using linkage and linkage disequilibrium analyses with these markers. Twelve families were analysed for informativeness and of these, seven went on to have prenatal testing (a total of eight fetuses tested). All of these fetuses were predicted to be heterozygous unaffected (FD carriers). Seven fetuses have come to term and are normal. In the absence of a recombinant proband, a panel of three proximal and three distal markers is sufficient to provide informative flanking markers and an 87-96 per cent likelihood of a highly predictive test. In an additional family at 1:4 risk for FD, no DNA was available from the propositus. This family was analysed using linkage disequilibrium to the #18 allele of the tightly linked marker D9S58 in conjunction with linkage analysis using data from two unaffected children. Prenatal diagnosis in this family indicated an affected fetus.

Base Sequence↗

Structure and function of microplasminogen: I. Methionine shuffling, chemical proteolysis, and proenzyme activation.

We have cloned and expressed microplasminogen (mPlg), consisting of the N-terminal undecapeptide of human glu-Plg spliced to its proenzyme domain. This truncated (approximately 28 kDa) proenzyme retained the distinctive catalytic activities of the larger parent. Replacement of M residues followed by M shuffling permitted subsequent scission by site-directed chemical proteolysis (in CNBr/formic acid) without impairing any of the protein's characteristic properties. Activation of chymotrypsinogen-related zymogens occurs by limited proteolysis; the newly liberated, highly conserved N-terminus (VVGG) forms a salt bridge with an aspartyl residue immediately upstream of the active site serine. The role of both of these elements in mPlg activation was probed using protein engineering and site-directed proteolysis to alter the length and amino acid composition of the N-terminus, and to replace the aspartate. All modifications affected both Km and Kcat. The results identify some structural parameters of the N-terminus required for proenzyme activation.

Amino Acid Sequence↗

Structure and function of microplasminogen: II. Determinants of activation by urokinase and by the bacterial activator streptokinase.

We have used a group of human microplasminogens (mPlg), modified by residue substitutions, insertions, deletions, and chain breaks (1) to study the determinants of productive interactions with two plasminogen activators, urokinase (uPA), and streptokinase (SK); (2) to explore the basis of species specificity in the zymogen-SK complex activity; and (3) to compare active SK complex formation in mPlg and microplasmin (mPlm). Modifications within the disulfide-bonded loop containing the activation site and the adjacent hexadecapeptide upstream sequence showed that uPA recognition elements encompassed R29 at the activation site and multiple elements extending upstream to perhaps 13 residues, all maintained in specific conformational register by surrounding pairs of disulfide bonds. A generally parallel pattern of structural requirements was observed for active zymogen-SK complex formation. Changes within the loop downstream of the activation site were tolerated well by uPA and poorly by SK. The introduction of selected short bovine (Plg) sequences in human mPlg reduced the activity of the resulting SK complexes. The requirements for active SK complex formation are different for mPlg and mPlm.

Amino Acid Sequence↗

Photodynamic efficiency of liposome-administered tetramethyl hematoporphyrin in two human bladder cancer cell lines.

The main problems presented by superficial bladder carcinoma, its high recurrence rate and multifocal appearance, require treatment of the bladder as a whole. Photodynamic therapy (PDT) is one such experimental treatment for superficial bladder carcinoma, involving the administration of a photosensitizer that accumulates in the tumor tissue, and subsequent irradiation of the tumor with light. Since the photosensitizers used in PDT suffer from several drawbacks, new photosensitizers are being sought. Drug delivery systems are also being investigated for the administration of hydrophobic photosensitizers and enhancement of photodynamic efficiency and tumor selectivity. In this study we examined a new photosensitizer, tetramethyl hematoporphyrin (TMHP), in two human bladder cancer cell lines. In the first pair of the experiments, TMHP was bound to unilamellar liposomes. Cellular uptake, dark toxicity and photodynamic efficiency were then studied. Fluorescence microscopy showed TMHP localization in the cytoplasm in a perinuclear region, sparing the nucleus. Dark toxicity occurred after incubation of cells with TMHP above a concentration of 20 micrograms/ml. Irradiation was carried out using an argon-pumped dye laser emitting a wavelength of 630 nm at a fluence of 3.6 and 7.2 J/cm2. Before irradiation, cells were incubated with TMHP at concentrations of 2.5 and 5 micrograms/ml for 1 h. Cell survival rates after incubation with 5 micrograms/ml TMHP and irradiation at 7.2 J/cm2 were 15.7% of control cells for Rec and 4.5% for Waf cells. Uptake studies showed a higher intracellular TMHP concentration in Waf than in Rec cells. This correlates with the higher PDT efficiency seen in Waf cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Transitional Cell↗

Effect of dimension of luting space and luting composite on marginal adaptation of a class II ceramic inlay.

This study evaluated the in vitro marginal quality at the interproximal cervical margin of class II Cerec restorations. Marginal quality was evaluated separately by (1) SEM analysis before and after simultaneous thermocycling and mechanical loading for the integrity of the restoration surface and (2) dye penetration after thermocycling and mechanical loading to evaluate the strength of the bond within the depth of the cavity. The results reveal that marginal integrity is influenced by the width of the luting space and the luting composite. With a luting space of 100 microns, marginal quality with as little as 3% to 14% loss of adhesion can be obtained. Luting spaces greater than 100 microns can partially be compensated by the luting composite. For Cerec inlays, highly filled luting composites with a high viscosity are recommended.

Adhesiveness↗