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Biomedical subjects

R Eckert

Publications and source records attributed to R Eckert.

At least 19 recordsLinked to original sources

[Recommendations for follow-up of malignant diseases].

The follow up of patients after primary therapy of cancer is being made difficult by the large number of sometimes contradictory recommendations, but also by a lack of evidence in many diseases. A chance for cure or a longer life expectancy with earlier diagnosis of an asymptomatic recurrence exists in only a few diseases. For far more diseases, a symptom oriented follow-up with an emphasis on quality of life, treatment associated complications, evaluation of symptoms, and psychological support is recommended. In patients with a dislike of regular visits or in diseases without options for further treatment it will be sufficient if the patient returns in the case of complaints. Recommendations for follow-up in selected frequent cancers will be given, based mainly on the latest recommendations by the European Society for Medical Oncology (ESMO).

Breast Neoplasms↗

[Treatment, follow up and prevention of colorectal carcinoma: strategies and controversies in the year 2005].

Colorectal carcinomas are common, so efficient strategies in their prevention and treatment will significantly lower mortality. Life style changes are an efficient primary prevention, but mass screening of the population can reduce morbidity and mortality even further. Complete resection is the clue to cure in colorectal carcinomas. Total mesorectal excision and laparoscopic operations have been established as effective interventions that may also improve patients' quality of life. Adjuvant therapy reduces recurrence rates and is recommended in stage II and III rectal carcinoma and stage III colon carcinoma. The optimal strategies for adjuvant therapy are currently being redefined. The availability of new drugs has extended the therapeutic options in metastatic disease, improving response rates and overall survival in the palliative situation. Sequential therapies with different drug combinations seem to be the most effective strategy. The operative resection of localized metastases to the liver or lung can cure some patients with metastatic disease.

Antineoplastic Agents↗

Missense mutations in exon 4 of the porcine LEPR gene encoding extracellular domain and their association with fatness traits.

Three non-synonymous single nucleotide polymorphisms (T221C, T232A and C233T) were detected in exon 4 of the porcine leptin receptor (LEPR) gene. The T232A substitution could be identified as a (Tsp509I) restriction fragment length polymorphism. The frequency of genotype TT varied in six genetic groups from 0.62 (Duroc) to 0.99 (Polish Large White). Sequencing of exon 4, performed for 30 animals, revealed that only two intragenic haplotypes (TC and AT at nucleotide position 232-233) were present. The phenotypic effect of the Tsp509I polymorphism was tested for the Polish Landrace (n = 241) and a synthetic line 990 (n = 243). There was no statistical evidence for the direct effect of the LEPR polymorphisms on fatness traits. However, in Polish Landrace allele A at position 232 was associated with thicker backfat over shoulder.

Animals↗

Diffusion behavior of gap junction hemichannels in living cells.

Due to its non-invasive character, fluorescence correlation spectroscopy (FCS) is particularly suited for the investigation of diffusion behavior of proteins in living cells. In this study we have investigated the diffusion properties of CFP-labeled gap junction hemichannels in the plasma membrane of living HeLa cells. Gap junction hemichannels or connexons are the precursors for the cell-cell- or gap junction channels that form large plaques at the contact areas between two adjacent cells. It has been proposed that new channels are recruited into a gap junction structure from a pool of hemichannels that can freely diffuse over the entire plasma membrane. The statistical approach shows that the geometry of the membrane within the focus is the most important property for the form of the autocorrelation curve and in turn for the determination of the diffusion coefficient. On the other hand binding-unbinding events which lead to anomalous diffusion have only a minor effect to the position and shape of the correlation curve compared to the geometry of the membrane.

Cell Membrane↗

In vitro internalization, intracellular transport, and clearance of an anti-CD11a antibody (Raptiva) by human T-cells.

Efalizumab (Raptiva) is a humanized CD11a-specific monoclonal antibody that was recently approved for the treatment of moderate to severe psoriasis. In psoriasis patients, the rate of efalizumab clearance from serum is related to T-cell surface expression of CD11a, suggesting a receptor-mediated clearance model for efalizumab (Bauer et al., 1999). However, limited experimental data are available to explain how the interaction with CD11a results in the systemic clearance of efalizumab. The following studies were designed to test the hypothesis that one mechanism of anti-CD11a antibody clearance is mediated in part by cellular internalization. This was tested in vitro using purified mouse and human T-cells as a model to study the cellular uptake and clearance of anti-CD11a antibodies. Data from these studies suggest that anti-CD11a antibodies are internalized by purified T-cells. Upon internalization, the antibodies appeared to be targeted to lysosomes and were cleared from within the cells in a time-dependent manner. CD11a-mediated internalization and lysosomal targeting of efalizumab may constitute one pathway by which this antibody is cleared in vivo.

Animals↗

Characterization and fabrication of fully metal-coated scanning near-field optical microscopy SiO2 tips.

The fabrication of silicon cantilever-based scanning near-field optical microscope probes with fully aluminium-coated quartz tips was optimized to increase production yield. Different cantilever designs for dynamic- and contact-mode force feedback were implemented. Light transmission through the tips was investigated experimentally in terms of the metal coating and the tip cone-angle. We found that transmittance varies with the skin depth of the metal coating and is inverse to the cone angle, meaning that slender tips showed higher transmission. Near-field optical images of individual fluorescing molecules showed a resolution < 100 nm. Scanning electron microscopy images of tips before and after scanning near-field optical microscope imaging, and transmission electron microscopy analysis of tips before and after illumination, together with measurements performed with a miniaturized thermocouple showed no evidence of mechanical defect or orifice formation by thermal effects.

Aluminum↗

Functional rescue of defective mutant connexons by pairing with wild-type connexons.

We have compared the functional and structural integrity of gap junction channels assembled from a Cx45 truncation mutant with those of gap junction channels assembled from wild-type (wt) Cx45 and Cx43. These channel-forming proteins are constitutively expressed in HeLa cells. The truncation mutant lacks the last 26 amino acids of the COOH-terminus, including nine serine phosphorylation sites that are associated with regulatory processes of these channels. We determined the presence of gap junction plaques in these cells with the immunogold freeze fracture technique, which showed that plaque formation is similar in all the clones investigated. Junctional permeability was probed with calcein transfer and flow cytometry analyses and junctional conductance was measured in cell pairs with double whole-cell patch-clamp techniques. For homotypic pairing only the truncated mutant did not form permeable channels. However, coupling was restored for heterotypic channels (pairing wtCx45- or wtCx43- with mutant-connexons), whose junctional communication was not different from that of the homotypic channels. Our results indicate that the presence of gap junction plaques does not warrant functional coupling and that heterotypic trCx45/wtCx45 channels can be regulated by the intact wtCx45 connexons. This dominant-positive effect is also operative when wtCx43 are paired with trCx45 connexons.

Connexin 43↗

Near-field optical microscopy based on microfabricated probes.

We demonstrate high resolution imaging with microfabricated, cantilevered probes, consisting of solid quartz tips on silicon levers. The tips are covered by a 60-nm thick layer of aluminium, which appears to be closed at the apex when investigated by transmission electron microscopy. An instrument specifically built for cantilever probes was used to record images of latex bead projection patterns in transmission as well as single molecule fluorescence. All images were recorded in constant height mode and show optical resolutions down to 32 nm.

Journal Article↗

[Pathogenetic and clinical aspects of low estradiol level in the man].

To elucidate a possible role of low estradiol (E2) levels in blood serum of men, normal values were determined in 91 healthy men (age 20-75 years), classified as high or low complaint-index due to a psychological questionnaire. Statistical analysis gave no correlation of estradiol levels to age or complaint index in normal men whereas testosterone (T) could be significantly correlated to complaint-index (p < 0.01) and free testosterone (fT) could be significantly correlated to age (p < 0.001) and complaint-index (p < 0.01). T and E2 were determined in 1370 clinical patients with various urological diseases, T, fT and E2 in 1261 ambulant patients. In 72/1370 (5.2%) and 76/1261 (6%) patients, low E2-levels (< 10 pg/ml) were found in blood serum. In 56/76 (74%) patients with low E2-levels, T or fT was simultaneously low. Isolated low E2-levels were found in 20/1261 (1.6%) patients. In clinical patients, no special urological disease correlated to low E2-levels. Due to these results, low E2 levels in men are in most cases the result of low testosteron levels. The adequate hormonal treatment in men is therefore the replacement of testosteron. Substitution of E2 in men is at that time an experimental therapy, that is limited on selected cases.

Adult↗

Quantitative determination of gap junctional permeability in the lens cortex.

We have developed a simple dye transfer method, which allows the gap junction permeability of lens fiber cells to be quantified. Two fixable fluorescent dyes (Lucifer yellow and rhodamine-dextran) were introduced into peripheral lens fiber cells via mechanical damage induced by removing the lens capsule. After a defined incubation period, lenses were fixed, sectioned, and the distribution of the dye recorded using confocal microscopy. Rhodamine-dextran and Lucifer yellow both labeled the extracellular space between fiber cells and the cytoplasm of fiber cells that had been damaged by capsule removal. For the gap junctional permeable dye Lucifer yellow, however, labeling was not confined to the damaged cells and exhibited intercellular diffusion away from the damaged cells. The extent of dye diffusion was quantified by collecting radial dye intensity profiles from the confocal images. Effective diffusion coefficients (D eff) for Lucifer yellow were then calculated by fitting the profiles to a series of model equations, which describe radial diffusion in a sphere. D eff is the combination of dye diffusion through the cytoplasm and through gap junction channels. To calculate the gap junctional permeability (Pj) an estimate of the cytoplasmic diffusion coefficient (Dcyt = 0.7 x 10(-6) cm2/sec) was obtained by observing the time course of dye diffusion in isolated elongated fiber cells loaded with Lucifer yellow via a patch pipette. Using this approach, we have obtained a value for Pj of 31 x 10(-5) cm/sec for fiber-fiber gap junctions. This value is significantly larger than the value of Pj of 4.4 x 10(-6) cm/sec reported by Rae and coworkers for epithelial-fiber junctions (Rae et al., 1996. J. Membrane Biol. 150:89-103), and most likely reflects the high abundance of gap junctions between lens fiber cells.

Animals↗

Number, size, and composition of cell clusters as related to breast FNA adequacy.

The question of what types of parameters should be employed in establishing adequacy guidelines for fine-needle aspirations (FNAs) of palpable breast lesions remains without consensus opinion among cytopathologists. Although some investigators have suggested guidelines, based largely upon cellularity standards, these have been somewhat conflicting, and the overall issue of whether or not cellularity is an appropriate measure of adequacy is controversial. This study examines the number, size, and composition of cell clusters in FNAs of palpable breast masses which had surgical follow-up, in an effort to identify characteristics of diagnostic aspirates which could contribute to adequacy guidelines. Seventy-seven consecutive cases with surgical follow-up were selected from four quality-assurance (QA) categories (true positive, TP; true suspicious, TS; true negative, TN; and false negative, FN). All cases were evaluated in terms of overall cellularity (numbers of cell clusters), proportions of different-sized epithelial clusters (small, medium, and large), and proportions of epithelial to fibrofatty elements. Although the FN category showed the lowest average cellularity, the application of cellularity cutoff values as a condition of adequacy would have resulted in conspicuous numbers of cases from the "true" categories (TN, and to a lesser degree TS and TP) being rendered inadequate. For example, if cases with fewer than six epithelial clusters were excluded by virtue of inadequacy, 69% of the FN cases would have been affected, along with 40% of the TN, 6% of the TS, and 4.5% of the TP. In terms of composition of the aspirate, neither proportions of the differently-sized epithelial clusters nor ratios of epithelial:fibrofatty elements reliably distinguished the "true" from the "false" diagnoses. This study concluded that quantitative parameters alone are insufficient measures for determining specimen adequacy in FNA of palpable breast lesions. Rather, adequacy remains based upon factors such as confidence of needle placement, cell preservation, and correlation with clinical and mammographic findings. Diagn. Cytopathol. 1999;21:105-111.

Biopsy, Needle↗

Connexins in the lens: are they to blame in diabetic cataractogenesis?

The pathohistology of the diabetic lens is an enigma. Under normal conditions the lens behaves as a functional syncitium, whereas the diabetic lens exhibits a localized zone of fibre cell swelling and rupture that is confined to the lens outer cortex. Because the lens fibre cells are extensively coupled by gap junction channels, it is believed that the abnormal closure of these channels is responsible for this phenomenon. New evidence concerning regional differences in gap junction gating supports this contention, and it is used to propose a new hypothesis that may explain the cellular changes observed in the diabetic lens.

Animals↗

Changes of intracellular calcium concentrations by phenylephrine in renal arterial smooth muscle cells.

In smooth muscle cells isolated from swine renal interlobar arteries, phenylephrine (PE) at concentrations of 1-10 microM produced biphasic increases of the intracellular calcium concentration. An early transient rise was followed by a maintained plateau. The maintained component was sensitive to extracellular calcium, in contrast to the early transient, which was still observed in nominally calcium-free solution. Nifedipine (1 microM) and NiCl2 (100 microM) only weakly affected the calcium signal, suggesting that voltage-sensitive calcium channels play only a minor role in the PE-induced changes in intracellular calcium. Thapsigargin (0.5 microM) elevated the intracellular calcium concentration and depressed both the early transient and the maintained component of the PE response. In calciumfree medium PE induced a transient rise of the intracellular calcium concentration with a depressed plateau. Readmission of calcium elevated the intracellular calcium concentration above the baseline. Both components of the PE-induced calcium signal were completely abolished when the cells were pretreated with the phospholipase C (PLC) inhibitor U73122 (2 microM). LaCl3 (100 microM, 1 mM), an inhibitor of calcium-release-activated current (ICRAC), had no effect on the PE-induced calcium signal. GdCl3 (50 microM), SKF 96365 (10 microM) and flufenamic acid (100 microM), reported to inhibit nonselective cation channels, blocked or transiently reduced the maintained calcium signal. Several protein kinase inhibitors such as genistein (10 microM), H7 (50 microM), H89 (1 microM) and bisindolylmaleimide (0.2 microM) reduced the maintained calcium signal. We conclude that the initial transient spike of the PE-induced calcium signal is due to release of calcium from inositol 1,4,5-trisphosphate-sensitive calcium stores evoked by alpha 1-adrenoceptor-coupled stimulation of PLC and that the maintained component is due to capacitative calcium entry, which is modulated by protein kinases.

Adrenergic alpha-Agonists↗

MAPKAP kinase 2 is essential for LPS-induced TNF-alpha biosynthesis.

MAPKAP kinase 2 (MK2) is one of several kinases that are regulated through direct phosphorylation by p38 MAP kinase. By introducing a targeted mutation into the mouse MK2 gene, we have determined the physiological function of MK2 in vivo. Mice that lack MK2 show increased stress resistance and survive LPS-induced endotoxic shock. This is due to a reduction of approximately 90% in the production of tumor necrosis factor-alpha (TNF-alpha) and not to a change in signalling from the TNF receptor. The level and stability of TNF-alpha mRNA is not reduced and TNF-alpha secretion is not affected. We conclude that MK2 is an essential component in the inflammatory response which regulates biosynthesis of TNF-alpha at a post-transcriptional level.

Animals↗

Localised fibre cell swelling characteristic of diabetic cataract can be induced in normal rat lens using the chloride channel blocker 5-nitro-2-(3-phenylpropylamino) benzoic acid.

Chloride channels are known to be involved in the regulated volume decrease that occurs when the rat lens is exposed to hypotonic challenge. We now report that chloride channel blockage makes the rat lens gain water under isotonic conditions, suggesting that chloride and water fluxes may also play an important role under resting conditions. Histological comparison of hypotonically and isotonically swollen rat lenses revealed a significant difference: in the former, fibre cells were swollen from the periphery inwards, while in the latter, swollen fibre cells were confined to a discrete cortical zone which was located 150-200 micron from the lens surface with cells on either side of this zone appearing unaffected. This localised fibre cell swelling is remarkable because of its similarity to the situation in the diabetic rat lens.

Animals↗

Severe hemolytic reaction due to anti-JK3.

A 35-year-old gravida 3, para 3 Filipino woman with a negative antibody screen, no prior history of transfusion, and no hemolytic disease of the newborn in her children suffered a massive postpartum hemorrhage requiring transfusion. A severe hemolytic transfusion reaction occurred 5 days after delivery. Subsequently, a panagglutinin on a routine antibody identification panel was identified as anti-Jk3. The patient's red blood cell phenotype was Jk(a-b-) and all of her children were Jk(a-b+), yet the antibody that formed reacted with equal strength against all Jk(a)- or Jk(b)-positive cells. The rare Jk(a-b-) phenotype is more common in Polynesians. Anti-Jk3, like other Kidd system antibodies, is difficult to detect because in vivo production may be absent between provocative episodes and because these antibodies often show weak in vitro reactions. The increasing numbers of Pacific Islanders in the United States could result in more frequent encounters with this rare phenotype. Increased awareness of ethnic variability in blood phenotypes and of the capricious nature of Kidd antibodies can help pathologists and technologists deal more effectively with these cases.

Adult↗

Role of intracellular Ca2+ stores in smooth muscle of human penile erectile tissue.

OBJECTIVE: In human erectile tissue smooth muscle contraction and detumescence are highly dependent on an increase in cytosolic [Ca2+]. The Ca2+ influx can be derived from the extracellular space or from intracellular sarcoplasmic stores. The role of both pathways was evaluated in an organ bath study on human cavernosal strips. PATIENTS AND METHODS: The tissue was obtained from 12 patients with chronic erectile dysfunction. The effects of Ca2+-free solution, ryanodine, caffeine and of nifedipine on electrically and adrenergically induced contractions were evaluated. RESULTS: Following an incubation period of 10 min in Ca2+-free solution the electrically induced contraction was reduced to 20%, whereas the contraction induced by phenylephrine (PE) was only reduced to 64 +/- 6% (mean +/- SEM). Ryanodine inhibited the PE-contraction to 30 +/- 6% and the additional application of caffeine or nifedipine further reduced the contraction to 11% and 8%. CONCLUSION: The results give evidence for a role of intracellular Ca2+-stores in human cavernosal tissue. Whether the more marked effect of ryanodine in tissue from patients with erectile failure in comparison with similar experiments in rabbit cavernosal tissue might be a sign of an increased cavernosal contractility in these patients remains to be shown in future experiments with normal erectile tissue.

Animals↗