Biomedical subjects

H Niemann

Publications and source records attributed to H Niemann.

At least 19 recordsLinked to original sources

PubMed2007

Altered mRNA expression patterns in bovine blastocysts after fertilisation in vitro using flow-cytometrically sex-sorted sperm.

Sperm-sexing has been used to produce embryos and offspring of a pre-determined sex in a number of species. However, the fertility of sex-sorted sperm is reduced and the full effects of sperm-sexing remain to be elucidated. The purpose of the present study was to investigate the potential effects of sex-sorted sperm on mRNA expression patterns of developmentally important genes employing in vitro produced bovine embryos. Bovine embryos were produced in vitro with unsorted and sex-sorted sperm and mRNA expression patterns were determined for glucose-3 transporter (Glut-3), glucose-6-phosphate dehydrogenase (G6PD), X-inactive specific transcript (X-ist) and Heat shock protein 70.1 (Hsp) using semi-quantitative endpoint reverse transcriptase-PCR in male and female, day-7 and 8 embryos. The relative abundance (RA) of Glut-3 was higher for day-7 male than female embryos, and day-7 embryos derived from unsorted compared with sex-sorted sperm. The RA of G6PD was higher for embryos derived from unsorted than sex-sorted sperm, and for day-8 female compared with male embryos. The RA of Xist was higher for female than male embryos, and for day-7 female embryos derived from unsorted than sex-sorted sperm. Hsp RA was higher for female compared with male embryos, was similar for day-7 and 8 embryos, and unsorted and sex-sorted sperm derived embryos. These results demonstrate differential expression of developmentally important genes between male and female embryos, and embryos derived from unsorted and sex-sorted sperm.

Animals↗

PubMed2006

Inhibition of porcine endogenous retroviruses (PERVs) in primary porcine cells by RNA interference using lentiviral vectors.

A potential risk in pig-to-human xenotransplantation is the transmission of PERVs to human recipients. Here we show for the first time the inhibition of PERV expression in primary porcine cells by RNA interference using lentiviral vectors. Cells were transduced with lentiviral vectors coding for short hairpin (sh) RNAs directed against PERV. In all primary porcine cells studied and in the porcine kidney cell line PK-15, expression of PERV-mRNA was significantly reduced as measured by real-time PCR. Most importantly, expression of PERV proteins was almost completely suppressed, as shown by Western blot analysis. Thus, lentiviral shRNA vectors could be used to knockdown PERV expression and create transgenic pigs with a reduced risk of PERV transmission during xenotransplantation.

Animals↗

PubMed2006

Performances under representative pressure and temperature conditions of the gas chromatography-mass spectrometry space experiment to investigate Titan's atmospheric composition.

In the frame of the calibration of the aerosol collector and pyrolyser, and gas chromatography-mass spectrometry experiments of the Huygens probe arrived at Titan, systematic experimental studies were led to estimate the influence of the operating conditions on the analyses that should have been achieved in the Titan's atmosphere. The primary objective of this study was to estimate the influence of operating conditions variations induced by (i) instrumental modifications made shortly before the probe launch which can have changed the operating pressures; (ii) the change of the probe environmental conditions (pressure, temperature) during its descent in the atmosphere; (iii) a possible deviation of pressure and temperature regulations from their nominal values because of the long journey of the instrument in space, or of other external events. The secondary objective of this work was to create an analytical database that can be used as a reference to treat the chromatograms obtained in situ, and help to identify chromatographically the analyzed species, complementary to mass spectrometry. Beyond the application to a specific instrument, this work was also useful to experimentally estimate the fundamental evolution of the separation as a function of the changes of operating conditions with time. The obtained results show (i) the significant influence of inlet and outlet pressure variation on the time of analysis, but not on the separation power. It thus enables to significantly shorten the analysis duration, and thus to analyze more compounds within the fixed time of analysis of the instrument; (ii) the significant influence of temperature on the retention. In this frame, the enthalpies of exchange between the gas phase and the stationary phase of the species were determined to be used to retrieve the analyzed species in case of deviation of the operating temperature; (iii) that the possible aging of the columns does not have influence on the columns efficiency and separation power; (iv) the analytical capabilities of the gas chromatography-mass spectrometry experiment within operating conditions representative of those encountered in situ. Finally, in spite of possible operating condition changes, it is shown that results coming from the gas chromatograph-mass spectrometer experiment, which are currently under analysis, could bring important information on the Titan's atmosphere and its history.

Atmosphere↗

PubMed2006

Utilization of frozen-thawed epididymal ram semen to preserve genetic diversity in Scrapie susceptible sheep breeds.

The European Union has introduced transmissible spongiform encephalopathy (TSE) resistance breeding programmes for several sheep breeds to cope with the genetic susceptibility to Scrapie infections. Due to the different allele frequencies among breeds, strong selection for ARR alleles is associated with a loss of genetic diversity in small populations and in larger populations with unfavourable ARR allele frequencies. To ensure maintenance of genetic diversity, an adhoc cryopreservation programme was initiated employing epididymal sperm from 109 rams representing 16 different breeds within one breeding season. Epididymal semen was chosen for this adhoc programme because time consuming training of rams for ejaculated semen collection via an artificial vagina was not possible. Prior to freezing, average sperm motility was 79.7% and acrosome integrity was 93.7%. After freezing, these levels were decreased to 60.5 and 72.8%, respectively. An insemination trial using frozen-thawed epididymal semen resulted in a lambing rate of 87.5%. Results show that this semen preservation method is robust and efficient and associated with high fertility. It may also be useful for other animal species.

Acrosome↗

PubMed2006

Isolation of bovine cardiomyocytes for reprogramming studies based on nuclear transfer.

The goal of this study was to establish and validate a protocol for preparing bovine cardiomyocytes from slaughterhouse material for nuclear transfer experiments. The cardiomyocyte was selected because it is a terminally differentiated cell and strongly expresses a unique subset of genes which can be monitored during the reprogramming period. A total of 39 trials were conducted, and an optimized protocol was developed yielding individual contractile cardiomyocytes from 3-5-month-old bovine fetuses The basic protocol involves stabilization of bovine heart tissue for transportation from the slaughterhouse to the laboratory by perfusion with Custodiol. This was followed by an enzymatic dissociation with collagenase in calcium-free medium and yielded individual contractile rod-shaped cardiomyocytes. Subsequent addition of Ca2+ caused the cardiomyocytes to round up which was an essential pre-condition for drawing them into glass transfer pipettes for delivery into the perivitelline space and for efficient electrofusion with cytoplasts derived from in vitro matured bovine oocytes. The use of cardiomyocytes maintained at 37 degrees C in nuclear transfer, resulted in a significantly reduced proportion of blastocysts compared to adult fibroblasts (14.0% versus 32.7%). Storage of cardiomyocytes at 4 degrees C prior to nuclear transfer was not compatible with blastocyst development. It is expected that this system will be valuable for investigating the reprogramming of gene expression which occurs after somatic cell nuclear transfer.

Animals↗

PubMed2006

High incidence of single nucleotide polymorphisms in the prion protein gene of native Brazilian Caracu cattle.

Different alleles of the human and ovine prion protein gene correlate with a varying susceptibility to transmissible spongiform encephalopathies. However, the pathogenic implications of specific polymorphisms in the bovine prion protein gene (PRNP) are only poorly understood. Previous studies on the bovine PRNP gene investigated common European and North American cattle breeds. As a consequence of decades of intensive breeding for specific traits, these modern breeds represent only a small fraction of the bovine gene pool. In this study, we analysed PRNP polymorphisms in the native Brazilian Caracu breed, which developed in geographical isolation since the 16th century. A total of 10 single nucleotide polymorphisms (SNPs) were discovered in the coding region of the Caracu PRNP gene. Eight of the SNPs occurred at high frequencies in Caracu cattle (variant allele frequencies = 0.10-0.76), but were absent or only rarely observed in European and North American breeds. One of the Caracu SNPs was associated with an amino acid exchange from serine to asparagine (f = 0.17). This SNP was not detected in Holstein-Friesian, Simmental and German Gelbvieh and was only rarely detected in beef cattle (f = 0.01). We found 17 haplotypes for PRNP in the Caracu breed.

Animals↗

PubMed2005

Automated segmentation of the optic nerve head for diagnosis of glaucoma.

Glaucoma is the second most common cause of blindness worldwide. Low awareness and high costs connected to glaucoma are reasons to improve methods of screening and therapy. A well-established method for diagnosis of glaucoma is the examination of the optic nerve head using scanning-laser-tomography. This system acquires and analyzes the surface topography of the optic nerve head. The analysis that leads to a diagnosis of the disease depends on prior manual outlining of the optic nerve head by an experienced ophthalmologist. Our contribution presents a method for optic nerve head segmentation and its validation. The method is based on morphological operations, Hough transform, and an anchored active contour model. The results were validated by comparing the performance of different classifiers on data from a case-control study with contours of the optic nerve head manually outlined by an experienced ophthalmologist. We achieved the following results with respect to glaucoma diagnosis: linear discriminant analysis with 27.7% estimated error rate for automated segmentation (aut) and 26.8% estimated error rate for manual segmentation (man), classification trees with 25.2% (aut) and 22.0% (man) and bootstrap aggregation with 22.2% (aut) and 13.4% (man). It could thus be shown that our approach is suitable for automated diagnosis and screening of glaucoma.

Algorithms↗

PubMed2005

Effect of stage of follicular growth during superovulation on developmental competence of bovine oocytes.

The final steps of oocyte capacitation and maturation are critical for embryonic development but detailed information is scarce on how the oocyte is affected during this period. In this study, 2033 oocytes were collected from 106 superovulated cattle at four different time points before ovulation. Follicular characteristics were measured and oocyte quality was assessed by morphology, mRNA expression of eight marker genes or developmental ability after in vitro/in vivo maturation and subsequent in vitro fertilization and culture. Approaching ovulation, expected increases in follicular size and cumulus expansion suggested progression of oocyte maturation. No differences were found in the expression patterns of analyzed genes, except for heat-shock-protein (Hsp) that was lower in in vivo matured oocytes collected shortly before ovulation. Oocytes collected at this time also had higher developmental ability measured as blastocyst rates (57.6%) after in vitro production while no differences were found between oocytes recovered earlier at the first three time points (39.3-41.5%). We conclude that oocytes recovered late in the preovulatory period are more developmentally competent than oocytes recovered at the pre-capacitation and the capacitation period, probably due to the former having matured in vivo. However, a precisely defined time for aspirating immature oocytes for subsequent in vitro development seems not to be crucial.

Animals↗

PubMed2005

[Noise induced annoyance and morbidity. Results from the pan European LARES-survey].

Traffic noise (road, train and flight noise, and the noise of parking cars), is the dominant source of annoyance in the living environment of many European countries. This is followed by neighbourhood noise (neighbouring apartments, staircases and noise within the apartment). The subjective experience of noise stress can, through central nervous processes, lead to an inadequate neuro-endocrine reaction and finally to regulation diseases. Within the context of the LARES-survey, noise annoyance within the everyday living environment was collected and evaluated in connection with medically diagnosed illnesses. Adults who indicated chronically strong annoyance due to neighbourhood noise were found to have an increased health risk in the cardiovascular system, the movement apparatus as well as depression and migraine. For adults with chronically strong annoyance caused by traffic noise, the risks to the respiratory system also increased. In older people, both neighbourhood and traffic noise indicated, in general, a lower risk of noise annoyance induced illness than in adults. It can be assumed that the effects of noise induced annoyance in older people is concealed by the physical consequences of age (with a strong increase in illnesses). With children, the effects of noise induced annoyance from traffic as well as neighbourhood noise is evident in the respiratory system. The increased illness risks in the respiratory system in children do not seem to be caused primarily by air pollutants but rather, as with case of neighbourhood noise, though emotional stress.

Adolescent↗

PubMed2005

Transgenic farm animals: present and future.

Until recently, pronuclear microinjection of deoxyribonucleic acid (DNA) was the standard method for producing transgenic animals. This technique is now being replaced by more efficient protocols based on somatic nucleartransferthat also permit targeted genetic modifications. Lentiviral vectors and small interfering ribonucleic acid technology are also becoming important tools for transgenesis. Transgenic farm animals are important in human medicine as sources of biologically active proteins, as donors in xenotransplantation, and for research in cell and gene therapy. Typical agricultural applications include improved carcass composition, lactational performance and wool production, as well as enhanced disease resistance and reduced environmental impact. Product safety can be ensured by standardisation of procedures and monitored by polymerase chain reaction and array technology. As sequence information and genomic maps of farm animals are refined, it becomes increasingly practical to remove or modify individual genes. This approach to animal breeding will be instrumental in meeting global challenges in agricultural production in the future.

Animals↗

PubMed2004

Improvement of the developmental capacity of oocytes from prepubertal cattle by intraovarian insulin-like growth factor-I application.

The developmental potential of oocytes from prepubertal cattle is decreased, compared with those from their adult counterparts. The aim of the present study was to improve the developmental capacity of oocytes from prepubertal cattle by either systemic application of recombinant bovine somatotropin (rbST) or intraovarian injection of insulin-like growth factor-I (IGF-I). Blastocyst yields and the mRNA expression pattern (relative abundance, RA) of three putative marker genes (i.e., glucose transporter-1, Glut-1; eukaryotic translation initiation factor-1A, eIF1A, and upstream binding factor, UBF) were selected as criteria to determine the success of the treatments. At 6-7 mo of age, 30 healthy Holstein calves were randomly assigned to three experimental groups. The first group served as control and received an intraovarian injection of 0.6 ml acetic acid. The second group received a single s.c. injection of 500 mg of rbST. The third group received an intraovarian injection of 6 microg recombinant human IGF-I. During the following 2 wk, follicles were aspirated four times via transvaginal ultrasound-guided technology. All animals were i.m. injected with 60 mg FSH 48 h prior to each aspiration. The treatments were repeated with the same animals at 9-10, 11-12, and 14-15 mo of age. For comparison, five adult cows were each i.m. injected with 100 mg FSH and underwent oocyte retrieval. The proportion of oocytes considered to be developmentally competent was higher in cows than calves (65% vs. 58%, 50%, 52%) for the control, rbST, and IGF-I groups, respectively. The rate of blastocysts was similar in IGF-I-treated calves and cows (28% and 25%) and was higher (P </= 0.05) than in the controls and the rbST group (11% and 16%). The RA for Glut-1 was lower (P </= 0.05) in two- to four- cell embryos from calves, compared with cows. At the 8- to 16- cell stage, Glut-1 RA was similar in IGF-I-treated calves and cows. The RA for eIF1A was higher (P </= 0.05) in 8- to 16-cell embryos derived from cows than those from the control group. Results show that IGF-I intraovarian injection increased blastocyst yields and mRNA expression of Glut-1 and eIF1A to levels found in embryos produced from adult cows. This treatment may at least partially overcome the developmental deficiency of oocytes derived from calves and could be a step forward toward the use of prepubertal animals in breeding programs aimed at shortening the generation interval.

Age Factors↗

PubMed2004

Nucleolar ultrastructure and protein allocation in in vitro produced porcine embryos.

The nucleolus formation was studied as an indirect marker of the ribosomal RNA (rRNA) genes activation in porcine embryos following oocyte maturation, fertilization, and culture in vitro. Nucleologenesis was assessed by transmission electron microscopy (TEM), light microscopical autoradiography following 20 min of 3H-uridine incubation, and immunocytochemical localization of key nucleolar proteins involved in rRNA transcription (upstream binding factor (UBF), topoisomerase I, and RNA polymerase I) and processing (fibrillarin, nucleophosmin, nucleolin) by confocal laser scanning microscopy. During the first four post-fertilization cell cycles, TEM revealed spherical nucleolus precursor bodies (NPBs), consisting of densely packed fibrils, as the most prominent intra-nuclear entities of the blastomeres. Fibrillo-granular nucleoli were observed in some blastomeres in a single embryo during the 5th cell cycle, i.e., the tentative 16-cell stage, where formation of fibrillar centres (FC), a dense fibrillar component, and a granular component on the surface of the NPBs was seen. In this embryo, autoradiographic labeling was detected over the nucleoplasm and in particular over the nucleoli. Fibrillarin was immunocytochemically localized in the presumptive NPBs of the pronuclei. This protein was again localized to the presumptive NPBs together with nucleolin from late during the 3rd cell cycle, i.e., the four-cell stage in some embryos. UBF, RNA polymerase I, and nucleophosmin were localized to the presumptive NPBs in a proportion of the embryos at the 4th cell cycle, i.e., the tentative eight-cell stage and onwards. Toposiomerase I was not localized to intra-nuclear entities even during the 5th post-fertilization cell cycle. Moreover, a considerable proportion of the blastomere nuclei apparently did not show localization of other nucleolar proteins. In conclusion, porcine embryos produced in vitro display a substantial delay in or even lack of the development of functional nucleoli.

Animals↗

PubMed2004

Gene expression patterns in in vitro-produced and somatic nuclear transfer-derived preimplantation bovine embryos: relationship to the large offspring syndrome?

A considerable proportion of the offspring born from somatic nuclear transfer (sNT)-derived and in vitro-produced (IVP) embryos, particularly in ruminants and mice, is affected by multiple abnormalities of which a high birth weight is the predominant feature; a phenomenon that has been called "large offspring syndrome (LOS)". The underlying mechanisms are largely unknown at present, but changes in epigenetic modifications occurring during preimplantation development resulting in perturbed embryonic and fetal gene expression patterns are thought to be involved in the syndrome. This review summarizes results from studies comparing mRNA expression patterns from IVP and sNT-derived embryos to those of their in vivo counterparts, which are regarded as the "gold standard". Numerous aberrations have been observed ranging from suppression of expression to de novo overexpression or more frequently to a significant up- or down-regulation of a specific gene. These observations emphasize the need for further studies during preimplantation embryo development to gain insight in the molecular, preferentially epigenetic, mechanisms regulating embryonic and fetal development. Understanding these mechanisms will help to improve biotechnologies applied to early embryos in all species including humans.

Animals↗

PubMed2004

Impaired declarative memory in depressed patients is slow to recover: clinical experience.

INTRODUCTION: The temporal course of recovery of depressed patients' cognitive impairment is not fully understood. METHODS: We used the California Verbal Learning Test (CVLT) to test declarative memory in 24 depressed patients before and after 35 days of antidepressive treatment as well as after long-term follow-up (> 12 months) in order to relate improvement of depression to recovery of cognitive impairment. RESULTS: Patients with complete remission after 35 days had generally been less impaired at baseline. The disturbance of declarative memory in treatment responders as well as in non-responders did not change from baseline to end of treatment (day 35). However, our results revealed normal values in the CVLT sum score as well as in measures of short- and long-delay free-recall measures in both groups after long-term full remission. DISCUSSION: We conclude that clinical response to antidepressive treatment precedes improvement of declarative memory. A low degree of impairment of declarative memory is associated with early complete remission of depression.

Amitriptyline↗

PubMed2004

Multimodal retinal image registration for optic disk segmentation.

OBJECTIVES: The analysis of the optic disk morphology with the means of the scanning laser tomography is an important step for glaucoma diagnosis. A method we developed for optic disk segmentation in images of the scanning laser tomograph is limited by noise, non-uniform illumination and presence of blood vessels. Inspired by recent medical research, we wanted to develop a tool for improving optic disk segmentation by registration of images of the scanning laser tomograph and color fundus photographs and by applying a method we developed for optic disk segmentation in color fundus photographs. METHODS: The segmentation of the optic disk for glaucoma diagnosis in images of the scanning laser tomograph is based on morphological operations, detection of anatomical structures and active contours and has been described in a previous paper. The segmentation of the optic disk in the fundus photographs is based on nonlinear filtering, Canny edge detector and a modified Hough transform. The registration is based on mutual information using simulated annealing for finding maxima. RESULTS: The registration was successful 86.8% of the time when tested on 174 images. Results of the registration have shown a very low displacement error of a maximum of about 5 pixels. The correctness of the registration was manually evaluated by measuring distances between the real vessel borders and those from the registered image. CONCLUSIONS: We have developed a method for the registration of images of the scanning laser tomograph and fundus photographs. Our first experiments showed that the optic disk segmentation could be improved by fused information from both image modalities.

Fundus Oculi↗

PubMed2004

Evaluation of computer-assisted image enhancement in minimal invasive endoscopic surgery.

OBJECTIVES: This paper focuses on the evaluation of the usage of computer-aided image processing methods for minimal invasive surgery. During video endoscopy of visceral cavities the images are displayed directly on the monitor without further processing. In the course of the operation the former good quality of the images decreases due to typical disturbances like bleeding, smoke or flying particles. These disturbances can be reduced by using image processing methods like color normalization, temporal filtering or equalization. METHODS: In this double-blinded analysis, 14 surgeons with different levels of experience evaluated 120 image pairs and 5 image sequences, directly comparing original and processed images or movies. RESULTS: Color normalization and equalization proved to significantly enhance video endoscopic images. With regard to temporal filtering, an improvement could be seen in the image sequences with filter size 5 being a greater enhancement than filter size 3. Comparing the state of experience and its influence on the results, it occurred that the experienced surgeons preferred the original color while altogether agreeing that the color-normalized images were better. CONCLUSIONS: The results obtained in the present evaluation show that the image processing methods which were used can significantly improve the quality of video endoscopic images. As a result of this, necessary lavages of the operated area are reduced and a better overview and orientation for the surgeon can be reached.

Humans↗

PubMed2004

Light fields for minimal invasive surgery using an endoscope positioning robot.

OBJECTIVES: To generate a fast and robust 3-D visualization of the operation site during minimal invasive surgery. METHODS: Light fields are used to model and visualize the 3-D operation site during minimal invasive surgery. An endoscope positioning robot provides the position and orientation of the endoscope. The a priori un-known transformation from the endoscope plug to the endoscope tip (hand-eye transformation) can either be determined by a three-step algorithm, which includes measuring the endoscope length by hand or by using an automatic hand-eye calibration algorithm. Both methods are described in this paper and their respective computation times and accuracies are compared. RESULTS: Light fields were generated during real operations and in the laboratory. The comparison of the two methods to determine the unknown hand-eye transformation was done in the laboratory. The results which are being presented in this paper are: rendered images from the generated light fields, the calculated extrinsic camera parameters and their accuracies with respect to the applied hand-eye calibration method, and computation times. CONCLUSION: Using an endoscope positioning robot and knowing the hand-eye transformation, the fast and robust generation of light fields for minimal invasive surgery is possible.

Algorithms↗

PubMed2003

Timing of blastocyst expansion affects spatial messenger RNA expression patterns of genes in bovine blastocysts produced in vitro.

Blastocyst formation and expansion are dependent on the differentiation and function of a proper transport of nutrients through the trophectoderm (TE) enclosing the inner cell mass (ICM). Coincident with compaction and cavitation, glucose becomes the preferred energy substrate of the early embryo. These hallmarks in early development require well-orchestrated gene expression patterns specifically with regard to timing and localization. The present study investigated the relative abundance (RA) of gene transcripts in the two lineages of in vitro-produced expanded bovine blastocysts in relation to timing of development, i.e., blastocyst expansion and localization of specific mRNAs. Expanded blastocysts from either Day 7 or Day 8 or isolated ICMs derived thereof were analyzed with the aid of a semiquantitative reverse transcriptase-polymerase chain reaction assay for gene transcripts, which are thought to play a pivotal role in blastocyst expansion, i.e., Na/K-ATPase alpha1 subunit (Na/K), E-cadherin (E-cad), zonula occludens protein-1 (ZO-1), desmocollin II (Dc II), plakophilin (Plako), trophoblastic function (interferon tau [IFtau]), and glucose transport (glucose transporter-1, -3, -4 [Glut-1, -3, -4]). Total cell number, ICM cell number, or ICM/total cells ratio were similar in Day 7 and Day 8 expanded blastocysts. Significant differences were determined in the RA for Na/K, E-cad, Dc II, Plako, and ZO-1 transcripts between TE cells of expanded blastocysts derived from either Day 7 or Day 8. The RA of Dc II, Glut-1, and Glut-4 was significantly decreased in the ICM compared with the TE at Day 7. Similarly, the RA of Na/K, Dc II, Glut-1, and Glut-4 at Day 8 of development was significantly decreased in the ICM compared with the TE. Interestingly, no differences were observed when comparing ICMs originating from blastocysts expanded at either Day 7 or Day 8. Plako and IFtau transcripts were not detected in isolated ICMs, indicating that expression of these mRNAs is restricted to the TE. In contrast, similar expression patterns within the ICM and TE were determined for Na/K, E-cad, ZO-1, and Glut-3 mRNA. Dc II, Glut-1, and Glut-4 were more abundant in the TE than in ICM. Results show that expression of developmentally important genes is related to the two cell lineages in the early embryo and emphasize the critical role of a well controlled spatial gene expression pattern for regular preimplantation development.

Animals↗