Unkonventionelle Heilmethoden von Knie- und Sprunggelenkverletzungen bei Sportlern

Eine richtige Behandlung von Sportverletzungen und Sportschäden ist für den Sporttreibenden sehr wichtig und beeinflusst in vielen Fällen entscheidend die weitere sportliche Karriere. Daher sollten Verletzungen und Schäden möglichst rasch, schonend und umfassend behandelt werden. Eine Reihe von Behand-lungsmöglichkeiten für Sportverletzungen steht zur Verfügung. An erster Stelle steht oft die Schulmedizin, die dank der Spitzenleistungen der modernen Chirurgie in der Sportmedizin oft zum Einsatz kommt. Die konventionelle Medizin geht dabei im allgemeinen symptomorientiert vor, d.h. sie befasst sich nur mit dem kranken Organ; dabei wird der Mensch nicht als eine Einheit aus Körper, Seele und Geist betrachtet. Mens sana in corpore sano kannten schon die alten Römer – ein gesunder Geist ruht in einem gesunden Körper. Griechi-sche und römische Mediziner behandelten daher bereits vor Tausenden von Jahren nach ganzheitlichen Konzepten. Schon in der frühen Lehre von Hippo-krates und später bei Paracelsus spielten die Heilkräfte der Natur bei der Heilung des Kranken eine wesentliche Rolle. Viele internationale Studien haben ergeben, dass Patienten mit der modernen wissenschaftlichen Schulmedizin nicht mehr zufrieden sind. Sie suchen nach Alternativen, die auf natürliche Steigerung der Leistung und ursächliche Aus-heilung von Krankheiten und Körperschäden abzielt und nicht auf die vorder-gründige Beseitigung von Symptomen. Solche Behandlungen bietet die unkonventionelle Medizin an, indem sie auf individuelle, qualitative und effektive Diagnose und Therapie setzt, die Geist, Körper, Seele und soziales Umfeld des Patienten berücksichtigt. Dies erscheint besonders für die sportmedizinische Betreuung von Bedeutung. Die Anwendung der unkonventionellen Heilmetho-den kann dem Behandler eine funktionelle Denkweise, die eine ganzheitliche naturheilkundlich-psychologisch-sportmedizinische Betreuung ermöglicht, eröff-nen. Die Akzeptanz unkonventioneller Heilmethoden in der Medizin ist aller-dings nach wie vor unterschiedlich, und zwar sowohl auf der Ebene der Behandler, als auch auf der Ebene der Behandelten. Die vorliegende Arbeit beschäftigte sich daher mit der Problematik, unkonventi-onelle Heilmethoden bei der Behandlung von Sportverletzungen einzusetzen. Ziel dieser Untersuchung war es, die gebräuchlichen unkonventionellen Heilmethoden und ihre Anwendungen bei Sportverletzungen, aufzuzeigen, die Beweggründe, warum sich Sportler mit den unkonventionellen Heilmethoden behandeln lassen, festzustellen, sowie eine repräsentative Umfrage unter ärzt-lichen und nichtärztlichen Behandlern zu machen, um den Einsatz von unkon-ventionellen vs. konventionellen Heilmethoden zu untersuchen. Dazu wurden zunächst acht zentrale Fragen zu vorgegebenem Thema formuliert, die im Laufe der Studie beantwortet werden sollten. Die Beantwortung der anfangs formulierten Problematik wurde im Raum Baden-Württemberg durchgeführt. An der Studie nahmen 274 Sportler aus der Region Karlsruhe und Heidelberg im Zeitraum März 2005 bis September 2007 sowie 67 Behandler aus dem Region Baden Württemberg in der Periode August 2006 bis September 2007 teil. Bei den statistischen Auswertungen der Fragebögen ergab sich, dass aus den 274 Probanden 128 Personen (46,7 Prozent) nur schulmedizinisch, 80 Per-sonen (29,2 Prozent) nur unkonventionell und 66 Personen (24,1 Prozent) mit beiden Heilrichtungen behandelt worden waren. Von 128 Personen, die nur schulmedizinisch behandelt wurden, zeigten 82 Personen (64,1 Prozent) ein Interesse, künftig mit alternativen Therapien behandelt zu werden. Der Altersdurchschnitt der Anwender alternativer Therapien betrug 34,2 Jahre, der der Nichtanwender 36,1 Jahre. Frauen wandten die unkonventionellen Heil-methoden mit 59,6 Prozent (N=87) häufiger als Männer an. Die Mehrheit der Probanden, sowohl in der Anwender-Gruppe (41,1 Prozent mit Abitur; 39 Prozent mit einem Hochschulabschluss) wie auch in der Nichtanwender-Gruppe (33,6 Prozent mit Abitur; 34,4 Prozent mit einem Hochschulabschluss), wies ein höheres Bildungsniveau auf. In der Behandler Gruppe waren 47 Männer und 20 Frauen, darunter 34 Ärzte, 26 Heilpraktiker und fünf Physiotherapeuten. Das durchschnittliche Alter betrug 41,1 Jahre. Bei den Behandlern waren gute Verträglichkeit (55,2 Prozent), Individualität (43,3 Prozent) und Ganzheitlichkeit (40,3 Prozent) für die Anwendung unkon-ventioneller Heilmethoden bestimmend. Allgemein boten Behandler am häufigsten Akupunktur (11 Prozent) an gefolgt von Osteopathie (7,2 Prozent) und Homöopathie, sowie Sauerstofftherapie (mit jeweils 6,5 Prozent). Bei Knie- und Sprunggelenksverletzungen wurden vor allem Bandverletzungen mit 30,8 Prozent bei Kniegelenk (davon 14,4 Prozent Bandläsion; 14,1 Prozent Kreuzbandruptur und 2,3 Prozent Seitenbandruptur) bzw. mit 47,5 Prozent bei Sprunggelenk (davon 25,4 Prozent Banddistorsion und 22,1 Prozent Band-ruptur) und Prellungen (13,1 Prozent bei Knie- bzw. 20,1 Prozent bei Sprungge-lenk) mit alternativen Heilverfahren behandelt. Dabei dominierten Akupunktur (25,7 Prozent) und Homöopathie (25,3 Prozent); aber auch Ozontherapie (12,1 Prozent) und Manuelle Therapie (11,3 Prozent) wurden angewandt. Die Behandler wiesen auf eine positive Wirkung unkonventioneller Therapien bei der Prophylaxe hin. Sportler waren sich darunter in ihrer Mehrheit bewusst (91,8 Prozent), aber nur weniger als die Hälfte (39,7 Prozent) der Anwender nutzten diese Methoden auch tatsächlich für die Prophylaxe. Die Behandler waren mehrheitlich (82,1 Prozent) der Ansicht, dass sich die beiden Heilrichtungen sinnvoll ergänzen. Entscheidend dabei waren die Fest-stellungen, dass die Schulmedizin überfordert ist (31,5 Prozent), dass Schul-mediziner zu schnell zu operativen Methoden greifen (26,9 Prozent) und dass sie sich zu wenig Zeit für die Patienten nehmen (22,4 Prozent). Von daher er-scheint der Einsatz unkonventioneller Heilmethoden komplementär zur den schulmedizinischen Heilverfahren sinnvoll und komplementär zu sein, um eine rasche und vollständige Wiederherstellung bei Sportverletzungen von Knie- und Sprunggelenk zu erziehen Die in dieser Studie präsentierten Ergebnisse erlaubten neue Einblicke in die Problematik des Einsatzes unkonventionelle Heilmethoden bei der Behandlung von Sportverletzungen. Damit schließen sie eine Lücke in der Wissenschaft und erfordern zugleich eine weitere Betrachtung dieses relevanten Themas

Sperm parameters and DNA fragmentation of balanced chromosomal rearrangements carriers

Introduction. Somatic chromosomal rearrangements that occur in infertile males are thought to be one of the major genetic factors influencing male infertility. The objective of this retrospective study was to evaluate sperm parameters in a group of patients with balanced translocations. Material and methods. We analyzed semen of 84 balanced somatic translocation carriers [35 Robertsonian translocation (RT group) and 49 reciprocal translocation (RCT group)] and 57 men with normal karyotype (control group). Semen samples were evaluated for sperm concentration, its motion parameters and vitality, round cell number (CASA) and DNA fragmentation index (TUNEL). Cytogenetic evaluation was also performed for each study participant. Results. Sperm concentrations were lower when comparing the RT group to the control (p < 0.001) and RCT groups (p < 0.05). Occurrence of abnormal sperm concentration was more common among RT carriers (74.3%) than in the other groups (42.9% in RCT group and 28.1% in control group). The sperm progressive motility and vitality in RT carriers (21.53% and 62.17%) were lower than in control group (39.77% and 77.47%, p < 0.001, respectively) and RCT carriers (31.47% and 76.17%, p < 0.001, respectively). The RCT carriers and the control group did not differ in regard to sperm concentration, progressive sperm motility, motility grade D and sperm vitality. There were no significant differences in DNA fragmentation in carriers of both studied structural chro­mosomal rearrangements in comparison to subjects with normal karyotype. Conclusions. RT carriers had significantly lower semen parameters in comparison to not only the subjects with normal karyotypes but also the RCT carriers

Application of FISH method for preimplantation genetic diagnostics of reciprocal and Robertsonian translocations

Introduction. Carriers of reciprocal (RCP) and Robertsonian (RT) translocations are known to be at risk for reproductive difficulties. Preimplantation genetic diagnosis (PGD) is one of the options these carriers have to try to fulfill their desire to have a child. The FISH technique is one of the best method to detect RCPs, and, together with the Next Generation Sequencing, to diagnose RTs. The aim of the present study was to assess the usefulness of the FISH method for rapid diagnosis of translocations in our center to improve the reproductive counseling. Material and methods. From 2008 to 2012 one hundred and twenty seven fresh cycles of the in vitro fertilization (IVF; without freezing embryos) were performed in 42 couples with an RCP and 35 couples with an RT translocations. The patients were diagnosed before IVF as translocation carriers and therefore they opted for PGD. The classical FISH protocol has been applied with specific oligonucleotide probes. Results. In total 521 blastomeres were tested in order to determine the presence or absence of genetic anomalies resulting from one of the parents being a translocation carrier. Despite the large number of abnormal embryos (407 embryos — 78.1% of all examined embryos), 19.4% of blastomeres appeared to come from a normal or balanced embryos that may have been transferred to the uterus. In 63 of the 127 cycles embryo transfer (ET) was feasible and 24 women had a successful singleton or twin pregnancy. Thus, a live delivery rate of 18.9% per started cycles and 38.1% per cycle with ET was obtained. Conclusion. FISH should be regarded as an optimal preimplantation genetic diagnosis method for specific RCP and RT translocation carriers to increase the chance of successful IVF procedure

WNT5A gene and protein expression in endometrial cancer

Introduction. WNT5A (Wnt family member 5A) belongs to the WNT family of secreted signaling glycoproteins that play essential role in developmental, physiological and pathological processes. WNT5A was shown to take part in carcinogenesis process playing both oncogenic and suppressor functions in various types of human malignancies. This study aimed to assess the expression of the WNT5A gene at the mRNA and protein levels in the specimens derived from endometrial cancer (EC) or unchanged control endometrium. The associations between the WNT5A expression levels and clinicopathological characteristics and survival of EC patients were evaluated. Materials and methods. Total RNA was isolated in order to assess the relative amounts of WNT5A mRNA by quantitative polymerase chain reaction (QPCR) in samples of unchanged endometrial control (n = 8) and tumor samples of EC patients (n = 28). Immunohistochemistry (IHC) was used to determine the presence of WNT5A protein in the sections of formalin-fixed, paraffin-embedded tissue specimens derived from unchanged endome­trial controls (n = 6) and EC tumors (n = 19). Significance of differences in WNT5A expression levels between the studied groups of EC patients and correlations between the WNT5A and demographic data, pathological features, hematological parameters and overall survival of the patients were evaluated by statistical analysis. Results. The level of WNT5A mRNA was decreased in EC in comparison to unchanged endometrium. WNT5A expression was associated with primary tumor invasion status exhibiting reduced level of transcripts in EC that involved organs beyond the uterus when compared to the uterus-confined cancers. WNT5A immunoreactivity was visualized in the cytoplasm and nuclei of EC cells as well as in the luminal and glandular epithelial cells of unchanged endometrium. WNT5A mRNA expression levels correlated negatively with cytoplasmic, and positively with nuclear immunoreactivity of the WNT5A protein in the EC cells. In addition, the relationships between blood leucocyte count (in particular granulocytes and lymphocytes) of patients with EC and their WNT5A mRNA and protein expression levels were established. A positive correlation between the nuclear immunoexpression of WNT5A protein in the cancer cells in cell nuclei and mean platelet volume in blood was also found. Conclusions. The results of the first study of WNT5A expression at the transcript and protein levels indicate that it could be considered as a potential marker of molecular changes that take place during EC development

WNT4 Expression in Primary and Secondary Kidney Diseases: Dependence on Staging

Background/Aims: WNT4 protein is important for kidney development. Its expression was found to be altered in experimental models of chronic kidney disease (CKD). However, the expression of the WNT4 gene has yet not been studied in human renal biopsy samples from patients with broad spectrum of glomerular disease and at different stages of CKD. Thus, the aim of the study was to assess the WNT4 gene expression in renal biopsies of 98 patients using the real-time PCR technique. Materials: In order to assess the relative amounts of mRNA, in samples of patients with manifestation of different renal diseases and separately at different stages of CKD, by QPCR, total RNA was isolated from human kidney tissues collected during renal biopsies. Results of blood and urine samples assessment were used to calculate the correlations of biochemical parameters with WNT4 gene expression in both studied groups. Results: After pathomorphological evaluation, 49 patients were selected as presenting the most common cases in the studied group. Among the patients who developed focal segmental glomerulosclerosis (FSGS; n = 13), IgA nephropathy (IgAN; n = 10), IgAN with morphological presentation of focal segmental glomerulosclerosis (IgAN/FSGS; n = 8), membranous nephropathy (MN; n = 12), and lupus nephritis (LN; n = 6) were included in the analysis. We found that the level of WNT4 mRNA was higher in kidney specimens obtained from patients with MN as compared to those diagnosed with LN or IgAN. A correlation between WNT4 gene expression and serum albumin and cholesterol levels was observed in patients with FSGS, while WNT4 mRNA levels correlated with plasma sodium in patients diagnosed with LN. After consideration of 98 patients, based on the KDIGO classification of CKD, 20 patients were classified as CKD1 stage, 23 as stage 2, 13 as stage 3a, 11 as stage 3b, 13 as stage 4, and 18 as stage 5. WNT4 gene expression was lower in the CKD patients in stage 2 as compared to CKD 3a. Correlations of WNT4 mRNA level at different stages of CKD with indices of kidney function and lipid metabolism such as serum levels of HDL and LDL cholesterol, TG, urea, creatinine, sodium, and potassium were also found. Conclusions: Our results suggest that altered WNT4 gene expression in patients with different types of glomerular diseases and patients at different stages of CKD may play a role in kidney tissue disorganization as well as disease development and progression

Seminal plasma modifies the transcriptional pattern of the endometrium and advances embryo development in pigs

Research Areas: Veterinary SciencesABSTRACT - Background: Seminal plasma (SP) promotes sperm survival and fertilizing capacity, and potentially affects embryo development, presumably via specific signaling pathways to the internal female genital tract. Objectives: This study evaluated how heterologous SP, infused immediately before postcervical artificial insemination (AI) affected embryo development and the transcriptional pattern of the pig endometria containing embryos. Materials and Methods: Postweaning estrus sows (n = 34) received 40-mL intrauterine infusions of either heterologous pooled SP or Beltsville Thawing Solution (BTS; control) 30 min before AI of semen extended to 10% of homologous SR. Embryos (all sows) and endometrium samples (3 sows/group) were removed during laparotomy 6 days after the infusion of SP or BTS to morphologically evaluate the embryos to determine their developmental stage and to analyze the endometrial transcriptome using microarrays (PORGENE 1.0 ST GeneChip array, Affymetrix) followed by qPCR for further validation. Results: Embryo viability was equal between the groups (similar to 93%), but embryo development was significantly (P < 0.05) more advanced in the SP-treated group compared to control. A total of 1,604 endometrium transcripts were differentially expressed in the SP group compared to the control group. An enrichment analysis showed an overrepresentation of genes and pathways associated with the immune response, cytokine signaling, cell cycle, cell adhesion, and hormone response, among others. Conclusions: SP infusions prior to AI positively impacted the preimplantation embryo development and altered the expression of the endometrial genes and pathways potentially involved in embryo development.info:eu-repo/semantics/publishedVersio

Ultrastructural characterization of mammalian k-fibers by large-scale electron tomography

Eukaryotic cells have to divide constantly in order to promote the growth of certain organs, to replace dying or damaged cells, or to set up an entire organism. These essential processes are called mitosis in the case of somatic cell division. Mitotic cell division is the process during which chromosomes, centrosomes, and microtubules (MTs) are involved to set up a bipolar structure called the “mitotic spindle”. This bipolar spindle is formed by MTs, which are presumably mainly organized from the centrosomes. However, more data are being published that suggest MTs nucleation can occur from other MTs or even a chromosome surface. These biopolymers are built from α/β-tubulin heterodimers and can dynamically grow and shrink to exert forces necessary for chromosome segregation. Previous studies of spindles during mitosis have allowed the identification of different MT classes based on their plus-ends interaction with different cellular target sites. One of the MT classes is the kinetochore microtubules (KMTs), which physically connect chromosomes and centrosomes (i.e. spindle pole) via a specialized protein structure termed the “kinetochore”. This kinetochore-to-spindle pole connection has been studied in many organisms. In budding yeast, this connection is established by only a single KMT. In contrast, multiple KMTs bind to each mammalian kinetochore and form an MT bundle also called “k-fiber”. The ultrastructural architecture of the mammalian k-fiber connection is not well documented. Currently, different models concerning the nature of the kinetochore-to-spindle pole connection via k-fibers are discussed in the literature, i.e. a direct, semi-direct or indirect connection. The widely accepted ‘direct’ model proposes that all k-fibers of the mammalian spindle are formed through tight bundles of up to 20 KMTs, with all MT minus ends associated with the centrosome. However, it is necessary to understand the k-fibers structure in order to interpret its role during chromosome segregation. Here the architecture of the k-fiber was studied in human HeLa, U2OS and RPE-1 cell lines, as these different types of cells have been widely used in studies of mitosis. This thesis aimed to systematically investigate the characteristics of mammalian k-fibers and their attachment to the kinetochore within mammalian metaphase spindles. For that, the ultrastructure of mitotic spindles and k-fibers were analyzed using serial-section electron tomography primarily in HeLa cells. Furthermore, the spindle ultrastructure was compared by electron tomography to metaphase spindles in both U2OS and RPE-1 cells. Electron tomographic analysis of the mitotic spindle in HeLa cells revealed that the kinetochore-to-spindle pole connection is formed by k-fibers consisting of ~9 KMTs. Moreover, the data revealed that not all KMTs in k-fibers are directly associated with one of the spindle poles. Instead, KMT ends were located along the length of k-fibers indicating strongly for a semi-direct connection between the kinetochores and the spindle poles. Unexpectedly, by correlating the k-fiber ultrastructure with its position in the mitotic spindle, it can be demonstrated that the k-fiber structure varied depending on the position on the metaphase plate. It can also be shown that k-fibers located in the center of the metaphase plate had a tendency to form straighter and more bundled k-fibers. In contrast, k-fibers associated with the periphery of the metaphase plate had a more loose and disorganized structure resembling a fusiform shape. Furthermore, additional analysis of U2OS and RPE-1 cells indicated ultrastructural differences between the different cell lines. Mainly, differences between HeLa and RPE-1 cells were observed. K-fibers observed in RPE-1 cells showed a lower curvature and overall a more bundled ultrastructure compared to HeLa or U2OS cells. However, due to the small sample size for U2OS and RPE-1 cells, the results have to be confirmed in future experiments to conclude that there are indeed functional and structural differences in the k-fiber organization in different mammalian cell lines. Taken together, this work presents the first detailed quantitative ultrastructural analysis of KMTs in whole spindles in three different human cell lines. The data revealed that the currently favored direct model of k-fiber ultrastructure is oversimplified and needs to be corrected in terms of the k-fibers interaction with the spindle pole and the surrounding MT network within the mitotic spindle. The data here will serve as a structural basis for further analyses of mutant situations and contribute to our understanding of the overall organization and function of MTs in mitotic spindles.:Summary 6 Zusammenfassung 8 List of figures 10 List of tables 13 List of abbreviations and symbols 14 1 Introduction 19 1.1 The morphology of the mitotic spindle 21 1.1.1 Centrosomes 22 1.1.2 Microtubules 23 1.2 Kinetochores, KMTs and k-fibers 28 1.2.1 A brief history of k-fiber formation in mammalian cells 30 1.2.2 Models of the k-fiber ultrastructure in mammalian cells 32 2 Aims of this thesis 35 3 Materials and methods 36 3.1 Materials 37 3.1.1 Mammalian cell lines 37 3.1.2 Chemicals 38 3.1.3 Instrumentation and materials 40 3.1.4 Solutions and buffers 44 3.1.5 Software 46 3.2 Methods 47 3.2.1 Handling of cell cultures 47 3.2.2 Custom-designed incubation chambers 49 3.2.3 Specimen preparation for electron microscopy 51 3.2.4 Quality assessment of samples, acquisition of the tomographic data, and the 3D reconstruction 59 3.2.5 Ultrastructural analysis of MTs in mitotic spindles 62 3.2.6 Ultrastructural analysis of the k-fiber organization 70 4 Results 76 4.1 Initial characterization of mammalian mitotic spindles 77 4.2 Ultrastructure of KMTs 84 4.3 Curvature and tortuosity of KMTs 91 4.4 Ultrastructure of k-fibers 98 4.5 Effect of metaphase position on the k-fiber ultrastructure 102 5 Discussion 110 5.1. Comparison of data sets from different cell lines 111 5.2. Establishing a data analysis pipeline for the analysis of KMTs 113 5.3 Ultrastructural characterization of KMTs and k-fibers in HeLa cells 114 5.3.1 K-fibers have an unexpectedly low number of KMTs 115 5.3.2 Semi-direct kinetochores-to-spindle pole connection 117 5.3.3 Shape of k-fibers 121 5.4 Positional effect on the k-fiber shape 124 5.5 Comparison of k-fiber ultrastructure in different mammalian cells 127 5.6 Outlook 130 References 133 Appendix 1 149 Appendix 2 150 Appendix 3 151 Appendix 4 152 Acknowledgments 15

RRobert92/MT_Analysis: The Custer for Spatial-Graph Analysis

Added: performance improvement Fiber area analysis Neighborhood density analysis Minus end as seed along KMT analysi

RRobert92/MT_Analysis V1.21

Bug fix from version 1.