Andrology: Application of different in-situ hybridization detection methods for human sperm analysis

The detection of some types of aneuploidy in human spermatozoacan be based on the use of the fluorescence in-situ hybridizationtechnique (FISH). One of the crucial steps for FISH is to achievea proper decondensation and denaturation of the DNA in the specimen,so as to obtain efficient hybridization results. However, afterDNA decondensation the morphology of sperm heads is partly distortedand the majority of the tails is lost. This situation leadsto problems in the distinction between disomic and diploid spermatozoa,as well as between abnormal spermatozoa and somatic cells. Double-and triple-target FISH can partly solve this discriminationproblem. To improve these procedures we adapted the steps ofdecondensation and visualization of the single sperm cells.Firstly, DNA decondensation with 25 mM dithiothreitol in 1 MTris at pH 9.5 resulted in sperm cells with intact morphologyof both the head and the tail, and allowed efficient single-,double- and triple-target ISH to be performed. Secondly, weapplied a novel detection method, based on enzyme immunocyto-chemicalreactions, with coloured precipitation products. Thirdly, thisISH procedure was combined with Diff-Quik staining and bright-fieldmicroscopy. This absorption method has the advantage of a permanentsignal, and the adapted cytoplasmic staining of the sperm plasmamembrane allows the visualization of the outline of the singlespermatozoon. Using this approach, therefore, it is possibleto discriminate between disomic, diploid and abnormal spermatozoa,somatic cells and spermatozoa that overlap, because the morphologyof the cells is not distorted and the tails of the spermatozoaare intact and properly visualized.     

Restricted field of view magnetic resonance imaging of a dynamic time series.

A restricted field of view (rFOV) approach for imaging a dynamic time series of volumes of limited spatial extent within a larger subject is described. The shorter readout with rFOV-MRI can be exploited to either limit image artifacts or increase spatial resolution. To accomplish rFOV imaging of a multislice volume for a dynamic series, an outer volume suppression (OVS) preparation that saturates signal external to a cylinder through the subject is followed by slice-selective excitation and a spiral readout. The pass- and stopband efficiencies of the OVS in an agar gel phantom were 97% (+/-1.5%) and 3% (+/-1%), respectively. Profiles of the temporal signal-to-noise ratio (SNR) were measured in a phantom and an adult brain. The rFOV sequence reduced distortions from off-resonance signal and T2*-induced blurring compared to a conventional sequence. Sequence utility is demonstrated for high-resolution rFOV functional MRI (fMRI) in the visual cortex. The rFOV sequence may prove to be useful for other multislice dynamic and high-resolution imaging applications.     

Sister-Sister in vitro fertilization surrogate pregnancy with donor sperm: The case for surrogate gestational pregnancy

A case of surrogate pregnancy is described in the sister of a 40-year-old hysterectomized woman where two oocytes of the latter were fertilized in vitro with known donor sperm and transferred into the surrogate. A normal singleton pregnancy developed which was complicated after 24 weeks of gestation with recurrent antepartum hemorrhages due to grade 3 placenta praevia. A healthy female baby was delivered by elective cesarean section at 36 weeks of gestation. The legal, social, psychological, and ethical issues of surrogacy remain unsettled and are discussed in this case report.     

Neurogenic muscle hypertrophy in radiculopathy

The course of radiculopathy is sometimes associated with weakness and wasting of muscles. Very rarely in such cases, however, is hypertrophy of muscle fibres observed. Three cases are presented of sciatica with enlarged calves caused by hypertrophy of type 1 or types 1 and 2 muscle fibres. In light of the literature and the results obtained, an attempt is made to explain the cause of rare clinical symptoms and draw attention to diagnostic and therapeutic difficulties.     

Effect of meal volume on gastric emptying

Abstract While the volume of a liquid meal has been identified as the principal accelerator of gastric emptying of liquids, the relationship between meal volume and gastric emptying of solids has been controversial. With solid foods, the need to reduce solid foods into small particles (trituration) before passage might obscure the effect of meal volume on solid propulsion. To distinguish trituration from driving force as the rate-limiting factor for emptying, 75 (1.6 mm) nylon spheres were fed along with different amounts of steak meals (150, 300 and 600 g), or alternatively, 50, 100 or 200 (1.6 mm) nylon spheres were fed to six dogs with 300 g steak meals. To examine the effect of meal volume on gastric emptying, we studied the effect of different meal volumes on the speed of gastric emptying of liquids (150, 300, 600 and 1200 ml of phosphate buffer) and solids (150, 300 and 600 g of cooked beef steak) in five dogs with duodenal fistulas. Intestinal inhibition was eliminated by diverting all chyme through the fistulas. In the absence of intestinal feedback, we found that gastric emptying of steak and spheres were different in that steak emptying was independent of meal volume (g min^-1 was constant across 150–600 g) while sphere emptying was affected by the number of spheres in the stomach and that liquid emptying was dependent on the meal volume (ml min^-1 increased across 150–1200 ml). Thus, meal volume accelerated gastric emptying provided the process is not rate-limited by trituration.     

Evaluation of computerized analysis of the propagation of human duodenojejunal contractions

Abstract Detection of peristalsis in the human small intestine has been limited in the past by both the available measurement techniques and the complexity of this activity. Recent developments in ambulant recording have provided a means of monitoring the occurrence of intestinal contractions at multiple sites in the small bowel, but the problem of complexity remains. Using digital data recorded from an intra-luminal strain-gauge transducer in the proximal gut, an algorithm was implemented to identify and classify contractile events within the small bowel. By modelling propagated activity the effect of varying transducer spacing and the number of transducers used was assessed. The question of variability of apparent velocity of peristaltic contractions was examined using successive cross-correlation calculations to extract underlying phase differences between samples of 512 minutes of manometric recording over 150 mm of human small bowel. The effective velocity was found to have a median value of 14 mm sec^-1 and an inter quartile range of 12–18 mm see^-1). It is proposed that, in dynamically tracking variations in phase difference between adjacent recording sites, cross-correlation techniques should be used to control the parameters used for the recognition of propagated contractile events and thereby improve the specificity of this process.     

A 4-year controlled clinical study into the use of a ceramic hydroxylapatite implant material for the treatment of periodontal bone defects

10 patients with chronic adult periodontitis who had greater than 1 tooth with infra-bony pockets were treated at the test defects by periodontal flap procedures with implantation of hydroxylapatite particles; the control defects were treated by the same surgical procedures but without the implant. A total of 58 test defects and 59 control defects were treated. Each defect had measurements carried out at given sites on the involved tooth surfaces, the sites being considered for subsequent tabulation purposes under the category of shallow (less than 3 mm) moderate (3-6 mm) and deep (greater than 6 mm) initial pocket depths. There were 146 and 152 shallow sites, 216 and 241 moderate sites and 140 and 133 deep sites, at test and control sites, respectively. Measurements of recession, probing pocket depths and probing attachment levels were made at 6 months and 1, 2, 3 and 4 years. At all sites over the period of the study, for the moderate and deep initial pockets there was a significant reduction in probing depths and an increase in the probing attachment levels. At the 4th year of assessment for the initially deep pockets, the reduction in probing depths was significantly greater for the sites treated with the implant material. In view of the difficult clinical problem posed by the treatment of teeth with deeper periodontal bone defects, further research using either this type of implant material or similar material should be considered.     

Astrocyte and microglial motility in vitro is functionally dependent on the hyaluronan receptor RHAMM

RHAMM (Receptor for Hyaluronic Acid Mediated Motility) has been identified as a receptor for the extracellular matrix component hyaluronan (HA) and was recently shown to be essential for the locomotion of normal and transformed peripheral cells. Until now the potential role of RHAMM in the motility of neural-derived cells has not been investigated. Here, we report that cultured primary astrocytes, astrocyte cell lines, and microglia express this receptor and exhibit RHAMM-dependent motility. Immunocytochemical localization of RHAMM showed that it was often present as aggregates at the periphery of cells in contact with one another or concentrated on protruding processes of isolated cells. Glial cells contained 50 and 72 kDa forms of RHAMM, and both of these forms were found to have HA binding capacity. Time lapse imaging of cell locomotion revealed a significant inhibition of motility and process elongation by neutralizing anti-RHAMM antibodies and by peptides corresponding to the HA binding domains of RHAMM. These results demonstrate that RHAMM serves a role in glial cell locomotion in vitro and provide the basis for investigations of the motile behavior of glial cells in vivo after CNS injury.     

Bone Replacement Materials in Hand Surgery

Abstract Background: In recent years, biomaterials are being found and frequently utilized in bone defects. They have also gained significant precedence in hand surgery. Objectives: The respective requirements for such replacement material will be cited and acknowledged in this article. The individual material groups will also be referred to in this review. An introduction to some of the customary bone replacement materials will be cited and concluded with a corresponding recommendation. Conclusion: The implantation of autologous cancellous bone is still regarded today as the “gold standard”. Nevertheless, the usage of bone replacement material can be an enormous advantage in certain indications. The original article can be found online at There was an error in the author’s affiliation and the address for correspondence was incomplete. Please note the correct institution and complete address: Department of Orthopedics and Traumatology, Hand-, Foot- and Reconstructive Surgery, Kreiskrankenhaus Gummersbach GmbH, Germany. Alexander von Friesen, MD Department of Orthopedics and Traumatology, Hand-, Foot- and Reconstructive Surgery Kreiskrankenhaus Gummersbach GmbH Wilhelm-Breckow-Allee 20 51643 Gummersbach Germany Phone (+49/2261) 171-575, Fax -449 e-mail: Friesen@kkh-gummersbach.de