The use of granulocyte colony stimulating factor to enhance oocyte release in women with the luteinized unruptured follicle syndrome.

PURPOSE: To determine if an injection of granulocyte colony stimulating factor (G-CSF) prevoulatory can enable oocyte release from the follicle in women who have failed to release in natural cycles despite an endogenous luteinizing hormone (LH) surge, and also despite treatment with human chorionic gonadotropin (hCG) or a gonadotropin releasing hormone agonist (GnRHa). MATERIALS AND METHODS: A single injection of 100 mg G-CSF was given in the late follicular phase followed by hCG 10,000 units at peak follicular maturation in women with at least three consecutive cycles of luteinization without oocyte release. RESULTS: Six women had ten cycles with G-CSF and hCG. Definite release occurred in four, inconclusive in four, and definitely the luteinized unruptured follicle in two. Biochemical pregnancies occurred in two of the cycles where oocyte release occurred and a live delivered pregnancy in another cycle of release. CONCLUSIONS: Without controls one cannot state with certainty that G-CSF enabled oocyte release when hCG and leuprolide failed. Nevertheless, the data do support a trial with G-CSF before proceeding to IVF-ET.

In non-transformed cells Bak activates upon loss of anti-apoptotic Bcl-XL and Mcl-1 but in the absence of active BH3-only proteins.

Mitochondrial apoptosis is controlled by proteins of the B-cell lymphoma 2 (Bcl-2) family. Pro-apoptotic members of this family, known as BH3-only proteins, initiate activation of the effectors Bcl-2-associated X protein (Bax) and Bcl-2 homologous antagonist/killer (Bak), which is counteracted by anti-apoptotic family members. How the interactions of Bcl-2 proteins regulate cell death is still not entirely clear. Here, we show that in the absence of extrinsic apoptotic stimuli Bak activates without detectable contribution from BH3-only proteins, and cell survival depends on anti-apoptotic Bcl-2 molecules. All anti-apoptotic Bcl-2 proteins were targeted via RNA interference alone or in combinations of two in primary human fibroblasts. Simultaneous targeting of B-cell lymphoma-extra large and myeloid cell leukemia sequence 1 led to apoptosis in several cell types. Apoptosis depended on Bak whereas Bax was dispensable. Activator BH3-only proteins were not required for apoptosis induction as apoptosis was unaltered in the absence of all BH3-only proteins known to activate Bax or Bak directly, Bcl-2-interacting mediator of cell death, BH3-interacting domain death agonist and p53-upregulated modulator of apoptosis. These findings argue for auto-activation of Bak in the absence of anti-apoptotic Bcl-2 proteins and provide evidence of profound differences in the activation of Bax and Bak.

Comparison of reconstruction nails for high subtrochanteric femur fracture fixation.

Subtrochanteric osteotomies were created in 18 matched pairs of embalmed cadaveric femora. The femora were stabilized with a Synthes, Zimmer, or Richards second generation femoral reconstruction nail with retrograde blade or screws. The femoral pairs were randomly assigned to groups based on nails used: Synthes versus Zimmer, Synthes versus Richards, and Zimmer versus Richards. The reconstructions were cyclically loaded in bending for 2000 cycles and then loaded to failure. The mean stiffness of the Synthes, Zimmer, and Richards reconstructions was 17%, 40%, and 40% of the intact femora, respectively. The Richards construct was the strongest, and predominately failed by fracture at the distal interlocking screw hole. The Zimmer construct failed by bending of the nail at the osteotomy site and fracture of the proximal femoral shaft. The Synthes construct was the most flexible and least strong and failed by bending of the spiral, retrograde blade with concomitant fracture of the femoral neck. This study indicates that fixation of subtrochanteric femur fractures with a Synthes spiral blade or Richards or Zimmer reconstruction nails provides stable fixation for postoperative loading conditions. However, the Richards and Zimmer nails were able to withstand higher loads than was the Synthes nail before failure.

Effect of periosteal stripping on cortical bone perfusion: a laser doppler study in sheep.

The goals of internal fixation are an accurate reduction and stable fixation in the presence of adequate bony vascularity. This can be achieved by a variety of means including plate fixation. A certain amount of periosteal stripping is necessary for proper open reduction of a fracture and for proper plate application. With displaced diaphyseal fractures, cortical bone perfusion (CBP) is already compromised. Further damage, in terms of periosteal stripping for plate fixation, may not be acceptable. Little information is available as to what extent the periosteum contributes to cortical bone perfusion. The purpose of this study was to determine the acute effects of periosteal stripping on cortical bone perfusion in a sheep tibia model. Twenty-three sheep were operated on and had the medial aspect of their right tibia exposed. Cortical bone perfusion measurements were obtained using laser Doppler flowmetry prior to periosteal stripping and after periosteal stripping. The results of this study show that the cortical bone perfusion significantly decreased by 20% after periosteal stripping over the entire length of the tibia. We therefore conclude that the periosteum contributes to diaphyseal bone perfusion and that it is important to preserve this source with fractures where blood supply is already significantly compromised.

Cortical bone perfusion in plated fractured sheep tibiae.

The limited contact dynamic compression plate and partial contact plate were designed to decrease contact with cortical bone in an attempt to decrease cortical ischemia, remodeling, and eventual porosis under the plate after use of standard dynamic compression plates. This study quantified cortical bone blood flow beneath the plate with these three different designs in a sheep tibia fracture model. In 18 skeletally immature sheep, the right tibia was fractured and then was internally fixed with an interfragmentary screw and a dynamic compression plate, limited contact dynamic compression plate, or partial contact plate. At 12 weeks, cortical bone perfusion was assessed with laser Doppler flowmetry in nine areas beneath the plate. The baseline (before fracture) cortical bone cell flux averaged 100 +/- 60 mV. After fracture, this decreased to 60 +/- 48 mV (p < 0.0003); immediately after plating, the perfusion averaged 29 +/- 25 mV (p < 0.01). Cortical bone perfusion then increased to 106 +/- 52, 165 +/- 71, and 163 +/- 71 mV at 2, 6, and 12 weeks after fracture (p < 0.001 for all when compared with values after plating). No significant differences in cortical perfusion were seen between the types of plate. Cortical porosity under the plate was assessed with digital density analysis of microradiographs of this region. No significant difference was seen between the types of plate in this analysis or in biomechanical and disulphine blue perfusion analysis. Thus, no significant advantage was seen for the new plate designs used in this model. This lack of advantage may be a result of the immature animals used in the study, the protocol for blood flow measurement, the invasive periosteal stripping employed to create the fracture, or all three. However, as advantages with the new plate designs have been seen in other studies, this area warrants further investigation.

Cortical bone blood flow in reamed and unreamed locked intramedullary nailing: a fractured tibia model in sheep.

We compared the effects of reamed versus unreamed locked intramedullary nailing on cortical bone blood flow in a fractured sheep tibia model. A standardized spiral fracture was created by three-point bending with torsion, and each tibia was stabilized by insertion of a locked intramedullary nail. Eleven animals were randomized into two groups: one that had reaming before nail insertion and one that did not. Blood flow was measured in real time using laser Doppler flowmetry. Cortical bone perfusion measurements were made at three locations (proximal diaphysis, fracture site, distal diaphysis) and at eight time intervals (prefracture, postfracture, postreaming, postnail insertion, postlocking, and at 2-, 6-, and 12-week follow-up). All animals were killed at 12 weeks postoperatively. After reamed nail insertion, cortical bone perfusion was significantly decreased (p < 0.0009). After unreamed nail insertion, perfusion was decreased less (p < 0.003). Insertion of locking screws did not affect blood flow. Cortical bone perfusion was greater in the unreamed group at completion of the procedure (p < 0.011), at 2-week follow-up (p < 0.006) and at 6-week follow-up (p < 0.027). The findings suggest that cortical revascularization had occurred by 6 weeks in the unreamed group but not until 12 weeks in the reamed group. The study demonstrates that cortical circulation is spared to a greater degree by unreamed nailing. This may be advantageous in severe open tibial fractures where blood supply is already significantly compromised.

Geometric changes in the cervical spinal canal during impact.

SUMMARY OF BACKGROUND DATA: Although the extent of injury after cervical spine fracture can be visualized by imaging, the deformations that occur in the spinal canal during injury are unknown. STUDY DESIGN: This study compared spinal canal occlusion and axial length changes occurring during a simulated compressive burst fracture with the residual deformations after the injury. METHODS: Canal occlusion was measured from changes in pressure in a flexible tube with fluid flowing through it, placed in the canal space after removal of the cord in cadaver specimens. To measure canal axial length, cables were fixed in C1 and led through the foramen transversarium from C2-T1, then out through the base, where they were connected to the core rods of linearly variable differential transformers (LVDT). Axial compressive burst fractures were created in each of ten cadaveric cervical spine specimens using a drop-weight, while force, distraction, and occlusion were monitored throughout the injury event. Pre- and post-injury radiographs and computed tomography scans compared transient and post-injury spinal canal geometry changes. RESULTS: In all cases, severe compressive injuries were produced. Three had an extension component in addition to compression of the vertebra and retropulsion of bone into the canal. The mean post-injury axial height loss measured from radiographs was only 35% of that measured transiently (3.1 mm post-injury, compared with 8.9 mm measured transiently), indicating significant recovery of axial height after impact. Post-injury and transient height loss were not significantly correlated (r2 = 0.230, P = 0.16) demonstrating that it is not a good measure of the extent of injury. Similarly, mean post injury canal area was 139% of the minimum area measured during impact, indicating recovery of canal space, and post-injury and transient values were not significantly correlated (r2 = 0.272, P = 0.12). Mean post-injury midsagittal diameter was 269% of the minimum transient diameter and showed a weak but significant correlation (r2 = 0.481, P = 0.03). CONCLUSIONS: Two potential spinal cord injury-causing mechanisms in axial bursting injuries of the cervical spine are occlusion and shortening of the canal. Post-injury radiographic measurements significantly underestimate the actual transient injury that occurs during impact.

Cortical bone microperfusion: response to ischemia and changes in major arterial blood flow.

In order to investigate the effects of short-term ischemia on cortical bone microperfusion, an isolated porcine tibia diaphyseal preparation based on intact nutrient vessels was developed. Laser Doppler flowmetry (LDF) was utilized to assess continuously the cortical microcirculation and the response to short-term ischemia. The femoral artery was isolated and clamped to develop the condition of bone ischemia. On release of the clamp, reactive hyperemia was documented in all animals. Using a roller pump connected to a segment of femoral artery, the same preparation was utilized to investigate the effect of a changing femoral artery flow on the cortical microcirculation. A positive correlation between LDF output and change in arterial inflow (r = 0.64) was defined. This model has the potential for studying the effect of ischemia on bone cell viability.

Influence of hycanthone on morphology and serotonin uptake of Schistosome mansoni.

The intrinsic content of serotonin (5HT) and the uptake of 5HT by Schistosoma mansoni taken from mice which were given a single intramuscular therapeutic dose of hycanthone were studied. Drug-exposed worms were found to have intrinsic values of 5HT which were similar or slightly less than controls. Uptake measurements were made on single and on paired worms recovered from mesenteric, portal, or hepatic sites and incubated in 75% horse serum or in Fischer's medium. All groups of treated worms were found to take up, on average, similar or lower amounts of 5HT compared to controls. These findings are in contrast to a recent report of very considerable increases in content or in 5HT acquisition in vitro by hycanthone-exposed parasites. This communication suggests that the mode of action of hycanthone cannot be explained as being due to increased 5HT uptake. Morphological changes in hycanthone-treated worms include loss of body weight and size, loss of hemoglobin pigment from the gut, deterioration of the tegument, and derangement of the vitellaria. The loss of gut contents occurs early after exposure to hycanthone and may indicate that interference with gut physiology and the nutritional state of the worms is one consequence of the drug, although the mechanism of these changes has not yet been elucidated.