Factors affecting cryopreservation-associated damages in sperm motility of cockerels (Gallus gallus domesticus).

1. Sperm are exposed to severe osmotic stress during cryopreservation, which results in impairment of fertilisation ability, including motility and viability, in poultry. Sperm osmotolerance is regulated by many extracellular factors and varies widely in birds, leading to uncertainty in the nature of the osmotic injury.2. Tail bending is a primary response resulting from cell swelling from excessive osmotic stress. However, the underlying mechanism responsible for tail bending is largely unknown. This study examined the relationship between osmotic stress and post-thaw motility, with a particular focus on the role of Na+/K+ ATPase (NKA) in the tail bending response.3. Cryopreserved sperm exhibited rapidly reduced motility when maintained at 37°C. The combination of temperature change and osmotic stress was a primary factor responsible for tail bending. This work tested a hypothesis known to be associated with post-thaw tail abnormality in other species and found that cold shock, that is not accompanied by an apoptotic response, may occur. Ouabain inhibition of Na+/K+ ATPase activity alleviated the tail bending response in fresh and post-thaw sperm.4. These results demonstrated that the combination of temperature change and osmotic stress has a primary impact on the reduction of post-thaw motility, with a particular role in NKA activity, in the tail bending response of chicken sperm. These results provide a foundation for establishing cryopreservation methodology to ensure the optimal fertilisation potential of cryopreserved chicken sperm.

A surgical navigation system for aortic vascular surgery: a practical approach.

In aortic vascular surgery, a navigation system must represent the anatomical map of individual patient in order to detect the important artery. To provide a proper fit for positions along the dorsoventral axis, the spinous process was added to a currently used anatomical point set consisting of four anterior body landmarks. In addition, we attempted to reduce the registration error by compensating for alignment errors resulting from variations in tissue thickness at each landmark. The alignment values were examined using a human phantom consisting of a skeleton model with subcutaneous tissue in the semilateral position. Using this method, a phantom simulation and five clinical trials were performed. Target errors were evaluated at the orifice of the intercostal artery. In the phantom simulation, the error at the target point was 4.1 ± 2.7 mm. However, for one patient undergoing thoracoabdominal aortic aneurysm replacement surgery, the target error was 8.0 mm using the proposed method.

Usefulness of 4D-CTA in the detection of cerebral dural sinus occlusion or stenosis with collateral pathways.

In time-resolved CT angiography (4D-CTA), it is of substantial merit to detect the veins and sinuses of the whole brain with the simultaneous demonstration of the natural drainage flow in order to find occlusion or stenosis of the dural sinuses with collateral pathways. As preoperative information for patients with brain tumors, it is important to detect feeding arteries, incidentally found aneurysms or other vascular lesions, and to detect patency of the dural sinuses and the important cortical veins, whether they are compressed by tumors or not. On the other hand, cerebral venous thrombosis (CVT) may occur in patients due to various causes, which has not been unusual in recent years. For patients with acute symptomatic or chronic non-symptomatic CVT, identification of dural sinus occlusion (DSO) or dural sinus stenosis (DSS) and compensatory collateral pathways is necessary for suitable thrombolytic therapy or careful investigation to avoid further CVT. This study reviews our experiences in 116 cases of 4D-CTA for 90 patients with brain tumors and 26 other patients including 11 with arteriovenous malformation, and four with acute CVT and other conditions. 4D-CTA presented DSO/DSS with compensatory venous collateral pathways, which was helpful to detect the severity of the venous abnormality, and see whether it was compressed by brain tumors, or due to other causes in patients with symptomatic or non-symptomatic CVT. 4D-CTA is a useful non-invasive diagnostic tool to detect cerebral venous abnormalities as an alternative to DSA.

Interlaboratory validation of in vitro eye irritation tests for cosmetic ingredients. (2) Chorioallantoic membrane (CAM) test.

A chorioallantoic membrane (CAM) assay evaluates the blood vessel reaction and damage to the CAM of a fertilized hen's egg. Two types of CAM assays, the hen's egg test-chorioallantoic membrane (HET-CAM) method and the chorioallantoic membrane-trypan blue staining (CAM-TB) method, were evaluated as alternative methods to the Draize eye irritation test (Draize test). The validation project was composed of three test phases in which 10, 15 and 14 test chemicals, respectively, were evaluated. The test procedure of the five independent laboratories was controlled under the same standard operating procedure (SOP). The interlaboratory variation was relatively high for both methods. However, the rank correlation was relatively high among the values obtained by the five laboratories. The variation associated with the CAM-TB method was smaller than that of the HET-CAM method, which requires macroscopic observation, suggesting that the objectivity and quantitativeness differs between the assay systems. The average values using these two methods were compared with the maximum average Draize total score (MAS). The correlation coefficient (r) between the HET-CAM scores and the MAS was 0.688. This suggests that a simple linear regression may not be appropriate for HET-CAM. However, the Spearman's rank correlation coefficient (rs) was relatively high (rs=0.802). In contrast, the CAM-TB test results showed a good correlation with the MAS when the test chemicals were classified according to their physical properties (r=0.801, liquid and r=0.926, powder). These results suggest that both the HET-CAM and CAM-TB methods may present alternative method of evaluation of eye irritation despite problems of interlaboratory reproducibility.