Determining Direction of Axonal Flow in the Equine Ramus Communicans by Ultrastructural Examination of the Plantar Nerves 2 Months after Transecting the Ramus.

The ramus communicans, neural connection between medial and lateral plantar nerves of the horse, was transected to determine the degree to which medial and lateral plantar nerves contribute to the plantar ramus. After 2 months, sections of plantar nerves immediately proximal and distal to the communicating branch were collected and processed for electron microscopy. All examined nerves had undergone Wallerian degeneration and contained regenerating and mature fibers. Layers of the myelin sheath were separated by spaces and vacuoles, indicating demyelination of medial and lateral plantar nerves. Shrunken axons varied in diameter and were surrounded by an irregular axolemma. Shrunken axoplasm of both myelinated and non-myelinated fibers contained ruptured mitochondria and cristae, disintegrating cytoskeleton, and vacuoles of various sizes. The cytoplasm of neurolemmocytes contained various-sized vesicles, ruptured mitochondria within a fragile basal lamina and myelin whorls of multilayered structures indicative of Wallerian degeneration. These ultrastructural changes, found proximal and distal to the ramus in medial and lateral plantar nerves, suggest that axonal flow is bi-directional through the ramus communicans of the pelvic limbs of horses, a previously unreported finding. As well, maturity of nerves proximal and distal to the ramus indicates that all nerve fibers do not pass through the ramus.

Excision of the deep branch of the lateral palmar nerve of horses to resolve lameness caused by proximal suspensory desmitis.

OBJECTIVE: To assess outcome after neurectomy of the deep branch of the lateral palmar nerve (DBLPaN) as a treatment for horses with persistent lameness associated with chronic proximal suspensory desmitis (PSD) of the thoracic limb. STUDY DESIGN: Case series. ANIMALS: Adult, mixed-breed horses (n = 4), weighing 510-585 kg, used for amateur show-jumping. METHODS: Records of 4 horses chronically lame because of PSD of one or both thoracic limbs that were treated by neurectomy of the DBLPaN were reviewed. The site of pain causing lameness was localized using regional anesthesia. The proximal aspect of the suspensory ligament of the affected limb(s) of all horses were enlarged on ultrasonographic examination, but fiber disruption was not observed. All horses remained lame after conservative therapy. Neurectomy was performed with the horses anesthetized and positioned in dorsal recumbency. RESULTS: All 4 horses were sound at 6 weeks and remained sound for at least 12 months after neurectomy. CONCLUSION: Lameness in horses caused by chronic PSD can be resolved by neurectomy of the DBLPaN in horses that are refractory to conservative management.

A proposed new classification for the renal collecting system of cattle.

OBJECTIVE: To evaluate the intrarenal anatomy of kidneys obtained from cattle and to propose a new classification for the renal collecting system of cattle. SAMPLE POPULATION: 37 kidneys from 20 adult male mixed-breed cattle. PROCEDURES: Intrarenal anatomy was evaluated by the use of 3-D endocasts made of the kidneys. The number of renal lobes and minor renal calyces in each kidney and each renal region (cranial pole, caudal pole, and hilus) was quantified. RESULTS: The renal pelvis was evident in all casts and was classified into 2 types (nondilated [28/37 {75.7%}] or dilated [9/37 {24.3%}]). All casts had a major renal calyx associated with the cranial pole and the caudal pole. The number of minor renal calices per kidney ranged from 13 to 64 (mean, 22.7). There was a significant correlation between the number of renal lobes and the number of minor renal calices for the entire kidney, the cranial pole region, and the hilus region; however, there was not a similar significant correlation for the caudal pole region. Major and minor renal calices were extremely narrow, compared with major and minor renal calices in pigs and humans. CONCLUSIONS AND CLINICAL RELEVANCE: The renal collecting system of cattle, with a renal pelvis and 2 major renal calices connected to several minor renal calices by an infundibulum, differed substantially from the renal collecting system of pigs and humans. From a morphological standpoint, the kidneys of cattle were not suitable for use as a model in endourologic research and training.

The bovine kidney as an experimental model in urology: external gross anatomy.

The objective of this work was to obtain and record detailed and accurate measurements of the bovine kidney and to compare these new data with findings in humans. Thirty-eight bovine kidneys were used. The total number of lobes, along with the number of lobes located in the cranial polar, caudal polar and hilar regions, were recorded. Several measurements of the kidneys were made and evaluated. The hilar region presents the greatest length (mean of 76.87 mm) of the 3 renal regions of the kidney. The large area of the bovine renal hilus could make access to hilar structures easier than in the human kidney. The coefficient of variation for renal length was small (8.14%), while the coefficient of variation for the lobar number was high (26.82%). The number of renal lobes ranged from 13 to 35, with a mean of 20.62. The hilar region presents the highest number of lobes, while the cranial pole presents the lowest. The number of lobes in the cranial and caudal poles increases with the width of these regions. This is different from the hilar region, in which the lobar number increases with the length of the hilus. These data indicate that the adult bovine kidney can be used as a model for certain urologic procedures, but researchers must be aware that there are some major differences between the adult bovine kidney and the human kidney, as indicated by the data reported in this paper.

Room temperature/Corcoran/Dow Corning™-Silicone plastination process.

For 20 years, the cold temperature/S10/von Hagens' plastination technique was used to preserve biological specimens without challenge. It became the "gold standard" for preservation of beautiful, dry biological specimens. Near the end of the 21st century, a group from the University of Michigan and environs and Dow Corning™, USA, combined silicone ingredients, similar to the von Hagens' plastination products, however in a different sequence. The new polymer (Cor-tech) was combined with the cross-linker to design the "impregnation mix" which would invade the cellular structure of the specimen and yet was stable at room temperature. Later, curing would be by application of the catalyst onto the impregnated specimen. This unique sequencing of products would become the "Room temperature/Dow Corning™/Corcoran-Silicone plastination technique." The results of this room temperature technique provided similar plastinates, beautiful and practical for demonstration, containing no toxic chemical residues and forever preserved. As the name implies, impregnation of this silicone mix could be done at room temperature, without having to be kept cold. Both processes (cold and room temperature) required the same four basic steps for plastination. As well, both processes used similar basic polymers and additives to produce plastinates. However, they were combined in a different sequence. Cold temperature combines polymer and catalyst/chain extender, which is not stable and therefore must be kept colder than -15°C, while room temperature combines polymer with cross-linker which is stable, and likely forever.

Plastination of larger and massive specimens-With Silicone.

In 1977, plastination was unveiled, which replaced tissue fluid with a curable polymer. Today preservation via plastination of various animal and plant tissues, organs, and whole bodies is an extremely useful technique to display such and help educate vast arrays of both allied science students and the lay public across the planet. The diversity of applications of plastination techniques seems to be without limits. In fact, the only real limitation to plastination is one's imagination! The size of plastinates during the early years of plastination was comparatively small and dictated primarily by the size of the available plastination kettle/chamber, 35 L Heidelberg plastination kettle (49 cm H × 34.5 cm diam.). In the 1990s larger chambers were designed and slowly became available:150-210 cm (long) × 65-80 cm (wide) and 83-92 cm (high). Today a few large vacuum chambers are in service which will accommodate whole bodies of man and domestic or exotic animals. Today, at least two gigantic chambers are available to impregnate massive specimens. These are 3.5 m × 2 m × 1.5 m (Dalian) and 4 m × 3 m × 2.2 m (Guben). Also, the need for larger quantities of acetone and impregnation mix, not to mention the great increase in specimen preparation time, makes this a major investment. The "cold temperature process" is used to impregnate these massive creations. The room temperature technique could be used. The same four plastination steps are necessary for larger and massive specimens. Besides their tremendous size, the slippery silicone polymer is a reckoning force.

Cold temperature/Biodur® /S10/von Hagens'-Silicone plastination technique.

Plastination is a late 20th century preservation methodology which replaces tissue fluid within a specimen with a curable polymer, such as silicone. Plastination yields superb, beautiful, well-preserved specimens each with their own unique qualities. Silicone polymer is used around the world to preserve macroscopic cadavers or portions/organs thereof. Plastination was conceived by Dr. Gunther von Hagens, Universität Heidelberg, Heidelberg, Germany prior to 1977. Silicone polymer was the primary polymer which emerged initially for plastination. The Biodur® line of silicone polymer and additives was chosen and manufactured because it has consistently produced the best plastinates since the inception of plastination. Since the discovery of silicone, generic and similar silicone polymers are known and used around the World by many industries and used in numerous products. The plastination process has four steps: Specimen preparation, Specimen dehydration and degreasing, Vacuum-forced impregnation of specimens and Specimen hardening.

Coloring plastinated specimens.

In the early days of plastination, plastinate Color was the usual grey/brown familiar to formalin-fixed biological specimens. Initially, trials with Kaiserling's, Klotz, Jore's and McCormick's fixative solutions were disappointing. Vascular injections with Colored epoxy were a great breakthrough in the 1980s. Biodur AC10® stain was the first stain of note to be applied to gross specimens to be plastinated and was applied in the last acetone bath. As plastination became more popular, specimen Color became an important and necessary aspect. Reactivation of the normal Color of red blood cells within a formalin-fixed specimen was introduced as a mechanism to restore Color to plastinated specimens. Painting of plastinated vessels was tried with some success, and finally, a superior new proprietary type of silicone coloration was developed. More recently, a versatile red pigment stain was developed. All of these have added aesthetically to the plastination processes and will certainly be a reality in the years to come. The various methodologies to Color plastinates are presented. Time will tell how effective these may or may not be.

History and development of plastination techniques.

Plastination was a game-changing invention for macroscopic anatomical preparation. The method yielded dry, odourless, tangible and durable specimens which allowed new exhibition and teaching set-ups and paved the way for sophisticated preparations and spectacular positioning of specimens. Despite the impact of the new method, there have been similar techniques in place before. Exsiccation techniques, polymer embeddings and specimen impregnation with hardening substances were earlier methods which already included the main concepts that were later combined and refined in plastination. S10 silicone plastination, the technique most commonly known and applied, was followed by plastination methods suitable for research and sectional anatomy teaching. Numerous variations of sheet plastination techniques allow research applications and new ways of presenting topographic relations and mesoscopic insights. Besides the development of plastination techniques in sensu stricto, related techniques had a renaissance with new applications and developments, including corrosion casting and diaphonization methods. This brief review shall provide a historical context of plastination including some anecdotal spotlights on the ideas and innovations that lead to nowadays plastination techniques.

Proportional analysis of the pig renal parenchyma and sinus structures.

AIMS: This study was performed to determine the proportion of the parenchyma and sinus structures of pig kidneys and the distance between the collecting system and the kidney surface. METHODS: Forty-one pig kidneys were analyzed. Cavalieri's principle was used to obtain the volume of the cortex, medulla and sinus, as well the proportions of the arteries, veins and collecting system in the sinus. RESULTS: The volume of the renal parenchyma varied from 129 to 488.4 cm(3). The renal cortex was 83.79% and the medulla 16.21%. The collecting system occupied the greatest part of the sinus, ranging from 34.78 to 71.91% of the renal sinus. The collecting system was closer to the dorsal than to the ventral surface in the cranial pole (p < 0.001) and the hilar zone (p < 0.01). The distance from the collecting system to the medial border was shorter than that to the lateral border in the caudal pole (p < 0.001). CONCLUSION: With this new information about the variation in thickness of the pig renal parenchyma, continued studies using the pig model are needed to support the use of radiofrequency ablation and cryoablation in deep and large renal tumors with a component in the renal sinus.

Characterization of a novel anthropomorphic plastinated lung phantom.

Phantoms are widely used during the development of new imaging systems and algorithms. For development and optimization of new imaging systems such as tomosynthesis, where conventional image quality metrics may not be applicable, a realistic phantom that can be used across imaging systems is desirable. A novel anthropomorphic lung phantom was developed by plastination of an actual pig lung. The plastinated phantom is characterized and compared with reference to in vivo images of the same tissue prior to plastination using high resolution 3D CT. The phantom is stable over time and preserves the anatomical features and relative locations of the in vivo sample. The volumes for different tissue types in the phantom are comparable to the in vivo counterparts, and CT numbers for different tissue types fall within a clinically useful range. Based on the measured CT numbers, the phantom cardiac tissue experienced a 92% decrease in bulk density and the phantom pulmonary tissue experienced a 78% decrease in bulk density compared to their in vivo counterparts. By-products in the phantom from the room temperature vulcanizing silicone and plastination process are also identified. A second generation phantom, which eliminates most of the by-products, is presented. Such anthropomorphic phantoms can be used to evaluate a wide range of novel imaging systems.

How useful is plastination in learning anatomy?

In recent years plastination has begun to revolutionize the way in which human and veterinary gross anatomy can be presented to students. The study reported here assessed the efficacy of plastinated organs as teaching resources in an innovative anatomy teaching/learning system. The main objective was to evaluate whether the use of plastinated organs improves the quality of teaching and learning of anatomy. For this purpose, we used an interdepartmental approach involving the departments of Veterinary Anatomy, Human Anatomy, Veterinary Surgery, and Education Development and Research Methods. The knowledge base of control and experimental student groups was examined before and after use of the fixed or plastinated resources, respectively, to gather information evaluating the effectiveness of these teaching resources. Significant differences (p < 0.001) between control and experimental groups of Human and Veterinary Anatomy were observed in the post-test results. The Veterinary Surgery students had the most positive opinion of the use of plastinated specimens. Using these data, we were able to quantitatively characterize the use of plastinated specimens as anatomy teaching resources. This analysis showed that all the plastinated resources available were heavily used and deemed useful by students. Although the properties of plastinated specimens accommodate student needs at various levels, traditional material should be used in conjunction with plastinated resources.

Acute lymphoblast leukemia in sheep induced by inoculation of bovine leukemia virus: diagnostic electron microscopic morphological study.

Six control lambs were inoculated with Tris buffer, 7 lambs were inoculated with an early passage of bovine leukemia virus (B.L.V.) culture, and 7 lambs with a late passage B.L.V. All experimental lambs were positive with the agar gel immunodiffusion assay (AGID) within 3 months of inoculation and remained positive throughout the 8-year duration of the experiment. The earliest onset of leukemia was at 14 months and the latest was at 44 months after inoculation. Five lambs died with leukemia, two were inoculated with early passage, and three were inoculated with late passage of B.L.V. Eight years after the inoculation, the remaining nine inoculated lambs were clinically normal. The diagnostic ultrastructural morphology of the leukemic lymphoblasts in this study were characterized by hand-mirror cells, multiple nucleoli, irregular nuclear contour with deep indentions, electron-dense granules in the euchromatin, and nuclear cytoplasmic pockets, nuclear myelin figures, mitochondrial variation in size and density, disruption of rough endoplasmic reticulum, and increased ribosomal density. This study shows abundant cytoplasmic processes of hairy cell leukemia. The nuclei of the leukemia lymphoblasts showed electron-dense granules of varying sizes, which were not seen in any of the normal lymphoblasts.

Correlation of magnetic resonance images with anatomic features of the equine tarsus.

OBJECTIVE: To correlate anatomic features of the equine tarsus identified in plastinated sections with images obtained via magnetic resonance imaging (MRI). ANIMALS: 4 horses. PROCEDURE: MRI (1.5-Tesla magnet) of the tarsus was performed on the pelvic limbs of 4 clinically normal horses following euthanasia. After imaging, tarsocrural joint spaces and vasculature were injected with colored latex. Sagittal and transverse sections of the tarsi were plastinated to facilitate interpretation of MR images. RESULTS: Relevant anatomic structures were identified and labeled on the plastinated tissue slices and corresponding MR images. Results indicated high correlations between MRI findings and those of plastinated sections. CONCLUSIONS AND CLINICAL RELEVANCE: The data obtained provided certain reference standards for normal anatomic structure sizes and positions in the equine tarsus. This information may aid future physiologic or clinical studies of this joint.

Correlation of computed tomographic images with anatomic features of the abdomen of ringed seals (Phoca hispida).

OBJECTIVE: To correlate anatomic features of the abdomen of the ringed seal (Phoca hispida) identified in plastinated cross-sections with images obtained via computed tomography (CT) and thereby establish reference standards for normal abdominal organ size and position in this species. SAMPLE POPULATION: 2 adult male ringed seal cadavers. PROCEDURE: With the seal in sternal recumbency, CT images of the abdomen were acquired by use of a 4th-generation CT scanner. Image slice thickness was 1 cm, with no interslice gap. After imaging, the abdominal region was sectioned transversely into 4-cm slices, which were plastinated and photographed. Plastinated slices were matched to their corresponding CT images in preparation for anatomic descriptions. RESULTS: Relevant anatomic features were identified and labeled on both the plastinated tissue slice and corresponding CT image. Normal abdominal organ size and position were assessed, and topographic relationships among organs were ascertained. CONCLUSIONS AND CLINICAL RELEVANCE: The data obtained provide some reference standards for normal abdominal organ size and position in ringed seals. This information may aid researchers of future physiologic and clinical studies in this species.

Cumulative human impacts on marine predators.

Stressors associated with human activities interact in complex ways to affect marine ecosystems, yet we lack spatially explicit assessments of cumulative impacts on ecologically and economically key components such as marine predators. Here we develop a metric of cumulative utilization and impact (CUI) on marine predators by combining electronic tracking data of eight protected predator species (n=685 individuals) in the California Current Ecosystem with data on 24 anthropogenic stressors. We show significant variation in CUI with some of the highest impacts within US National Marine Sanctuaries. High variation in underlying species and cumulative impact distributions means that neither alone is sufficient for effective spatial management. Instead, comprehensive management approaches accounting for both cumulative human impacts and trade-offs among multiple stressors must be applied in planning the use of marine resources.

Cranial pole nephrectomy in the pig model: anatomic analysis of arterial injuries in tridimensional endocasts.

PURPOSE: To assess the intrarenal arteries injuries after cranial pole nephrectomy in a pig model to compare these findings with those in humans. MATERIALS AND METHODS: Polyester resin was injected through the ureter and the renal artery to make three-dimensional casts of 61 pig kidneys. The cranial pole of the kidneys was sectioned at four different sites before the solidification of the resin, and the casts were examined for arterial damage. RESULTS: Section performed through the hilus (15 kidneys): The cranial division of the renal artery was sectioned in two (13.33%) cases, the ventral branch of the cranial division of the renal artery was sectioned in 13 (86.7%) cases, and the dorsal branch of the cranial division of the renal artery was sectioned in 11 (73.34%) cases. Section at 0.5 cm cranial to the hilus (16 kidneys): The cranial division of the renal artery was sectioned in 1 (6.25%) case, the ventral branch of the cranial division of the renal artery was sectioned in 14 (87.5%) cases, and the dorsal branch of the cranial division of the renal artery was sectioned in 13 (81.25%) cases. Section at 1.0 cm cranial to the hilus (15 kidneys): The ventral branch of the cranial division of the renal artery was sectioned in five (33.33%) cases, and the dorsal branch of the cranial division of the renal artery was injured in five (33.33%) cases. Section at 1.5 cm cranial to the hilus (15 kidneys): No lesions were found in the main arteries, only in the interlobular branches. CONCLUSIONS: As previously demonstrated in humans, sections at 1.0 cm or more cranially to the hilus in pigs also showed a significant decrease in damage to the major intrarenal arteries. Therefore, as regards arterial damage, the pig kidney is a useful model for partial nephrectomy in the cranial (upper) pole.

Plastinated heart slices aid echocardiographic interpretation in the dog.

Our aim was to compare plastinated sections of the canine heart with corresponding two-dimensional (2D) echocardiographic images. Thirteen dog hearts were fixed by dilation and then processed by the S10 silicon plastination method (Biodur). Two dogs without evidence of cardiac disease were imaged using 2D echocardiography so as to obtain a complete series of the standard right and left parasternal images, which were compared with corresponding plastinated slices obtained by knife sectioning of the hearts. The plastinated slices revealed the internal anatomy of the heart with great detail and were particularly useful to display the spatial relationship between complex anatomic structures. The plastinated slices corresponded accurately with the echocardiographic images. Because of the dilation of the right heart during the fixation process, it was not possible to obtain plastinated specimens in ventricular systole. This paper may be a reference atlas for assisting 2D echocardiography interpretation.

Legacy and contemporary persistent organic pollutants in North Pacific albatross.

Here we report the first measurements of polybrominated diphenyl ethers (PBDE 47, 99, and 153) alongside 11 organochlorine pesticides (OCPs) and 28 polychlorinated biphenyls (PCBs) in the plasma of albatross from breeding colonies distributed across a large spatial east-west gradient in the North Pacific Ocean. North Pacific albatross are wide-ranging, top-level consumers that forage in pelagic regions of the North Pacific Ocean, making them an ideal sentinel species for detection and distribution of marine contaminants. Our work on contaminant burdens in albatross tissue provides information on transport of persistent organic pollutants (POPs) to the remote North Pacific and serves as a proxy for regional environmental quality. We sampled black-footed (Phoebastria nigripes; n = 20) and Laysan albatross (P. immutabilis; n = 19) nesting on Tern Island, Hawaii, USA, and Laysan albatross (n = 16) nesting on Guadalupe Island, Mexico. Our results indicate that North Pacific albatross are highly exposed to both PCBs and OCPs, with levels ranging from 8.8 to 86.9 ng/ml wet weight and 7.4 to 162.3 ng/ml wet weight, respectively. A strong significant gradient exists between Laysan albatross breeding in the Eastern Pacific, having approximately 1.5-fold and 2.5-fold higher levels for PCBs and OCPs, respectively, compared to those from the Central Pacific. Interspecies levels of contaminants within the same breeding site also showed high variation, with Tern black-footed albatross having approximately threefold higher levels of both PCBs and OCPs than Tern Laysan albatross. Surprisingly, while PBDEs are known to travel long distances and bioaccumulate in wildlife of high trophic status, we detected these three PBDE congeners only at trace levels ranging from not detectable (ND) to 0.74 ng/ml wet weight in these albatross.