Effects of adenosine pretreatment on detection of free radicals in ischemic and reperfused canine gracilis muscle flaps by use of spin-trapping electron paramagnetic resonance spectroscopy

OBJECTIVE: To determine whether adenosine pretreatment attenuates free radical production and muscle damage in ischemic and reperfused canine skeletal muscle. ANIMALS: 9 healthy mixed-breed dogs. PROCEDURE: Dogs were anesthetized, and both gracilis muscles were isolated, leaving only the major vascular pedicle intact. Saline (0.9% NaCl) solution was injected into the artery supplying the control flap, whereas adenosine (10 mg) was injected into the contralateral artery. Ischemia was induced in both flaps for 4 hours. alpha-Phenyl-N-tert-butylnitrone was administered IV to each dog 1 hour prior to reperfusion. Following 15 minutes of reperfusion, effluent blood samples from each muscle flap were obtained and processed for spin-trapping electron paramagnetic resonance (EPR) spectroscopy. Muscle biopsy specimens were obtained for histologic evaluation, and dogs were euthanatized. RESULTS: EPR spectra of strong intensity were obtained from analysis of 5 of 9 paired samples. Signals identified were characteristic of oxygen- and carbon-centered free radical adducts. Signal intensity of spectra from adenosine-treated flaps was significantly less than that of control flaps; mean signal attenuation was 36% in the adenosine-treated group. Histologic evaluation of muscle flaps did not reveal significant differences between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of canine muscle flaps with adenosine prior to a period of ischemia reduced but did not completely attenuate free radical production after reperfusion. However, adenosine pretreatment did not affect histologic abnormalities.     

Myoperitoneal Microvascular Free Flaps in Dogs: An Anatomical Study and a Clinical Case Report

The purpose of the anatomical study was to identify potential myoperitoneal microvascular free flaps, in dogs, that are based on a single artery and vein. The angiosomes of the right deep circumflex iliac artery and left phrenicoabdominal (cranial abdominal) artery were evaluated in six medium-sized canine cadavers. The right deep circumflex iliac artery and left phrenicoabdominal (cranial abdominal) artery were injected with a mixture of barium and latex (equal parts). The entire right and left transversus abdominis muscles were dissected from the abdominal wall and radiographed. The angiograms of the deep circumflex iliac artery showed poor arborization of the vessels within the transversus abdominis muscle in all six cadavers. The angiograms of the phrenicoabdominal (cranial abdominal) artery showed consistent filling of the vascular bed of the cranial half of the transversus abdominis muscle flap in all six dogs. The vascular pedicle lengths and the diameter of the arteries and veins of both the deep circumflex iliac and phrenicoabdominal (cranial abdominal) myoperitoneal free flaps were found to be acceptable for microvascular anastomosis. The deep circumflex iliac flap was unacceptable because of inadequate vascular perfusion. The cranial abdominal artery had a consistent, large branch that supplied the cranial half of the transversus abdominis muscle, thereby making a myoperitoneal flap supplied by this vessel a potentially useful free flap. An 8-year-old male, neutered, mixed-breed dog was evaluated for possible repair of a large defect of the hard palate. Previous operations, using local tissue flaps, had been unsuccessful. A myoperitoneal free flap, based on the right cranial abdominal artery, and consisting of the cranial portion of the transversus abdominis muscle, was used successfully to reconstruct the hard palate. Migrating epithelium from the edges of the wound covered the myoperitoneal flap by 10 weeks after surgery. Therefore, the cranial abdominal myoperitoneal free flap can be considered for reconstruction of intra-oral defects that cannot be repaired using conventional local flap techniques.     

Spin-trapping detection of superoxides in polymorphonuclear leukocytes stimulated with serum-opsonized zymosan

To clarify where oxygen radicals are generated in polymorphonuclear leukocytes (PMNs) during phagocytosis, superoxides (O2-) from opsonized symosan (OZ)--stimulated human PMNs were detected by the ESR and spin-trapping methods. PMNs were preactivated with OZ for the indicated periods of time at 37 degrees C. Then a spin-trapping agent, 5, 5-dimethyl-1-pyrroline N-oxide (DMPO), was added to them, and they were further incubated for 30 sec for ESR observations. The ESR spectra consisted of two components due to the DMPO-OOH and DMPO-OH spin adducts. To clarify where these spin-adducts were present, cells were separated from extracellular fluid by brief centrifugation and resuspended in Hanks' balanced salt solution. ESR examination of two fractions showed that the DMPO-OOH adducts was present in the cell fraction, whereas the DMPO-OH adducts were present in the extracellular fluid. When DMSO was used as a scavenger of hydroxyl radicals (.OH), DMPO-CH3 adducts were observed in the fluid fraction but not in the cell fraction. Both spin adducts were completely abolished by Cu, Zn-SOD but not catalase. These results indicated that O2- were produced inside phagosomes of OZ-stimulated PMNs and .OH were produced outside them by spontaneous decomposition of the DMPO-OOH adducts.     

Caudal Sartorius Muscle Flap in the Dog

An anatomic study was performed on canine cadavers to define the blood supply to the caudal sartorius muscle. The vascular supply to this muscle was segmental with the saphenous artery and vein providing a distal vascular pedicle. Anastomotic channels existed between distal and proximal capillary beds within the muscle belly. This anatomic information was used to determine the feasibility of performing caudal sartorius muscle flaps in dogs. The caudal sartorius muscle was transposed to the medial tibial region in four dogs. The muscle flap was based on a singular vascular pedicle of the saphenous artery and vein. The muscle transpositions were all successful on day 14 as evidenced by gross appearance and results of histologic examination. Grossly, the muscles were well adhered to the recipient sites and were covered by connective tissue. Histologically, the specimens were characterized by viable skeletal muscle fibers, large amounts of granulation tissue, varying degrees of inflammatory response, and small foci of myocyte necrosis (2 cases). Seroma formation was a consistent postoperative complication.     

Effect of Ischemia and Reperfusion on Neutrophil Accumulation in Equine Microvascular Tissue Flaps

Objective— To investigate neutrophil accumulation after ischemia and reperfusion (IR) in microvascular tissue flaps in horses.
Study Design— Randomized controlled experiment.
Sample Population— A total of 8 horses between 1 and 10 years of age, 4 of each sex.
Methods— Control and experimental myocutaneous island flaps based on the superficial branch of the deep circumflex iliac vessels were dissected on each horse. Atraumatic vascular clamps were applied to the pedicle of the experimental flap for 90 minutes and then removed to allow reperfusion. Based on the assumption that rapid infiltration of neutrophils into affected tissues is a hallmark of IR injury, radiolabeled autogenous leukocytes were used to indirectly quantify neutrophil accumulation in flap tissues. Labeled leukocytes were administered through a jugular catheter 30 minutes before flap reperfusion. Biopsies were collected from each flap over a 6 hour postischemia time period; in group 1 (  n = 4  ) from 0 to 6 hours postischemia, and in group 2 (  n = 4  ) from 24 to 30 hours postischemia. Biopsies were examined scintigraphically and histologically for evidence of neutrophil infiltration.
Results— All control flaps survived and 6 of 8 experimental flaps survived. There was no significant evidence of acute neutrophil infiltration into flap tissues after reperfusion in either group.
Conclusions— The results of this study suggest that equine myocutaneous flap tissues can survive a 90-minute ischemic period and reperfusion. No significant evidence of the occurrence of IR injury in flap tissues was found.
Clinical Relevance— The reasons for the previously reported failures of equine free tissue transfer remain uncertain, but they do not appear to be caused by neutrophil mediated injury associated with ischemia and reperfusion.     

Free Radical Formation after Intensive Exercise in Thoroughbred Skeletal Muscles

Although high oxygen consumption in skeletal muscle may result in severe oxidative stress, there are no direct studies that have documented free radical production in horse muscles after intensive exercise. To find a new parameter indicating the muscle adaptation state for the training of Thoroughbred horses, we examined free radical formation in the muscle by using electron paramagnetic resonance (EPR). Ten male Thoroughbred horses received conventional training for 18 weeks. Before and after the training period, all horses performed an exhaustive incremental load exercise on a 6% incline treadmill. Muscle samples of the middle gluteal muscle were taken pre-exercise and 1 min, 1 hr, and 1 day after exercise. Muscle fiber type composition was also determined in the pre-exercise samples by immunohistochemical staining with monoclonal antibody to myosin heavy chain. We measured the free radical in the muscle homogenate using EPR at room temperature, and the amount was expressed as relative EPR signal intensity. There was a significant increase in Type IIA muscle fiber composition and a decrease in Type IIX fiber composition after the training period. Before the training period, the mean value of the relative EPR signal intensity showed a significant increase over the pre-exercise value at 1 min after the exercise and an incomplete recovery at 24 hr after the exercise. While no significant changes were found in the relative EPR signal intensity after the training period. There was a significant relationship between percentages of Type IIA fiber and change rates in EPR signal intensity at 1 min after exercise. The measurement of free radicals may be useful for determining the muscle adaptation state in the training of Thoroughbred horses.     

The effects of allopurinol on the ultrastructure of ischaemic and reperfused large intestine of sheep

Objective To test the possible inhibitory effect of allopurinol on reperfusion injury, caused by oxygen-derived free radicals, of sheep large intestine.
Design An ultrastructural study on caecal tissues from control and treated groups.  

Animals

Fifty sheep in four ischaemic and reperfused (treatment) groups and one control group. Three of the treatment groups were subdivided for half to be injected with allopurinol and the other half with its solvent, potassium hydroxide (KOH).
Procedure Ischaemia of the caecum was induced in the four treatment groups for 60 minutes by clamping the apex. Allopurinol and its KOH solvent were injected intravenously in three treatment groups prior to ischaemia. Samples were collected before and 1 hour after induction of ischaemia and 1 min, 1 h and 8 h after reperfusion. Tissues were processed and examined with an electron microscope.
Results Untreated and solvent injected sheep showed minor ultrastructural changes following ischaemia. With reperfusion, there was severe mitochondrial, goblet cell and basement membrane damage. Tissues from allopurinoltreated sheep were preserved and appeared similar to tissues from the control group.  

Conclusion:

Pre-treatment with allopurinol prevented damage to tissues whereas untreated or allopurinol solventtreated showed severe damage following reperfusion. It is believed that allopurinol, an analogue of hypoxanthine and xanthine, prevents reperfusion injury by competitively binding with xanthine oxidase. This reduces or inhibits the xanthine oxidase mediated conversion of hypoxanthine to xanthine thereby preventing the formation of oxygen-derived free radicals.     

Evaluation of the Cranial Rectus Abdominus Muscle Pedicle Flap as a Blood Supply for the Caudal Superficial Epigastric Skin Flap in Dogs

This study evaluates the cranial rectus abdominus muscle pedicle flap as the sole blood supply for the caudal superficial epigastric skin flap. This flap was composed of a cranially based rectus abdominus muscle pedicle flap that was attached to the caudal superficial epigastric island skin flap (including mammary glands 2 to 5) via the pudendoepigastric trunk. Selective angiography of the cranial epigastric artery in eight cadaver dogs proved that the arterial vasculature in the cranial rectus abdominus was contiguous with that in the caudal superficial epigastric skin flap. In the live dog study, three of six of the flaps failed because of venous insufficiency. Necrosis of mammary gland 2 occurred in two of six flaps. One of six flaps survived with the exception of the cranial most aspect of mammary gland 2. Angiography of the cranial epigastric artery proved that arterial blood supply to these flaps was intact. Histological evaluation of the failed flaps showed full-thickness necrosis of the skin and subcutaneous tissues, the presence of severe congestion, and venous thrombosis. Retrograde venous blood flow through the flap was inconsistent, and hence resulted in failure of this myocutaneous flap. Use of this flap for clinical wound reconstruction cannot be recommended.     

Effects of Epidural Anesthesia on Microcirculatory Blood Flow in Free Medial Saphenous Fasciocutaneous Flaps in Dogs

OBJECTIVE: To assess the effects of epidural anesthesia using lidocaine on microcirculatory blood flow, volume, and velocity in free fasciocutaneous flaps in dogs. Study Design-In vivo experimental investigation. Animal Population-Ten adult dogs weighing 20 to 25 kg. METHODS: A medial saphenous fasciocutaneous free flap was removed and an orthotopic transfer was performed by anastomosing the primary flap vessels back to the medial saphenous vessels. Blood flow (mL(LD)/min/100 g), volume (%volume or tissue hematocrit) and velocity (mm/s) in the flap were recorded throughout the procedure. After epidural anesthesia, blood flow, volume, and velocity values were again recorded. RESULTS: Microcirculatory blood flow, volume, and velocity, as measured by a laser-Doppler flowmeter, failed to reveal any significant changes over time. Immediately after epidural anesthesia, mean arterial pressure was significantly reduced and remained depressed throughout the experimental procedure. CONCLUSIONS: Epidural anesthesia combined with general anesthesia does not improve microcirculatory flow in free flaps in the pelvic limbs of dogs. No significant change in blood flow to the medial saphenous fasciocutaneous free flap occurred after division and anastomosis of the vascular pedicle. CLINICAL RELEVANCE: We recommend that epidural anesthesia with 2% lidocaine be used with caution in dogs undergoing microvascular free-flap transfer.     

The Medial Saphenous Fasciocutaneous Free Flap in Dogs

The purpose of this study was to identify the angiosome of the medial saphenous artery and vein and to evaluate the use of this cutaneous angiosome as a free skin flap in the dog. In phase 1 of this study, selective angiography of the medial saphenous artery performed in six canine cadavers showed that the skin covering the entire medial femorotibial area, the distal half of the caudal head of the sartorius muscle, and the gracilis muscle were perfused by the medial saphenous artery. In phase 2, a medial saphenous fasciocutaneous island flap was raised and sutured back to the skin edges of the donor wound in three dogs. One hundred percent survival of all of the flaps occurred. In phase 3, a medial saphenous fasciocutaneous microvascular free flap was transferred to a wound that was created over the dorsal metacarpal (n = 3) or metatarsal region (n = 3). The mean length ± SD of the medial saphenous vascular pedicle was 80 ± 13 mm (n = 5); the mean diameter ± SD of the medial saphenous artery was 2.8 ± 0.2 mm (n = 5) and the mean diameter ± SD of the medial saphenous vein was 4.2 ± 0.2 mm (n = 5). One hundred percent of all flaps survived (n = 6). Selective angiography of the distal cranial tibial artery (metatarsal wounds, n = 3) and the median artery (metacarpal wound, n = 3) was performed 3 weeks after surgery. All of the vascular anastomoses were patent and neovascularization of the wound beds was present. This free flap was found to be acceptable for cosmetic reconstruction of wounds located on the distal extremity.     

Cranial Sartorius Muscle Flap in the Dog

An anatomic study was performed on canine cadavers to define the blood supply to the cranial sartorius muscle. The vascular supply to this muscle was found to be a single dominant pedicle branching from the femoral artery at the proximal portion of the muscle. This anatomic information was applied in designing a study to determine the feasibility of performing a cranial sartorius muscle flap in the dog. The cranial sartorius muscle was transposed to the caudal abdominal region in four dogs. The muscle flap was based on the singular vascular pedicle defined in the anatomic study. All muscle transpositions were successful on day 19 as evidenced by gross appearance and histologic examination. Grossly, the muscles were well adhered to the recipient sites and were covered by connective tissue. Histologically, the specimens were characterized by viable skeletal muscle fibers, proliferative and maturing granulation and fibrous connective tissue, and mild to moderate mononuclear inflammation. Seroma formation and infection were the two postoperative complications noted. The cranial sartorius muscle flap has potential clinical application for repair of traumatic caudal abdominal hernias and large inguinal hernias in the dog.     

Deep circumflex iliac cutaneous free flap in cats

OBJECTIVE: To develop and assess the survival of a microvascular cutaneous free flap based on the ventral branch of the deep circumflex iliac (DCI) artery and vein in cats. STUDY DESIGN: Experimental study. ANIMALS: Phase 1: 6 feline cadavers; Phase 2: 2 adult cats; Phase 3: 10 adult cats. METHODS: Phase 1: Selective angiographic study of the deep circumflex iliac artery was completed in 6 feline cadavers. After injection of the DCI artery with barium, high-detail radiographs were made of skin flaps harvested from the lateral flank and thigh region. The extent of the cutaneous angiosome was mapped with regard to the underlying anatomical landmarks. Phase 2: An island flap based on anatomic boundaries of the DCI angiosome derived from phase 1 of the study was elevated in 2 cats. Flaps were observed for 3 weeks for survival. Phase 3: Free skin flaps based on the DCI vessels were harvested in 10 cats and transferred to the dorsal interscapular region. Flaps were evaluated for 2 weeks for survival. Tissue samples were collected for histopathology, and angiograms of the flaps were completed. RESULTS: Phase 1: Angiograms revealed a large primary cutaneous angiosome of the DCI artery located over the lateral femoral region, which extended from the iliac crest to the level of the patella. Phase 2: All island flaps survived for 3 weeks. Phase 3: Six free flaps survived for 2 weeks, and 4 flaps failed completely. Failure of 1 flap occurred because of avulsion of the venous and arterial anastomosis postoperatively. Another cat had intraoperative hemorrhage, which resulted in anemia and hypovolemia and likely caused the flap to fail. The other 2 flaps that failed had poor perfusion intraoperatively and had the longest ischemia times. CONCLUSIONS: The cutaneous DCI free flap in cats may be clinically useful in reconstruction of large cutaneous wounds. The length of ischemia time for successful cutaneous free flap transfer in the cat may be shorter than in other species. CLINICAL RELEVANCE: Large wounds created by trauma or oncologic ablative surgery in cats could be reconstructed with cutaneous microvascular free flap. Additional studies assessing the critical ischemia time of cutaneous flaps in cats and evaluating the use of this flap clinically are needed.     

Free tissue transfer of the rectus abdominis myoperitoneal flap for oral reconstruction in a dog

A five-month-old intact/male Boxer dog was presented 5-days following bite wound trauma to the maxillary region resulting in an oronasal fistula extending from the maxillary canine teeth to the soft palate. Multiple surgical procedures using local, buccal mucosal flaps failed to repair the oronasal fistula. Free tissue transfer of the rectus abdominis myoperitoneal flap using microvascular surgical techniques was successful in providing soft tissue reconstruction of the hard palate area. Complications of these surgical techniques included muscle contraction and subsequent muzzle distortion. Small, refractory oronasal fistulae at the perimeter of the myoperitoneal flap were repaired by primary wound closure.     

Experimental definition of latissimus dorsi, gracilis, and rectus abdominus musculocutaneous flaps in the dog

Dissection and injection studies in canine cadavers and in anesthetized dogs were conducted to determine the feasibility of using the latissimus dorsi, gracilis, and rectus abdominus muscles as musculocutaneous free flaps. Lengths of vascular pedicles for the latissimus dorsi (2 +/- 0.8 cm), gracilis (1.8 +/- 0.8 cm), and rectus abdominus (1.9 +/- 0.9-cm cranial deep epigastric, 1.7 +/- 0.5-cm caudal deep epigastric), as well as arterial diameters (1.28 +/- 0.31-mm thoracodorsal for the latissimus dorsi, 1.10 +/- 0.33-mm muscular branch for the gracilis, 1.25 +/- 0.25-mm cranial deep epigastric and 1.26 +/- 0.32-mm caudal deep epigastric for the rectus abdominus) were considered satisfactory for microvascular transfer. Fluorometry demonstrated overlying cutaneous perfusion in all flaps based on their muscle vascular pedicles, with the exception of the rectus abdominus flap based on the caudal deep epigastric artery. In this instance, up to 20% of the cutaneous element had questionable or no perfusion.     

Methods to assess free radicals and oxidative stress in biological systems.

Oxidative stress results from a disruption of the prooxidant/antioxidant cellular balance and monitoring free radical status becomes an interesting challenge in animal and human nutrition. In the present work, merits and limitations of different analytical techniques (HPLC, GC-MS, fluorometric and colourometric assays, ELISA, gel electrophoresis) for the measurement of radical mediated alterations in the cellular integrity of lipids (malondialdehyde, hydrocarbon gases, F2-isoprostanes) proteins (protein carbonyls, 3-nitrotyrosine) and DNA (8-hydroxy-2'-deoxyguanosine) are discussed. Besides these indirect methods, owing to the fact that free radicals are paramagnetic, electron paramagnetic resonance spectroscopy combined with spin trapping has become a valuable tool to directly assess and to better understand the mechanisms of free radical reactions. With this approach a radical that is too short-lived to be detected, adds to a spin-trapping agent to form a relatively long-lived radical adduct. Information obtained from the hyperfine splitting of the spin-trapped adduct can provide identification and quantification of the originally generated free radicals.     

Definition of the gracilis musculocutaneous flap for distant transfer in cats.

Dissection, injection, and surgical studies in feline cadavers and in anesthetized cats were conducted to determine the feasibility of using the gracilis muscle as the basis for a free musculocutaneous flap. The vascular pedicle of the flap consisted of the femoral artery and vein. Mean length (1.6 +/- 0.2 cm) of the vascular pedicle and mean artery (1.33 +/- 0.19 mm) and vein (2.55 +/- 0.38 mm) diameters were satisfactory for microvascular transfer. Fluorometry revealed overlying cutaneous perfusion in the flaps on the basis of their muscle vascular pedicles. To ensure survival of the flap, the muscular branches of the femoral artery and vein supplying the gracilis muscle had to be carefully preserved during surgical elevation of the flap.     

Creatine kinase isoenzymes in serum of pigs having myocardial and skeletal muscle necrosis.

Serum creatine kinase (CK) and lactic dehydrogenase (LD) isoenzyme activities were measured in blood serum of pigs having myocardial damage and skeletal muscular lesions. Myocardial and muscular damage was induced by restraint stress provoked by intravenous infusion of a pharmacological restraint (succinylcholine-chloride) during 12 minutes. Pigs of Swedish Landrace and Swedish Landrace X Yorkshire breed, stress-susceptible (halothane-sensitive) and nonreacting pigs were studied. Severe myocardial damage and slight to moderate skeletal muscle necrosis were found 24 hours after restraint stress in the stress-susceptible pigs whereas in nonreacting pigs generally only myocardial lesions of moderate extent were registered. No significant increase was detected in the serum CK-BB (CK-1) or CK-MB (CK-2) activity whereas a pronounced elevation of the CK-MM (CK-3) activity was found, particularly in the stress-sensitive animals. In the myocardial tissue of pigs only a low CK-MD activity was found (about 4-5% CK-MD in addition to CK-MM) and this may explain the low CK-MB activity in serum of pigs subjected to severe myocardial damage. This is further supported by the pronounced increase in the anodal serum fractions LD 1-2 in animals free from skeletal muscular lesions. In the halothane-sensitive pigs skeletal muscle necrosis besides the myocardial lesions contributed to the high levels of CK-MM activity in serum.     

Development of a Free Latissimus Dorsi Muscle Flap in Cats

Anatomic and experimental evaluation of the feline latissimus dorsi muscle was performed to assess its potential use as a free muscle flap. In the anatomic study, nonselective angiography of the subscapular artery was performed in nine heparinized feline cadavers. The muscle dimensions and vascular anatomy of the dissected latissimus dorsi muscle were recorded. In the experimental study four cats underwent heterotopic transplantation of a partial latissimus dorsi flap, and three cats underwent orthotopic transplantation of a complete latissimus dorsi flap. The mean length and width of the latissimus dorsi muscle was 19.0 and 5.4 cm, respectively. The dominant vascular pedicle was the thoracodorsal artery and vein. The average length and diameter of the thoracodorsal artery was 2.7 cm and 0.6 mm, respectively. Minor vascular pedicles were provided by branches of the intercostal arteries. Numerous choke anastomoses existed between the two pedicle systems. Viability of muscle flaps based on subjective evaluation, angiography, and histopathology, was 66% and 100% in the heterotopic and orthotopic studies, respectively. Flap failure seemed to be caused by both arterial and venous thrombosis. The latissimus dorsi muscle flap met criteria required for application in microvascular reconstruction. The vascular pattern was appropriate and consistent. Donor site morbidity was low, whereas surgical accessibility was high. The muscle satisfied the physical criteria of a free flap. Long-term anastomotic patency and flap viability was shown.     

A Comparison of the Cutaneous Trunci Myocutaneous Flap and Latissimus Dorsi Myocutaneous Flap in the Dog

Latissimus dorsi and cutaneous trunci myocutaneous flaps of equal dimension and location were randomly elevated on opposite sides of the thorax in 10 dogs (group 1) and resutured to their respective bed. The procedure was repeated in four additional dogs (group 2); however, the short perforating branches of the thoracodorsal artery and vein were divided at the base of each cutaneous trunci myocutaneous flap, whereas the cutaneous pedicle and underlying cutaneous trunci muscle were divided in the latissimus dorsi myocutaneous flaps to determine subsequent skin survivability and the major source of circulation of each myocutaneous flap. There was little difference in the percentage of skin survival between the latissimus dorsi and cutaneous trunci myocutaneous flaps in group 1 dogs. Circulation to the "skin island" of group 2 latissimus dorsi myocutaneous flaps originated from intramuscular anastomotic connections between the major branch of the thoracodorsal artery entering the latissimus dorsi muscle and the proximal lateral intercostal arteries perforating the muscle. Ligation of the short perforating branches of the thoracodorsal artery resulted in partial skin necrosis in all group 2 cutaneous trunci myocutaneous flaps. Results from this study indicate that it is unnecessary to elevate the latissimus dorsi muscle for major skin flap elevation and survival. The thicker latissimus dorsi myocutaneous flap is more difficult to develop surgically and appears to have no clinical major advantage over the more mobile cutaneous trunci myocutaneous flap or the adjacent thoracodorsal axial pattern flap for closure of large skin defects within the radius of flap rotation.     

Temporal and topographic profiles of tissue hypoxia following transient focal cerebral ischemia in rats

Intravascular accumulation of blood cells after brain ischemia-reperfusion can cause obstruction of cerebral blood flow and tissue hypoxia/ischemia as a consequence. In the present study, we examined temporal and topographic changes of tissue hypoxia/ischemia after occlusion of the middle cerebral artery (MCA) for 60 min in rats with immunohistochemical staining for hypoxia (2-nitroimidazole hypoxia marker: hypoxyprobe-1 adducts). Our results showed that tissue hypoxia expressed as positive staining for hypoxyprobe-1 adducts preceded neuronal degeneration. Platelets and granulocytes were detected close to the hypoxyprobe-1 adducts positive area. These results suggested that the hypoxic environment could persist even after reperfusion of MCA, because of vascular obstruction with accumulation of platelets and granulocytes.