Bruce R. Pynn

A Histochemical Method for the Simultaneous Demonstration of Capillaries and Fiber Type in Skeletal Muscle

A modified ATPase method for the simultaneous demonstration of capillaries and fiber types in skeletal muscle is presented. Muscle biopsies were obtained from mice, hamsters, rats, cats, and dogs, quick frozen, and sectioned at 8 microns in a cryostat. The frozen slides were fixed in a neutral formalin solution at 4 C for 5 min, and then incubated at 37 C for 1 hr in a medium containing ATP, Pb2+, and Ca2+ in a tris-maleate buffer (pH 7.2). Dilute (NH4)2S was used as a developer. To test the reliability of the proposed method, serial sections of each biopsy were stained separately for capillaries (amylase-PAS method) and for fiber types by a standard myosin ATPase (m-ATPase) method. Fiber type percent and capillary parameters were determined for each biopsy. No difference in results was observed for parameters determined using the modified ATPase method compared to the standard capillary and fiber type staining methods. This modified technique is therefore suitable for the simultaneous demonstration of capillaries and fiber types in skeletal muscle.

An isolated skeletal muscle model suitable for acute ischemia studies

A modified isolated canine gracilis model of acute complete muscle ischemia was developed and then tested metabolically and histologically in 25 animals to assess its validity. In each dog, both gracili were isolated on their major vascular pedicles. One muscle underwent ischemia and reperfusion by placing and removing microvascular clips on the artery and vein. The other gracilis muscle was used as a control. Total muscle blood flow measurements, blood samples, and muscle biopsies were taken every other hour for up to 11 hr after preparation. The fiber-type profile of the gracilis was determined bilaterally using a myosin ATPase stain (n = 10). The results verified these hypotheses: after surgical preparation, the right and left muscles in the same dog are equivalent metabolically, after a 2-hr stabilization period, gracilis blood flow, oxygen and glucose uptake, lactate release, and tissue glycogen, lactate, phosphocreatine, and ATP levels remain within normal limits and unchanged for the next 9 hr, the surgical isolation of the gracilis muscle on a single vascular pedicle does not result in significant metabolic changes, in this model, a 2-hr ischemia is reversible, but a 7-hr ischemia results in irreversible ischemic injury. As well, fiber-type profile, muscle blood flow, and metabolic parameters can very significantly among animals supporting the necessity of a contralateral control. Therefore, this modified gracilis muscle model with its contralateral muscle as a control is suitable for acute skeletal muscle ischemia experiments of at least 9-hr duration.

Isometric contractile function recovery following tourniquet ischemia

The purpose of this study was to document the recovery of isometric contractile function following tourniquet ischemia. Male Wistar rats (N = 27) were subjected to unilateral hindlimb tourniquet ischemia of 0 hr (control, N = 6), 1 hr (N = 5), 2 hr (N = 5), 3 hr (N = 5) and 4 hr (N = 3). Following a 2-week recovery period, isometric force measurements were made from both gastrocnemii of each rat with the contralateral limb acting as the control side. Each muscle was analyzed for maximal twitch (Pt, N/g), maximal rate of rise of twitch tension (DP/dt, N/sec), time to peak tension (TPT, msec), half relaxation time (RT 1/2, msec), maximal tetanus (P0, N/g, at 100 Hz), and fatigue (Burke Fatigue Protocol). Pt, P0, and DP/dt were significantly different from control values (P less than 0.05) for all hours of tourniquet ischemia. A strong negative correlation (P less than 0.001) was found for twitch (R = -0.84), tetanus (R = -0.78), and maximal rate of force development (R = -0.83) with respect to increasing hours of ischemia. The recovery of isometric twitch and tetanic function following tourniquet ischemia is inversely related to the ischemic interval. This study quantified the relationship between muscle ischemia and recovery of function following a 2-week interval and stresses the functional physiological changes which occur in skeletal muscle following tourniquet ischemia.

Calcium pyrophosphate dihydrate deposition disease of the temporomandibular joint: A case report and review of the literature

An asymptomatic preauricular swelling in a 58-year-old man appeared as a parotid mass and was diagnosed as calcium pyrophosphate dihydrate crystal deposition disease of the temporomandibular joint. This article describes the diagnostic criteria and management of this rare condition and reviews the 13 cases that have been reported to date.

Delayed neovascularization in free skin flap transfer to irradiated beds in rats.

A model for the study of neovascularization with a normal epigastric free flap set into an irradiated defect in the Fischer F344 rat is presented. In this model, both the administration of radiation and the flap transfer mimic the clinical situation. Significantly less tissue survives loss of the complete vascular pedicle at the second to fourth days following flap creation in rats with an irradiated bed. Later survival is not different from controls. Delayed neovascularization is proposed as the mechanism responsible for this effect during the period corresponding to the onset of the late phase of the response to skin radiation in rats. That neovascularization does occur, although delayed, suggests that the induced endarteritis may not be as important as previously suggested.

The effect of intraoperative ischemia on the recovery of contractile function after free muscle transfer

Isometric contractile function was studied after recovery in free, vascularized muscle transfer subjected to graded periods of intraoperative ischemia. Fifteen dogs had orthotopic replantation of their left gracilis muscles, with intraoperative ischemia times grouped as 0 (n = 3), 1 to 2 (n = 3), 2 to 3 (n = 4), or 3 to 4 (n = 5) hours. After recovery (mean 61.8 weeks), isometric twitch and tetanic tension and fatigue measurements were made in the replants and in the contralateral, control gracilis. On the average, replants were found to produce significantly less twitch (0.32 +/- 0.13 versus 0.49 +/- 0.24 N/g) and 75 Hz tetanic tension (2.2 +/- 0.9 versus 3.4 +/- 0.5 N/g) than controls. However, in several individual replants, 100% of control maximal tetanic tension was observed. Intraoperative ischemia time of up to 4 hours was not correlated with functional return. It is concluded that (1) full recovery is possible after free muscle transfer; (2) intraoperative ischemia, if less than 4 hours long, is not the primary determinant of functional recovery; and (3) factors besides intraoperative ischemia must be operative in producing the variability in recovery seen in this setting.

An apparatus for the calibration of electromagnetic flowprobes on small veins in situ

Electromagnetic flowprobe calibration must be done under controlled conditions similar to those encountered experimentally. This in situ calibration apparatus is simple in design, inexpensive, and provides pressure and flow conditions analogous to those found in small veins in vivo.

Parasymphyseal Fracture with an Associated Temporomandibular Joint Dislocation: Case Report

A case of traumatic mandibular fracture with associated unilateral anterior dislocation of the temporomandibular joint in a child is described. Although anterior dislocation is common, this combination of fracture and separate dislocation to our knowledge has not been reported in a young child. The unusual management required in this case is discussed.

Metabolic characteristics of experimental free vascularized canine gracilis muscle transfers.

A canine gracilis model was used to study muscle energy metabolism and enzyme activities after free vascularized muscle transfer. Fifteen male mongrel dogs underwent orthotopic, free transfer of the left gracilis with microneurovascular anastomosis. After a minimum of 10 months’ recovery, muscle biopsy specimens were obtained from the transfers and the contralateral controls and analyzed for relative fiber type areas and maximum activities of phosphorylase, hexokinase, phosphofructokinase, glycerol-3-phosphate dehydrogenase (GPDH), pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, 3-hydroxyacyl coenzyme A dehydrogenase (HAD), and creatine phosphokinase. Biopsy specimens obtained before and after a-10 minute, 20-Hz contraction were analyzed for glucose, glycogen, glycolytic intermediates, phosphocreatine, total creatine, and adenine nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine monophosphate, and inosine). There was no significant transfer versus control difference in type I relative fiber area (45 ± 4 percent versus 44 ± 3 percent). Total creatine was significantly reduced in the transferred muscles relative to control (83.1 ± 3.0 mmol/kg versus 100.6 ± 5.1 mmol/kg dry weight). Maximal activities of phosphorylase, pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, HAD, and creatine phosphokinase were diminished in transfers relative to controls, although hexokinase activity was significantly higher in the freely transferred gracilis muscles. During the 20-Hz contraction, muscle transfers produced less force initially, although the force/time integral over the 10-minute stimulation was similar in transfers (277 ± 25 N/g/second) and controls (272 ± 24 N/g/second). The contraction was associated with significant glycogen use and lactate accumulation in both transfers and controls, although this was less pronounced for the transfers. Glycolytic flux appeared muted in the transfers relative to controls. Significant, similar high-energy phosphagen reductions and inosine monophosphate accumulation were noted during the contraction in both groups. Contractile activity is associated with the expected pattern of muscle metabolite changes following free vascularized transfer, indicating the components of cellular energy metabolism are not qualitatively altered after microneurovascular muscle transfer. In contrast, quantitative differences suggest that free vascularized muscle transfer can be associated with a muscle enzyme profile consistent with deconditioning and the presence of denervated muscles fibers in the absence of fiber type profile changes.