Spinal cord injury in the rat.

Only limited therapeutic measures are currently available for the treatment of spinal cord injury. This review describes the pathologic mechanisms of trauma-induced spinal cord injury in rats, which will contribute to new understanding of the pathologic process leading to spinal cord injury and to further development of new therapeutic strategies. Spinal cord injury induced by trauma is a consequence of an initial physical insult and a subsequent progressive injury process that involves various pathochemical events leading to tissue destruction; the latter process should therefore be a target of pharmacological treatment. Recently, activated neutrophils have been shown to be implicated in the latter process of the spinal cord injury in rats. Activated neutrophils damage the endothelial cells by releasing inflammatory mediators such as neutrophil elastase and oxygen free radicals. Adhesion of activated neutrophils to the endothelial cell could also play a role in endothelial cell injury. This endothelial cell injury could in turn induce microcirculatory disturbances leading to spinal cord ischemia. We have found that some therapeutic agents that inhibit neutrophil activation alleviate the motor disturbances observed in the rat model of spinal cord injury. Methylprednisolone (MPS) and GM1 ganglioside, which are the only two pharmacological agents currently clinically available for treatment of acute spinal cord injury, do not inhibit neutrophil activation in this rat model. Taken together, these observations raise a possibility that other pharmacological agents that inhibit neutrophil activation used in conjunction with MPS or GM1 ganglioside may have a synergistic effect in the treatment of traumatic spinal cord injury in humans.

Contribution of CD4+ and CD8+ T cells and interferon-gamma to the progress of chronic rejection of kidney allografts: the Th1 response mediates both acute and chronic rejection.

T cells mediating chronic rejection (CR) of human kidney allografts were characterized by comparing them with those mediating acute rejection (AR). Two lines of analysis were performed using biopsy specimens (23 CR and 8 AR). First, the extent of infiltration of CD4+ and CD8+ T cells into allografts was assessed from mRNA expression of CD4 and CD8. The group of CR specimens was not significantly different from the group of AR specimens in terms of the extent of CD4+ and CD8+ T cell infiltration, underlining the importance of the immunological contribution to the progress of CR. Second, Th1/Th2 polarization in infiltrating T cells was investigated by measuring mRNA expression of interferon gamma (IFN-gamma; a Th1 cytokine) and interleukin 4 (IL-4; a Th2 cytokine). IFN-gamma expression was detected in most CR specimens, and was not significantly different between the group of CR specimens and the group of AR specimens. On the other hand, IL-4 expression was detected in only two CR specimens and one AR specimen; from its pathological features, the AR in this last case was concomitant with CR. These results suggest that most cases of CR and of AR are mediated by Th1 mechanisms, although some cases of CR show features of both Th1 and Th2.

Role of neutrophils in spinal cord injury in the rat.

Activated neutrophils are thought to be involved in tissue injury through the release of various inflammatory mediators. To understand the role of neutrophils in spinal cord injury, the effects of nitrogen mustard-induced leukocyte depletion and the administration of an anti-P-selectin monoclonal antibody on motor disturbances observed following spinal cord compression were examined in rats. Spinal cord injury was induced by applying a 20-g weight for 20 min at the level of the 12th thoracic vertebra, resulting in motor disturbances of the hindlimbs 24 h postcompression. Motor disturbances, evaluated using Tarlov's index, an inclined-plane test and climbing ability, were markedly attenuated in rats with nitrogen mustard-induced leukocytopenia. Administration of the anti-P-selectin monoclonal antibody, by which adhesion of activated neutrophils to endothelial cells may be inhibited, also attenuated motor disturbances. Histological examination revealed that intramedullary hemorrhages observed 24 h after compression at the 12th thoracic vertebra of the spinal cord were significantly attenuated in leukocytopenic animals and those which received the anti-P-selectin monoclonal antibody. The accumulation of neutrophils at the site of compression, as evaluated by measuring the tissue myeloperoxidase activity, significantly increased with time following the compression, peaking at 3 h postcompression. Spinal cord myeloperoxidase activity did not increase in sham-operated animals. Leukocyte depletion and administration of the anti-P-selectin monoclonal antibody both reduced the accumulation of neutrophils in the damaged spinal cord segment 3 h postcompression. These observations strongly suggest that activated neutrophils play an important role in compression-induced thoracic spinal cord injury and that a P-selectin-mediated interaction between activated neutrophils and endothelial cells may be a critical step in endothelial cell injury leading to spinal cord injury.

A biological method to evaluate bifunctional chelating agents to label antibodies with metallic radionuclides.

UNLABELLED: Since the lysosome is a common organelle for protein digestion, pursuing the fate of radiolabeled metabolites after lysosomal proteolysis in liver cells is ideal to evaluate bifunctional chelating agents (BCAs). METHODS: We used galactosyl-neoglycoalbumin (NGA) and mannosyl-neoglycoalbumin (NMA) as carrier proteins for hepatic parenchymal and nonparenchymal cells, respectively. These proteins were labeled with 111In using 1-(4-isothiocyanatobenzyl)ethylenediaminetetraacetic acid (SCN-Bz-EDTA) as a model. RESULTS: NGA-SCN-Bz-EDTA-111In exhibited rapid accumulation in the hepatic parenchymal cells, followed by hepatobiliary excretion of the metabolites with an elimination rate that was faster and much slower than that of NGA-DTPA-111In and NGA-131I, respectively. This metabolite represented all the radioactivity registered in the liver at 1 hr postinjection. Subcellular distribution studies indicated the metabolites were located only in the lysosome fraction, and the difference in elimination rates of the metabolites from the lysosome fraction was responsible for the variations in radioactivity clearance from the cells. CONCLUSION: The biological characteristics of radiolabeled metabolites play a critical role in eliminating the radiolabel from liver cells. The present method portrays a highly useful model to pursue the fate of radiolabeled metabolites in the liver.

Neuroprotection by recombinant thrombomodulin.

We examined whether recombinant human soluble thrombomodulin (rhs-TM) reduces compression trauma-induced spinal cord injury through protein C activation in rats. Administration of rhs-TM, either before or after the induction of spinal cord injury (SCI), markedly reduced the resulting motor disturbances. However, neither rhs-TM pretreated with an anti-rhs-TM monoclonal antibody (MAb) F2H5, which inhibits thrombin binding to rhs-TM, nor those pretreated with MAb R5G12, which selectively inhibits protein C activation by rhs-TM, prevented the motor disturbances. Intramedullary hemorrhages, observed 24 h after trauma, were significantly reduced in animals given rhs-TM. The increase in the tissue levels of tumor necrosis factor-alpha (TNF-alpha), TNF-alpha mRNA expression, and the accumulation of leukocytes in the damaged segment of the spinal cord were significantly inhibited in animals receiving rhs-TM, but these effects were not observed following administration of rhs-TM pretreated with MAb R5G12 or MAb F2H5. Leukocytopenia and activated protein C all produced effects similar to those of rhs-TM. These findings suggest that rhs-TM prevents compression trauma-induced SCI by inhibiting leukocyte accumulation by reducing the expression of TNF-alpha mRNA and such therapeutic effects of rhs-TM could be dependent on its protein C activation capacity. Findings further suggest that thrombomodulin can be implicated not only in the coagulation system but in regulation of the inflammatory response.

Role of leukocytes in spinal cord injury in rats.

Spinal cord injury (SCI) is a serious condition that produces life-long disabilities. Only limited therapeutic measures are currently available for its treatment. This review describes the role of leukocytes in pathologic mechanisms of trauma-induced SCI in rats, which contributes to new understanding of the pathologic process involved in SCI and could lead to the development of new therapeutic strategies by which leukocyte activation can be regulated. SCI induced by trauma is a consequence of an initial physical insult that is followed by a progressive injury process which involves various pathochemical events that lead to tissue destruction. Therapeutic intervention in SCI should therefore be directed at reducing or alleviating this secondary process. Although the mechanisms are not fully understood, progressive vascular events, especially activated neutrophil-induced endothelial cell damage, have been shown to be implicated. We have found that some therapeutic agents, which inhibit leukocyte activation directly or indirectly, alleviate the motor disturbances observed in a rat model of SCI. Methylprednisolone (MPS) and GM1 ganglioside, which are the only two pharmacological agents currently clinically available for treatment of acute SCI, do not inhibit neutrophil activation in this rat model. Taken together, these observations raise a possibility that pharmacological agents that inhibit leukocyte activation used in conjunction with MPS or GM1 may have a synergistic effect in the clinical treatment of traumatic SCI in humans.

Methylprednisolone reduces spinal cord injury in rats without affecting tumor necrosis factor-alpha production.

Methylprednisolone (MPS) is the only therapeutic agent currently available for traumatic spinal cord injury (SCI). However, little is known about its therapeutic mechanisms. We have demonstrated that tumor necrosis factor-alpha (TNF-alpha) plays a critical role in posttraumatic SCI in rats. Since MPS has been shown to inhibit TNF-alpha production in vitro, it is possible that MPS can reduce SCI by inhibiting TNF-alpha production. To examine this possibility, we investigated the effect of MPS on TNF-alpha production in injured segments of rat spinal cord. Leukocytopenia and high-dose intravenous administration of MPS markedly reduced the motor disturbances observed following spinal cord trauma. Both treatments also reduced the intramedullary hemorrhages observed histologically 24 hr posttrauma. Leukocytopenia significantly reduced tissue levels of both TNF-alpha mRNA and TNF-alpha, 1 and 4 hr posttrauma, respectively, and it also inhibited the accumulation of leukocytes in the injured segments 3 hr posttrauma, while MPS had no effects. Lipid peroxidation and vascular permeability at the site of spinal cord lesion were both significantly increased over time after the induction of SCI, peaking 3 hr posttrauma. These events were significantly reduced in animals with leukocytopenia and in those given anti-P-selectin monoclonal antibody compared to sham-operated animals. Administration of MPS significantly inhibited both the increase in lipid peroxidation and the vascular permeability. These findings suggested that MPS reduces the severity of SCI, not by inhibiting the production of TNF-alpha at the site of spinal cord trauma, but by inhibiting activated leukocyte induced lipid peroxidation of the endothelial cell membrane. This suggests that MPS may attenuate spinal cord ischemia by inhibiting the increase in endothelial permeability at the site of spinal cord injury.

Role of neutrophil elastase in compression-induced spinal cord injury in rats.

We have previously demonstrated the importance of activated neutrophils in compression-induced spinal cord injury (SCI) in rats. In the present study, we investigate the action of neutrophil elastase in posttraumatic SCI, using two neutrophil elastase inhibitors (Eglin C and L658,758). SCI was induced by applying a 20-g weight to the spinal cord for 20 min at the level of T12, resulting in hindlimbs motor disturbances, which, when evaluated using a inclined-plane test, were significantly attenuated by Eglin C or L658,758. Histologic examination revealed that intramedullary hemorrhages observed 24 h after trauma were markedly attenuated in these agents. These inhibitors also significantly decreased neutrophil accumulation as shown by myeloperoxidase activity in the damaged spinal cord segment. Induction of leukocytopenia had the same effects as Eglin C or L658,758. These findings implicated neutrophil elastase in SCI. The enzyme may induce vascular damage leading to spinal cord ischemia.

Reduction of spinal cord injury by administration of iloprost, a stable prostacyclin analog.

To investigate whether iloprost, a stable analog of prostacyclin, is useful for the prevention of posttraumatic spinal cord injury, we examined its effects on compression trauma-induced spinal cord injury in rats. Spinal cord injury was induced by applying a 20-g weight for 20 minutes to the spinal cord at the level of T-12, resulting in motor disturbances in the hindlimbs. These motor disturbances, evaluated using Tarlov's index, were markedly attenuated in rats with nitrogen mustard-induced leukocytopenia. Administration of iloprost also attenuated the motor deficits. Histological examination revealed that intramedullary hemorrhages observed 24 hours after trauma were significantly attenuated in leukocytopenic animals and in animals that received iloprost. The accumulation of leukocytes at the site of trauma, evaluated by measuring tissue myeloperoxidase activity, significantly increased with time following the trauma, peaking at 3 hours postinjury. Spinal cord myeloperoxidase activity in sham-operated animals did not increase postoperatively. Leukocyte depletion and administration of iloprost reduced the accumulation of leukocytes in the damaged spinal cord segment 3 hours posttrauma. These findings indicate that iloprost attenuates motor disturbances induced by spinal cord trauma and that its therapeutic efficacy can be partly explained by its inhibition of leukocyte accumulation at the traumatized site.

Magnetic resonance imaging study on spinal cord plasticity in patients with cervical compression myelopathy.

Spinal cord plasticity in 55 patients with cervical compression myelopathy was assessed with magnetic resonance imaging, by which the transverse area of the spinal cord was measured at the site of maximum compression before and after surgery and compared with the conventional modalities of computed tomographic myelography. A high correlation (r = 0.901, P less than 0.01) was observed between the preoperative measurements of magnetic resonance imaging and computed tomographic myelography. The preoperative transverse area was in good correlation with the preoperative Japanese Orthopaedic Association score (r = 0.466, P less than 0.01). In most patients with a spinal cord area of less than 0.45 cm2, the clinical results were poor despite considerable morphologic restoration of the spinal canal obtained after decompression surgery, reflecting an irreversible pathology developed in the spinal cord.

Clinical considerations and biochemical basis of prognosis of cervical spinal cord injury.

A total of 118 patients with cervical spinal cord injury were studied to determine the neurologic improvement achieved by either conservative or surgical treatment. Useful recovery was observed in 55% of the patients with incomplete cord injuries, but in none of those with complete cord injuries. There was no significant difference between the treatment groups regarding neurologic improvement. In experimental studies on rats, the increased levels of lipid peroxides and thromboxane after spinal cord injury were found to be proportional to the magnitude of injury. These evidences suggest that spinal cord injury is directly related with the magnitude of injury, and the prognosis is determined entirely at the time of injury.

Effects of EPC-K1 on lipid peroxidation in experimental spinal cord injury.

STUDY DESIGN: A study in which levels of lipid peroxidation were measured, the thiobarbituric acid-reactive substances were estimated in an experimental rat model, and the recovery was assessed. OBJECTIVE: To ascertain the occurrence of thiobarbituric acid-reactive substances in the damaged spinal cord, and to investigate the effectiveness of a hydroxyl radical scavenger EPC-K1, a phosphate diester linkage of vitamins E and C, in attenuating the severity of spinal cord injury. SUMMARY OF BACKGROUND DATA: Lipid peroxidation has been reported to play an important role in spinal cord injury. There is no report on the use of EPC-K1 to attenuate the severity of spinal cord injury in either animal or human studies. METHODS: Spinal cord injury was induced by placing a 25-g weight on T12, and the animals were divided into six groups. Group 1 (sham) received only laminectomy. Group 2 (control) received spinal cord injury. Group 3 received EPC-K1 5 minutes before injury. Group 4 received it 5 minutes after injury. Group 5 received it 3 hours after injury. Group 6 received it five times, respectively: at 5 minutes, then 1, 2, 3, and 4 hours after injury. The levels of thiobarbituric acid-reactive substances were measured in the spinal cord, and the recovery was assessed. RESULTS: The thiobarbituric acid-reactive substances content increased after injury, with two peaks, at 1 and 4 hours. Concentration at the 4-hour peak was lower in nitrogen mustard-induced leukocytopenia rats than in the control rats. The EPC-K1 injection reduced thiobarbituric acid-reactive substances content at 1 and 4 hours after injury in Group 3 (respectively, 34.3% and 42.7% vs. control) and only that at 4 hours in Group 6 (24.9% vs. control). Motor function recovery and histologic findings were better in these two groups than in Group 2. CONCLUSION: Repeated injection of EPC-K1 attenuated the severity of spinal cord injury.

Influence of dietary vitamin E deficiency on compression injury of rat spinal cord.

The influences of vitamin E deficiency on compression injury of the rat spinal cord associated with ischemia were investigated. Growing rats were divided into two groups and given a diet containing either 2 IU/100 g or less than 0.1 IU/100 g of alpha-tocopherol acetate, respectively, for 6-8 weeks before experiments. Motor disturbances induced by spinal cord injury were found to be enhanced by vitamin E deficiency. The spinal cord blood flow (SCBF) was reduced by compression and subsequently increased transiently and then decreased gradually in both groups, but the level was lower in the vitamin E-deficient group than in the control group. After injury, the vitamin E-deficient group showed lower recoveries than the control group in the amplitude and latency of spinal cord evoked potentials and greater pathological changes of the spinal cord, such as bleeding and edema. The increase in the level of TBA-reactive substances in the spinal cord after injury increased with decrease in the dietary level of vitamin E. These results suggest that vitamin E may have a protective effects against ischemic spinal cord injury by its antioxidant effect.

[Ecological approach to the patients with digestive diseases in Kitakyushu City and its suburbs].

Nine hundred and ninety nine patients were admitted in our Department (the Third Department of Internal Medicine, School of Medicine, UOEH) during the five years more since the opening date of the University Hospital (July, 9, 1979), and 864 cases in them (86.2%) suffered from the various digestive diseases. Most of the in-patients with digestive diseases in our Department are resident in Kitakyushu city and its suburbs, especially in Yahatanishi-ku, Wakamatsu-ku and Onga county, therefore, it may be possible to investigate the ecological characteristics of the in-patients of our Department in the relation to the outbreak, clinical course and outcome of the digestive diseases. Namely, it may be assumed that the incidence and prevalence of the idiopathic inflammatory bowel disease (IBD) including ulcerative colitis and Crohn's disease are relatively high in this area (Kitakyushu city and its suburbs) as compared with the average of all Japan. Although the true causes of these illness are still unknown, the inclination of haptoglobin phenotypes (HP) which include 2-2, 2-1 & 1-1 type 1-1 strongly suggests to the association with some genetical factors on the high incidence of these diseases (IBD). In this connection, Hp type 1-1 were recognized 4 in 11 cases (36.4%) with ulcerative colitis, and 3 in 7 cases (42.9%) with Crohn's disease in our Department whereas only 3-5% in normal controls. Secondly, the patients with carcinoma of the biliary tree (bile duct and gall bladder) are relatively more, namely, 17 cases of bile duct cancer and 3 cases of gall bladder cancer were admitted in our Department during this term. It is interesting to note that hepatohilar type of the bile duct cancer was observed comparatively high (4 in 17 cases, 52.9%) in the past five years-more although the etiology is unknown. Finally, several characteristics in liver diseases particularly in viral hepatitis were illustrated in this study, namely, the ratio of transient HBV infection to whole (transient and persistent) HBV infection in the patients with acute viral hepatitis (due to HBV) is high (80.9%), HBeAg positivity is high in chronic B-hepatitis (44.9%), the ratio of alcoholic cirrhosis to whole liver cirrhosis is relatively high (34.9%) and HBsAg positivity is lower in liver cirrhosis due to non-alcoholic origin (mainly due to hepatitis virus) than the average of this country, and also, hepatocellular carcinoma (HCC) without liver cirrhosis is higher (23.0%) than the average of whole Japan (less than 15%) statistically.

[A case of pancreatic cancer with invasion to the wall of the intrahepatic bile duct].

Presented here is the case of a 69-year-old woman with pancreatic cancer. Efforts were made to discover the origin of the cancer but without success. Tumor nodules were scarcely observed in the pancreas macroscopically at autopsy, while histological examination of the pancreas revealed the presence of a well-differentiated adenocarcinoma (duct cell carcinoma). The invasion to the wall of the intrahepatic bile duct was also observed microscopically.

[Two cases of severe acute pancreatitis-CT scan was useful to the assessment].

We report two cases of severe acute pancreatitis; a 53-year-old man (Case 1) and a 60-year-old woman (Case 2). Case 1 was classified as "severe" according to the Ranson's criteria and he died of MOF on the 21st hospital day. Case 2 was classified as "moderate", but a large pancreatic abscess was observed by CT scan. She died of this abscess complicated with duodenal perforation on the 33rd hospital day. CT findings showed that this case was not "moderate" but "severe". Therefore, we believe that the findings of CT scan are an important factor for assessment of the severity of acute pancreatitis.

[A case of Bartter-like syndrome with abdominal pain, paresis of the extremities and muscle weakness].

A 16-year-old woman was admitted to our hospital because of abdominal pain, paresis of extremities, and muscle weakness. Bartter's syndrome was suspected because of the features of the hypokalemia, hyperaldosteronism, hyperreninemia, increased concentration of plasma angiotensin I & II, the defect in distal fractional reabsorption of chloride and normotension. The concentrations of the plasma angiotensin II and aldosterone, however, were decreased to normal levels after admission probably due to a decrease in the salt intake because of the regular hospital diet. Furthermore, the hyperplasia of the juxtaglomerular apparatus was not found. Therefore, at first we thought we were dealing with another disease, that is pseudo-Bartter's syndrome, which is caused by different pathogenesis. Gill et al. reported that the defect in distal fractional reabsorption of chloride was a characteristic feature in the diagnosis of the Bartter's syndrome. Thus, we tried to explain the clinical symptoms and diagnosis of this case as the Bartter's syndrome according to the theory of Gill et al. If the defect of chloride reabsorption was the only pathogenesis of the Bartter's syndrome, other symptoms seen in our case could be thought of as secondary (or new) changes. Therefore, this case could be differentiated from the pseudo-Bartter's syndrome or the "true" Bartter's syndrome because of the clinical symptoms and the defect in chloride reabsorption.

[A case of adult Still's disease with cardial symptoms of remittent high fever and arthralgia for more than six months].

A 46-year-old woman was admitted to our hospital because of fever of unknown origin. Her fever continued with a daily rise to around 38 degrees C to 40 degrees C for more than six months, occasionally accompanied by polyarthralgia, erythematous rash and cervical lymphadenopathy. The erythrocyte sedimentation rate was 75 mm per hour and CRP was over 6+. The white-cell count was 15,100 with 21 percent band forms and the alpha 2-globulin was 18.8 percent of total protein. The tests for autoantibodies were negative. Clinical course and laboratory findings in this case were most compatible with adult Still's disease. A radionucleotide bone scan with 99mTc showed a greatly increased uptake of the radionucleotide at the large joints of the whole body, and a gallium scan also revealed accumulation of the radionucleotide at the bone marrow of the sternum. These findings were recognized during high fever and disappeared when the body temperature returned to normal. These examinations might be useful for investigating the active site of inflammation and for studying the pathogenesis of adult Still's disease.

[The difficulty of distinguishing retroperitoneal schwannoma from liver tumor in the caudate lobe].

A 68-year-old woman was admitted to our hospital for the evaluation of an abdominal tumor. Both ultrasonography and computerized tomography revealed a mass at the caudate lobe of the liver. However, it was difficult to distinguish the mass from a liver tumor in the caudate lobe. The laboratory data showed only a slight elevation of the serum gamma-GTP level and ICG retention. The abdominal arteriography revealed a vascular tumor located in the retroperitoneal space, which was receiving its blood supply from the right inferior phrenic artery. When a laparotomy was performed, a yellowish solid mass (8 X 6 X 4.5 cm) was found behind the lateral segment of the left liver lobe. Histological examination showed interlacing bands of uniform spindle cells of which elongated nuclei were arranged in a palisading pattern. The tumor was diagnosed as schwannoma. Literatures on retroperitoneal schwannoma are reviewed, as this tumor is relatively rare.

[A pregnant woman with active ulcerative colitis maintained on total parenteral nutrition].

A 28-year-old woman was admitted to our hospital with complaints of mucosanguinous stool and low grade fever. She was diagnosed as a typical chronic continuous type of ulcerative colitis by the findings of barium enema and colonoscopy. Since she had an allergy to sulfasalazine, prednisolone was chosen. She became pregnant during an active stage while being treated with 20 mg of prednisolone a day. Prednisolone was withdrawn to avoid the side-effects of the medicine on the fetal outcome. This resulted in her symptoms becoming far worse and the oral ingestion being discontinued. Total parenteral nutrition (TPN) was required under careful nutritional management. The TPN consisted of glucose, electrolytes, amino acids, vitamins, trace elements and intravenous lipid preparation. Her total energy intake was 2320 kcal a day. Vitamins were administered to her on the bases of the guideline of the American medical association. Rapid turnover proteins, transferrin, vitamins, trace elements and amino acids in addition to routine laboratory tests were measured to estimate her nutritional condition. The data showed that biotin was 10 times lower than the expected value and that other factors were within normal limits. This is the first case in Japan where a woman suffering from an active ulcerative colitis was treated with TPN and delivered of a healthy baby. We concluded that TPN under careful control was useful in the nutritional management and therapy of the pregnant patient who suffered from severe colitis. We believe that the amount of biotin's supplementation should be increased in this type of case because it was 10 times lower than the normal value, although the deficiency symptoms did not develop.