Ionotropic glutamate receptors and glutamate transporters are involved in necrotic neuronal cell death induced by oxygen-glucose deprivation of hippocampal slice cultures.

Organotypic hippocampal slice cultures represent a feasible model for studies of cerebral ischemia and the role of ionotropic glutamate receptors in oxygen-glucose deprivation-induced neurodegeneration. New results and a review of existing data are presented in the first part of this paper. The role of glutamate transporters, with special reference to recent results on inhibition of glutamate transporters under normal and energy-failure (ischemia-like) conditions is reviewed in the last part of the paper. The experimental work is based on hippocampal slice cultures derived from 7 day old rats and grown for about 3 weeks. In such cultures we investigated the subfield neuronal susceptibility to oxygen-glucose deprivation, the type of induced cell death and the involvement of ionotropic glutamate receptors. Hippocampal slice cultures were also used in our studies on glutamate transporters reviewed in the last part of this paper. Neurodegeneration was monitored and/or shown by cellular uptake of propidium iodide, loss of immunocytochemical staining for microtubule-associated protein 2 and staining with Fluoro-Jade B. To distinguish between necrotic vs. apoptotic neuronal cell death we used immunocytochemical staining for active caspase-3 (apoptosis indicator) and Hoechst 33342 staining of nuclear chromatin. Our experimental studies on oxygen-glucose deprivation confirmed that CA1 pyramidal cells were the most susceptible to this ischemia-like condition. Judged by propidium iodide uptake, a selective CA1 lesion, with only minor affection on CA3, occurred in cultures exposed to oxygen-glucose deprivation for 30 min. Nuclear chromatin staining by Hoechst 33342 and staining for active caspase-3 showed that oxygen-glucose deprivation induced necrotic cell death only. Addition of 10 microM of the N-methyl-D-aspartate glutamate receptor antagonist MK-801, and 20 microM of the non-N-methyl-D-aspartate glutamate receptor antagonist 2,3-dihyroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline to the culture medium confirmed that both N-methyl-D-aspartate and non-N-methyl-D-aspartate ionotropic glutamate receptors were involved in the oxygen-glucose deprivation-induced cell death. Glutamate is normally quickly removed, from the extracellular space by sodium-dependent glutamate transporters. Effects of blocking the transporters by addition of the DL-threo-beta-benzyloxyaspartate are reviewed in the last part of the paper. Under normal conditions addition of DL-threo-beta-benzyloxyaspartate in concentrations of 25 microM or more to otherwise untreated hippocampal slice cultures induced neuronal cell death, which was prevented by addition of 2,3-dihyroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline and MK-801. In energy failure situations, like cerebral ischemia and oxygen-glucose deprivation, the transporters are believed to reverse and release glutamate to the extracellular space. Blockade of the transporters by a subtoxic (10 microM) dose of DL-threo-beta-benzyloxyaspartate during oxygen-glucose deprivation (but not during the next 48 h after oxygen-glucose deprivation) significantly reduced the oxygen-glucose deprivation-induced propidium iodide uptake, suggesting a neuroprotective inhibition of reverse transporter activity by DL-threo-beta-benzyloxyaspartate during oxygen-glucose deprivation under these conditions. Adding to this, other results from our laboratory have demonstrated that pre-treatment of the slice cultures with glial cell-line derived neurotrophic factor upregulates glutamate transporters. As a logical, but in some glial cell-line derived neurotrophic factor therapy-related conditions clearly unwanted consequence the susceptibility for oxygen-glucose deprivation-induced glutamate receptor-mediated cell death is increased after glial cell-line derived neurotrophic factor treatment. In summary, we conclude that both ionotropic glutamate receptors and glutamate transporters are involved in oxygen-glucose deprivation-induced necrotic cell death in hippocampal slice cultures, which have proven to be a feasible tool in experimental studies on this topic.

Long-term results of periosteal transplantation in osteochondritis dissecans of the knee.

Between 1986 and 1991, a total of 18 patients (11 men and 7 women) with osteochondritis dissecans of the knee were treated with periosteal transplantation. Median patient age was 19 years (range: 16-45 years). Eight patients were reoperated up to 8 years postoperatively, due to reduced range of motion, synovitis, or formation of an exostosis in the transplanted area. Of 14 patients who were available for follow-up after 8 years (range: 5-10), 2 were completely pain free. Six patients had reduced range of motion, knee instability, or quadriceps muscle atrophy. The number of reoperations and the presence of continued knee pain in most patients does not justify the extensive procedure of periosteal transplantation.

Chronic hepatitis B virus infection in Asian countries.

Of the estimated 50 million new cases of hepatitis B virus (HBV) infection diagnosed annually, 5-10% of adults and up to 90% of infants will become chronically infected, 75% of these in Asia where hepatitis B is the leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma (HCC). In Indonesia, 4.6% of the population was positive for HBsAg in 1994 and of these, 21% were positive for HBeAg and 73% for anti-HBe; 44% and 45% of Indonesian patients with cirrhosis and HCC, respectively, were HBsAg positive. In the Philippines, there appear to be two types of age-specific HBsAg prevalence, suggesting different modes of transmission. In Thailand, 8-10% of males and 6-8% of females are HBsAg positive, with HBsAg also found in 30% of patients with cirrhosis and 50-75% of those with HCC. In Taiwan, 75-80% of patients with chronic liver disease are HBsAg positive, and HBsAg is found in 34% and 72% of patients with cirrhosis and HCC, respectively. In China, 73% of patients with chronic hepatitis and 78% and 71% of those with cirrhosis and HCC, respectively, are HBsAg positive. In Singapore, the prevalence of HBsAg has dropped since the introduction of HBV vaccination and the HBsAg seroprevalence of unvaccinated individuals over 5 years of age is 4.5%. In Malaysia, 5.24% of healthy volunteers, with a mean age of 34 years, were positive for HBsAg in 1997. In the highly endemic countries in Asia, the majority of infections are contracted postnatally or perinatally. Three phases of chronic HBV infection are recognized: phase 1 patients are HBeAg positive with high levels of virus in the serum and minimal hepatic inflammation; phase 2 patients have intermittent or continuous hepatitis of varying degrees of severity; phase 3 is the inactive phase during which viral concentrations are low and there is minimal inflammatory activity in the liver. In general, patients who clear HBeAg have a better prognosis than patients who remain HBeAg-positive for prolonged periods of time. The outcome after anti-HBe seroconversion depends on the degree of pre-existing liver damage and any subsequent HBV reactivation. Without pre-existing cirrhosis, there may be only slight fibrosis or mild chronic hepatitis, but with pre-existing cirrhosis, further complications may ensue. HBsAg-negative chronic hepatitis B is a phase of chronic HBV infection during which a mutation arises resulting in the inability of the virus to produce HBeAg. Such patients tend to have more severe liver disease and run a more rapidly progressive course. The annual probability of developing cirrhosis varies from 0.1 to 1.0% depending on the duration of HBV replication, the severity of disease and the presence of concomitant infections or drugs. The annual incidence of hepatic decompensation in HBV-related cirrhosis varies from 2 to 10% and in these patients the 5-year survival rate drops dramatically to 14-35%. The annual risk of developing HCC in patients with cirrhosis varies between 1 and 6%; the overall reported annual detection rate of HCC in surveillance studies, which included individuals with chronic hepatitis B and cirrhosis, is 0.8-4.1%. Chronic hepatitis B is not a static disease and the natural history of the disease is affected by both viral and host factors. The prognosis is poor with decompensated cirrhosis and effective treatment options are limited. Prevention of HBV infection thorough vaccination is still, therefore, the best strategy for decreasing the incidence of hepatitis B-associated cirrhosis and HCC.

Immunohistochemical study of P53, PCNA and AFP in hepatocellular carcinoma, a comparison between Indonesian and Japanese cases.

Mutations of p53 as a tumor suppressor gene in hepatocellular carcinoma (HCC) have been reported to occur with varying frequency in different geographic regions, which might be different etiology for HCC. Overexpressions of p53 (well known for its implications in mutations of the p53 gene), PCNA and alpha-fetoprotein (AFP) have been reported to be associated with carcinogenesis and/or tumor progression and poor prognosis in various types of cancer. To estimate the geographical difference of the p53 gene, PCNA and AFP in HCC, we examined 14 Japanese HCC cases, 8 Indonesian HCC cases, and 27 Indonesian chronic active hepatitis (CAH) or liver cirrhosis cases, using immunohistochemical approaches. Overexpression of p53 was identified in 37.5% of Japanese HCC, in 62.5% of Indonesian HCC and none in CAH. The mean PCNA Labeling Index of Japanese HCC, Indonesian HCC and CAH was detected in 48.6%, 30.4%, and 43.5%, respectively. AFP was detected in 35.7% of Japanese and 25% of Indonesian HCC. The rate of p53 overexpression in Indonesian HCC was as high as in HCC of southern part of China, which might share the similar etiology in both regions.

[Medial migration of a Hansson hook-pin after femoral neck fracture]. / Medial migration af Hansson-søm efter collum femoris-fraktur.

Medial migration of a LIH (Lars Ingvar Hansson) hookpin is a very rare complication to internal fixation of hip fractures. We report a case of medial migration of an LIH hook-pin into the acetabulum. The importance of using the correct length hook-pin and ensuring that the base of the hook-pin is outside the lateral cortex of the femur is stressed.

Effectiveness of the analogue of natural Schisandrin C (HpPro) in treatment of liver diseases: an experience in Indonesian patients.

OBJECTIVE: To determine the effect of dimethyl-4,4'-dimethoxy-5,6, 5',6-dimethylene dioxybiphenyl-2,2'-dicarboxylate (HpPro) on patients with acute and chronic liver diseases. METHODS: An open trial and a prospective randomized and controlled study were performed. The open trial consisted of 56 cases (16 cases of acute hepatitis, 20 cases of chronic hepatitis, 14 cases of liver cirrhosis and 6 cases of fatty liver). Controlled study consisted of 20 cases of Child A chronic hepatitis which were randomly treated with either HpPro or a mixture of known drugs which used as a liver protective agent in Indonesia as control for one week. The patients were then crossed over those two drugs in the next week. RESULTS: In the open trial, after 4 weeks' treatment with HpPro 7.5 mg orally three times daily, acute hepatitis, chronic hepatitis and fatty liver cases showed rapid decrease of SGOT and SGPT. In the liver cirrhosis cases, SGOT and SGPT were decreased slowly. In the controlled trial, nine patients received HpPro 7.5 mg three times daily orally and eleven were treated with a mixture of known drugs as the controls. After one week treatment, HpPro group clinically showed significant decrease of SGPT and SGOT levels compared to control group (P = 0.035). At the second week, HpPro group showed significant decrease of SGOT compared to control group (P = 0.038) but the decrease of SGPT was not significant (P = 0.096). CONCLUSION: Treatment with HpPro is effective to reduce liver impairment in acute and chronic liver diseases on Indonesian patients. No side effect of HpPro was observed.

Ethnicity, socioeconomic status, transfusions and risk of hepatitis B and hepatitis C infection.

This study identifies the risk factors for hepatitis B virus (HBV) and hepatitis C virus (HCV) and measures the prevalence of hepatitis B surface antigen (HBsAg) and antibody to hepatitis C (anti-HCV) in the general population of Jakarta. A population-based sample of 985 people aged 15 and above was surveyed. Risk factors were identified through questionnaires and home visits. Serum was analysed for HBsAg, antibody to hepatitis B surface antigen (anti-HBs), anti-HCV, aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The seroprevalence was: 4.0% (39/985) for HBsAg, 17.2% (170/985) for anti-HBs, and 3.9% (38/985) for anti-HCV. The risk factors for hepatitis B and hepatitis C infection had little in common. Low socioeconomic status was a strong risk factor for HBsAg (adjusted odds ratio (OR) 18.09; 95% confidence interval (CI) 2.35-139.50). In addition, the Chinese group has 2.97 higher risk of having HBV infection compared with the Malayan ethnic group (adjusted OR 2.97; 95% CI 1.22-7.83). There was moderate positive trend between family size and risk of HBsAg positivity (P = 0.130). Age over 50 (adjusted OR 14.72; 95% CI 4.35-49.89) and history of transfusion were significant risk factors for hepatitis C (adjusted OR 3.03; 95% CI 1.25-7.33). Hepatitis B and hepatitis C infections have different risk factors in Jakarta, a high risk in population for both diseases. Hepatitis B transmission is associated with low socioeconomic status, Chinese ethnic group and large family size, while hepatitis C is associated with an older age and a history of transfusions.

[Non-traumatic subcutaneous emphysema of the femur. A rare complication of colorectal cancer]. / Ikketraumatisk subkutant emfysem på femur. En sjaelden komplikation til kolorektal cancer.

A case of subcutaneous emphysema of the left thigh as the initial symptoms of a perforated cancer of the descending colon is presented The literature on the subject is briefly reviewed. The resemblance to nontraumatic gas gangrene, and the importance of early therapy is discussed.

[Acute pulmonary infections at autopsy. A study of clinical and macroscopic diagnoses at autopsy compared with microscopic autopsy findings]. / Akutte lungeinfektioner ved obduktion. Undersøgelse af klinisk og makroskopisk diagnostik ved obduktion sammenholdt med mikroskopiske obduktionsfund.

The clinical diagnoses and gross diagnoses of acute pulmonary infections were investigated for 100 consecutively performed hospital autopsies, and the diagnoses were compared with the histological findings. We found 34 cases of pneumonia and one case of tuberculosis. Of these, 29 infections represented principal diagnoses, i.e. as causative of or contributive towards death. The predictive values, the sensitivities and specificities were estimated. The predictive value for gross diagnostics was low with a value of 42.9% against 58.8% for clinical diagnostics. The sensitivities for both clinical diagnostics and gross diagnostics were about 30% and the specificities about 85%. The study shows that sampling for histology from all pulmonary lobes is essential for correct autopsy diagnoses, either from areas that appear to be infected on gross examination or from the peripheral parts. Furthermore autopsy performance is still of great value for clinical diagnostics and for medical statistics.

[Survey of clinical diagnoses and autopsy findings. Autopsy findings and sensitivity, specificity and predictive values for clinical diagnosis during the periods 1 July 1980-30 June 1081 and 1 July 1990-30 June 1993]. / Undersøgelse af kliniske diagnoser og obduktionsfund. Obduktionsfund og sensitivitet, specificitet og proediktive voerdier for klinisk diagnostik i perioderne 1.7.1980-30.6.1981 og 1.7.1990-30.6.1993.

The sensitivity, specificity and clinical accuracy of clinical diagnoses were determined and compared for two periods of time: 1.7.1980-30.6.1981 and 1.7.1990-30.6.1993 based on the analysis of 286 and 138 autopsies respectively. The autopsy rate decreased from 82.7% in the first period to 11.2% in the second. The first period shows a generally higher sensitivity and accuracy for positive diagnosis. Both periods reveal the lowest sensitivity for pulmonary embolism and the lowest accuracy for positive clinical diagnosis of pneumonia/bronchopneumonia. For malignancies and arteriosclerotic heart diseases significant discrepancy between the periods was demonstrated using the chi 2-test. The results are influenced by low autopsy rates causing fewer true-positive diagnoses and a declining sensitivity. This type of study is a useful tool for demonstrating changes in the diagnostic procedure. The present investigation demonstrates a need for further analysis of malignancies to explain the simultaneous decrease in sensitivity, specificity and accuracy in spite of an increasing number of malignancies in autopsy findings.

[Antibiotic prophylaxis in Danish orthopedic alloplastic surgery]. / Antibiotikaprofylakse i dansk ortopaedisk alloplastikkirurgi.

In order to assess the routine use of prophylactic antibiotics (AB) in arthroplastic surgery in Denmark, questionnaires were sent to all Danish orthopaedic departments and all general surgical departments that perform orthopaedic surgery. Fifty-six departments (93%) returned the questionnaires. All departments use prophylactic AB in primary knee and hip arthroplasty and in revision arthroplasty. In addition, all departments but one use prophylactic AB in arthroplasty secondary to osteosynthesis. The largest group of departments uses penicillinase-resistant penicillin (PRP) in their standard prophylaxis regimens. The second largest group uses second generation cephalosporins. With one exception, all use cefuroxime. A small group uses other types of AB. Fifteen percent of the departments combine systemic AB with gentamicin bone cement (GC) in primary hip arthroplasty, whereas 22% use this combination in primary knee arthroplasty. Significantly more departments use GC in revision arthroplasty (89%) and in arthroplasty secondary to osteosynthesis (63%). Prolonged antibiotic prophylaxis (beyond 24 hours) is practised to a significantly higher degree in revision arthroplastic surgery than in the primary arthroplasties. In conclusion, one of two homogeneous groups of prophylactic AB is used in arthroplastic surgery in Denmark as prescribed in the literature.

Release of trigger finger in children. Long term results.

During a 10 year period 58 patients had 80 trigger fingers operated on at this hospital. The male/female ratio was 32:26 and most children were operated on at between the ages of 2 and 4 years, and 15 months (range 2-99) after the lesion had been noticed. The operation was quick and effective with no complications. Only one patient was re-operated on, with a good result. Thirteen patients had bilateral lesions. Both hands were examined and in 11 patients (19%) we found that the thickening corresponding to the trigger finger was also present on the opposite side but giving no symptoms. There seems to be a predisposition to bilateral affections.

Hepatitis C virus genotypes in Singapore and Indonesia.

5' untranslated and partial core (C) region sequence of hepatitis C virus (HCV) in 21 Singaporean and 15 Indonesian isolates were amplified by reverse-transcription polymerase chain reaction and sequenced with the use of conserved primer sequences deduced from HCV genomes identified in other geographical regions. The HCV genotypes are predominantly that of Simmonds type 1 and less of type 2 and 3 with the latter genotype currently not detected in Indonesia. The 5' untranslated sequences are related to HCV-1. DK-7 (Denmark), US-11 (United States of America), HCV-J4, SA-10 (South Africa), T-3 (Taiwan), HCV-J6, HCV-J8, Eb-1 and Eb-8. When compared with the prototype HCV-1, insertions are found within the 5' untranslated region of Singaporean isolates and not in the Indonesians. There are Singaporean and Indonesian isolates that have sequences within the 5' untranslated region that differ slightly from each other. Microheterogeneity is observed in the core region of two Singaporeans and one Indonesian isolate. Finally, not all HCV isolates can be amplified with the conserved core sequence primers when compared with the ease with which these isolates can be amplified with 5' untranslated region conserved primers.

[Quality assurance in amputation of the lower extremities due to vascular insufficiency]. / Kvalitetssikring ved amputationer på underekstremiteterne grundet vasculaer insufficiens.

In this retrospective study we reviewed the clinical charts for 82 patients who underwent lower limb amputation at our department during 1990 and 1991. Our results are compared with the literature, and different aspects of the treatment are discussed. Subsequently a model for future quality assurance is presented.

[Importance of the autopsy rate. A comparison between clinical assessment and findings at autopsies during the periods: 1 July 1908--30 June 1981 and 1 July 1990--30 June 1991]. / Obduktionsfrekvensens betydning. Sammenligning mellem klinisk vurdering og obduktionsfund i perioderne 1.7.1980-30.6.1981 og 1.7.1990-30.6.1991.

On the 1.7.1990 a new law was introduced in Denmark whereby specific consent had to be obtained from the deceased's relatives before autopsy could be performed. Autopsy records from two periods before and after this date, namely from 1.7.1980 to 1.7.1981 and from 1.7.1990 to 30.6.1991, are examined here with respect to autopsy rate and agreement/disagreement between clinical assessment as to cause of death and findings on autopsy. With an autopsy rate of 82.7% before and 10.3% after the new law we found that there was significantly more disagreement between the clinical assessment and the autopsy findings in the 1990/1991 period, presumably because of a selection bias towards difficult cases. The autopsy material from 1990/1991 could only be used for statistics in a limited fashion. A very low autopsy rate of about 10% implies that 25-30% of all death certificates from hospitals must be assumed to be incorrect as concerning serious diseases. The study shows that there is a considerable need for the performance of autopsies for the sake of medical statistics.

Computer registration of infections used to measure the effect of prophylactic antibiotics on postoperative infections following osteosynthesis in hip fractures.

Continuous registration of operations and wound infections was used to demonstrate the effect of prophylactic antibiotics in hip fracture osteosynthesis. In order to monitor wound infections and other postoperative complications in an orthopaedic department, 2 years' data on 688 patients with hip fractures, were entered into a personal computer program. During 1990, there was no formal policy for antibiotic prophylaxis; during 1991 prophylactic cefuroxime was recommended for osteosynthesis of hip fractures. In 1990 56% of patients were given prophylaxis and in 1991 this rose to 79%. Overall, 68% of patients had prophylaxis. The overall rate of deep wound infections (DWI) was significantly lower in patients treated with prophylactic antibiotics (0.6%), compared with those without prophylaxis (4.6%). Patients with DWI were admitted to the hospital for an average of 43.7 days, compared with 14.6 days for patients without complications. We recommend the use of prophylactic cefuroxime in hip fracture osteosynthesis. Computer registration of complications is a useful method for clinical quality control in an orthopaedic department.

Ductal adenoma of the breast: the cytological features of six cases.

Ductal adenoma of the breast may simulate malignancy by both clinical, histological, and cytological examination. Fine-needle aspirations from breast lesions are now daily routine and preoperative recognition of this benign lesion is desirable. This study describes the cytologic features of six additional cases of ductal adenoma. The smears from all the lesions were highly cellular with epithelial cells in sheets. Numerous large fragments of purple stroma in tight connection with epithelial cells often making finger-like hyaline structures or globules between cells were seen. Naked oval nuclei in the background were observed in all cases indicating benignancy. We find the cytologic picture in fine-needle aspirations from ductal adenomas sufficiently characteristic for preoperative diagnosis. The differential diagnoses toward other benign and malignant conditions are discussed.