Osteoarthritis of the Knee in the Rabbit Produced by Immobilization:: Attempts to Achieve a Reproducible Model for Studies on Pathogenesis and Therapy

In order to obtain a reproducible experimental model of osteoarthritis a method of immobilizing the rabbit's knee in extension by means of a plastic splint was developed. The right knees of the rabbits were immobilized for periods varying from 4 days to 24 weeks. With the left knees as controls the knees were studied in a variety of ways among these being radiography (126 rabbits), histological sections stained with Alcian Blue (88 rabbits), analysis of ³⁵S-sulphate uptake (22 rabbits) and ³⁵S-autoradiography (6 rabbits). In 27 rabbits the regaining of mobility after immobilization was studied. After 5-6 weeks of immobilization most of the knees showed moderate or severe changes including loss of articular cartilage and osteophyte formation. Immobilization of the rabbit's knee by this method provokes a fairly easily reproducible type of degenerative joint disease showing similarities to advanced osteoarthritis as seen in humans.     

A mouse model for Stickler's syndrome: ocular phenotype of mice carrying a targeted heterozygous inactivation of type II (pro)collagen gene (Col2a1)

The influences of targeted heterozygous inactivation of type II (pro)collagen gene (Col2a1) on eye structures in the 15-month-old C57BL/6JOlaHsd mouse was studied. The eyes were collected from C57BL mice heterozygous for a targeted inactivation of one allele of the Col2a1 gene (Col2a1(+/-) mice). The eyes of C57BL mice with normal gene alleles were used as controls (Col2a1(+/+) mice). Ocular histology was analyzed from tissue sections, stained with hematoxylin and eosin, toluidine blue and alcian blue. Type II collagen was localized by immunohistochemistry. Hyaluronan (HA) was stained utilizing the biotinylated complex of the hyaluronan-binding region of aggrecan and link protein (bHABC). The anterior segment of the eye was well-formed in both genotypes, but typical folding of ciliary processes was decreased, while increased stromal extracellular matrix vacuolization was seen in the Col2a1(+/-) mice. In the lens of these mice, subcapsular extracellular matrix changes were observed. Differences in retinal structures or the number of the eyes with retinal detachment were not detected between the genotypes. In Col2a1(+/-) mice, staining for type II collagen was weaker in cornea, ciliary body, iris, lens, vitreous, retina, choroid and sclera than in the control mice. HA staining was detected in the extraocular tissues, ciliary body, iris and the choroid of both genotypes. HA staining was observed only in the vitreous body of the control animals. Heterozygous inactivation of Col2a1 gene causes structural defects in the murine eye. The observed structural changes in the ciliary body, lens and vitreous of the Col2a1(+/-) mice may represent ocular features found in the human Stickler syndrome, where the abnormalities result from COL2A1 gene mutations which lead to functional haploinsufficiency.     

Drug-related visits to a district hospital emergency room

The role of adverse drug reactions (ADRs) as a cause of hospital visits varies depending on the type of hospitals. Our aim was to determine the incidence of drug-related emergency department visits to a district hospital, and to identify the drugs and patient groups involved. All patient visits to the emergency department of a Finnish district hospital were evaluated prospectively for 6 months. The physician on duty and a clinical pharmacologist selected all possibly drug-related visits for further scrutinising. The causality assessment (drug-related or not) was judged according to WHO criteria, based on the patients' files, including laboratory and other data. Of the 7113 evaluated visits, 167 (2.3%) were "certainly" or "probably" drug-related; 102 (1.4% of all) were related to ADRs and 65 (0.9%) to intentional overdoses. The most common ADRs were gastrointestinal symptoms (n=17) caused by antibiotics, opioids, nonsteroidal antiinflammatory or cytostatic drugs. The International Classification of Disease (ICD-10) codes on patients' files were insensitive to disclose ADRs. The ADR patients were older (mean age 57 years) than the intentional overdose patients (38 years; P<0.001). Males predominated in the intentional overdose group (38 males, 27 females) but not in the ADR patients. The majority of intentional overdoses was caused by psychotropics. The ADRs lead to hospitalisation in a higher frequency (51%) than did the intentional overdoses (35%). In conclusion, the incidence of "certainly" or "probably" drug-related visits to the district hospital emergency room was relatively low. The ICH-10 codes on patients' files were found to be insensitive to disclose the ADRs, even when they lead to hospital admission, casting doubts on the usefulness of ICH codes alone in ADR evaluation.     

Mutation screening of the androgen receptor promoter and untranslated regions in prostate cancer

BACKGROUND: Mechanisms, other than gene amplification, leading to overexpression of AR in androgen ablation-resistant prostate cancer remain unknown and could include genetic alterations in the promoter or untranslated regions (UTR) of the AR gene. MATERIALS AND METHODS: DNAs from five prostate cancer cell lines, 19 LuCaP xenografts, 44 clinical tumors, and 36 non-malignant controls were used for screening mutations in the upstream regulatory region, promoter and the 5'- and 3'-UTRs of the AR gene with denaturating high performance liquid chromatography (DHPLC) and sequencing. RESULTS: Ten different sequence variations were found in prostate cancer cell lines and xenografts. However, none of them were recurrent or were found in clinical prostate cancer specimens or in normal controls. CONCLUSIONS: Recurrent mutations in the promoter or UTRs of AR seem to be rare, and thus not likely mechanisms for the increased expression of the gene in the androgen ablation-resistant prostate cancer.     

Amplification of the urokinase gene and the sensitivity of prostate cancer cells to urokinase inhibitors

OBJECTIVES: To evaluate the frequency of the urokinase-type plasminogen activator (uPA) gene amplification and the sensitivity of prostate cancer cells to uPA inhibition, as we previously found one hormone-refractory prostate tumour with high-level amplification of the uPA (alias PLAU) gene, and also showed that a uPA inhibitor, amiloride, can effectively reduce the invasion potential of the PC-3 prostate cancer cell line. MATERIALS AND METHODS: Sixty-three locally recurrent hormone-refractory tumours and 78 hormone-refractory metastases from 29 patients who died from prostate cancer were analysed for uPA gene-copy number using fluorescence in situ hybridization. The Matrigel invasion assay was used to study the influence of uPA inhibitors on the invasive potential of prostate cancer cell lines. RESULTS: Of the locally recurrent hormone-refractory tumours, 21% had an increased copy number of uPA, but no high-level amplifications were found; 31% of the metastases had increased copy number and one high-level amplification of the uPA. Matrigel invasion assays with two specific uPA inhibitors, B428 and p-aminobenzamidine, showed that invasion of a prostate cancer cell line containing uPA gene amplification was inhibited by these small-molecule uPA inhibitors, while invasion of prostate cell lines without uPA gene amplification were not. CONCLUSION: These results suggest that selective inhibition of the uPA pathway in individuals whose tumours contain uPA gene amplification may provide therapeutic benefit.     

Positive genetic test led to an early diagnosis of myxoma in a 4-year-old boy

Less than 10% of cardiac myxomas are familial. These familial cases are related to Carney complex, a multiple neoplasia and lentiginosis syndrome. Mutations in the PRKAR1alpha gene are the cause of Carney complex in most patients. We report a boy, who had PRKAR1alpha gene mutation, and atrial myxoma that was diagnosed in a routine echocardiographic study at the age of four years. Surgical excision of myxoma was performed. This case demonstrates the benefit of screening genetically the kindreds of patients with familial myxomas, and the importance of close follow-up of individuals affected with this mutation irrespective of age.     

Influenza A(H1N1) viruses of the 1977/78 outbreak: isolation and haemagglutination properties.

During the H1N1 outbreak of 1977/8, the virus was isolated in embryonated eggs from 59 out of 76 patients (78%) with the serologically confirmed infection. A similar isolation frequency has been achieved during a period of six H3N2 outbreaks since 1972/3. The H1N1 strains were isolated less frequently from late specimens (collected 4--6 days from the onset of illness) and more often only in the second passage compared with the H3N2 viruses. The new H1N1 strains resembled those prevalent in the 1950s with respect to their ability to agglutinate erythrocytes of certain laboratory animals and to be eluted from these, and thus differed from the H3N2 viruses.