Increased calpain expression in activated glial and inflammatory cells in experimental allergic encephalomyelitis

In demyelinating diseases such as multiple sclerosis (MS), myelin membrane structure is destabilized as myelin proteins are lost. Calcium-activated neutral proteinase (calpain) is believed to participate in myelin protein degradation because known calpain substrates [myelin basic protein (MBP); myelin-associated glycoprotein] are degraded in this disease. In exploring the role of calpain in demyelinating diseases, we examined calpain expression in Lewis rats with acute experimental allergic encephalomyelitis (EAE), an animal model for MS. Using double-immunofluorescence labeling to identify cells expressing calpain, we labeled rat spinal cord sections for calpain with a polyclonal millicalpain antibody and with mAbs for glial (GFAP, OX42, GalC) and inflammatory (CD2, ED2, interferon gamma) cell-specific markers. Calpain expression was increased in activated microglia (OX42) and infiltrating macrophages (ED2) compared with controls. Oligodendrocytes (galactocerebroside) and astrocytes (GFAP) had constitutive calpain expression in normal spinal cords whereas reactive astrocytes in spinal cords from animals with EAE exhibited markedly increased calpain levels compared with astrocytes in adjuvant controls. Oligodendrocytes in spinal cords from rats with EAE expressed increased calpain levels in some areas, but overall the increases in calpain expression were small. Most T cells in grade 4 EAE expressed low levels of calpain, but interferon gamma-positive cells demonstrated markedly increased calpain expression. These findings suggest that increased levels of calpain in activated glial and inflammatory cells in EAE may contribute to myelin destruction in demyelinating diseases such as MS.     

Localization and possible gene expression of proteoglycan decorin in the trabecular meshwork

It is known that trabecular meshwork cells produce proteoglycans and that local production may be associated with aqueous outflow resistance. In an attempt to identify intraocular production of proteoglycan decorin in the anterior chamber angle of mammalian eyes, we conducted a Northern blot analysis and immunohistochemical studies. Northern blot analysis suggested gene expression of proteoglycan decorin in trabecular meshwork cells. Also, immunohistochemical studies using anti-decorin antibody demonstrated decorin-like immunoreactivity in the trabecular meshwork and around the Schlemm's canal. Our data demonstrate the presence of proteoglycan decorin in the outflow pathway, suggesting that decorin is a component of extracellular matrices in these regions and may be associated with outflow resistance.     

Increased diameters of collagen fibrils precipitated in vitro in the presence of decorin from various connective tissues

Proteoglycans were isolated from bovine skin, sclera, deep flexor tendon and the periphery of the temporomandibular joint disc with urea. Decorin was purified from each of these extracts by ion-exchange, hydrophobic-interaction and gel-filtration chromatography. Purities were assessed by amino acid analysis and by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) of the protein cores released by digestion with chondroitin-ABC-lyase. In these respects the decorins were indistinguishable. However the glycosaminoglycan chains released by digesting the proteoglycans with papain varied widely in mobility on SDS-PAGE: that from skin decorin migrating fastest and that from tendon decorin slowest. The effects of each of the decorins on collagen fibrillogenesis in vitro were similar, all reducing the rate of fibril growth (by 55 to 71%, depending on the source of the proteoglycan) and increasing the diameters of the fibrils formed (by 27 to 66%). Core protein alone, isolated from skin decorin, reduced the rate of fibril growth as effectively as intact decorin, but had no effect on the diameter of fibrils formed. The dermatan sulphate chain and the protein thus appear to play different roles in the interaction of intact decorin with collagen. These data suggest that decorin found in fibrous connective tissues may increase Type I collagen fibril diameters, resulting in tissues that are better able to withstand tensile forces.     

Gene therapy by skeletal muscle expression of decorin prevents fibrotic disease in rat kidney

This experiment was an attempt to use the soybean residue derived from the production of "soy milk". The residue contains about 18% protein, 70% carbohydrates and 7.5% lipid as fish feed for rearing common carp, Cyprinus carpio. There were 4 types of diets: (1) soybean residue, (2) soybean residue digested with Papain, (3) soybean residue (64%) mixed with beef liver (34%) and (4) same mixture as (3) but digested with Papain. The results indicate that the percentage increase in weight and length of fish feeding with beef liver supplemented diets was higher than those feeding with soybean residue alone. This was possibly due to the fact that beef liver was able to supplement the nutrient deficiency in soybean. The two types of feeds (2 and 4) digested with Papain also yielded significantly better fish growth in terms of weight and length gains, than their counterparts without digestion. Furthermore, the water turbidity of the tanks added with digested feeds was significantly less, as Papain was able to hyrolyse the protein substrates suspended in the water, and thus lowered the turbidity.     

Increased kidney xanthine oxidoreductase activity in salt-induced experimental hypertension

OBJECTIVES: To assess the efficacy and safety of low molecular weight heparin (LMWH) as deep venous thrombosis (DVT) prophylaxis following total knee arthroplasty. DATA SOURCES: Medline 1986 to June 1997, Embase, and manufacturers were used to identify randomized controlled trials. REVIEW METHODS: Trials included were randomized studies of LMWH with routine radiological screening for DVT. Placebo or active controls were included. Two reviewers independently screened trials for inclusion, and assessed their quality. Pooled relative risk estimates of DVT and proximal DVT rates were calculated using a DerSimonian and Laird random effects model. Sensitivity of the results to the type of control used and the quality of the trial was assessed. RESULTS: The relative risk of DVT for a patient given LMWH is 0.73 (95% CI 0.66 to 0.80) when compared with patients treated with adjusted dose heparin or warfarin controls. The relative risk for proximal DVT is 0.58 (95% CI 0.38 to 0.90). The relative risk of pulmonary emboli in the LMWH group was 0.55 (95% C.I. 0.20 to 1.57). No excess of bleeding was recorded in the LMWH group. CONCLUSIONS: Low molecular weight heparin is more efficacious than either adjusted dose heparin or adjusted dose warfarin, when used to prevent DVT and proximal DVT following total knee arthroplasty.     

Suppression of decorin expression and partial induction of anchorage-independent growth by the v-src oncogene in human fibroblasts

OBJECTIVE: To develop and test the intra-rater reliability of an interview-administered questionnaire that assesses lifetime patterns of total physical activity including occupational, household, and exercise/sports activities. METHODS: The questionnaire was developed and pretested using cognitive interviewing techniques on a sample of women with and without previous breast cancer diagnoses. A pilot study was conducted with 115 women who were interviewed twice, 6 to 8 wk apart by interviewers trained in cognitive interviewing methods. Respondents used recall calendars to record their education, occupations, life events, and physical activity patterns before the interviews. Interviewers helped respondents recall their lifetime exposures, including their occupational, household, and exercise/sports activities, using these calendars and memory-probing strategies. Activity levels were estimated as the average number of hours of activity per week over different time periods. Means and correlation coefficients were estimated and compared for the two time periods. RESULTS: The questionnaire was found to be highly reliable. The test-retest correlations for hours per week spent in total lifetime physical activity was 0.74, for lifetime occupational activity was 0.87, for household activity was 0.77, and for exercise/sports activities was 0.72. CONCLUSIONS: This is the first questionnaire to measure lifetime physical activity by collecting data on each type of physical activity separately over lifetime and by measuring frequency, intensity, and duration of each activity. It is also the first physical activity questionnaire to be developed, refined, and administered using cognitive-based methods employed in survey research. Respondents were able to reliably recall their lifetime physical activity patterns. This instrument can be used for any disease outcome for which physical activity may be a risk factor.     

Topographic variation in biglycan and decorin synthesis by articular cartilage in the early stages of osteoarthritis: an experimental study in sheep

From October 1988 to January 1992, nine isolates of Pseudomonas aeruginosa carrying transferable plasmids encoding imipenem-hydrolyzing beta-lactamase (pI = c. 9.5) were recovered from nine different patients in a neurosurgical ward of a hospital in Japan. The beta-lactamase activities of the sonicated extracts from the transconjugants were inhibited by EDTA and this was partially reversible by the addition of zinc cation. The substrate specificity and pI of the beta-lactamase were similar to those of the metallo beta-lactamases from P. aeruginosa and Serratia marcescens TN9106. All strains were resistant to imipenem, carbenicillin and antipseudomonal cephems including ceftazidime, cefsulodin, cefpirome, while four and five strains were susceptible to piperacillin and aztreonam, respectively. Both low level imipenem resistance and high level cephem resistance were co-transferred with the production of metallo beta-lactamase, while resistance to piperacillin, aztreonam, and high level imipenem-resistance were not selected. Production of chromosomal cephalosporinase in piperacillin resistant strains was derepressed, and production of outer membrane protein of D2 was diminished in highly imipenem resistant strains. Six strains were isolated in 1991, and the amounts of antipseudomonal agents, especially imipenem, used in the neurosurgical ward increased markedly in this year. Only three of the nine isolates had the same serotype, pyocin type and phage type. Our results suggest that the repeated isolation of imipenem and cephem-resistant P. aeruginosa producing metallo beta-lactamase was related to the high usage of antipseudomonal beta-lactam antibiotics such as imipenem, and was exacerbated by the dissemination of a plasmid.     

Congenital giant hydronephrosis in adults

Giant hydronephrosis is the term designating the presence of more than 1,000 ml. of fluid in the collecting system. Of 4 cases of congenital hydronephrosis found over a three-year period at the Ochsner Medical Institutions, 2 were associated with unsuspected carcinomas. Hydronephrosis may first be detected as a mass palpable in the abdomen and displacing the intestines on gastrointestinal films. Excretory urography, retrograde pyelography, and angiography confirm the diagnosis. The treatment of choice is nephrectomy.     

Increase of decorin content in articular cartilage following running

The effect of long distance running exercise (40 km/day for 15 weeks, five days a week) on the decorin content of articular cartilage from the knee joint was studied in female beagle dogs. Samples from load bearing sites on the lateral plateau of the tibia (TL), and pooled material from two minimum load bearing sites on the posterior section of lateral (FLP) and medial (FMP) femoral condyles were analyzed. The running exercise protocol did not lead to significant changes in the overall glycosaminoglycan content of the cartilage. However, the amount of decorin significantly increased in the TL samples, and also in the FMP pool. These results support earlier in vitro observations that decorin synthesis is stimulated by loading, independent of the synthesis of aggrecan.     

Increased expression of human histocompatibility leukocyte antigen-G in colorectal cancer cells

Human histocompatibility leukocyte antigen (HLA)-G is a nonclassical major histocompatibility complex class I molecule. HLA-G is known to provide tolerance from recognition by natural killer cells. We studied HLA-G expression in 39 human colorectal cancers and 23 extra-neoplastic colon tissue samples by RT-PCR. The expression of HLA-G mRNA was significantly more frequent in colorectal cancer (34 of 39 cases) than in the extraneoplastic tissue (10 of 23 specimens; chi2 test, p = 0.0003). HLA-G expression was also confirmed on the cancer cells immunohistochemically. These results suggested that HLA-G on colorectal cancer cells may be correlated with escape from immunological surveillance during colon cancer development.     

Increased epidermal growth factor receptor expression in metaplastic bronchial epithelium

Epidermal growth factor receptor (EGFr) is expressed in human bronchial epithelial cells, and non-small cell lung cancers express increased EGFr. Squamous metaplasia of the bronchial epithelium occurs in chronic smokers and is considered an early premalignant change. In this study, EGFr expression was examined in biopsies of histologically normal and metaplastic bronchial tissues obtained from 69 smokers who were enrolled in a randomized placebo-controlled chemoprevention trial. This trial tested the effects of 6 months of treatment with 13-cis retinoic acid (13cRA) on bronchial metaplasia. EGFr expression was examined as a marker of bronchial metaplasia and response to 13cRA treatment. In bronchial biopsies obtained from patients in this study, EGFr expression was higher in metaplastic biopsies than in normal biopsies (P = 0.02). Smoking cessation during treatment correlated with reduced metaplasia (P < 0.001) and EGFr expression (P = 0.02), but multivariate analysis suggested that this effect of smoking cessation on EGFr expression was dependent upon reversal of bronchial metaplasia. 13cRA treatment did not alter EGFr expression (P = 0.23). Baseline EGFr expression levels in metaplastic biopsies did not predict metaplasia reversal. This study demonstrated that increased EGFr expression is a biomarker of bronchial metaplasia, but it did not support the hypothesis that EGFr is a biomarker of retinoid response in lung cancer chemoprevention trials.     

Increased heat shock protein expression after stress in Japanese quail

The health care received from first admission to a pediatric rheumatology clinic to 9-year follow-up was assessed in 109 patients with chronic inflammatory rheumatic diseases or chronic idiopathic musculoskeletal pain. Ninety-five of the patients had received hospital care after the first admission, of whom 53 patients > or = 18 years, 21 patients < 18 years, and 33 parents of patients < 18 years rated their degree of satisfaction with the health care from 0 to 10. Mean scores of satisfaction with different aspects of care ranged from 6.0 to 9.6. Among patients > or = 18 years, those with idiopathic pain were less satisfied than those with inflammatory rheumatic diseases on the availability of care (mean 6.4 vs. 8.5, p < 0.001), continuity of care (mean 6.5 vs. 8.4, p < 0.001), and empathy of the health care providers (mean 6.7 vs. 7.9, p <0.05). The diagnostic group and the occurrence of remission predicted the level of global satisfaction in patients > or = 18 years. In patients < 18 years, chronic family difficulties predicted patient satisfaction and physical disability and chronic family difficulties predicted parent satisfaction. In conclusion, most parents and patients with inflammatory rheumatic diseases were satisfied with the health care. However, some patients with idiopathic pain had unmet needs for care.     

Effects of angiotensin converting enzyme inhibition on glomerular number, juxtaglomerular cell activity and renin content in experimental unilateral hydronephrosis

OBJECTIVE: To determine the effects of hydronephrosis on glomerular number and juxtaglomerular cell synthetic activity and the protective influence of angiotensin converting enzyme inhibition. DESIGN: A comparison of sham and contralateral kidneys with 8-week ipsilateral ureteral ligated hydronephrotic kidneys in BALB/c mice. Enalapril was administered from 5 weeks in additional sham and hydronephrotic kidney groups. METHODS: Renin and prorenin immunohistochemistry was applied to sections of perfusion-fixed kidneys at the light and electron microscope level. Glomerular number was estimated by a physical disector-fractionator stereological method. An enzyme kinetic renin assay was performed in kidney tissue and plasma. RESULTS: Glomerular number in hydronephrotic kidneys decreased significantly compared with sham and contralateral kidneys. Renin content in hydronephrotic kidneys did not change compared with sham or contralateral kidneys, but the renin content in the glomerulus was significantly greater in hydronephrotic than in contralateral kidneys and similar to in sham kidneys. Contralateral kidneys enlarged significantly and their total renin content decreased significantly compared with hydronephrotic and sham kidneys. Plasma renin was unchanged. Fewer juxtaglomerular cells were labelled for renin and prorenin in contralateral than in hydronephrotic or sham kidneys. Granulopoiesis and exocytotic profiles were markedly greater in hydronephrotic than in contralateral or sham kidneys. Following enalapril, glomerular number was significantly higher in hydronephrotic kidneys and renin content increased proportionally more in contralateral than in hydronephrotic or sham kidneys. CONCLUSION: Hydronephrosis for 8 weeks results in atrophy of 50% of glomeruli and exerts an inhibitory influence on contralateral juxtaglomerular cells while augmenting ipsilateral renin production per remaining glomerulus with maintenance of plasma renin. Enalapril preserves glomeruli and reverses the contralateral inhibitory influence, suggesting an angiotensin-related mechanism.     

Increased ipsilateral expression of Fos following lateral hypothalamic self-stimulation

Immunohistochemical labeling of Fos protein was used to visualize neurons activated by rewarding stimulation of the lateral hypothalamic level of the medial forebrain bundle (MFB). Following training and stabilization of performance, seven rats were allowed to self-stimulate for 1 h prior to anesthesia and perfusion. Brains were then processed for immunohistochemistry. Two control subjects were trained and tested in an identical manner except that the stimulator was disconnected during the final 1 h test. Among the structures showing a greater density of labeled neurons on the stimulated side of the brains of the experimental subjects were the septum, lateral preoptic area (LPO), medial preoptic area, bed nucleus of the stria terminalis, substantia innominata (SI), and the lateral hypothalamus (LH). Several of these structures, the LPO, SI, and LH, have been implicated in MFB self-stimulation by the results of psychophysical, electrophysiological, and lesion studies.