Correlation between leukotriene D4-induced contraction and cytosolic calcium elevation: a quantitative and simultaneous evaluation in smooth muscle

The role of Ca++ as an intracellular messenger in leukotriene (LT)D4-induced muscle contraction was investigated by measuring force development and elevation in cytosolic Ca++ concentration simultaneously in strips of guinea pig ileal longitudinal muscle loaded with the fluorescent calcium indicator Fura 2. Upon addition of LTD4, a simultaneous increase in tension and cytosolic calcium concentration, [Ca++]i, was observed. Cumulative applications of LTD4 induced concentration-dependent increases in both muscle tension and [Ca++]i, being the half-maximal effect reached at approximately 6 to 9 nM. A statistically significant positive correlation (r = 0.993, P < .001) exists between the two parameters examined. Removal of calcium in the bathing solution, accompanied by addition of 7.5 mM EGTA, completely prevented any increase in either calcium levels or force development, thus indicating a role for Ca++ influx, rather than a release from intracellular stores. All of the LTD4 antagonists tested were able to counteract the effect of the leukotriene on both [Ca++]i and tension increase. However, although LY171883 shifted both of the LTD4 curves to the right in a parallel fashion, FPL 55712 and ICI 198,615 behaved as non-competitive antagonists in reversing the effect of LTD4 on [Ca++]i and tension. Thus, these results strongly suggest that changes in muscle tension induced by LTD4 are attributable to changes in cytosolic free Ca++ concentrations in guinea pig ileum.     

Epstein-Barr virus nuclear antigen 2 transactivates the long terminal repeat of human immunodeficiency virus type 1

Human immunodeficiency virus type 1 (HIV-1)-infected subjects show a high incidence of Epstein-Barr virus (EBV) infection. This suggests that EBV may function as a cofactor that affects HIV-1 activation and may play a major role in the progression of AIDS. To test this hypothesis, we generated two EBV-negative human B-cell lines that stably express the EBNA2 gene of EBV. These EBNA2-positive cell lines were transiently transfected with plasmids that carry either the wild type or deletion mutants of the HIV-1 long terminal repeat (LTR) fused to the chloramphenicol acetyltransferase (CAT) gene. There was a consistently higher HIV-1 LTR activation in EBNA2-expressing cells than in control cells, which suggested that EBNA2 proteins could activate the HIV-1 promoter, possibly by inducing nuclear factors binding to HIV-1 cis-regulatory sequences. To test this possibility, we used CAT-based plasmids carrying deletions of the NF-kappa B (pNFA-CAT), Sp1 (pSpA-CAT), or TAR (pTAR-CAT) region of the HIV-1 LTR and retardation assays in which nuclear proteins from EBNA2-expressing cells were challenged with oligonucleotides encompassing the NF-kappa B or Sp1 region of the HIV-1 LTR. We found that both the NF-kappa B and the Sp1 sites of the HIV-1 LTR are necessary for EBNA2 transactivation and that increased expression resulted from the induction of NF-kappa B-like factors. Moreover, experiments with the TAR-deleted pTAR-CAT and with the tat-expressing pAR-TAT plasmids indicated that endogenous Tat-like proteins could participate in EBNA2-mediated activation of the HIV-1 LTR and that EBNA2 proteins can synergize with the viral tat transactivator. Transfection experiments with plasmids expressing the EBNA1, EBNA3, and EBNALP genes did not cause a significant HIV-1 LTR activation. Thus, it appears that among the latent EBV genes tested, EBNA2 was the only EBV gene active on the HIV-1 LTR. The transactivation function of EBNA2 was also observed in the HeLa epithelial cell line, which suggests that EBV and HIV-1 infection of non-B cells may result in HIV-1 promoter activation. Therefore, a specific gene product of EBV, EBNA2, can transactivate HIV-1 and possibly contribute to the clinical progression of AIDS.     

Finishing of glass-ionomer cement (SEM study)

A total of 12 glass-ionomer cement specimens were utilized in the present study. The specimens were divided into two equal groups. The first group was used after 10 minutes from setting, while the second was utilized after 24 hours from setting. Each group was divided into three equal subgroups (2 specimens each). The first subgroups were finished under wet condition (wet finished). The second subgroups were dry finished. On the other hand, the third subgroups were kept undisturbed (as set) under mylar strips. The specimens surfaces were then examined by a Scanning Electron Microscope (SEM). It was found that, finishing of the specimens after 24 hours from setting demonstrated more acceptable surface topography either in wet or dry conditions than finishing after 10 minutes from setting. Moreover, the dry finished specimens displayed more acceptable surface topography than the wet finished specimens. On the other hand, the as set (undisturbed) specimens the most acceptable surface topography.     

An improved system for the control of dissolved oxygen in freshwater aquaria

A system for the removal and control of dissolved oxygen (DO) from freshwater was designed and constructed with aquarium-type fish studies in mind. Degassed water was obtained using a partial vacuum of -14 psi, and DO regulated at an aquarium scale using electronically controlled aeration with timed partial water renewal. The degassing system was capable of producing water with approximately 1.7 mg L(-1) DO within 10 min of operation, and 0.55 mg L(-1) after 2h. The control system was capable of maintaining DO levels of ca 0.8 mg L(-1) over 48 h in the absence of aeration and further capable of precisely controlling DO levels as low as 1.16+/-0.002 mg L(-1) (mean+/-SEM) with aeration over a 48 h period.     

Effect of profound hearing loss on a central auditory nucleus

The present study was designed to investigate the type and extent of degeneration occurring in the human central auditory system subsequent to profound hearing loss. The authors have examined the size of one population of neurons in the ventral cochlear nucleus in seven subjects with profound hearing loss (audiometric responses poorer than 90-100 dB HL). Six normal subjects, ages 35-78, were used as controls. Cell size in the hearing-impaired subjects ranged from normal to reduced by more than 50 percent. Two factors appear to contribute to the variability in cell size reduction. The correlation coefficient (Spearman rs) of cell size with duration of profound deafness was -0.48, indicating a moderate tendency for neurons to become smaller with longer periods of deafness. The correlation coefficient of cell size with number of surviving cochlear ganglion cells was 0.73, indicating a stronger tendency for neurons to be larger with greater eighth nerve innervation of the cochlear nucleus. Two cases of Scheibe degeneration showed the most severe degenerative change in the central auditory system.     

Active surveillance of health and safety in microbiology laboratories

In microbiology laboratories highly infectious material is handled alongside complex and potentially dangerous equipment, and staff are therefore at risk of infections and accidents. Acts of parliament and regulations exist to protect staff in the workplace, including those exposed to biological agents. The current monitoring of health and safety in laboratories seeks to ensure that employers and employees comply with existing regulations, but this form of passive surveillance is of limited value because it does not highlight shortcomings in techniques, equipment, premises, or personnel. We propose a scheme for the surveillance of health and safety in microbiology laboratories that will actively seek information about laboratory incidents and practices, in order to enable appropriate preventive measures to be instituted.     

Esterified estrogens with and without methyltestosterone decrease arterial LDL metabolism in cynomolgus monkeys

OBJECTIVES: To assess retinal complications and to identify risk factors for retinal complications following aqueous shunt procedures. MATERIALS AND METHODS: Records of 38 consecutive aqueous shunt procedures that were performed on 36 patients at the Eye Institute of the Medical College of Wisconsin, Milwaukee, from June 1993 to March 1995 (minimum follow-up, 6 months) were reviewed. The mean +/- SD follow-up was 11.4 +/- 5.2 months (median, 10.5 months). RESULTS: Twelve patients (32%) had the following retinal complications: 4 serous choroidal effusions (10%) that required drainage, 3 suprachoroidal hemorrhages (8%), 2 vitreous hemorrhages (5%), 1 rhegmatogenous retinal detachment (3%), 1 endophthalmitis (3%), and 1 scleral buckling extrusion (3%). Surgical procedures for retinal complications were required in 8 (67%) of these 12 patients. Visual acuity decreased 2 lines or more in 9 (75%) of these 12 patients. The median onset of a postoperative retinal complication was 12.5 days, with 10 patients (83%) experiencing complications within 35 days. Serous choroidal effusions developed in 10 other patients (26%), and these effusions resolved spontaneously. Visual acuity decreased 2 lines or more in 2 (20%) of these additional 10 patients. Patients who experienced serious retinal complications were significantly older, had a higher rate of hypertension, and postoperative ocular hypotony. Serious retinal complications were distributed evenly among patients with Krupin valves with discs and Molteno and Baerveldt devices. Experience with the Ahmed glaucoma valve implant was limited. CONCLUSION: Aqueous shunt procedures may be associated with significant retinal complications and subsequent visual loss.     

Association between oxygen delivery and consumption in patients undergoing cardiac surgery. Is there supply dependence?

Numerous investigators have suggested that cell glycoconjugates are modified by the development of cancer and the progression of this to a malignant form. Accordingly, in the present work, beta-D-galactosidase, alpha-L-fucosidase, beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-galactosaminidase activities were studied in human thyroid and gastric tumours. Samples were obtained from human gastric mucosa and thyroid gland tumours together with a part of the surrounding normal tissue (control). Enzyme activity was determined spectrophotometrically based on the release of p-nitrophenol from suitable p-nitrophenyl-derivative substrates. Results showed that beta-D-galactosidase, alpha-L-fucosidase, beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-galactosaminidase activities were detected in tumour and control samples from thyroid and gastric tissues. The gastric mucosa also showed alpha-L-mannosidase activity. The specific activities of these glycosidases were higher (two- or three-fold) in tumour tissues as compared with their controls. beta-D-galactosidase, beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-galactosaminidase activities from thyroid and gastric tumours showed a significant increase in V(max) as compared with their respective controls (P < 0.05 or P < 0.001). Thyroid alpha-L-fucosidase activity showed a statistically and significantly increased affinity (lower K(m)) in tumour samples as compared to normal tissue. In conclusion both gastric and thyroid tumours showed enhanced glycosidase activity as compared with enzyme activity observed in normal tissue. These results are in agreement with the notion of a markedly raised degradation within lysosomes of tumour cells.