Muscular dystrophy. Brain, as well as brawn?

Mutations in the dystrophin gene can lead to muscular dystrophy. The dystrophin-associated complex of proteins that was first characterized at the muscle cell membrane is now also being found in other cell types.

Renal cell carcinoma-derived gangliosides suppress nuclear factor-kappaB activation in T cells.

Activation of the transcription factor nuclear factor-kappaB (NFkappaB) is impaired in T cells from patients with renal cell carcinomas (RCCs). In circulating T cells from a subset of patients with RCCs, the suppression of NFkappaB binding activity is downstream from the stimulus-induced degradation of the cytoplasmic factor IkappaBalpha. Tumor-derived soluble products from cultured RCC explants inhibit NFkappaB activity in T cells from healthy volunteers, despite a normal level of stimulus-induced IkappaBalpha degradation in these cells. The inhibitory agent has several features characteristic of a ganglioside, including sensitivity to neuraminidase but not protease treatment; hydrophobicity; and molecular weight less than 3 kDa. Indeed, we detected gangliosides in supernatants from RCC explants and not from adjacent normal kidney tissue. Gangliosides prepared from RCC supernatants, as well as the purified bovine gangliosides G(m1) and G(d1a), suppressed NFkappaB binding activity in T cells and reduced expression of the cytokines IL-2 and IFN-gamma. Taken together, our findings suggest that tumor-derived gangliosides may blunt antitumor immune responses in patients with RCCs.

Lung cancer mortality in workers exposed to sulfuric acid mist and other acid mists.

Mortality patterns were studied in 1,165 workers exposed to sulfuric acid mist and other acid mists (primarily hydrochloric acid mist) in steel-pickling operations. Standardized mortality ratio (SMR) analysis of the full "any acid exposure" cohort (n = 1,165), with the use of U.S. death rates as a standard, showed that lung cancer was significantly elevated, with a mortality ratio of 1.64 [95% confidence interval (CI) = 1.14-2.28, based on 35 observed deaths]. The lung cancer mortality ratio for workers exposed only to sulfuric acid (n = 722) was lower (SMR = 1.39), but further restriction to the time 20 years and more from first employment in a job with probable daily sulfuric acid exposure (approximately equal to 0.2 mg/m3) yielded a mortality ratio of 1.93 (95% CI = 1.10-3.13). An excess lung cancer risk was also seen in workers exposed to acids other than sulfuric acid (SMR = 2.24; 95% CI = 1.02-2.46). When comparison was made to other steel workers (rather than to the U.S. general population) to control for socioeconomic and life-style factors such as smoking, the largest lung cancer excess was again seen in workers exposed to acids other than sulfuric acid (SMR = 2.00; 95% CI = 1.06-3.78). Adjustment for potential differences in smoking habits showed that increased smoking was unlikely to have entirely explained the increased risk. Mortality from causes of death other than lung cancer was unremarkable, with the exception of significantly low rates for deaths due to digestive system diseases.

Signal transduction abnormalities in T lymphocytes from patients with advanced renal carcinoma: clinical relevance and effects of cytokine therapy.

Studies have demonstrated abnormalities of the CD3/T-cell antigen receptor (TCR) and pathways of signal transduction in T lymphocytes from animals and patients with advanced malignancy. Diminished expression of TCRzeta and p56(lck) that are associated with the TCR and reduced nuclear localization of RelA containing nuclear factor kappaB (NFkappaB) complexes have been noted. These defects have been described in T cells from patients with malignant melanoma, renal cell carcinoma (RCC), ovarian cancer, and colorectal cancer. Preliminary observations also indicate possible correlation with clinical variables such as stage in selected instances. To further characterize altered expression of TCRzeta, p56(lck), and impaired activation of NFkappaB, T lymphocytes were obtained from 65 patients with RCC, the majority of whom were receiving combination cytokine therapy [interleukin (IL)-2, IFN alpha-containing regimens] and 37 control individuals. In 29 of these patients, levels of TCRzeta and p56(lck) were determined by Western blots of T-cell lysates and semiquantitated using densitometry. Relative levels were then correlated with a series of clinical variables including response to therapy, performance status, survival, disease sites, age, and others. In another group of 28 patients (three individuals from the first group), the frequency of abnormal NFkappaB activation was studied using electrophoretic mobility shift assays after activation of T cells with phorbol myristate acetate/ionomycin or anti-CD3 monoclonal antibody. Changes in these signaling molecules during cytokine treatment were also investigated. TCRzeta and p56(lck) were detected in the peripheral blood T cells in 27 of 29 patients, and overall, reduced levels were noted visually in 12 of 29 (41%) and 13 of 29 (45%) individuals, respectively. When levels were semiquantitated using densitometry, significant decreases of TCRzeta (P = 0.029) and p56(lck) (P = 0.029) but not CD3epsilon (P = 0.131), compared with control levels, were found. In patients treated with IL-2/IFN alpha-based therapy, relative levels of TCRzeta increased significantly (P = 0.002) on day 15 of cycle one compared with the baseline. Correlations of TCRzeta or p56(lck) levels with response or disease variables, except for lower TCRzeta levels (P < 0.001) in the presence of bone metastases, were not found. Abnormal NFkappaB activation after stimulation with phorbol myristate acetate/ionomycin and/or anti-CD3 monoclonal antibody was found in 59% of patients (17 of 28) and was not accounted for by the advanced age of the study cohort. Activation of NFkappaB in peripheral blood T cells was inducible during cytokine therapy in four of six individuals who displayed impaired NFkappaB activity prior to therapy. Moreover, impaired activation of NFkappaB does not appear linked to a reduction of TCRzeta expression, because in five patients, normal TCRzeta levels were present although kappaB binding was not inducible. In the majority of patients with advanced RCC, peripheral blood T cells express TCRzeta and p56(lck), and in a subset, reduced levels of these TCRzeta associated molecules are seen that may increase during cytokine-based therapy. Abnormal activation of NFkappaB is also present in >50% of patients and may also revert to normal during IL-2/IFN alpha-based treatment. This alteration in NFkappaB activation occurred in the presence of normal expression of TCRzeta-associated signaling elements. The clinical significance of these findings remains unclear.

Mechanisms of apoptosis in T cells from patients with renal cell carcinoma.

Tumors may escape immune recognition and destruction through the induction of apoptosis in activated T lymphocytes. Results from several laboratories suggest that FasL (L/CD95L) expression in tumors may be responsible for this process. In this study of patients with renal cell carcinoma (RCC), we provide evidence for two mechanisms of T-cell apoptosis. One mechanism involves the induction of apoptosis via FasL expression in tumor cells. This is supported by several observations, including the fact that tumor cells in situ as well as cultured cell lines expressed FasL mRNA and protein by a variety of techniques. The FasL in RCC is functional because in coculture experiments, FasL+ tumors induced apoptosis in Fas-sensitive Jurkat T cells and in activated peripheral blood T cells but not in resting peripheral blood T cells. Most importantly, antibody to FasL partially blocked apoptosis of the activated T cells. Moreover, Fas was expressed by T cells derived from the peripheral blood (53% median) and tumor (44.3% median) of RCC patients. Finally, in situ staining for DNA breaks demonstrated apoptosis in a subset of T cells infiltrating renal tumors. These studies also identified a second mechanism of apoptosis in RCC patient peripheral T cells. Whereas these cells did not display DNA breaks when freshly isolated or after culture for 24 h in medium, peripheral blood T cells from RCC patients underwent activation-induced cell death after stimulation with either phorbol 12-myristate 13-acetate/ionomycin or anti-CD3/CD28 antibodies. Apoptosis mediated by exposure to FasL in tumor cells or through T-cell activation may contribute to the failure of RCC patients to develop an effective T-cell-mediated antitumor response.

Laser interferometric visual acuity in senile macular degeneration.

Twenty-nine eyes from 15 patients were affected by senile macular degeneration and had clear ocular media. Tests with retinal interference fringe patterns produced by a low-energy helium-neon laser and Snellen's charts showed that in 13 eyes, interferometric and Snellen's acuities were within one line. In the remaining 16 eyes, laser interferometric acuities were notably better than Snellen's acuities. Our results suggest that laser interferometric visual acuities, prior to lens extraction, will prove inconsistent in accurately predicting postoperative Snellen's acuities in patients with senile macular degeneration.

Healing of diabetic foot ulcers and pressure ulcers with human skin equivalent: a new paradigm in wound healing.

HYPOTHESIS: In patients with diabetic foot and pressure ulcers, early intervention with biological therapy will either halt progression or result in rapid healing of these chronic wounds. DESIGN: In a prospective nonrandomized case series, 23 consecutive patients were treated with human skin equivalent (HSE) after excisional debridement of their wounds. SETTING: A single university teaching hospital and tertiary care center. PATIENTS AND METHODS: Twenty-three consecutive patients with a total of 41 wounds (1.0-7.5 cm in diameter) were treated with placement of HSE after sharp excisional debridement. All patients with pressure ulcers received alternating air therapy with zero-pressure alternating air mattresses. MAIN OUTCOME MEASURE: Time to 100% healing, as defined by full epithelialization of the wound and by no drainage from the site. RESULTS: Seven of 10 patients with diabetic foot ulcers had complete healing of all wounds. In these patients 17 of 20 wounds healed in an average of 42 days. Seven of 13 patients with pressure ulcers had complete healing of all wounds. In patients with pressure ulcers, 13 of 21 wounds healed in an average of 29 days. All wounds that did not heal in this series occurred in patients who had an additional stage IV ulcer or a wound with exposed bone. Twenty-nine of 30 wounds that healed did so after a single application of the HSE. CONCLUSIONS: In diabetic ulcers and pressure ulcers of various durations, the application of HSE with the surgical principles used in a traditional skin graft is successful in producing healing. The high success rate with complete closure in these various types of wounds suggests that HSE may function as a reservoir of growth factors that also stimulate wound contraction and epithelialization. If a wound has not fully healed after 6 weeks, a second application of HSE should be used. If the wound is not healing, an occult infection is the likely cause. All nonischemic diabetic foot and pressure ulcers that are identified and treated early with aggressive therapy (including antibiotics, off-loading of pressure, and biological therapy) will not progress.

Morphometric analysis of spermatozoa in the assessment of human male fertility.

A videomicrographic system was developed for measurement of morphometric parameters of human spermatozoa. Contours of the images of spermatozoa on a video monitor are digitized by manually tracing them with the cursor of an electromagnetic digitizer integrated to a microcomputer. The accuracy and precision of the methodology were evaluated. A comparison of human sperm heads in shallow wet preparations and in dried, stained preparations indicated that the latter were smaller in length, width, projected area, and circumference, but that the ratio length/width was not different. An analysis was made of 457 ejaculates from 16 fertile donors. The variation between ejaculates within donors was similar in magnitude to the variation between donors. A study was performed comparing seminal sperm morphometry in single specimens from 30 fertile and 30 infertile men. The sperm head length/width ratio was the parameter that differed the most between these two groups. Moreover, it was the per-ejaculate variability of this parameter, rather than the central tendency, that maximized the difference.

Position of the wrist associated with the lowest carpal-tunnel pressure: implications for splint design.

Increased carpal-tunnel pressure has been implicated in the pathophysiology of carpal tunnel syndrome, but it is not known whether splints that immobilize the wrist in a functional position of extension minimize carpal tunnel pressure. To determine the position of the wrist that results in the lowest carpal-tunnel pressure, twenty control subjects and four patients who had carpal tunnel syndrome were evaluated with use of a new, dynamic method that continuously measures carpal tunnel pressure throughout the range of motion of the wrist. The pressure was measured by means of a pressure transducer connected to a flexible catheter that had been inserted into the carpal canal. The position of the wrist was measured simultaneously with use of a two-axis electrogoniometer. Aided by a computer monitor that displayed a moving line of real-time carpal-tunnel pressure, each subject was instructed to move the wrist throughout the range of motion and to adjust it to the position that corresponded to the lowest carpal-tunnel pressure. For the control subjects, the lowest carpal-tunnel pressure averaged 8 +/- 4 millimeters of mercury (1.07 +/- 0.53 kilopascals), and the average position of the wrist associated with the lowest pressure was 2 +/- 9 degrees of extension and 2 +/- 6 degrees of ulnar deviation.(ABSTRACT TRUNCATED AT 250 WORDS)

Mortality among a cohort of uranium mill workers: an update.

AIMS: To evaluate the mortality experience of 1484 men employed in seven uranium mills in the Colorado Plateau for at least one year on or after 1 January 1940. METHODS: Vital status was updated through 1998, and life table analyses were conducted. RESULTS: Mortality from all causes and all cancers was less than expected based on US mortality rates. A statistically significant increase in non-malignant respiratory disease mortality and non-significant increases in mortality from lymphatic and haematopoietic malignancies other than leukaemia, lung cancer, and chronic renal disease were observed. The excess in lymphatic and haematopoietic cancer mortality was due to an increase in mortality from lymphosarcoma and reticulosarcoma and Hodgkin's disease. Within the category of non-malignant respiratory disease, mortality from emphysema and pneumoconioses and other respiratory disease was increased. Mortality from lung cancer and emphysema was higher among workers hired prior to 1955 when exposures to uranium, silica, and vanadium were presumably higher. Mortality from these causes of death did not increase with employment duration. CONCLUSIONS: Although the observed excesses were consistent with our a priori hypotheses, positive trends with employment duration were not observed. Limitations included the small cohort size and limited power to detect a moderately increased risk for some outcomes of interest, the inability to estimate individual exposures, and the lack of smoking data. Because of these limitations, firm conclusions about the relation of the observed excesses in mortality and mill exposures are not possible.

Mortality of workers employed in shoe manufacturing.

A retrospective cohort mortality study was conducted among 7814 white shoe manufacturing workers followed from 1940 through 1982. The workers were potentially exposed to solvents (including toluene) and solvent-based adhesives. Benzene may have been present as an impurity of toluene. Mortality due to leukemia and aleukemia was not statistically significantly elevated. Statistically significant excess mortality due to cancer of the trachea, bronchus and lung was observed in the total cohort [standardized mortality ratio (SMR) 147, 95% confidence interval (95% CI) 120-180] and showed a statistically significant trend in standardized relative risk with increasing potential latency, but not with increasing duration of employment. Chronic nonmalignant respiratory disease was significantly elevated among the men (SMR 158, 95% CI 114-217) but was less than expected among the women (SMR 79), a finding suggesting a possible contribution of smoking to the mortality from respiratory cancer. However, adjustment for the potential effects of smoking did not completely eliminate the increased risk for lung cancer.

Cardiovascular mortality among munitions workers exposed to nitroglycerin and dinitrotoluene.

A retrospective cohort mortality study with 5529 nitroglycerin, 4989 dinitrotoluene, and 5136 unexposed workers compared the mortality of the exposed groups with that of the United States population and that of the unexposed group with life-table analysis and Poisson regression. Mortality from ischemic heart disease was close to that expected, and mortality from cerebrovascular disease was slightly less than that expected, for the workers with both nitroglycerin and dinitrotoluene exposure and for those with dinitrotoluene exposure only. A significant interaction between age and nitroglycerin exposure was detected in the Poisson regression analyses for ischemic heart disease, particularly for workers actively exposed to nitroglycerin. The rate ratio for the workers under 45 years of age and actively exposed to nitroglycerin was 3.30 (95% confidence interval 129-8.48). This study did not show a chronic effect of nitroglycerin or dinitrotoluene exposure on cardiovascular disease risk. Potential biases related to the company's medical screening program may have limited the ability to detect chronic cardiovascular effects.

Mortality of industrial workers exposed to acrylonitrile.

OBJECTIVES: This study was designed to evaluate the relationship between occupational exposure to acrylonitrile and cancer mortality. MATERIALS AND METHODS: Workers (18079 white men, 4293 white women, 2191 nonwhite men, and 897 nonwhite women) employed in acrylonitrile production or use in the 1950s through 1983 were followed through 1989 for vital status and cause of death. Exposure-response relationships were evaluated from quantitative estimates of historical exposures. Tobacco use was determined for a sample of workers to assess potential confounding. Mortality rates between the exposed and unexposed workers in the cohort were compared using the Poisson regression. RESULTS: Analyses by cumulative, average, peak, intensity, duration, and lagged exposure revealed no elevated risk of cancers of the stomach, brain, breast, prostate or lymphatic and hematopoietic systems. Mortality from lung cancer was elevated for the highest quintile of cumulative exposure. When the decile categories were used, the relative risk did not continue to increase at higher levels. Adjustment for cigarette use reduced the risk for lung cancer only slightly. Separate analyses for wage and salaried workers, long-term and short-term workers, fiber and nonfiber plants, and individual plants revealed no clear exposure-response patterns. CONCLUSIONS: The results indicate that exposure to acrylonitrile at the levels studied is not associated with an increased relative risk for most cancers of a priori interest. The excess of lung cancer in the highest quintile of cumulative exposure may indicate carcinogenic activity at the highest levels of exposure, but analyses of exposure-response do not provide strong or consistent evidence for a causal association.

Exposure assessment for a study of workers exposed to acrylonitrile.

Procedures used to develop estimates of exposure to acrylonitrile for a cohort study (>25000 workers in 8 monomer, fiber, and resin companies from 1952 to 1983) are presented. Visits to the companies were made, interviews of workers were conducted, historical records were made, and measurements were taken. On the basis of similar tasks, locations, other exposures, and a similar distribution of exposures to acrylonitrile, 3600 exposure groups were formed. Special procedures were used to reduce the misclassification of workers performing tasks that varied in time but that were inadequately reflected in the job title. A software program organized and retained all exposure information on each exposure group. Quantitative estimates of acrylonitrile exposure were developed using a hierarchical approach in a software program that documented the derivation of each estimate and facilitated data review. Two of the estimation methods were evaluated in a comparison with measurement data.

The NIOSH/FAA Working Women's Health Study: evaluation of the cosmic-radiation exposures of flight attendants. Federal Aviation Administration.

Air crew are exposed to elevated levels of cosmic ionizing radiation of galactic and solar origin and are among the more highly exposed occupational groups to ionizing radiation in the United States. Depending on flight route patterns, the annual dose may range from 0.2 to 5 mSv. By comparison, the average annual radiation dose equivalent of occupationally exposed adults in the United States is estimated to be 1.1 mSv. Cosmic-radiation dose depends primarily on altitude and geomagnetic latitude and to a lesser degree on solar activity. Although the International Commission on Radiological Protection has recommended that air crew exposures to natural radiation in-flight be treated as occupational exposures, United States flight crew exposures to natural cosmic radiation are not regulated or typically monitored. There are approximately 148,000 air crew (flight deck crew and flight attendants) in the United States.

Aggrecanolysis in human osteoarthritis: confocal localization and biochemical characterization of ADAMTS5-hyaluronan complexes in articular cartilages.

OBJECTIVE: Human osteoarthritis (OA) is characterized by aggrecanase-mediated depletion of cartilage aggrecan. We have examined the abundance, location and some biochemical properties of the six known aggrecanases (A disintegrin and metalloproteinase with thrombospondin-like motifs 1 (ADAMTS1) 4, 5, 8, 9 and 15) in normal and OA human cartilages. METHODS: Formalin-fixed, ethylenediamine tetraacetic acid (EDTA)-decalcified sections of full-depth cartilage from human OA tibial plateaus and normal control samples were studied by confocal imaging. Probes included specific antibodies to aggrecanases and two aggrecan epitopes, as well as biotinylated hyaluronan binding protein (HABP) for hyaluronan (HA) visualization. Cartilage extracts were analyzed by Western blot for the individual proteinases and aggrecan fragments. RESULTS: ADAMTS5 was present in association with cells throughout normal cartilage and was markedly increased in OA, particularly in clonal groups in the superficial and transitional zones, where it was predominantly co-localized with HA. Consistent with the confocal analysis, a high molecular weight complex of ADAMTS5 and HA was isolated from human OA cartilage by isotonic salt extraction and chromatography on Superose 6. The complex eluted with an apparent molecular size of about 2x10(6) and contained major ADAMTS5 forms of 150, 60, 40 and 30kDa. The yield of most forms on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was markedly enhanced by prior digestion of the complex with either Streptomyces hyaluronidase or chondroitinase ABC. CONCLUSION: ADAMTS5 abundance and distribution in human OA cartilages is consistent with a central role for this enzyme in destructive aggrecanolysis. HA-dependent sequestration of ADAMTS5 in the pericellular matrix may be a mechanism for regulating the activity of this proteinase in human OA cartilage.

Experimental manipulation of compaction of the mouse embryo alters patterns of protein phosphorylation.

Compaction, occurring at the eight-cell stage of mouse development, is the process of cell flattening and polarisation by which cellular asymmetry is first established. Changes in the pattern of protein phosphorylation have been correlated with this early event of development (TL Bloom, J McConnell: Mol Reprod Dev 26:199-210, 1990). In the study reported here, groups of embryos were treated in ways known to affect particular features of compaction and were then labeled with [32P]orthophosphate; the phosphoproteins obtained were examined following electrophoresis in one and two dimensions. Four-cell embryos were treated with protein synthesis inhibitors, which advance cell flattening. This treatment resulted in only minor differences from the phosphoprotein profile of untreated four-cell embryos. Inhibition of protein synthesis at the eight-cell stage has little effect on cell flattening or polarisation. However, some phosphoproteins that are observed normally in eight-cell but not in four-cell embryos were no longer detectable if labeling took place in the presence of protein synthesis inhibitors. Eight-cell embryos incubated in phorbol 12-myristate 13-acetate, which disrupts various features of compaction, showed a relative increase in the phosphorylation of a group of phosphoprotein spots associated with the eight-cell but not with the four-cell stage. Embryos incubated in Ca2(+)-free medium, which prevents intercellular flattening and delays polarisation, showed a relative decrease in the phosphorylation of the same group of phosphoprotein spots. The behaviour of these phosphoproteins may therefore be correlated with some of the features of compaction.

The effects of phorbol ester on mouse blastomeres: a role for protein kinase C in compaction?

The effects of phorbol myristate acetate (PMA) and other activators of protein kinase C on the cytoskeletal organization of mouse oocytes and early embryos have been examined. The effects observed depended on the developmental stage on exposure to PMA. PMA had little effect on the cytoskeletal or microvillous organization of unfertilized oocytes. Interphase cells from embryos prior to compaction showed limited disruption and loss of microvilli when exposed to PMA and foci of polymerized actin remained visible in the cytocortex of embryos up to the early 8-cell stage. When compacted late 8-cell embryos were exposed to PMA, most microvilli were lost and little polymerized actin remained in the cytocortex. PMA also caused loss of microtubules from compact 8-cell embryos under some experimental conditions. Intercellular flattening was both prevented and reversed. The relevance of these observations to the rearrangement of cell-cell contacts and cytoskeletal organization seen during compaction at the 8-cell stage is discussed and a possible role for protein kinase C in the generation of cell polarity proposed.