Structural domains of the muscle-tendon junction. 1. The internal lamina and the connecting domain

The force generated within skeletal muscle fibers of vertebrates is transmitted to the tendon at the muscle-tendon junction. Ultrastructural analysis of the murine muscle-tendon junction following a variety of experimental manipulations has produced evidence that the muscle-tendon junction can be described in terms of four principal domains (Trotter and Eberhard, 1983), two of which are discussed in the present report. Each domain is defined by the shape and orientation of its principal components, and by its position with respect to the plasma membrane. The internal lamina is composed of actin filaments, with a center to center spacing of approximately 12 nm, oriented mainly parallel to the principal vector of contractile force, and to the plasma membrane. These filaments are cross-linked into a structural unit, perhaps by the electron-dense structures which are associated with them. The internal lamina is morphologically connected to the external lamina (lamina densa) by a population of fine filaments oriented approximately perpendicular to the principal vector of contractile force. These filaments which constitute the connecting domain, are approximately 2–8 nm in diameter and are at least 50 nm long. They pass through three separate regions: the sarcoplasm between the internal lamina and the plasma membrane; the plasma membrane proper; and the extracellular space between the plasma membrane and the lamina densa. This third region is often referred to as the lamina lucida. These filaments are composed of at least three separate components in series, linked together by noncovalent interactions. The existence of these discrete structural domains implies that each has a different molecular composition and different mechanical properties.     

Suppression of Leukocyte-Enhanced Cold Ischemia/Reperfusion Injury of Liver Endothelium with the Benzoquinone Antioxidant Idebenone

Objective: Despite the large body of evidence for a major role of neutrophils and oxidant stress, the exact pathogenesis of the early ischemia/reperfusion injury after cold preservation of the liver is not well understood. The potential benefit of an antioxidant on metabolic liver function during reperfusion has been demonstrated in several studies.

Materials and Methods: We describe a cold storage/reperfusion damage model with isolated perfused pig livers, where the effects of neutrophils and idebenone, a recently developed benzoquinone antioxidant were studied. The integrity of sinusoidal endothelial cells (SEC) was estimated by hyaluronic acid concentration in perfusate and the expression of endothelial constitutive nitric oxide synthase (ecNOS) after reperfusion and compared to lipid peroxidation and antioxidant content.

Results: Hyaluronic acid displayed the highest levels and ecNOS mRNA was most depressed in livers reperfused with neutrophils after 20 h cold storage; this was accompanied by an increase in lipid peroxidation (TBARS) and a breakdown of endogenous lipophilic antioxidants (-tocopherol and coenzyme Q-10). These effects were attenuated, when neutrophils were excluded from reperfusion and almost completely abolished by the addition of 200 μmol/L idebenone.

Conclusions: These data suggest that a leukocyte-mediated damage based on reactive oxygen species markedly contributes to the reperfusion injury of SEC after cold preservation of the liver. Therefore, the presence of effective antioxidants in the early reperfusion phase may be beneficial for liver graft integrity.     

Initial severity of metabolic acidosis predicts the development of acute lung injury in severely traumatized patients

OBJECTIVES: First, to determine whether the severity of shock, as measured by systemic hypotension and metabolic acidosis, is significantly associated with a higher risk of acute lung injury in patients with severe trauma. Second, to determine whether the volumes of blood and crystalloid solutions administered in the early posttrauma period are independent risk factors for acute lung injury in severely traumatized patients. DESIGN: Prospective observational study. SETTING: Level I urban trauma center in a university hospital. PATIENTS: A total of 102 severely injured, mechanically ventilated trauma patients with an Injury Severity Score > or =16 and aged between 18 and 75 yrs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Initial clinical and laboratory data were collected in the emergency department, and on a daily basis thereafter during the patient's intensive care unit stay. Of the 102 severely injured patients enrolled, 42 developed acute lung injury (41%) and 60 did not (59%). A total of 93% of the trauma patients who developed acute lung injury during the 17-month study period were included in the study. Initial base deficit was significantly lower in patients who developed acute lung injury than in those who did not (-8.8+/-4.5 vs. -5.6+/-5.1, p<.01). The difference in systolic blood pressure between the two groups was not significant. CONCLUSIONS: In this group of severely injured trauma patients, the degree of metabolic acidosis at the time of admission identified those patients with the highest probability of developing acute lung injury. In addition, the volume of crystalloid solution administered during the first 24 hrs was significantly greater in patients who later developed acute lung injury. Finally, there was a significantly higher morbidity in patients who developed acute lung injury, whereas mortality did not differ between the two groups.     

Epidemiology of Cyclospora cayetanensis and other intestinal parasites in a community in Haiti

We conducted an exploratory investigation in a community in Haiti to determine the prevalence of Cyclospora cayetanensis infection and to identify potential risk factors for C. cayetanensis infection. In 2001, two cross-sectional stool surveys and a nested case-control study were conducted. In 2002, a follow-up cross-sectional stool survey was conducted among children < or =10 years of age. Stool specimens from study participants and water samples from their wells were examined for Cyclospora and other intestinal parasites. In stools, the prevalence of infection with Cyclospora in persons of all ages decreased from 12% (20 of 167 persons) in February 2001 to 1.1% (4 of 352 persons) in April 2001, a 90.8% decrease. For children < or =10 years of age, the prevalence rates were 22.5% (16 of 71 children) in February 2001, 3.0% (4 of 135 children) in April 2001, and 2.5% (2 of 81 children) in January 2002. Use of the water from the artesian well in the northern region of the community versus the one in the south was the only risk factor associated with Cyclospora infection in multivariate analyses (odds ratio, 18.5; 95% confidence interval, 2.4 to 143.1). The water sample from one of the nine wells or water sources tested (one sample per source) in January 2001, shortly before the investigation began, was positive for Cyclospora by UV fluorescence microscopy and PCR. None of the water samples from the 46 wells or water sources tested during the investigation (one sample per source per testing period, including the artesian wells) were positive for Cyclospora. Further studies are needed to assess the role of water as a possible risk factor for Cyclospora infection in Haiti and other developing countries.