Leucine transport in Xenopus laevis oocytes: functional and morphological analysis of different defolliculation procedures

L-leucine uptake in stage V Xenopus laevis oocytes was affected by the specific methods used to remove the follicle cells. In the presence of 100 mM NaCl, L-leucine uptake was reduced by 67.5% +/- 5.7 when defolliculation was performed enzymatically by collagenase treatment, whereas the reduction was 30.5% +/- 6.4 after mechanical defolliculation. The Na(+)-dependent uptake of 0.1 mM L-leucine was 18.6 +/- 4.6 pmol oocyte-1 40 min-1 in folliculated oocytes and 5.6 +/- 1.9 in collagenase defolliculated oocytes (means +/- SE). L-leucine uptake was not affected by the removal of the follicular layer if defolliculation occurred after the transport period; radiolabeled L-leucine is therefore not taken up into a compartment that is removed by the defolliculation process. The different L-leucine uptake rates observed in folliculated and defolliculated oocytes were not due to non-specific L-leucine binding to membranes. L-leucine kinetics showed that the L-leucine Vmax and Km values were lower in oocytes deprived of the follicular layer than in control oocytes enveloped in intact follicular layers. The Vmax and Km values of Na(+)-dependent L-leucine transport, calculated from data obtained the day after defolliculation by collagenase treatment, were: 16 +/- 1.5 pmol oocyte-1 40 min-1 and 57 +/- 21 mumol (mean +/- SD). The Na(+)-activation curve of 0.1 mM L-leucine was hyperbolic in folliculated oocytes and sigmoidal in defolliculated oocytes. The morphological analysis performed in parallel with the transport experiments showed that after defolliculation, the fibers forming the vitelline membrane tended to be arranged in a more regular orthogonal array, and the number of oocyte microvilli was reduced after collagenase treatment. Mechanical defolliculation did not appreciably affect the oocyte microvilli, however this procedure did not completely remove all follicle cells. The damage to collagenase treated oocytes was reversible, and the functional and structural features of most oocytes improved upon subsequent in vitro incubation. The recovery process seemed to involve protein synthesis in view of the increased value of L-leucine Vmax, and microscopic observation showing recovery of the microvillar apparatus.     

Monitoring eosinophil activation and liver function after liver transplantation

BACKGROUND: Portal tract eosinophil infiltration and an increase in the blood eosinophil count (EOS) have been shown to be specific markers of liver allograft rejection. The graft eosinophil infiltration is associated with the local release of eosinophil cationic protein. Therefore, serum eosinophil cationic protein concentration (ECP) is a potential marker for acute allograft rejection. We investigated the chronological relationship among and diagnostic value of serial changes in EOS, ECP, and liver function tests (LFTs) following liver transplantation. METHODS: EOS, ECP, serum alpha-glutathione S-transferase concentration, and conventional LFTs were measured in serial samples collected over the first 3 postoperative months following 58 liver transplants. The diagnostic potential of each test, alone or in combination, was reviewed over the entire follow-up period. RESULTS: EOS and ECP increased at a median period of 3.5 and 4 days, respectively, before the diagnosis of acute rejection, and this increase was significantly earlier than the corresponding changes in LFTs (P<0.05). There was a significant correlation between the day of the first increase in EOS and alpha-glutathione S-transferase (rs=0.535; P=0.009) and EOS and alanine transaminase (rs=0.629; P=0.004). The optimum combination of tests for the diagnosis of acute rejection was an increase in both EOS and GST with a predictive efficiency of 84%. CONCLUSIONS: Increases in EOS and ECP are early indicators of acute liver allograft rejection and precede evidence of hepatocellular damage. However, an increase in ECP was also frequently associated with infection. Therefore, we recommend the regular monitoring of EOS in conjunction with routine LFTs after liver transplantation as an aid to the early diagnosis of acute rejection.     

Community respiratory viral infection in adult lung transplant recipients

STUDY OBJECTIVE: To define the epidemiology, clinical manifestations, and long-term complications of respiratory viral infections in adult lung transplant recipients. DESIGN: Retrospective review of the records of 122 adult lung transplant recipients over a 5-year period at one institution. RESULTS: Ten episodes of infection with respiratory syncytial virus, parainfluenza, influenza, or adenovirus were identified. All patients presented with symptoms of respiratory tract infection. Two patients died acutely and four patients subsequently had development of obliterative bronchiolitis (OB). CONCLUSIONS: These data suggest community respiratory viral infections cause significant morbidity and mortality in lung transplant recipients. Further prospective studies are warranted to clarify the relationship between respiratory viral infection and OB and to define the optimal therapy for these viral infections.     

DNA sequence recognition by bis-linked netropsin and distamycin derivatives

The dihydropyridine receptor (DHPR), a voltage-gated L-type Ca2+ channel, and the Ca2+ release channel/ryanodine receptor isoform-1 (RyR1) are key molecules involved in skeletal muscle excitation-contraction coupling. We have reported age-related decreases in the level of DHPR expression in fast- and slow-twitch muscles from Fisher 344 cross Brown Norway (F344BNX) rats (Renganathan, Messi and Delbono, J. Membr. Biol. 157 (1997) 247-253). Based on these studies we postulate that excitation-contraction uncoupling is a basic mechanism for the decline in muscle force with aging (Delbono, Renganathan and Messi, Muscle Nerve Suppl. 5 (1997) S88-92). In the present study, we extended our studies to older ages and we intended to prevent or retard excitation-contraction uncoupling by restricting the caloric intake of the F344BNX rats from 16 weeks of age. Three age groups, 8-, 18-, and 33-month old caloric restricted rats, were compared with ad libitum fed animals. The number of DHPR and RyR1 and DHPR/RyR1 ratio (an index of the level of receptors uncoupling) in skeletal muscles of 8-month and 18-month rats was not significantly different in either ad libitum fed or caloric restricted rats. However, the age-related decrease in the number of DHPR, RyR1 and DHPR/RyR1 ratio observed in 33-month old ad libitum fed rats was absent in 33-month old caloric restricted rats. These results suggest that caloric restriction prevents age-related decreases in the number of DHPR, RyR1 and DHPR/RyR1 ratio observed in fast- and slow-twitch rat skeletal muscles.     

A three-dimensional computerized mesh diagram analysis and its application in soft tissue facial morphometry

A modified computerized mesh diagram analysis that allows rapid and independent quantifications of soft tissue facial size and shape in the three-dimensional space is presented. Normal references are provided, and the application of the method is also exemplified by the analysis of two maxillofacial surgical patients. The Three-Dimensional Facial Morphometry method has been used for the collection of the x, y, z coordinates of 22 soft tissue landmarks in 50 men and 50 women (all healthy young white adults). The method detects the three-dimensional coordinates of retroreflective, wireless markers positioned on selected facial landmarks with two charge-coupled device cameras, working in the infrared field. The midpoint between the right and left tragus landmarks served as the origin of the coordinate axes, and the landmark coordinates were rotated, setting the intercantheal line horizontal on both the frontal and the horizontal planes, and the Camper's plane inclined at -7.5 degrees on the sagittal plane. A standardized mesh of equidistant horizontal (dimension: half the upper face width), vertical (half the vertical projection of upper face height), and anteroposterior (half the horizontal projection of upper face depth) lines was consequently constructed. The lattice was replicated on the entire face and comprised 84 parallelepipeds. Both male and female reference meshes had a harmonious and symmetric appearance, with gender differences in facial size but not in facial shape. The standard normal reference was superimposed on the patient's tracing, and the global (size plus shape) difference was then evaluated by the calculation of the relevant displacement vectors for each soft tissue landmark. A global difference factor was calculated as the sum of the modules of all the displacement vectors. Consequently, a size normalization was performed, and the shape difference (size-standardized) was then evaluated by the calculation of new relevant displacement vectors for each landmark, as well as a shape--global difference vectors.     

Plutonium anomalies in attic dust and soils at locations surrounding the Nevada Test Site

Attic dust and soil samples were collected during the Summer of 1996 and the Spring of 1997 in southern Nevada and southern Utah. Analysis of the samples for radiocesium and plutonium give activity ratios of radiocesium/plutonium that range from 0.7 +/- 0.1 to 27 +/- 2, well below the world-wide fallout ratio of 34 +/- 4 (as of 1 July 1997). This indicates anomalous plutonium throughout the region, including areas to the south of the Nevada Test Site (NTS), generally believed to have received only world-wide fallout. Safety tests and above-ground detonations that resulted in incomplete fission, conducted at the NTS during the 1950's and 1960's, are likely sources of this excess plutonium.     

Dietary fish oil enhances macrophage production of nitric oxide

Melanotic neuroectodermal tumor of infancy (MNTI) is a rare but well-documented lesion of neuroectodermal derivation. Maturation of the neural elements has been reported only occasionally. We report a case of MNTI of the maxilla showing maturation of neural elements to ganglionic cells.     

Regression of established mouse leukemia by GM-CSF-transduced tumor vaccine: implications for cytotoxic T lymphocyte responses and tumor burdens

This study investigated the therapeutic effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on a mouse leukemia model. By using a retroviral vector, mouse GM-CSF cDNA was transduced into a highly tumorigenic T leukemia cell line, RL male 1. Injection of GM-CSF-secreting RL male 1 cells into syngeneic BALB/c mice elicited protective immunity in the animals, which could regress preestablished tumors introduced either by a subcutaneous or in an intravenous route. However, the therapeutic effects were less prominent in the mice inoculated with a large tumor load or in mice treated later. Winn tests further demonstrated that the splenocytes from the late-treated group conferred poorer protective effects in terms of reducing the growth of parental RL male 1 cells in naive mice than the splenocytes from the early-treated group. Nonetheless, upon stimulation in vitro, the activity of tumor-specific cytotoxic T lymphocytes (CTL) was comparable in the splenocytes of both groups of mice. Histological analysis also indicated that the CD8+ T cells appeared as early as 3 days following vaccination at the vaccine sites and at the tumor sites in both groups of mice. Above observations implied that the T cells in the animals bearing large tumors appeared to be in a state of suppression or anergy. Systematic histological analyses for 2 weeks provided further insight into various infiltrates at the vaccine sites and at the tumor sites in response to the inoculation of GM-CSF-secreting tumor vaccine.     

The characteristics of the clinical course and treatment of suppurative wounds of the abdominal wall after the removal of a destructively altered appendix

Under analysis are specific features of the clinical course and treatment of infectious complications of the abdominal wall wounds in 209 patients after destructive forms of appendicitis. The microflora of the purulent wound contained gram-negative bacteria in combination with anaerobic nonclostridial flora. The electrochemically activated solution of potassium chloride EHAR-anolit and the endoscopic method were used in complex treatment of the purulent wounds. The recommended method of treatment makes the period of healing the wounds shorter and improves the results of treatment.