Inhibition of mitogen-activated protein kinase kinase blocks T cell proliferation but does not induce or prevent anergy

Three mitogen-activated protein kinase pathways are up-regulated during the activation of T lymphocytes, the extracellular signal-regulated kinase (ERK), Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase pathways. To examine the effects of blocking the ERK pathway on T cell activation, we used the inhibitor U0126, which has been shown to specifically block mitogen-activated protein kinase/ERK kinase (MEK), the kinase upstream of ERK. This compound inhibited T cell proliferation in response to antigenic stimulation or cross-linked anti-CD3 plus anti-CD28 Abs, but had no effect on IL-2-induced proliferation. The block in T cell proliferation was mediated by down-regulating IL-2 mRNA levels. Blocking Ag-induced proliferation by inhibiting MEK did not induce anergy, unlike treatments that block entry into the cell cycle following antigenic stimulation. Surprisingly, induction of anergy in T cells exposed to TCR cross-linking in the absence of costimulation was also not affected by blocking MEK, unlike cyclosporin A treatment that blocks anergy induction. These results suggest that inhibition of MEK prevents T cell proliferation in the short term, but does not cause any long-term effects on either T cell activation or induction of anergy. These findings may help determine the viability of using mitogen-activated protein kinase inhibitors as immune suppressants.     

The barrier domain for solute permeation varies with lipid bilayer phase structure

Resting blood pressure in both white and black families participating in the HERITAGE Family Study was analyzed using a simple familial correlation model to assess familial influences. The two samples of black and white families were analyzed separately and together, providing an opportunity to test for heterogeneity in the familial resemblance. Maximal heritability was 46% for systolic blood pressure (SBP) and 31% for diastolic blood pressure (DBP) in the pooled sample. Noticeably higher heritabilities were found in the black sample (68% for SBP and 56% for DBP) than in the white sample (43% for SBP and 24% for DBP). The patterns of familial correlations were similar in blacks and whites, with the exception that spouse resemblance was significant in white families but not in black families. These results along with the finding that the magnitude of the familial correlations was higher in the black sample than in the white sample suggest that the effects of host and familial environmental factors differ between the races.     

Studies of bystander effects in human fibroblasts using a charged particle microbeam

PURPOSE: To determine the role of cell-to-cell mediated effects in the response of cells to different radiations by using a charged particle microbeam capable of targeting individual cells with counted particles. MATERIALS AND METHODS: Primary human fibroblasts were irradiated conventionally with X-rays or alpha-particles and scored for the induction of micronucleated or apoptotic cell formation at various times after irradiation. Cells were also individually irradiated with helium-3 particles from a microbeam and the distribution of damaged cells between hit and non-hit cells scored. RESULTS: Conventionally exposed fibroblasts showed a dose-dependent production of micronucleated and apoptotic cells 3 days after irradiation for both X-rays and alpha-particles, with a high RBE value for alpha-particles at low doses. Targeting individual alpha-particles, using a microbeam, to four cells within a population produced more micronucleated and apoptotic cells than expected on the basis of a direct effect only. CONCLUSIONS: This study provides direct evidence for the production of transmissible, cell-to-cell effects between hit and non-hit cells individually exposed to charged particles.     

Disulfide bond mapping and structural characterization of spruce budworm antifreeze protein

We have previously reported that bryostation 1 (Bryo 1) induces differentiation of chronic lymphocytic leukemia (CLL) in vitro to a hairy cell (HC) stage. This study tests the hypothesis that Bryo 1-differentiated CLL cells are more susceptible to 2-chlorodeoxyadenosine (2-CdA) than parent CLL cells. A recently established EBV-negative CLL line (WSU-CLL) from a patient resistant to chemotherapy including fludarabine was used to test this hypothesis. Both Bryo 1 (10-1000 nM) and 2-CdA (5.6-22.4 microM) exhibited a dose-dependent growth inhibitory effect on the WSU-CLL cell line. In vitro, the sequential exposure to Bryo 1 (100 nM for 72 h) followed by 2-CdA (11.2 microM) resulted in significantly higher rates of growth inhibition than either agent alone. Changes in immunophenotype, enzymes, lipids, proteins, and the DNA of WSU-CLL cells were studied before and after Bryo 1 treatment. Bryo 1 induced a positive tartrate-resistant acid phosphatase reaction and two important markers, CD11c and CD25, after 72 h of culture, confirming the differentiation of CLL to HC. The Fourier transformation infrared spectroscopic analysis showed that the amount of membrane lipids significantly increased in Bryo 1-treated cells compared to controls after 24 h, whereas the protein content, as well as the DNA content, decreased. This finding supports the change of CLL to HC. To evaluate the in vivo efficacy of Bryo 1 and 2-CdA, we used a xenograft model of CLL in WSU-CLL-bearing mice with severe combined immune deficiency. s.c. tumors were developed by injection of 10(7) WSU-CLL cells, and fragments were then transplanted into a new batch of severe combined immunodeficient mice. Bryo 1 and 2-CdA at the maximum tolerated doses (75 micrograms/kg i.p. and 30 mg/kg s.c., respectively) were administered to the mice at different combinations and schedules. The survival in days, the tumor growth inhibition ratio, the tumor growth delay, and the log10 kill of the mice treated with Bryo 1 followed by 2-CdA were significantly better than the control and other groups. We conclude that the sequential treatment with Bryo 1 followed by 2-CdA resulted in higher antitumor activity and improved animal survival.     

Predictive value of monitoring parameters in fetal surgery

BACKGROUND/PURPOSE: The choice of monitoring parameters in fetal surgery has thus far been based on feasibility rather than on predictability. Ideally, monitoring should be noninvasive, have a rapid response time and high sensitivity, and be applicable to open and endoscopic techniques. Herein, the authors studied the response of several parameters to standardized episodes of fetal ischemia and stress. METHODS: Eight time-dated fetal lambs (110 days, term, 145 days) were used. Under general anesthesia, a balloon occluder was placed around the umbilical cord. Pulse oximetry (POx + heart rate, HR), electrocardiography (ECG), direct oximetry (DOx), and blood pressure (BP) were recorded continuously. After stabilization, the umbilical cord was completely occluded for 5 seconds, then released. False-negative recordings were defined as failure of a parameter to respond to umbilical occlusion; false-positive episodes were defined as 10% change in value over < or = 10 seconds during stabilization (baseline) period. RESULTS: The fetuses were monitored for an aggregate of 358 minutes. Baseline DOx was 64%+/-5%, POx, 66%+/-16%; HR, 141+/-18 beats per minute (bpm); systolic BP (SBP), 51+/-3 torr; and diastolic BP (DBP), 38+/-2 torr. During umbilical occlusion (n=15), SBP increased to 56+/-3 torr and DBP to 43+/-2 torr at 0.5 seconds, then returned to baseline at 8.0 seconds. A decrease was seen in DOx (start at 3.5s, maximum delta 9.9+/-1.5% at 10.5 seconds) and POx (start at 4.2 seconds, maximum delta 7.3+/-2.4% at 20.5 seconds). Heart rate showed <10% decrease (start at 8.5 seconds, nadir 131+/-14 bpm at 19.5 seconds). No ECG changes were noted. Sensitivity was 100% for DOx, POx, and BP, but only 14% for HR; specificity was 97% for DOx and 88% for POx; positive predictive value was 58% for DOx and 37% for POx; negative predictive value was 100% for DOx and POx. CONCLUSIONS: Direct intravascular oximetry and blood pressure provide a prompt and reliable response to acute fetal stress, but are too invasive for routine use. Bradycardia is an insensitive and late sign of fetal distress. Pulse oximetry has a rapid response time (<5 seconds), high sensitivity, and negative predictive value. In addition, its application is noninvasive and has proven to be feasible in open and endoscopic fetal surgical procedures. It therefore appears to be the monitoring parameter of choice for fetal surgery.     

Management and after-care of extensor tendon injuries of the hand

The investigation and management of extensor tendons play a minor role compared to those of flexor tendons. The finger extension does not seem to be as important as the flexion. But the practical value of the hand is determined by both. Different managements are used depending on the level of extensor tendon injury. More distal located injuries require a longer restraint than those, which are located more proximal. On the one hand this depends on the blood supply of the extensor tendon, which is by far better in the lower arm and dorsal hand than peripheral. On the other hand the reason is the different amplitude of gliding of extensor tendons, which decreases from proximal to distal to lower than 1 mm. Therefore the tension on extensor tendon sutures increases from proximal to distal. The varies techniques of extensor tendon reconstruction will be described.     

Effects of hypesthesia on oral behaviors of the orthognathic surgery patient

PURPOSE: The purpose of this study was to compare orthognathic surgery patients with and without significant hypesthesia with respect to perceived problems with specific oral behaviors. PATIENTS AND METHODS: Data from 116 patients 6 months after bilateral sagittal split osteotomy (BSSO) and mandibular advancement were analyzed. Tactile sensation in the right and left mental nerve areas was determined using monofilaments and brush strokes (von Frey hairs). The right infraorbital region was used as a control. A difference of 450 mg of force between the control and test sites was considered significant hypesthesia. Patients rated their level of subjective problems with swallowing liquids or solids, smiling, spitting, kissing, speaking, eating, and drooling on a scale from 1 (none to mild) to 7 (extreme). A value of 5 or greater was considered significant impairment. RESULTS: Hypesthesia was shown in 23 patients (19.8%) with the monofilaments and in 29 patients (25.0%) using brush stroke direction. In each of these two groups, a significant correlation was observed between hypesthesia and difficulty in chewing and kissing. No correlation was observed between any of the remaining seven oral behaviors and hypesthesia. CONCLUSION: These findings suggest that only certain oral behaviors are affected by hypesthesia of the mental nerve.     

Energy substrates and amino acids provided during in vitro maturation of bovine oocytes alter acquisition of developmental competence

Clinicians commonly include an assessment of leg length inequality (LLI) as a component of a musculoskeletal examination. Little research is available, however, documenting reliability and validity of clinical methods for assessing LLI. The purpose of this study was to determine the reliability and validity of assessing functional LLI using a pelvic leveling device. Subjects were 19 women and 13 men between the ages of 18 and 55 who reported having a diagnosed or suspected LLI. Clinical determination of LLI was made by placing rigid lifts under the suspected shorter lower extremity until the leveling device indicated that the iliac crests were level. This measurement was made twice by one investigator and once by a second investigator. Standing radiographic measurements of LLI using rigid lifts were used to establish validity of the clinical method. Intraclass correlation coefficients [ICC(2,1)] and absolute difference values were computed to assess reliability and validity. The mean absolute difference between the two clinical measurements of LLI by the same investigator was 0.29 cm (+/- 0.52), with an ICC = 0.84. The mean absolute difference between clinical measurements of LLI by the two investigators was 0.49 cm (+/- 0.46), with an ICC = 0.77. The ICC and mean absolute difference reflecting agreement between radiographic measurements and clinical measurements of LLI was 0.64 and 0.58 cm (+/- 0.58), respectively, for one investigator and 0.76 and 0.55 cm (+/- 0.37), respectively, for the second investigator. The intratester reliability, intertester reliability, and validity assessments included instances in which paired observations disagreed regarding which lower extremity was the shorter lower extremity. Factors that may be associated with the unacceptable reliability and validity of the clinical assessment method include asymmetric positioning of the ilia, body composition of the patient, and design of the clinical instrument. The authors discuss clinical implications related to assessment of LLI.     

Enterogastric bile reflux during technetium-99m-sestamibi cardiac imaging

Twenty-six patients with squamous cell cancer of the cervix were treated with i.v. paclitaxel, 250 mg/m2 over 3 h every 21 days. They received steroid, H1 and H2 blocker premedications, and granulocyte-colony-stimulating factor (G-CSF) support (5 microgram/kg/day). No prior chemotherapy, except as a radiation sensitizer, was allowed. The median age was 50 (range, 36-81) years, and performance status Zubrod was 1 (range, 0-2). Eight (33%) patients had prior surgery, and 22 (92%) had prior radiation therapy. Twenty-four patients were evaluable for response; 2 were later found to be ineligible. Five patients had partial responses (21%; 95% confidence interval, 6-40%), and 14 (58%; 95% confidence interval, 35-78%) had stable disease. The median duration of response was 10 (range, 3-27+) weeks. The responses were within the radiation port (four responses) and outside of it (one response). The median interval from the start of irradiation to the start of paclitaxel in responding patients was 94 weeks, whereas in patients with stable disease it was 68 weeks, and in patients whose disease progressed it was 46 weeks. Eighty-eight percent of the 105 cycles of paclitaxel were administered at a dose of 250 mg/m2 or higher. Granulocytopenia was brief and noncumulative, with grades 3 and 4 experienced by 5 and 3 patients, respectively. G-CSF was used for a median of 7 (range, 2-14) days/cycle. Anemia was mild, with G3 noted in 3 patients, and thrombocytopenia was not significant. Infections and musculoskeletal pain were mild and infrequent. Sensory (14 patients G1 or G2 and 2 patients G3) and motor (4 patients G1 or G2 and 1 patient G3) neurotoxicity was noted. There was no significant cardiovascular toxicity. Paclitaxel is active in patients with squamous cell cancer of the cervix and is well tolerated at this dose schedule with G-CSF support.     

Effect of tissue-plasminogen activator on leukocyte-endothelial interactions at the microcirculatory level

In free tissue transfer and replantation surgery, there is a debate over whether any pharmacologic agents should be used to improve vessel patency and tissue survival. Because tissue-plasminogen activator (t-PA) is a highly effective and safe fibrinolytic, it may be useful in obtaining and maintaining vessel patency. The direct effects of t-PA on skeletal muscle hemodynamics and leukocyte activation at the microcirculatory level were investigated. Male Sprague-Dawley rats (n = 20) were divided into three experimental groups: control (n = 8), vehicle (n = 6), and t-PA (n = 6). Using the cremaster muscle flap model and intravital microscopy, red blood cell velocity, vessel diameter, capillary perfusion, endothelial edema index, and leukocyte-endothelial interactions (rolling, adhering, and transmigrating leukocytes) in postcapillary venules were measured. In the vehicle and t-PA groups, vehicle or t-PA was infused by means of a catheter inserted into the lower abdominal aorta for local infusion. Except for a significant reduction in the diameter of the first order arterioles from 117 microm to 82 microm (medians; p = 0.026), t-PA did not significantly affect red blood cell velocity, vessel diameter, or capillary perfusion compared with vehicle. However, leukocyte-endothelial interactions did differ significantly in postcapillary venules. Adhering leukocytes counted per visual field decreased from 4.67 in the vehicle group and 3.50 in the control group to 1.67 in the t-PA group (medians; p = 0.015 and p = 0.005, respectively); transmigrating leukocytes in the t-PA group decreased from 4.75 in the vehicle group and 3.50 in the control group to 1.67 in the t-PA group (medians; p = 0.002 and p = 0.043, respectively). t-PA treatment significantly decreased the number of both adhering and transmigrating leukocytes. These novel findings on leukocyte-endothelial interactions suggest that t-PA has anti-inflammatory effect.