Epithelial antibiotics induced at sites of inflammation

The role of antimicrobial peptides in epithelial defense is not fully understood. An epithelial beta-defensin, lingual antimicrobial peptide (LAP), was isolated from bovine tongue and the corresponding complementary DNA cloned. LAP showed a broad spectrum of antibacterial and antifungal activities. LAP messenger RNA abundance was markedly increased in the epithelium surrounding naturally occurring tongue lesions. This increase coincided with the cellular hallmarks of acute and chronic inflammation in the underlying lamina propria, supporting a role for epithelial antimicrobial peptides as integral components of the inflammatory response.     

Flow-cytometric detection of the RI alpha subunit of type I cAMP-dependent protein kinase in human cells

cAMP-dependent protein kinase (PKA) is composed of two genetically distinct catalytic (C) and regulatory (R) subunits. There are two different classes of PKA, designated as type I and type II, which contain distinct R subunits (RI or RII, respectively) but share a common C subunit. Enhanced expression of type I PKA has been correlated with cell proliferation and neoplastic transformation. Detection of the different PKA subunits is usually performed by photoaffinity labeling with 8-N3-32P-cAMP or by radioimmunolabeling techniques. Both techniques are time consuming and require a high number of cells and the use of radioactive reagents. Using the MCF-10A normal human mammary cell line infected with a recombinant retroviral vector containing the human RI alpha gene (MCF-10A RI alpha), we have developed a flow-cytometric assay to detect the intracellular content of RI alpha protein in human cells. MCF-10A and MCF-10A RI alpha cells were fixed in 1.5% paraformaldehyde at 37 degrees C for 15 min and permeabilized by methanol and acetone (1:1) at -20 degrees C for 5 min before staining with a specific IgG2a MoAb followed by a FITC-conjugate rabbit-anti mouse IgG. This procedure was also successfully utilized to recognize RI alpha protein content in human peripheral blood lymphocytes. Flow-cytometric detection of the RI alpha subunit in human cells is feasible and allows the study of the role of type I PKA in cell growth and neoplastic transformation.     

Open pore biodegradable matrices formed with gas foaming

The aim of the present study was to investigate the efficacy and clinical tolerability of the specific leukotriene B4 receptor antagonist VML295 in the treatment of stable plaque psoriasis. Immunohistochemical and flow cytometrical methods were used to assess the effects on inflammation and epidermal proliferation. VML295 in the treatment of chronic plaque psoriasis was shown to be safe and well tolerated. After treatment, there was a statistically significant difference between patients treated with VML295 and patients treated with placebo with respect to the leukotriene B4-induced CD11b up-regulation on the cell surface of polymorphonuclear leukocytes derived from peripheral blood. Ex vivo CD11b up-regulation in the VML295-treated group was completely inhibited after 7 days of treatment (P = 0.001). This effect persisted until the end of the treatment period (P = 0.004 on day 15 and P < 0.0001 after 4 weeks), whereas CD11b up-regulation in the placebo group remained unaffected. There was no statistically significant difference in the median psoriasis area and severity index between the treatment groups at the end of the treatment period. During treatment, no significant histological changes were observed in the markers for cutaneous inflammation and epidermal proliferation. Although not statistically significant, a tendency for the increased expression of some markers of cutaneous inflammation and epidermal proliferation was observed after 1 week of treatment with VML295, and a decreased expression of these markers was seen after 4 weeks of treatment with VML295. This observation could indicate anti-inflammatory effects of VML295 appearing between 2 and 4 weeks after the start of treatment.     

Immunological and virological analyses of persons infected by human immunodeficiency virus type 1 while participating in trials of recombinant gp120 subunit vaccines

We have studied 18 participants in phase I/II clinical trials of recombinant gp120 (rgp120) subunit vaccines (MN and SF-2) who became infected with human immunodeficiency virus type 1 (HIV-1) during the course of the trials. Of the 18 individuals, 2 had received a placebo vaccine, 9 had been immunized with MN rgp120, and seven had been immunized with SF-2 rgp120. Thirteen of the 18 infected vaccinees had received three or four immunizations prior to becoming infected. Of these, two were placebo recipients, six had received MN rgp120, and five had received SF-2 rgp120. Only 1 of the 11 rgp120 recipients who had multiple immunizations failed to develop a strong immunoglobulin G antibody response to the immunogen. However, the antibody response to rgp120 was transient, typically having a half-life of 40 to 60 days. No significant neutralizing activity against the infecting strain was detected in any of the infected individuals at any time prior to infection. Antibody titers in subjects infected despite vaccination and in noninfected subjects were not significantly different. Envelope-specific cytotoxic T-lymphocyte responses measured after infection were infrequent and weak in the nine vaccinees who were tested. HIV-1 was isolated successfully from all 18 individuals. Sixteen of these strains had a non-syncytium-inducing (NSI) phenotype, while two had a syncytium-inducing (SI) phenotype. NSI strains used the CCR5 coreceptor to enter CD4+ cells, while an SI strain from one of the vaccinees also used CXCR4. Viruses isolated from the blood of rgp120 vaccinees were indistinguishable from viruses isolated from control individuals in terms of their inherent sensitivity to neutralization by specific monoclonal antibodies and their replication rates in vitro. Furthermore, genetic sequencing of the env genes of strains infecting the vaccinees did not reveal any features that clearly distinguished these viruses from contemporary clade B viruses circulating in the United States. Thus, despite rigorous genetic analyses, using various breakdowns of the data sets, we could find no evidence that rgp120 vaccination exerted selection pressure on the infecting HIV-1 strains. The viral burdens in the infected rgp120 vaccine recipients were also determined, and they were found to be not significantly different from those in cohorts of placebo-vaccinated and nonvaccinated individuals. In summary, we conclude that vaccination with rgp120 has had,to date, no obvious beneficial or adverse effects on the individuals we have studied.     

Protease inhibitors induce specific changes in protein tyrosine phosphorylation that correlate with inhibition of apoptosis in myeloid cells

To investigate early signaling events responsible for regulation of programmed cell death or apoptosis, we studied campothecin (a topoisomerase I inhibitor)-mediated apoptosis in the human promyelocytic leukemia cell line HL60. We demonstrate a tight correlation between protection of HL60 cells from apoptosis-associated internucleosomal DNA fragmentation by specific protease inhibitors or protein phosphatase inhibitors, with early tyrosine phosphorylation of a single protein substrate with a molecular weight of approximately 42,000. Exposure to protease inhibitors that did not protect HL60 cells from DNA fragmentation did not result in phosphorylation of this substrate. Likewise, a protein tyrosine kinase inhibitor that did not interfere with specific phosphorylation did not prevent DNA fragmentation. Taken together, these results suggest that phosphorylation of a Mr 42,000 substrate constitutes an important signaling event that may participate in regulation of the apoptotic response.     

Transdural spinal cord herniation: imaging and clinical spectra

OBJECTIVE: Many diseases induce asymmetric delays in the visual pathway, resulting in a spontaneous Pulfrich phenomenon (PP). The PP is a visual stereoillusion that may cause difficulties in persons when traveling in cars, crossing the road, or playing ball games. The authors developed and tested a simple new bedside procedure to detect PP. DESIGN: A case series. PARTICIPANTS: Disease simulation in 2 normal subjects and 18 patients with optic neuritis (ON) was examined. Ninety normal subjects were studied to determine normal range of PP. INTERVENTION: The new test, called swinging pen test (SPT), is performed by oscillating a pen by hand. The SPT was compared to a gold standard, a mechanical pendulum (MP). MAIN OUTCOME MEASURES: The authors measured simulated PP in two normal subjects and PP in 18 patients with ON and 90 normal control subjects. The Pearson product-moment correlation (r) and the Spearman rank correlation (rs) between SPT and MP were calculated. RESULTS: The magnitudes of simulated PP determined with the SPT and the MP correlated well (r = 0.92, P < 0.005, and r = 0.96, P < 0.001). Correlation also was good in patients with ON (rs = 0.90, P < 0.05). The positive predictive value of the SPT was 100%, and the negative predictive value was 92%. The PP was absent in all control subjects testing with either pendulum. The normal range for PP varied from -1.40 to 1.52 msec. For the SPT, the intraobserver variability coefficient was 8.2%, and the interobserver variability coefficient was 10.5%. CONCLUSIONS: The authors believe that SPT will be of value to clinicians on bedside evaluation of motion stereopsis dysfunctions. The normal range of PP was approximately +/- -1.5 msec (approximately +/- -1.5 cm), corresponding to a 0.3-log unit neutral density filter).     

The Health Effects of Low-Contact Water Activities in Fresh and Estuarine Waters

Four studies were carried out at separate locations to investigate the relationship between health effects and low-contact water sports, and intensive microbiological sampling was conducted in parallel to the health studies at each site. The two sports examined were marathon canoeing and rowing.
The extremes of water quality were at the estuarine sites on the River Torridge, where pollution levels varied from a geometric mean faecal coliform value of 62/100 ml at the Appledore/Instow site to 4613/100 ml at the Bideford site.
A comparison of 'exposed' and 'unexposed' groups, 5–7 days after exposure, showed that the health effects of low-contact water sports are minimal, within the water quality ranges which were studied.     

Qigong and L-1 straining maneuver oxygen system requirements with and without positive pressure breathing

Based on the characteristics of respiration and the intrathoracic pressure in Qigong (Q-G) maneuvering, it has been theorized that the Q-G maneuver may lessen the lack of coordination between aircraft oxygen apparatus and anti-G maneuvers and may be more compatible with positive pressure breathing (PPB). In an experiment intended to test this hypothesis, 5 male volunteers, trained in Q-G and L-1 maneuvers, performed the Q-G and the L-1 maneuvers without and with (PPB) at 4 and 6 kPa, respectively, with 14 respiratory parameters being measured. The results demonstrated that, when performing Q-G maneuver, the maximal expiratory flow rate averaged 1.175-1.645 L.s-1, the inspiratory peak flow, 1.003-1.297 L.s-1. Both these values were markedly lower than those of the L-1 maneuver, and matched well the performance of current aircraft oxygen apparatus. From the blood pressure and heart rate values, it is evident that PPB can further promote the blood pressure-raising effect of the Q-G maneuver, and alleviate pilots' fatigue.     

Tissue factor in the pathogenesis of atherosclerosis

The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese NIDDM. We performed a genome wide scan in F2 progenies obtained by crossing OLETF rats with two control strains, Long-Evans Tokushima Otsuka (LETO) and Fisher-344(F-344) rats. Since diabetes develops only in male progenies, we used only male F2 rats for the linkage studies.Highly significant linkage was observed between the phenotype, postprandial hyperglycemia and P-450ald locus on chromosome 1 and D7Mit 11 locus on chromosome 7. In addition, suggestive linkage was found between fasting glucose level and body weight and these two loci. Four other regions (D1Mit12, D2Mit11, D5Mgh14, and D17Arb1) on chromosome 1, 2, 5, and 17 were detected to influence body weight, fasting glucose level or postprandial hyperglycemia independently. We concluded that non-insulin-dependent diabetes mellitus(NIDDM) in OLETF rats is regulated by multiple genes which affect fasting, postprandial hyperglycemia, and obesity differently.