Extracorporeal photochemotherapy (photopheresis) induces apoptosis in lymphocytes: a possible mechanism of action of PUVA therapy

The mechanism of action of psoralen plus UVA (PUVA) and photopheresis is not entirely understood. These therapies are assumed to be immunomodulating partly by gradually decreasing leukocyte viability. We investigated whether this delayed form of cell death was due to apoptosis. Untreated and treated (PUVA exposed) leukocytes obtained from six patients with systemic sclerosis and (untreated) leukocytes from healthy control individuals were studied. Qualitative gel electrophoresis and quantitative in situ nick translation analysis of DNA fragmentation was performed. Apoptosis of the treated cells did occur (gel electrophoresis) after 24 h. At t = 0 h, immediately after exposure to PUVA, there was no evidence of DNA fragmentation in the treated cells. The percentage of treated cells undergoing apoptosis was 20-55% at t = 24 h (in situ nick translation). The untreated leukocytes of the patients and the healthy individuals showed no distinctive rise in apoptotic cells. Apoptosis of the leukocytes after PUVA or photopheresis treatment might be a mechanism of action and might explain the therapeutic response.     

The mechanism of cyclosporine toxicity induced by clarithromycin

Tsetse fly-transmitted trypanosomes (Trypanosoma spp.) cause "sleeping sickness' in man and have a serious impact on livestock-based agriculture in large areas of Africa. Multigene control of variation in susceptibility to trypanosomiasis is known to occur in mice, where the C57BI/6 (B6) strain is relatively resistant and the A/J (A) and Balb/c (B) strains are susceptible. Such resistance is also well described among several types of west African cattle. We report here the results of genome-wide scans for genes controlling this trait in the B6 mouse using crosses with two different susceptible strains. Regions on mouse chromosomes 5 and 17 were found to be important in determining resistance in both crosses while an additional region on chromosome 1 showed evidence of involvement in only one cross. We confirmed the size of the effect due to chromosome 17 in F3 intercross populations fixed for alternative parental chromosomes. The three loci are of large effect and account for most of the genetic variation in both F2 populations. We propose that they be designated Tir1, Tir2 and Tir3.     

Immune response against the non-repeat region (293-310) of the circumsporozoite protein of Plasmodium vivax

Immunization by peptides based on the repeat sequences of Plasmodium falciparum or P. vivax antigen(s) have shown inconsistent results during clinical trials in humans. This could be attributed to the lack of T-cell help or antigenic polymorphism. Thus, attention has been focused towards the more conserved non-repeat regions. The present study was undertaken to map the antigenic determinant in the vicinity of region II (outside the repeat) of CS protein of P. vivax. The immunogenicity of the peptide was studied alone and after linking with polytuftsin (PT), using alum and Freund's adjuvant, in inbred strains of mice with different genetic backgrounds. The humoral response and antigen induced T-cell proliferation assays clearly demonstrated the immunomodulatory activity of PT. Comparable results were observed with antigen(s) administered either in alum or Freund's adjuvant. The induction of IgG2a and IgG2b antibody isotypes by both, peptide as well as the conjugate, may indicate that the T-helper response involved is of Th1 type. Further the immunofluorescence studies have shown that antibodies recognized the air dried sporozoites of P. cynomolgi. The results thus show that the above sequence has overlapping B and T-cell determinants and that alum can be substituted for Freund's adjuvant in generating an effective immune response.     

Predischarge arrhythmia induction testing of implantable defibrillators may be unnecessary in selected cases

Complete postoperative evaluation of implantable cardioverter-defibrillators (ICDs) before discharge, including arrhythmia induction, has been the standard since their introduction. Whereas the original ICDs provided little telemetered information and used separate pace-sense and defibrillation leads, modern, third-generation devices provide pace-sense function information in addition to other data and are used in conjunction with integrated transvenous endocardial leads that combine pace-sense and defibrillation function. Changes in lead position, which can potentially result in either an inability to detect fibrillation or to terminate it, should be mirrored by changes in resting pace-sense function. Thus, for newer ICDs implanted with integrated endocardial lead systems, it is possible that in at least some cases predischarge arrhythmia inductions can be avoided. Two hundred patients receiving third-generation ICDs in conjunction with integrated transvenous leads were evaluated before discharge. Defibrillation detection or termination problems were seen in 8. Declines in resting R-wave amplitude and pacing impedance were significantly associated with such complications (-7 +/- 5 vs -0.3 +/- 2.3 mV [p <0.0001] and -158 +/- 138 vs -93 +/- 76 omega [p <0.05], for those with vs without complications, respectively), as were gross right ventricular lead migrations on chest x-ray. No patient with a defibrillation complication had an R-wave change of <3 mV. However, 13% of patients without complications had R-wave changes of >3 mV. It is concluded that a pace-sense evaluation of ICDs may be a satisfactory screen to determine those who need to go on to complete testing with arrhythmia induction in selected cases.     

Kinematic analysis of shear displacement as a means for operating mechanotransduction channels in the contact region between adjacent stereocilia of mammalian cochlear hair cells

In sensory hair cells of the cochlea, deflection of the stereociliary bundle results in direct mechanical gating of mechanoelectrical transduction channels, a function generally attributed to the tip link running between the tips of short stereocilia and the sides of adjacent taller ones. However, immunocytochemical experiments indicate that the channels may not be associated with the tip link but occur just below it in a region of contact between the stereocilia. To determine whether transduction channels in this location could be operated during physiologically appropriate deflections as effectively by shear displacement as if they were associated with the tip link, a two dimensional kinematic analysis of relative motion between stereocilia has been performed assuming contact between stereocilia is maintained during deflection. Bundle geometry and dimensions were determined from transmission electron micrographs of hair cells from several frequency locations between 0.27 and 13.00 kHz in the guinea-pig cochlea. The analysis indicates that for a 10 nm deflection of the tallest stereocilia of both inner and outer hair cells, i.e. within the range of the maximum sensitivity of mammalian hair bundles, the average shear displacement in the contact region would be 1.6 nm, but that it increases systematically towards higher frequency regions for outer hair cells. This displacement is comparable in magnitude to tip-link elongation for individual stereociliary pairs.     

Differential recruitment of B and T cells in coxsackievirus B4-induced pancreatitis is influenced by a capsid protein

Two genetically similar variants of coxsackievirus B4, CB4-P and CB4-V, cause distinct disease syndromes in mice. A multidisciplinary approach was used to examine the events occurring in situ. The CB4-P variant induced acute pancreatitis, followed by repair of the exocrine tissues, while the CB4-V variant induced chronic pancreatitis, characterized by extensive destruction of the exocrine tissues. Since CB4-V replicated more efficiently than CB4-P in vivo, the more extensive tissue injury associated with CB4-V infection could be explained as the result of a higher level of viral replication. However, the fact that CB4-V replicated more efficiently in a mouse strain that survives infection than in a strain that succumbs to infection suggests that immune-mediated mechanisms as well as viral cytolysis may contribute to pancreatic tissue injury. To address the role of the immune system in virus-induced pancreatitis, the cell types within the inflammatory infiltrate were analyzed by flow cytometry. B cells (34 to 75%) were the most abundant, followed by T cells (10 to 30%), natural killer cells (4 to 8%), and macrophages (0 to 6%). Recruitment (and perhaps proliferation) of B and T cells to the pancreatic tissues was influenced by viral strain. Differential recruitment of T and B cells may reflect altered antigenic sites between CB4-P and CB4-V. The viral sequence that affected T- and B-cell recruitment was identified as a threonine residue at position 129 of the VP1 capsid protein.     

Relationship of preoperative antiendotoxin core antibodies and adverse outcomes following cardiac surgery

This study assesses the reliability of a self-reported health questionnaire completed by 413 subjects aged 25-74 yr in the Erie County Periodontal Disease (ECPD) Study. Specific questions on general and oral health conditions were completed by each subject during a first visit and at a follow-up examination 2 yr later, and the two compared. Results showed that the overall measure of agreement between the two visits is substantial (average kappa, kappa = 0.80). Variation by gender and age were minimal. Questions regarding allergy to medications, oral treatment, reason for tooth extraction, health symptoms and history of systemic diseases exhibited high levels of agreement (kappa ranged from 0.71-0.90). Information on vitamin and mineral intake yielded kappa = 0.63. Oral conditions scored the lowest but were still acceptable (kappa = 0.57). These findings indicate that there were no significant discrepancies in self-reported responses to the health questionnaire used in the ECPD Study. Although the information provided by the subject may not be as accurate as compared to laboratory testing, it is nevertheless a reliable source of information which can be utilized cost-effectively in research studies.     

Differential transforming growth factor-beta secretion in adenocarcinoma and squamous cell carcinoma of the uterine cervix

Tumor cells from eight freshly isolated cervical cancers (i.e., four adenocarcinomas and four squamous carcinomas) were analyzed for their production of the immune-inhibitory cytokine transforming growth factor-beta (TGF-beta) in vitro. All fresh adenocarcinomas secreted significant levels of TGF-beta (mean 397, range between 207 and 782 pg/ml/10(5) cells/48 hr). In contrast, no detectable TGF-beta was present in the supernatants from the four fresh squamous carcinoma cultures (P < 0.001). These data suggest that major differences in the secretion of the immunoinhibitory cytokine TGF-beta exist between squamous cell carcinomas and adenocarcinomas of the uterine cervix. Furthermore, these findings suggest that at least some of the differences in the natural biologic behavior, as well as in the response to radiation treatment, between these two histologic types of cervical cancer could be related to differences in secretion of this immune-inhibitory cytokine.     

Ex vivo culture of peripheral blood CD34+ cells: effects of hematopoietic growth factors on production of neutrophilic precursors

A major potential application for ex vivo culture of hematopoietic progenitor cells is the treatment of cytopenia following high-dose chemotherapy and hematopoietic transplantation. We have previously postulated that infusion of a sufficient number of neutrophil postprogenitor cells generated by ex vivo culture of CD34+ cells may be able to abrogate neutropenia. In this article, we describe further development of an efficient stromal-free, cytokine-dependent, static culture system for generation of these cells. Our previous studies indicated that maximal production of nucleated cells and myeloid progenitor cells from PB CD34+ cells occurred with multiple hematopoietic growth factor (HGF), notably the 6-HGF combination of interleukin (IL)-1, IL-3, IL-6, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-CSF (GM-CSF), and stem cell factor (SCF). In the present study, we determine the contribution of each of these 6 HGF in generation of neutrophilic precursors. SCF, G-CSF, and IL-3 were found to be the most important HGF for production of neutrophilic cells. The 4-HGF combination of IL-3, IL-6, G-CSF, and SCF was optimized by performing dose-response experiments and shown to be as potent as 6 HGF for production of nascent CFU-GM and neutrophilic precursors.     

Axis II psychopathology as a function of Axis I disorders in childhood and adolescence

OBJECTIVE: To examine the occurrence of elevated personality disorder (PD) dimensional scores in a community sample of young adults as a function of the occurrence of Axis I disorders through age 18 years. METHOD: 299 individuals who had been interviewed regarding Axis I disorders twice while in adolescence (first when 14 through 18 years of age) were carefully assessed regarding Axis I and II psychopathology at age 24. RESULTS: The prevalence of PD diagnoses was relatively low (3.8% in participants with a history of Axis I versus 1.7% in participants with no Axis I history). The occurrence of all four Axis I diagnostic categories (major depression, anxiety disorders, disruptive behavior disorders, substance use disorders) in childhood and adolescence was associated with elevated PD dimensional scores. The likelihood of elevated PD dimensional scores increased as a function of the number of Axis I disorders. Elevated PD scores were significantly associated with a negative course of major depression. CONCLUSIONS: Although the rates of PDs were low, the findings suggest a substantial degree of association between early-onset Axis I disorders and Axis II psychopathology in young adulthood. More research is needed to develop assessment and treatment recommendations addressing the early manifestations of PDs.