Vaccination with plasmid DNA encoding mycobacterial antigen 85A stimulates a CD4+ and CD8+ T-cell epitopic repertoire broader than that stimulated by Mycobacterium tuberculosis H37Rv infection

Vaccination of mice with plasmid DNA carrying the gene for the major secreted mycobacterial antigen 85A (Ag85A) from Mycobacterium tuberculosis is a powerful technique for generating robust specific Thl helper T-cell responses, CD8+-mediated cytotoxicity, and protection against M. tuberculosis challenge (K. Huygen et al., Nat. Med. 2:893-898, 1996). We have now analyzed in more detail the antigen-specific immune CD4+- and CD8+-T-cell responses induced in BALB/c mice vaccinated with Ag85A DNA and have compared these responses to those generated by intravenous infection with M. tuberculosis. T-cell-epitope mapping, as measured by interleukin-2 and gamma interferon secretion from splenic T cells restimulated in vitro with synthetic 20-mer peptides spanning the complete mature sequence of Ag85A, demonstrated that DNA vaccination stimulated a stronger and broader T-cell response than did M. tuberculosis infection. Moreover, elevated cytotoxic T lymphocyte (CTL) activity against Ag85A-transfected and peptide-pulsed P815 target cells could be generated exclusively by vaccination with plasmid DNA, not following M. tuberculosis infection. By using DNA vaccination, three Ag85A CTL epitopes with predicted major histocompatibility complex class I binding motifs were defined. One of them was previously reported as a dominant, promiscuously recognized T-cell epitope in healthy humans with primary infections. These data strengthen the potential of DNA vaccination with respect to inducing antituberculous immunity in humans.     

Cataract surgery combined with transpupillary silicone oil removal through planned posterior capsulotomy

OBJECTIVE: The purpose of the study was to evaluate transpupillary removal of silicone oil combined with cataract surgery in patients after pars plana vitrectomy. DESIGN: A prospective case-control study. PARTICIPANTS: Fifty consecutive patients underwent cataract surgery combined with removal of silicone oil, which had served as intraocular tamponade after pars plana vitrectomy. In 28 patients, silicone oil was removed through a planned posterior capsulotomy, and in 22 patients, silicone oil was removed through pars plana sclerotomies. All patients were operated on by the same surgeon. INTERVENTIONS: Pars plana vitrectomy, cataract surgery, and silicone oil removal were performed. MAIN OUTCOME MEASURES: Frequencies of retinal redetachment, secondary cataract, cystoid macular edema, and vitreous hemorrhage; visual acuity; intraocular pressure; and duration of surgery and visual rehabilitation were measured. RESULTS: Frequencies of postoperative vitreous hemorrhage (1 of 28 [4%] vs. 10 of 22 [45%]) and secondary cataract (0 of 28 vs. 6 of 22 [27%]) were significantly lower (P < 0.05; chi-square test), and duration of surgery and visual rehabilitation were significantly shorter (P < 0.01) for patients with transpupillary silicone oil removal than for patients with drainage of silicone oil through pars plana sclerotomies. Rate of retinal redetachment (4 of 28 [14%] vs. 4 of 22 [18%]), time of retinal redetachment (36 +/- 32 postoperative days vs. 54 +/- 65 days), frequency of dislocated intraocular lenses (1 of 28 vs. 0 of 22), and postoperative visual acuity did not vary significantly between the two groups. Persisting comeal endothelial decompensation and clinically significant cystoid macular edema due to cataract surgery were not observed in any patient. CONCLUSIONS: Silicone oil removal can be combined with cataract surgery. In view of a decreased frequency of postoperative vitreous hemorrhage, reduced rate of secondary cataract, and shorter duration of surgery and visual rehabilitation, transpupillary drainage of silicone oil through a planned posterior capsulotomy compares favorably with removal of silicone oil through pars plana sclerotomies. Retinal redetachment usually occurs within the first 3 postoperative months.     

Molecular cloning of the cDNA encoding pp36, a tyrosine-phosphorylated adaptor protein selectively expressed by T cells and natural killer cells

Activation of T and natural killer (NK) cells leads to the tyrosine phosphorylation of pp36 and to its association with several signaling molecules, including phospholipase Cgamma-1 and Grb2. Microsequencing of peptides derived from purified rat pp36 protein led to the cloning, in rat and man, of cDNA encoding a T- and NK cell-specific protein with several putative Src homology 2 domain-binding motifs. A rabbit antiserum directed against a peptide sequence from the cloned rat molecule recognized tyrosine phosphorylated pp36 from pervanadate-treated rat thymocytes. When expressed in 293T human fibroblast cells and tyrosine-phosphorylated, pp36 associated with phospholipase Cgamma-1 and Grb2. Studies with GST-Grb2 fusion proteins demonstrated that the association was specific for the Src homology 2 domain of Grb-2. Molecular cloning of the gene encoding pp36 should facilitate studies examining the role of this adaptor protein in proximal signaling events during T and NK cell activation.     

Cloning, characterization, and chromosomal localization of human superillin (SVIL)

The specific aim of this study was to determine the response of alveolar bone after it was augmented vertically using distraction osteogenesis and subsequently loaded with implant restorations. Four dogs each had four implants placed horizontally into an edentulous mandibular quadrant and, after integration, a distraction osteogenesis device was fabricated in the laboratory. An osteotomy was made to allow the crest of the alveolar ridge to be distracted vertically. After 10 mm of vertical distraction, the device was stabilized with light cured resin. Following bone fill confirmation of the distraction gap at 10 weeks, two implants were placed into the ridges, one in distracted bone and one in nondistracted bone. After 4 months for implant integration, freestanding prostheses were fabricated. Crestal bone levels were evaluated throughout the period of function. Animals were sacrificed after 1 year of loading, for histologic evaluation of the bone. The vertical ridge augmentation averaged 8.85 +/- 1.05 mm after 10 weeks of healing following distraction, without change over 1 year of implant loading. Histologic examination showed that bone had formed between the distracted segments, creating an augmented ridge. The average thickness of the labial cortex in the distraction gap was significantly thinner than the lingual cortex in distracted bone and the lingual and labial nondistracted cortical bone. The presence of the dental implant did not significantly affect cortical bone thickness. Serial sections showed that implants remained integrated and functional without soft tissue inflammation. Dental implants placed into alveolar ridges augmented with the technique of distraction osteogenesis maintained bone and were functional for the length of this study.     

New p-terphenyl and polyketide metabolites from the sclerotia of Penicillium raistrickii

Three new p-terphenyls (1-3), a new xanthone derivative (4), and two known fungal metabolites (5 and 6) have been isolated from the sclerotia of Penicillium raistrickii (NRRL 2039). The structures for 3,3"-dihydroxy-6'-desmethylterphenyllin (1); 3'-demethoxy-6'-desmethyl-5'-methoxycandidusin B (2); 6'-desmethylcandidusin B (3); and 1,3,5, 6-tetrahydroxy-8-methylxanthone (4) were determined on the basis of HRMS and NMR data. Although compounds 1 and 4 exhibited mild antiinsectan and antibacterial activity, griseofulvin (5) was responsible for most of the activity of the sclerotial extract in dietary assays against the corn earworm Helicoverpa zea.     

Specific requirement for CD3epsilon in T cell development

T cell antigen receptor (TCR) and pre-TCR complexes are composed of clonotypic heterodimers in association with dimers of signal transducing invariant subunits (CD3gamma, -delta, -epsilon, and zeta). The role of individual invariant subunits in T cell development has been investigated by generating gene-specific mutations in mice. Mutation of CD3gamma, -delta, or zeta results in an incomplete block in development, characterized by reduced numbers of mature T cells that express low levels of TCR. In contrast, mature T cells are absent from CD3epsilon-/- mice, and thymocyte development is arrested at the early CD4(-)CD8(-) stage. Although these results suggest that CD3epsilon is essential for pre-TCR and TCR expression/function, their interpretation is complicated by the fact that expression of the CD3gamma and CD3delta genes also is reduced in CD3epsilon-/- mice. Thus, it is unclear whether the phenotype of CD3epsilon-/- mice reflects the collective effects of CD3gamma, -delta, and -epsilon deficiency. By removing the selectable marker (PGK-NEO) from the targeted CD3epsilon gene via Cre/loxP-mediated recombination, we generated mice that lack CD3epsilon yet retain normal expression of the closely linked CD3gamma and CD3delta genes. These (CD3epsilonDelta/Delta) mice exhibited an early arrest in T cell development, similar to that of CD3epsilon-/- mice. Moreover, the developmental defect could be rescued by expression of a CD3epsilon transgene. These results identify an essential role for CD3epsilon in T cell development not shared by the CD3gamma, CD3delta, or zeta-family proteins and provide further evidence that PGK-NEO can influence the expression of genes in its proximity.     

Cyclosporine dosage can be reduced when used in combination with an anti-intercellular adhesion molecule-1 monoclonal antibody in rats undergoing heterotopic heart transplantation

OBJECTIVE: Although occupationally embedded exercise is becoming a recognized topic in occupational therapy research, study of this phenomenon in children has been limited. The present study examined whether play elicits therapeutic patterns of movement through the use of two ABA single-case experiments. It was predicted that in two children with hypotonic cerebral palsy, the addition of a favorite game to the occupational form, or treatment environment, would increase functional vertical neck and back extension. METHOD: Each subject was positioned in prone on a wedge. During the A phases, each was given verbal directions to hold her head up. During the B phase, a favorite game was introduced. It was predicted that each subject would extend her back and neck in a functional way in order to manipulate and observe the game. The dependent variables were the mean range of neck extension and the mean range of back extension, which were measured by a videotaped, two-dimensional kinematic analysis. RESULTS: The addition of a favorite game to the occupational form improved vertical neck and back extension while discouraging nonfunctional neck hyperextension and fixing postures in both children. CONCLUSION: Embedding exercise within a play occupation enhanced the prone extension of two children with hypotonic cerebral palsy.     

The validity of the Tritrac-R3D activity monitor for the assessment of physical activity: II. Temporal relationships among objective assessments

Today the organ donor operation in brain dead donors is mostly projected as multiple organ procurement (MOP). Not only the kidneys, but heart and liver or additionally lungs and pancreas are removed in MOP. A good synchronization between the collaborating transplantation groups (thoracic and abdominal surgeon, urologist) is essential to prevent loss of an organ because of technical problems. In Germany urologists perform more than 40% of kidney transplantations. These urologic institutions perform the cadaver kidney retrievals and participate on MOP. If possible, an urologist experienced in renal transplantation should contribute to the care for quality of the kidneys. Since 1987 the transplant center of Bremen obtained 390 donor registrations. 202 organ donor operations have been performed (106 MOP and 96 exclusive kidney retrievals). 398 donor kidneys have been collected and 382 could be transplanted.     

Application of cloud-point extraction-reversed-phase high-performance liquid chromatography. A preliminary study of the extraction and quantification of vitamins A and E in human serum and whole blood

Acute intestinal graft-versus-host disease (GVHD) develops in about 30-50% of allogeneic bone-marrow transplant recipients: 10-20% have gastrointestinal emergencies (hemorrhage or perforation). Mortality reaches 30-60% in patients with acute, grade 2-4 GVHD. We studied 36 bone marrow recipients in whom acute intestinal GVHD developed. Seven had gastrointestinal emergencies: 4 severe gastrointestinal bleeding and 3 acute peritonitis. Three patients with gastrointestinal bleeding and one patient with peritonitis responded to medical therapy. Three needed surgery: one with bleeding and two with peritonitis, while 1 patient had embolization. Of the 7, two patients died, one after embolization and one after surgery. Two of the three surgically-treated cases are still alive several years after operation. From this experience we feel that surgery for gastrointestinal bleeding in acute GVHD is indicated only when medical treatment fails. Severe neutropenia, thrombocytopenia (<10.000 x mm3) and blood cultures positive for CMV have an unfavorable prognostic value.     

Differentiation and reversal of malignant changes in colon cancer through PPARgamma

PPARgamma is a nuclear receptor that has a dominant regulatory role in differentiation of cells of the adipose lineage, and has recently been shown to be expressed in the colon. We show here that PPARgamma is expressed at high levels in both well- and poorly-differentiated adenocarcinomas, in normal colonic mucosa and in human colon cancer cell lines. Ligand activation of this receptor in colon cancer cells causes a considerable reduction in linear and clonogenic growth, increased expression of carcinoembryonic antigen and the reversal of many gene expression events specifically associated with colon cancer. Transplantable tumors derived from human colon cancer cells show a significant reduction of growth when mice are treated with troglitazone, a PPARgamma ligand. These results indicate that the growth and differentiation of colon cancer cells can be modulated through PPARgamma.