Therapeutic vaccination for closed head injury

Closed head injury often has a devastating outcome, partly because the insult, like other injuries to the central nervous system (CNS), triggers self-destructive processes. During studies of the response to other CNS insults, it was unexpectedly discovered that the immune system, if well controlled, provides protection against self-destructive activities. Here we show that in mice with closed head injury, the immune system plays a key role in the spontaneous recovery. Strain-related differences were observed in the ability to harness a T cell-dependent protective mechanism against the effects of the injury. We further show that the trauma-induced deficit could be reduced, both functionally and anatomically, by post-traumatic vaccination with Cop-1, a synthetic copolymer used to treat patients with multiple sclerosis and found (using a different treatment protocol) to effectively counteract the loss of neurons caused by axonal injury or glutamate-induced toxicity. We suggest that a compound such as Cop-1 can be safely developed as a therapeutic vaccine to boost the body's immune repair mechanisms, thereby providing multifactorial protection against the consequences of brain trauma.     

Parametric description of cardiac vagal control

Heart rate variability (HRV) indices that reflect the magnitude of respiratory sinus arrhythmia (RSA) are commonly applied as non-invasive measures of cardiac vagal control. Recently, however, serious doubts have been raised about the accuracy and validity of such assessments. To evaluate these methods, we derived a theoretical model for the dependence of mean heart rate and RSA on gradual vagal blockade by atropine, and compared its predictions to actual experimental results. The experiment involved the injection of nine consecutive intravenous bolus doses of atropine to eight young healthy male subjects. Seven-minute recordings of ECG and respiration were made for each atropine dose. The heart rate (HR) signal was derived from the ECG recording, and mean heart rate and the power of the high frequency peak of HRV (which measures the magnitude of RSA) were computed. The experimental data were fitted to the model's equations, and optimal values were obtained for the model's parameters. A tight agreement is observed between the theoretical fitted curves and the experimental data. The parameters that were computed from fitting the experimental data to the mean heart rate equation display a surprisingly small variance among the different subjects. The parameters that were computed from fitting the experimental data to the RSA equation, and the resulting shape of these fitted curves, explain many of the conflicting results previously published, and provide a new quantitative insight to cardiac vagal activity.     

Current status of sentinel lymph node mapping and biopsy: facts and controversies

Lymphatic mapping and sentinel lymph node biopsy were first reported in 1977 by Cabanas for penile cancer. Since that time, the technique has become rapidly assimilated into clinical practice. Morton first described the application of lymphatic mapping for melanoma only a decade ago, and this technique is now accepted as the standard of care. The application for lymphatic mapping and sentinel lymph node biopsy in breast cancer remains approximately 5 years behind its utilization in melanoma. This technique has the potential to be utilized in all solid tumors. The rapid assent of this technique in clinical practice is the result of multiple factors, including accuracy, decreased morbidity, and supplying the pathologist with only a few nodes to allow a more focused and sensitive pathologic evaluation. Despite the success and acceptance of lymphatic mapping, many controversies remain. We have attempted to clearly highlight these controversies in this review.     

Profile of cytokine production within the periparasitic granuloma in human alveolar echinococcosis

Th2 responses, especially IL-10 secretion by circulating mononuclear cells are associated with the progressive form of AE and Th1 responses with resistance. The HLA B8, DR3, DQ2 haplotype is associated with the severity of AE in humans through immune-mediated mechanisms including an elevated production of Interleukin-10 (IL-10). Granulomatous infiltration of mononuclear cells around the parasitic vesicles is a hallmark of this disease; however, cytokine production by granuloma cells has never been studied. Tissue samples were taken in the periparasitic area and in the central area of the periparasitic granulomatous lesions from a patient with a progressive AE at surgery. Six pieces for each zone were incubated in culture medium with antibiotics and IL-2, together with irradiated autologous peripheral blood mononuclear cells as feeder cells. After four days the dead feeder cells were removed by density gradient centrifugation. Lymphocytes were stimulated with Echinococcus multilocularis vesicular fluid antigen (Emf) or PHA to study IL-10, IFN-, and IL-4 production in the supernatant. Emf-stimulated mononuclear cells from the central part of the lesions secreted more IL-10 and less IFN-gamma than cells from the periphery of the granuloma. At the basal level, IL-10 secretion by the locally infiltrating cells was also high and this confirms at the local granuloma level our previous results obtained from cultures of circulating mononuclear cells. The present study confirms that IL-10 secretion is a key feature of the immune response against E. multilocularis in humans. The location of the cells which produce the highest amount of IL-10, those in contact with parasitic structures, suggests that the parasite itself is able to modulate the immune response of the host so that the infiltrating cells cannot participate in the effector phase of the cellular immune response. The nature of the parasitic structures involved and the mechanisms which lead to an imbalanced cytokine production remain to be elucidated.     

Os odontoideum.

Between January 1981 and December 1985 at the Rizzoli Orthopaedic Institute 8 patients affected with atlo-axial instability secondary to os odontoideum were submitted to surgery: all of the patients were characterized by persistent cervical pain and neurological deficit, which was still at an initial stage in 5 cases, and more severe in the remaining 3 (severe spastic tetraparesis). All of the patients were submitted to a similar treatment protocol in three stages: 1) preoperative reduction in halo-plaster, 2) surgical stabilization of C1-C2 by posterior vertebral fusion (limited to C1-C2 in 6 cases and extended as far as the occiput in the remaining 2), 3) postoperative immobilization in halo-plaster for an average of 12 weeks. At follow-up obtained after an average of 4 years and 2 months there was consolidation of the fusion in all of the cases treated: in 1 case, however, revision of the fusion was required 3 and 1/2 months after surgery, whereupon fusion was finally obtained. The total regression of pain was obtained in all 8 of the cases treated and there was evident neurological recovery in the 3 patients who had been characterized by severe spastic tetraparesis prior to surgery. The treatment protocol used in this series of patients proved to be a reliable one. Thus, it was possible to avoid the onset of chronic atlo-axial instability and its severe neurological sequelae in those cases where neurological deficit was still at an initial stage; furthermore, effective recovery was obtained in patients in whom neurological deficit was severe.     

Study of lymphocyte subpopulations in bone marrow in a model of protein–energy malnutrition

ObjectiveProtein–energy malnutrition (PEM) is an important public health problem affecting millions of people worldwide. Hematopoietic tissue requires a high nutrient supply, and a reduction in leukocytes, especially lymphocytes, suggests that some nutritional deficiencies might be altering bone marrow function and decreasing its ability to produce lymphocytes. In this study, we evaluated the effect that PEM has on lymphocyte subtypes and the cell cycle of CD5⁺ cells.MethodsSwiss mice were subjected to PEM using a low-protein diet containing 4% protein. When the experimental group had lost about 20% of their original body weight, we collected blood and bone marrow cells and evaluated the hemogram, the myelogram, bone marrow lymphoid markers using flow cytometry, and the cell cycle in CD5⁺ bone marrow.ResultsMalnourished animals presented anemia, reticulocytopenia, and leukopenia with lymphopenia. The bone marrow was hypocellular, and flow cytometric analyses of bone marrow cells showed cells that were CD45⁺ (91.2%), CD2⁺ (84.9%), CD5⁺ (37.3%), CD3⁺ (23.5%), CD19⁺ (43.3%), CD22⁺ (34.7%), CD19⁺/CD2⁺ (51.2%), CD19⁺/CD3⁺ (24.0%), CD19⁺/CD5⁺ (13.2%), CD22⁺/CD2⁺ (40.1%), CD22⁺/CD3⁺ (30.3%), and CD22⁺/CD5⁺ (1.1%) in malnourished animals and CD45⁺ (97.5%), CD2⁺ (42.9%), CD5⁺ (91.5%), CD3⁺ (92.0%), CD19⁺ (52.0%), CD22⁺ (75.6%), CD19⁺/CD2⁺ (62.0%), CD19⁺/CD3⁺ (55.4%), CD19⁺/CD5⁺ (6.7%), CD22⁺/CD2⁺ (70.3%), CD22⁺/CD3⁺ (55.9%), and CD22⁺/CD5⁺ (8.4%) in control animals. Malnourished animals also presented more CD5⁺ cells in the G0 phase of cell cycle development.ConclusionMalnourished animals presented bone marrow hypoplasia, maturation interruption, prominent lymphopenia with depletion in the lymphoid lineage, and changes in cellular development. We suggest that these changes are some of the primary causes of lymphopenia in cases of PEM and partly explain the increase in susceptibility to infections found in malnourished individuals.     

Preschool Worth 4-Shape test: testability, reliability, and validity.

PURPOSE: The Worth 4-Dot is used to assess binocular fusion, but it is difficult to use with young children. We modified the Worth 4-Dot by replacing the circles with shapes while maintaining the same color configuration. The purpose of this study was to determine the testability, reliability, and validity of the Worth 4-Shape test. METHODS: Subjects aged 2 to 8 years and 4 patients aged over 8 years with best-corrected visual acuity of 20/40 or greater (n = 131 patients, n = 123 normals) attempted test and retest at 35 cm and 3m using the Worth 4-Shape and Worth 4-Dot. To provide a gold standard, medical history, bifoveal fixation, and stereoacuity were reviewed. RESULTS: Testability of the Worth 4-Shape was significantly higher than the Worth 4-Dot both in children aged less than 4 years (95.9% vs 79.5% at 35 cm, P <.001; 79.5% vs 55.1% at 3 m, P <.001) and older than 4 years (98.5% vs 88.6% at 35 cm, P <.001; 90.9% vs 82.4% at 3m, P =.04). Test-retest analysis found comparable concordance for the Worth 4-Shape and Worth 4-Dot tests (P >.3). The sensitivity and specificity of the Worth 4-Shape (92%, 97%) and Worth 4-Dot (90%, 94%) were comparable. Between-test analysis found 96% agreement between both tests at 35 cm and 97% agreement at 3m. CONCLUSIONS: The success rate for the Worth 4-Shape is higher than the Worth 4-Dot, especially in children aged less than 4 years, and has equivalent accuracy. The Worth 4-Shape test-retest reliability is high supporting its validity for use with young children.     

KIT is highly expressed in adenoid cystic carcinoma of the breast, a basal-like carcinoma associated with a favorable outcome.

Recent biological studies have classified breast carcinomas into HER2-overexpressing, estrogen receptor-positive/luminal, basal- and normal-like groups. According to this new biological classification, the objectives of our study were to assess the clinical, morphologic and immunophenotypic characteristics of adenoid cystic carcinoma of the breast in order to classify this subtype of breast carcinoma. A total of 18 cases of adenoid cystic carcinoma were identified from the Institut Curie files. Clinical information was available for 16 patients with a median follow-up of 6.5 years. Morphologically, all tumors were graded according to the system defined by Kleer and Oberman (histologic and nuclear grade). Immunophenotype was assessed with anti-ER, PR, HER-2, KIT, basal (CK5/6) and luminal cytokeratins (CK8/18) and p63 antibodies. One out of 18 tumors was nuclear grade 1 (16%), nine were nuclear grade 2 (50%) and eight were nuclear grade 3 (44%). All cases were estrogen receptor, progesterone receptor and HER-2 negative. Epithelial cells were strongly positive around glandular lumina with one or both cytokeratins, identifying the coexistence of CK5/6+ cells, CK5/6 and CK8/18+ cells, CK8/18+ cells and p63+ cells. All cases (100%) were also KIT positive. In all, 15 patients were treated by surgery. Nine of them received adjuvant radiotherapy. Follow-up was available for 16 patients. In all, 14 patients were alive. Two of them, initially treated by surgery only, presented a local recurrence. Two patients died (one of them treated by radiation therapy only died from her disease). Our study shows that adenoid cystic carcinoma of the breast is a special, estrogen receptor, progesterone receptor, HER-2 negative and highly KIT-positive, basal-like breast carcinoma, associated with an excellent prognosis. This highly specific immunophenotype could be useful to differentiate adenoid cystic carcinoma of the breast from other subtypes of breast carcinoma such as cribriform carcinoma.     

3-[4'-bromo-(1,1'-biphenyl)-4-yl]-N,N-dimethyl-3-(2-thienyl)-2-propen-1-amine: synthesis,cytotoxicity, and leishmanicidal,trypanocidal and antimycobacterial activities

Current therapies for Chagas' disease, leishmaniasis and tuberculosis are unsatisfactory because of the failure rates, significant toxicity and/or drug resistance. In this study, the compound 3-[4'-bromo-(1,1'-biphenyl)-4-yl]-N,N-dimethyl-3-(2-thienyl)-2-propen-1-amine (IV) was synthesized and its trypanocidal, leishmanicidal and antimycobacterial activities were investigated. The cytotoxicity was determined on V79 cells with three endpoints: nucleic acid content, 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide reduction and Neutral Red uptake. This compound was active against different species of mycobacteria and different life cycle stages of Trypanosoma cruzi. In experiments with trypomastigotes performed at 4 degrees C in the presence of blood, the activity was 8.8-fold more active than the standard drug, Crystal Violet. Higher activity was achieved against Leishmania amazonensis, with an ED(50)/24 h of 3.0 +/- 0.3 micro mol/L. The effect against trypanosomatids, which suggests high activity of compound IV against promastigotes of L. amazonensis and amastigotes of T. cruzi, stimulated further studies in vitro with amastigotes interiorized in macrophages and with in vivo models. Our results indicate that mammalian V79 cells are less susceptible to the action of compound IV than promastigotes of L. amazonensis (8.0-13.3-fold) and axenic amastigotes of T. cruzi (3.5-5.9-fold).