Adjuvant potential of vitamin E in the induction of experimental allergic encephalomyelitis: histological aspects

In this study we report the effect of Vit. E, as an immunostimulating factor, on the induction of Experimental Allergic Encephalomyelitis in Lewis rat. The animals were inoculated intracutaneously in the plantar areas with emulsion of isologous spinal cord suspensed in Incomplete Freund's Adjuvant (IFI) and Vit. E. Histologic examination revealed the basic lesion of a perivenous cuff of mononuclear cells and small areas of demyelinated axons. The clinical signes are graded in order to the time of induction. It is possible an action of "adjuvanticity" of Vit. E on the various cells involved in the immune response by cell transformation and moltiplication.     

COMPARISON OF PHYSIOLOGICAL REACTIONS AND PHYSIOLOGICAL STRAIN IN HEALTHY MEN UNDER HEAT STRESS IN DRY AND STEAM HEAT SAUNAS

The aim of the paper was to follow up major physiological reactions, provoked by heat stress during dry and wet sauna baths. A physical strain index and subjective estimation of heat comfort of subjects who had not taken sauna baths before was also evaluated. Ten healthy males aged 25-28 underwent a dry sauna bath and then after a one-month break they underwent a steam sauna bath. Each time, they entered the sauna chamber 3 times for 15 minutes with five-minute breaks. During breaks they cooled their bodies with a cold shower and then rested in a sitting position. Before and after the baths, body mass and blood pressure were measured. Rectal temperature and heart rate were monitored during the baths. The physiological strain index (PSI) and cumulative heat strain index (CHSI) were calculated. Subjects assessed heat comfort by Bedford''s scale. Greater body mass losses were observed after the dry sauna bath compared to the wet sauna (-0.72 vs. -0.36 kg respectively). However, larger increases in rectal temperature and heart rate were observed during the wet sauna bath (38.8% and 21.2% respectively). Both types of sauna baths caused elevation of systolic blood pressure, but changes were greater after the dry one. Diastolic pressure was reduced similarly. Subjective feelings of heat comfort as well as PSI (4.83 ± 0.29 vs. 5.7 ± 0.28) and CHSI (76.3 ± 18.4 vs. 144.6 ± 21.7) were greater during the wet sauna bath. It can be concluded that due to high humidity and reduction of thermoregulation mechanisms, the wet sauna is more stressful for the organism than the dry sauna, where the temperature is higher with low humidity. Both observed indexes (PSI and CHSI) could be appropriate for objective assessment of heat strain during passive heating of the organism.     

Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol

Background

Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells.

Results

The described protocol is able to provide osteoblastic cells producing nodules of calcium mineralization within 18 days from withdrawal of amniotic fluid samples. These cells display a complete expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2) within 30 days from withdrawal. In order to test the ability of these cells to proliferate on surfaces commonly used in oral osteointegrated implantology, we carried out cultures onto different test disks, namely smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium). Electron microscopy analysis evidenced the best cell growth on this latter surface.

Conclusion

The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.     

Complex formation between protactinium(V) and sulfate ions at 10 and 60 °C

Protactinium complexation with sulfate ions was studied with the element at tracer scale (C_Pa ∼ 10⁻¹² M) by solvent extraction method. The involved aqueous system was Pa(V)/H₂O/HClO₄/Na₂SO₄/NaClO₄ at 10 and 60 °C. The extraction experiments were conducted using the chelating agent thenoyltrifluoroacetone (TTA) in toluene. For both values of temperature, a systematic study was performed in order to determine the formation constants (β₁, β₂ and β₃) of sulfate complexes of Pa(V) at different ionic strength. For each temperature, the extrapolation of these constants to zero ionic strength was performed using the Specific Interaction Theory, leading to values of 2.8 ± 0.5, 6.5 ± 0.5, 7.8 ± 0.5 at 10 °C and 4.3 ± 0.3, 8.4 ± 1.3, 9.6 ± 0.4 at 60 °C. Interaction coefficients involving the sulfate complexes of protactinium(V) were also derived.     

A Novel,Live-Attenuated Vesicular Stomatitis Virus Vector Displaying Conformationally Intact,Functional HIV-1 Envelope Trimers That Elicits Potent Cellular and Humoral Responses in Mice

Though vaccination with live-attenuated SIV provides the greatest protection from progressive disease caused by SIV challenge in rhesus macaques, attenuated HIV presents safety concerns as a vaccine; therefore, live viral vectors carrying HIV immunogens must be considered. We have designed a replication-competent vesicular stomatitis virus (VSV) displaying immunogenic HIV-1 Env trimers and attenuating quantities of the native surface glycoprotein (G). The clade B Env immunogen is an Env-VSV G hybrid (EnvG) in which the transmembrane and cytoplasmic tail regions are derived from G. Relocation of the G gene to the 5′terminus of the genome and insertion of EnvG into the natural G position induced a ∼1 log reduction in surface G, significant growth attenuation compared to wild-type, and incorporation of abundant EnvG. Western blot analysis indicated that ∼75% of incorporated EnvG was a mature proteolytically processed form. Flow cytometry showed that surface EnvG bound various conformationally- and trimer-specific antibodies (Abs), and in-vitro growth assays on CD4+CCR5+ cells demonstrated EnvG functionality. Neither intranasal (IN) or intramuscular (IM) administration in mice induced any observable pathology and all regimens tested generated potent Env-specific ELISA titers of 10⁴–10⁵, with an IM VSV prime/IN VSV boost regimen eliciting the highest binding and neutralizing Ab titers. Significant quantities of Env-specific CD4+ T cells were also detected, which were augmented as much as 70-fold by priming with IM electroporated plasmids encoding EnvG and IL-12. These data suggest that our novel vector can achieve balanced safety and immunogenicity and should be considered as an HIV vaccine platform.     

Occurrence of adrenergic nerve fibers in human thymus during immune response

The adrenergic nerve fibers (ANF), the neuropeptide Y-like immunoreactive nerve fibers (NPY-NF) and the noradrenaline (NA) amount were studied in the human thymus in subjects previously treated or not treated with interferon therapy with the aim to identify the changes due to the interferon therapy. This therapy has been used in patients affected by multiple sclerosis (MS). Biochemical and morphological methods were used associated with quantitative analysis of images. The whole thymuses were removed during autopsies in young and adult patients not treated with interferon. Moreover, samples of thymus were removed from patients, either young or adult who had previously been treated with interferon therapy, and subjected, for diagnostic reasons, to thymic biopsy. All samples of thymus were weighed, measured and dissected. Thymic slices were stained with Eosin-orange for detection of the microanatomical details, or with Bodian's reaction for recognition of nervous structures. Histofluorescence microscopy was used for detection of ANF, and immunofluorescence microscopy for recognition of NPY-like immunoreactive structures. All morphological results were subjected to quantitative analysis of images. Noradrenaline contained in thymic structures was measured by biochemical methods. Our results only concerned the effects of the therapy and suggested that treatment with interferon therapy induces many changes in the thymic structures: (1) The protein content of thymus is significantly increased; (2) the NA content in the thymus is also significantly increased; (3) NPY-like immunoreactive structures in the thymus are significantly increased; (4) occurrence of NPY-like immunoreactivity is particularly and significantly increased both in thymic microenvironment and in structures resembling nerve fibers; (5) ANF are significantly increased in the same thymic structures in which NPY-like immunoreactivity is also increased (i.e. thymic microenvironment and structures resembling nerve fibers). The morphological and biochemical changes observed can also explain the immunological changes induced in the thymus after immunostimulating therapy.     

Validation of dHPLC for molecular diagnosis of beta-thalassemia in Southern Italy

Beta-thalassemia, the most common hereditary anemia in the Mediterranean area, results from over 200 causative mutations in the beta-globin locus. The aim of this study was to validate a denaturing high-performance liquid chromatography (dHPLC)-based assay for postnatal and prenatal molecular diagnosis of beta-thalassemia in Southern Italy. Sixty beta-thalassemic patients, affected either by thalassemia intermedia or thalassemia major, were analyzed in a blind study. We also carried out prenatal molecular diagnosis in 12 couples at-risk for having affected offspring. Chorionic villi samples were subjected to dHPLC analysis upon molecular characterization of the parental beta-globin alleles. Direct sequence analysis was used to validate each result, showing an accuracy rate of 100% for dHPLC. Overall, our protocol was able to identify the responsible mutations in all 96 analyzed subjects (including 12 prenatals in at-risk pregnancies), detecting the eight most common mutations in Southern Italy. Three rare mutations (one of which, reported here for the first time) that standard mutation detection methods failed to reveal, were also identified. dHPLC assay proved to be a reliable, rapid, and sensitive method for detecting both common and rare mutations within the beta-globin gene. Because of this property our protocol has the potential to be implemented for mutational screening in different areas of high prevalence for beta-thalassemia.