Femoral arteries energy dissipation and filtering function remain unchanged after cryopreservation procedure

The aim was to evaluate our cryopreservation method effects on the mechanical properties and filtering function of human superficial femoral arteries (SFA). SFA segments from 10 multiorgan donors were divided into two groups: fresh, tested 24-48 h after harvesting, and cryopreserved/defrosted, tested after 1 month of cryopreservation. The cooling process was carried out in three steps: 2 degrees C/min until -40 degrees C; 5 degrees C/min until -90 degrees C and finally a rapid cooling by transferring the bag to vapour phase of liquid nitrogen (-142 degrees C). Thawing was made in two steps, a slow warming time by exposing the bag to 20 degrees C during 20 min, followed by a rapid warming by immersion in a 40 degrees C warm bath until defrost. In a circulation mock, arterial pressure [Pressure signal (P)] and diameter [Diameter (D)] were registered at similar stretch-frequency, P and flow levels. A compliance transfer function (D/P) was used for the on-line assessment of the arterial wall elastic (E), viscous (eta), and inertial (M) properties. To evaluate the arterial wall filter function, the arterial wall D/P frequency response was characterized, the cut-off frequency (fc) was quantified, and the viscous energy dissipation (Weta) was calculated. After cryopreservation, there were not significant changes in E, eta, M, Weta, and fc.     

Glycan bioengineering in immunogen design for tumor T antigen immunotargeting

Bioengineering of Galβ3GalNAcα, known as Thomsen–Friedenreich disaccharide (TFD), is studied to promote glycan immunogenicity and immunotargeting to tumor T antigen (Galβ3GalNAcα-O-Ser/Thr). Theoretical studies on disaccharide conformations by energy minimization of structures using MM2 energy function showed that pentalysine (Lys5) linker and benzyl (Bzl) residue enhance TFD rigidity of the glycosidic bond. Antibodies raised against BzlαTFD-Lys5 immunogen recognize tumor T antigen. Competitive assays confirm that TFD-related structures are the main glycan epitope. Antibodies produced by glycan bioengineering recognize HT29, T47D, MCF7, and CT26 epithelial tumor cells. Epithelial tumor cell adhesion to T antigen-binding lectins and endothelial cells was lower in the presence of antibodies raised against the engineered immunogen. The immune response directed to the bioengineered glycoconjugate inhibited CT26 tumor cell proliferation and reduced tumor growth in an in vivo mouse model. These results show that TFD bioengineering is a useful immunogenic strategy with potential application in cancer therapy. The same approach can be extended to other glycan immunogens for immunotargeting purposes.     

Non-Invasive Biomechanical Evaluation of Implanted Human Cryopreserved Arterial Homografts

Native vessels–grafts biomechanical mismatch (BM) is related to graft failure. The BM could be reduced using human cryopreserved/defrosted arteries (cryografts), but post-thaw cryografts’ recovery could be associated with an impaired biomechanical behavior. In vitro, we demonstrated that our cryopreservation methods do not affect arteries’ biomechanics, but only post-implant studies would allow determining the cryografts’ biomechanical performance. Aim To characterize the biomechanical properties of implanted cryografts, and to compare them with cryografts pre-implant, recipients’ native arteries, and arteries from subjects with characteristics similar to those of the recipients and multiorgan donors (MOD) whose arteries were cryopreserved. Methods Native femoral arteries anastomosed to cryografts, implanted cryografts, and arteries from subjects, recipient-like and MOD-like, were studied. In vitro (pre-implant cryografts) and in vivo non-invasive studies were performed. Arterial pressure, diameter, and wall thickness were obtained to quantify local and regional biomechanical parameters, and to evaluate the arterial remodeling. Conclusion Implanted cryografts were remodeled, with an increased wall thickness, wall-to-lumen ratio, and wall cross-sectional area. The proximal–distal gradual transition in stiffness remained unchanged. Implanted cryografts were stiffer than MOD-like arteries, but more compliant than recipients’ arteries. The cryografts–native arteries biomechanical differences were lesser than those described for venous grafts or expanded polytetrafluoroethylene.     

Synthesis and pharmacological evaluation of fluorescent and photoactivatable analogues of antiplasmodial naphthylisoquinolines

The naphthylisoquinoline (NIQ) alkaloids from tropical Ancistrocladaceae and Dioncophyllaceae plants show high antiplasmodial activities in vitro and in vivo, even against chloroquine-resistant strains of the malaria pathogen. For the directed optimization of these activities, an investigation of the mode of action seems most rewarding. We have therefore embarked on the identification of the respective target protein in Plasmodium falciparum. For this purpose, we have developed a flexible pathway for the synthesis of a chemically divergent series of photoactive and fluorescent derivatives of such alkaloids and succeeded in preparing the first functionalized NIQ derivatives, 10, 12, and 35, suited for fluorescence and photoaffinity labeling experiments. Pharmacological investigations ensured that the modified alkaloid derivatives retained their antiplasmodial activity. The work may pave the way for a further improvement of the activity of these natural products and will thus increase their pharmacological potential as a valuable lead structure against the widespread tropical disease malaria.     

Gastrointestinal Kaposi's sarcoma associated to AIDS. Case report

In this paper, we describe a particular clinical case of gastrointestinal Kaposi's sarcoma associated with acquired immunodeficiency syndrome in a 51-year-old female patient who survived 6 years without anti-retroviral treatment after the diagnosis of HIV infection. The patient was admitted to our hospital with fever, skin lesions in both upper limbs and thighs (one day of evolution) and dry cough (about a month of evolution). She was admitted to the General Internal Medicine Service for control, diagnosis and treatment. An upper digestive bleeding was detected there. Once referred to our Gastroenterology Service, an appropriate selection of tests (anamnesis, gastric video endoscopy, histology) was carried out. In the upper gastric video endoscopy, several epithelial lesions were observed and a presumptive diagnosis of Kaposi's sarcoma was obtained. The diagnosis was finally confirmed by chain polymerase reaction (PCR) amplifcation of HHV8 DNA. This finding highlights the contribution of the molecular biology laboratory that allowed in this case the first molecular identification in our institution of the causative agent of Kaposi's syndrome with gastrointestinal manifestations.     

Linear and nonlinear viscoelastic modeling of aorta and carotid pressure-area dynamics under in vivo and ex vivo conditions

A better understanding of the biomechanical properties of the arterial wall provides important insight into arterial vascular biology under normal (healthy) and pathological conditions. This insight has potential to improve tracking of disease progression and to aid in vascular graft design and implementation. In this study, we use linear and nonlinear viscoelastic models to predict biomechanical properties of the thoracic descending aorta and the carotid artery under ex vivo and in vivo conditions in ovine and human arteries. Models analyzed include a four-parameter (linear) Kelvin viscoelastic model and two five-parameter nonlinear viscoelastic models (an arctangent and a sigmoid model) that relate changes in arterial blood pressure to the vessel cross-sectional area (via estimation of vessel strain). These models were developed using the framework of Quasilinear Viscoelasticity (QLV) theory and were validated using measurements from the thoracic descending aorta and the carotid artery obtained from human and ovine arteries. In vivo measurements were obtained from 10 ovine aortas and 10 human carotid arteries. Ex vivo measurements (from both locations) were made in 11 male Merino sheep. Biomechanical properties were obtained through constrained estimation of model parameters. To further investigate the parameter estimates, we computed standard errors and confidence intervals and we used analysis of variance to compare results within and between groups. Overall, our results indicate that optimal model selection depends on the artery type. Results showed that for the thoracic descending aorta (under both experimental conditions), the best predictions were obtained with the nonlinear sigmoid model, while under healthy physiological pressure loading the carotid arteries nonlinear stiffening with increasing pressure is negligible, and consequently, the linear (Kelvin) viscoelastic model better describes the pressure–area dynamics in this vessel. Results comparing biomechanical properties show that the Kelvin and sigmoid models were able to predict the zero-pressure vessel radius; that under ex vivo conditions vessels are more rigid, and comparatively, that the carotid artery is stiffer than the thoracic descending aorta; and that the viscoelastic gain and relaxation parameters do not differ significantly between vessels or experimental conditions. In conclusion, our study demonstrates that the proposed models can predict pressure–area dynamics and that model parameters can be extracted for further interpretation of biomechanical properties.     

Mucosal priming of newborn mice with S. Typhi Ty21a expressing anthrax protective antigen (PA) followed by parenteral PA-boost induces B and T cell-mediated immunity that protects against infection bypassing maternal antibodies

The currently licensed anthrax vaccine has several limitations and its efficacy has been proven only in adults. Effective immunization of newborns and infants requires adequate stimulation of their immune system, which is competent but not fully activated. We explored the use of the licensed live attenuated S. Typhi vaccine strain Ty21a expressing Bacillus anthracis protective antigen [Ty21a(PA)] followed PA-alum as a strategy for immunizing the pediatric population. Newborn mice primed with a single dose of Ty21a(PA) exhibited high frequencies of mucosal IgA-secreting B cells and IFN-γ-secreting T cells during the neonatal period, none of which was detected in newborns immunized with a single dose of PA-alum. Priming with Ty21a(PA) followed by PA-boost resulted in high levels of PA-specific IgG, toxin neutralizing and opsonophagocytic antibodies and increased frequency of bone marrow IgG plasma cells and memory B cells compared with repeated immunization with PA-alum alone. Robust B and T cell responses developed even in the presence of maternal antibodies. The prime-boost protected against systemic and respiratory infection. Mucosal priming with a safe and effective S. Typhi-based anthrax vaccine followed by PA-boost could serve as a practical and effective prophylactic approach to prevent anthrax early in life.