A Non-Covalent Antibody Complex for the Delivery of anti-cancer drugs

Antibody drug conjugates (ADCs), which are obtained by coupling a potent cytotoxic agent to a monoclonal antibody (mAb), are traditionally bound in a random way to lysine or cysteine residues, with the final product's heterogeneity having an important impact on their activity, characterization, and manufacturing. A new antibody drug delivery system (ADS) based on a non-covalent linkage between a Fc-binding protein, in this case Protein A or Protein G, and a mAb was investigated in the effort to achieve greater homogeneity and to create a versatile and adaptable drug delivery system. Recombinant staphylococcal Protein A and streptococcal Protein G were chemically PEGylated at the N-terminus with a 5 kDa and a 20 kDa PEG, respectively, yielding two monoconjugates with a mass of 48 50 and 48 45 kDa. Circular dichroism studies showed that both conjugates preserved secondary structures of the protein, and isothermal titration calorimetry experiments demonstrated that their affinity for mAb was approximately 107 M-1. Upon complexation with a mAb (Trastuzumab or Rituximab), in vitro flow-cytometry analysis of the new ADSs showed high selectivity for the specific antigen expressing cells. In addition, the ADS complex based on Trastuzumab and Protein G, conjugated with a heterobifunctional 20 kDa PEG carrying the toxin Tubulysin A, had a marked cytotoxic effect on the cancer cell line overexpressing the HER2/neu receptor, thus supporting its application in cancer therapy

Bioremediation Enhancement of Marine Sediments Contaminated by Crude Oil with Biogenic Pollutants Mobilizing Agents and Biosurfactants

One of the main limitations to the bioremediation of oil-contaminated marine sediments is the low hydrocarbons bioavailability. Aim of this PhD research project is to identify an environmental friendly approach to increase hydrocarbons bioavailability and biodegradation in oil-contaminated marine sediments. Several surfactants/pollutant mobilizing agents have been selected and applied to slurry microcosms: two microbial surfactants (sophorolipids and rhamnolipids), two types of cyclodextrins (hydroxypropyl- and randomly methylated- β-cyclodextrins; HPB-CD and RAMEB-CD), two commercial soy lecithin products (de-oiled and raw) and bile acids. Sophorolipids, cyclodextrins and to less extent, soy lecithins stimulated n-alkanes anaerobic degradation of actual oil-contaminated marine sediment from Gela (Sicily). Molecular analysis of 16S rRNA gene suggested that an Acidobacteria was probably responsible for the anaerobic biodegradation. Under aerobic condition, the same surfactants had inhibitor effects on n-alkanes degradation in Gela sediment, while HPB-CD and de-oiled soy lecithin were able to increase the degradation in crude oil-contaminated sediment from Ravenna. Increase of n-alkane bioavailability in oil-contaminated Gela sediment occurred in the presence of the two cyclodextrins and raw soy lecithin, both immediately after oil contamination as well as after the complete adsorption of hydrocarbons to sediment. Investigations of surfactant releasing formulations for the deployment of these agents to the sediments showed that HPB-CD could be efficiently encapsulated in agar hydrogels, while sophorolipids in polybutylene succinate (PBS) microspheres. The release rate of encapsulated HPB-CD was higher than encapsulated sophorolipids when formulations were incubated in marine water and similar in oil-contaminated sand slurries. Both encapsulated surfactants remarkably reduced adsorption of freshly spiked hydrocarbons to sand; conversely, only agar-encapsulated HPB-CD were able to desorb n-alkanes in weathered contaminated sand and increase their bioavailability and biodegradation similarly to free cyclodextrins. Therefore, encapsulation of HPB-CD in agar capsules might be the most promising solution for the enhancement of the bioremediation in marine sediment

Intrarenal arterial stiffness is increased in systemic sclerosis patients with anti-ribonucleic acid polymerase III antibodies

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Bosentan for digital ulcers prevention does not worsen cardiopulmonary exercise test parameters in SSc patients with interstitial lung disease

Bosentan for digital ulcers prevention does not worsen cardiopulmonary exercise test parameters in SSc patients with interstitial lung diseas

Angiogenic and angiostatic factors in renal scleroderma-associated vasculopathy

BACKGROUND: The angiogenesis in systemic sclerosis (SSc) is impaired. An imbalance of pro-angiogenic factors and angiogenesis inhibitors has been implicated in the progression of peripheral microvascular damage, defective vascular repair and fibrosis. Intrarenal resistance index are considered markers of renal vasculopathy. The aim of the study is to evaluate angiogenic and angiostatic factors (VEGF and endostatin) in SSc patients and to correlate with intrarenal hemodynamic parameters. METHODS: 91 SSc patients were enrolled in this study. Serum VEGF and endostatin levels were determined. All patients underwent a renal Doppler ultrasound RESULTS: A significant positive correlation was observed between endostatin and renal Doppler parameters (p<0.0001). A negative correlation was observed between serum levels of endostatin and eGFR (p<0.01). In SSc patients with high resistive index, serum levels of endostatin were significantly (p<0.01) higher than in SSc patients with normal resistive index. The serum levels of endostatin significantly increased with progression of nailfold videocapillaroscopy damage (p<0.01) and were significantly (p<0.05) higher in SSc patients with digital ulcers than in SSc patients without digital ulcers. CONCLUSION: This is the first study that assess in SSc patients intrarenal hemodynamic parameters and endostatin. In SSc patients, endostatin represents a marker of renal scleroderma-associated vasculopathy

A Large Number of T Lymphocytes Recognize Moloney-Murine Leukemia Virus-Induced Antigens, but a Few Mediate Long-Lasting Tumor Immunosurveillance

Abstract The CD8+ T cell response to Moloney-murine leukemia virus (M-MuLV)-induced Ags is almost entirely dominated by the exclusive expansion of lymphocytes that use preferential TCRVβ chain rearrangements. In mice lacking T cells expressing these TCRVβ, we demonstrate that alternative TCRVβ can substitute for the lack of the dominant TCRVβ in the H-2-restricted M-MuLV Ag recognition. We show that, at least for the H-2b-restricted response, the shift of TCR usage is not related to a variation of the immunodominant M-MuLV epitope recognition. After virus immunization, all the potentially M-MuLV-reactive lymphocytes are primed, but only the deletion of dominant Vβ rescues the alternative Vβ response. The mechanism of clonal T cell "immunodomination" that guides the preferential Vβ expansion is likely the result of a proliferative advantage of T cells expressing dominant Vβ, due to differences in TCR affinity and/or cosignal requirements. In this regard, a CD8 involvement is strictly required for the virus-specific cytotoxic activity of CTL expressing alternative, but not dominant, Vβ gene rearrangements. The ability of T cells expressing alternative TCRVβ rearrangements to mediate tumor protection was evaluated by a challenge with M-MuLV tumor cells. Although T cells expressing alternative Vβ chains were activated and expanded, they were not able to control tumor growth in a long-lasting manner due to their incapacity of conversion and accumulation in the T central memory pool

La tecnología y el rol del Contador Público en Argentina

El presente trabajo de investigación consiste en el análisis de como la evolución de la tecnología ha influido en los diferentes tareas que puede desarrollar un contador público y como varía también en función de la edad y facilidad que tenga el profesional para adaptarse a los constantes cambios que han surgido através del tiempo.La finalidad del mismo es analizar las ventajas y desventajas de la tecnología en dicha carrera y concientizar a los profesionales de la importancia de una actualización continua para ser útiles al momentode prestar servicios a los clientes.Para llevar a cabo dicho trabajo, se tomaron algunas entrevistas y cuestionarios a diferentes egresados variando el ámbito laboral y edad y se concluyó que la tecnología permite realizar de una manera más óptima y sujeta a menos riegos de error dicha labor, es decir que las evoluciones tecnológicas no deben ser tomadas como una amenaza de que pueda ser reemplazados por ella sino que al contrario, es el complemento esencial para explotar al máximo su potencial como profesional. También se observó que producto a esta evolución, han surgido nuevas oportunidades laborales ya que se necesita capital humano en otros puestos o realizando otro tipo de tareas; pero cabe aclarar que nada de esto tiene fruto si el contador no se esfuerza por encontrarse continuamente actualizado.Fil: Duboue, Camila. Universidad Nacional de Cuyo. Facultad de Ciencias Económicas.Fil: Duboue, Jorge. Universidad Nacional de Cuyo. Facultad de Ciencias Económicas.Fil: Rosato, Antonella. Universidad Nacional de Cuyo. Facultad de Ciencias Económicas

Serum uric acid as a marker of microvascular damage in systemic sclerosis patients

Background: Microvascular damage of skin and internal organs is a hallmark of systemic sclerosis (SSc). Serum uric acid (UA) represents a marker of inflammation and endothelial dysfunction. The aims of this study were to evaluate the correlation between serum UA and intrarenal arterial stiffness evaluated by Doppler ultrasound in SSc patients with normal renal function. We also evaluated the correlation between serum UA and other clinical variables of the disease. Methods: Forty-five SSc patients underwent clinical assessment, Doppler ultrasound of intrarenal arteries with evaluation of resistive index (RI), pulsatile index (PI), and systolic/diastolic ratio (S/D), echocardiography with systolic pulmonary artery pressure (PAPs), baseline pulmonary function tests, and nailfold videocapillaroscopy (NVC). In all patients serum UA was measured. Results: The serum UA showed a significant positive correlation with sCr (r = 0.33, p &lt; 0.0001) and PAPs (r = 0.38, p &lt; 0.01) &gt; and negative correlation with CKD-EPI (r = -0.35, p &lt; 0.01). The mean value of serum UA increased with severity of NVC damage. Using this cut-off value of 4.7 mg/dl, the mean value of Doppler indices of intrarenal stiffness is significantly different (p &lt; 0.05) in SSc patients with low normal or high normal serum UA. Conclusions: Serum UA concentration is higher in patients with high microvascular damage than in patients with low microvascular damage. These preliminary data must be confirmed in large prospective studies

Innate immune modulation induced by EBV lytic infection promotes endothelial cell inflammation and vascular injury in scleroderma

Microvascular injury is considered an initial event in the pathogenesis of scleroderma and endothelial cells are suspected of being the target of the autoimmune process seen in the disease. EBV has long been proposed as a trigger for autoimmune diseases, including scleroderma. Nevertheless, its contribution to the pathogenic process remains poorly understood. In this study, we report that EBV lytic antigens are detected in scleroderma dermal vessels, suggesting that endothelial cells might represent a target for EBV infection in scleroderma skin. We show that EBV DNA load is remarkably increased in peripheral blood, plasma and circulating monocytes from scleroderma patients compared to healthy EBV carriers, and that monocytes represent the prominent subsets of EBV-infected cells in scleroderma. Given that monocytes have the capacity to adhere to the endothelium, we then investigated whether monocyte-associated EBV could infect primary human endothelial cells. We demonstrated that endothelial cells are infectable by EBV, using human monocytes bound to recombinant EBV as a shuttle, even though cell-free virus failed to infect them. We show that EBV induces activation of TLR9 innate immune response and markers of vascular injury in infected endothelial cells and that up-regulation is associated with the expression of EBV lytic genes in infected cells. EBV innate immune modulation suggests a novel mechanism mediating inflammation, by which EBV triggers endothelial cell and vascular injury in scleroderma. In addition, our data point to up-regulation of EBV DNA loads as potential biomarker in developing vasculopathy in scleroderma. These findings provide the framework for the development of novel therapeutic interventions to shift the scleroderma treatment paradigm towards antiviral therapies

Left ventricular mass and intrarenal arterial stiffness as early diagnostic markers in cardiorenal syndrome type 5 due to systemic sclerosis

Background: Cardiorenal syndrome type 5 (CRS-5) includes a group of conditions characterized by a simultaneous involvement of the heart and kidney in the course of a systemic disease. Systemic sclerosis (SSc) is frequently involved in the etiology of acute and chronic CRS-5 among connective tissue diseases. In SSc patients, left ventricular mass (LVM) can be used as a marker of nutritional status and fibrosis, while altered intrarenal hemodynamic parameters are suggestive of early kidney involvement. Methods: Forty-two consecutive patients with a diagnosis of SSc without cardiac and/or renal impairment were enrolled to assess whether cardiac muscle mass can be related to arterial stiffness. Thirty subjects matched for age and sex were also enrolled as healthy controls (HC). All patients performed echocardiography and renal ultrasound. Results: Doppler indices of intrarenal stiffness and echocardiographic indices of LVM were significantly increased in SSc patients compared to HC. A positive correlation exists between LVM/body surface area and pulsatile index (p &lt; 0.05, r = 0.36), resistive index (p &lt; 0.05, r = 0.33) and systolic/diastolic ratio (p &lt; 0.05, r = 0.38). Doppler indices of intrarenal stiffness and LVM indices were significantly higher in SSc patients with digital ulcers than in SSc patients without a digital ulcer history. Conclusions: SSc is characterized by the presence of microvascular and multiorgan injury. An early cardiac and renal impairment is very common. LVM and intrarenal arterial stiffness can be considered as early markers of CRS onset. The clinical use of these markers permits a prompt identification of organ damage. An early diagnosis allows the appropriate setting of pharmacological management, by slowing disease progression