Immunologic Effects of Stereotactic Body Radiotherapy in Dogs with Spontaneous Tumors and the Impact of Intratumoral OX40/TLR Agonist Immunotherapy

Stereotactic body radiotherapy (SBRT) is known to induce important immunologic changes within the tumor microenvironment (TME). However, little is known regarding the early immune responses within the TME in the first few weeks following SBRT. Therefore, we used the canine spontaneous tumor model to investigate TME responses to SBRT, and how local injection of immune modulatory antibodies to OX40 and TLR 3/9 agonists might modify those responses. Pet dogs with spontaneous cancers (melanoma, carcinoma, sarcoma, n = 6 per group) were randomized to treatment with either SBRT or SBRT combined with local immunotherapy. Serial tumor biopsies and serum samples were analyzed for immunologic responses. SBRT alone resulted at two weeks after treatment in increased tumor densities of CD3+ T cells, FoxP3+ Tregs, and CD204+ macrophages, and increased expression of genes associated with immunosuppression. The addition of OX40/TLR3/9 immunotherapy to SBRT resulted in local depletion of Tregs and tumor macrophages and reduced Treg-associated gene expression (FoxP3), suppressed macrophage-associated gene expression (IL-8), and suppressed exhausted T cell-associated gene expression (CTLA4). Increased concentrations of IL-7, IL-15, and IL-18 were observed in serum of animals treated with SBRT and immunotherapy, compared to animals treated with SBRT. A paradoxical decrease in the density of effector CD3+ T cells was observed in tumor tissues that received combined SBRT and immunotherapy as compared to animals treated with SBRT only. In summary, these results obtained in a spontaneous large animal cancer model indicate that addition of OX40/TLR immunotherapy to SBRT modifies important immunological effects both locally and systemically.     

In vitro comparisons between Carica papaya and pancreatic lipases during test meal lipolysis: Potential use of CPL in enzyme replacement therapy

High levels of lipase activity are known to occur in Carica papaya latex, and this activity is being used in some biotechnological applications. The lipolytic activity of C. papaya lipase (CPL) on dietary triacylglycerols (TAG) has not yet been studied. Hence, the aim of this study was to characterise the specific activity of CPL on dietary TAG present in a crude preparation. Also, we have determined its stability during the lipolysis of a test meal at various pH values mimicking those occurring in the gastro-intestinal tract, with or without bile, and have compared these properties with those of porcine pancreatic extract (PPE) and human pancreatic lipase (HPL). CPL showed maximum stability at pH 6.0, both with and without bile. Some residual activity was still observed at pH 2 (20%), whereas the pancreatic lipases tested were immediately completely inactivated at this pH. In the absence of bile, the highest specific activities were measured at pH 6 in the case of CPL, PPE and HPL. Adding bile slightly decreased the CPL activity in the 4–6 pH range, thus shifting the optimum CPL activity to pH 7, where the presence of bile had no effect. Lipolysis levels decreased with the pH, but CPL was still more active than PPE at pH 5 on a relative basis. These results suggest that CPL might be a promising candidate for use as a therapeutic tool on patients with pancreatic exocrine insufficiency.     

Antimicrobial Activity of Nisin and Natamycin Incorporated Sodium Caseinate Extrusion‐Blown Films: A Comparative Study with Heat‐Pressed/Solution Cast Films

Antimicrobial edible films based on sodium caseinate, glycerol, and 2 food preservatives (nisin or natamycin) were prepared by classical thermomechanical processes. Food preservatives were compounded (at 65 °C for 2.5 min) with sodium caseinate in a twin‐screw extruder. Anti‐Listeria activity assays revealed a partial inactivation of nisin following compounding. Thermoplastic pellets containing food preservatives were then used to manufacture films either by blown‐film extrusion process or by heat‐press. After 24 h of incubation on agar plates, the diameters of K. rhizophila growth inhibition zones around nisin‐incorporated films prepared by solution casting (control), extrusion blowing or heat pressing at 80 °C for 7 min of nisin‐containing pellets were 15.5 ± 0.9, 9.8 ± 0.2, and 8.6 ± 1.0 mm, respectively. Since heat‐pressing for 7 min at 80 °C of nisin‐incorporated pellets did not further inactivate nisin, this indicates that nisin inactivation during extrusion‐blowing was limited. Moreover, the lower diameter of the K. rhizophila growth inhibition zone around films prepared with nisin‐containing pellets compared to that observed around films directly prepared by solution casting confirms that nisin inactivation mainly occurred during the compounding step. Natamycin‐containing thermoplastic films inhibited Aspergillus niger growth; however, by contrast with nisin‐containing films, heat‐pressed films had higher inhibition zone diameters than blown films, therefore suggesting a partial inactivation of natamycin during extrusion‐blowing.     

Comparative In Vitro Osteoinductivity Study of HA and α‐TCP/HA Bicalcium Phosphate

A bicalcium phosphate (BCP) bioceramic comprising α‐tricalcium phosphate (α‐TCP) and hydroxyapatite (HA) was prepared by a two‐step sintering. The microstructure of the BCP bioceramic was investigated using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X‐ray diffraction. The in vitro osteoinductivity was evaluated by culturing MG63 osteoblast‐like cells on the BCP bioceramic. Results showed that the BCP bioceramic comprising α‐TCP and HA in a moderate ratio possessed a hardness of 93.7 Hv. The cells spread faster on the BCP bioceramic than those on the commercial HA. It suggested that the BCP bioceramic can enhance osteoinductivity in vivo.     

Retention of Eucalyptol,a Natural Volatile Insecticide,in Delivery Systems Based on Hydroxypropyl‐β‐Cyclodextrin and Liposomes

Eucalyptol (Euc) is a natural monoterpene with insecticide effects. Being highly volatile and sensitive to ambient conditions, its encapsulation would enlarge its application. Euc‐loaded conventional liposomes (CL), cyclodextrin/drug inclusion complex, and drug‐in‐cyclodextrin‐in‐liposomes (DCL) are prepared to protect Euc from degradation, reduce its evaporation, and provide its controlled release. The liposomal suspension is freeze‐dried using hydroxypropyl‐β‐cyclodextrin (HP‐β‐CD) as cryoprotectant. The liposomes are characterized before and after freeze‐drying. The effect of Euc on the fluidity of liposomal membrane is also examined. A release study of Euc from delivery systems, in powder and reconstituted forms, is performed by multiple head extraction at 60 °C after 6 months of storage at 4 °C. CL and DCL suspensions are homogeneous, show nanometric vesicles size, spherical shape, and negative surface charge before and after freeze‐drying. Moreover, HP‐β‐CD does not affect the fluidity of liposomes. CL formulations present a weak encapsulation for Euc. The loading capacity of eucalyptol in DCL is 38 times higher than that in CL formulation. In addition, freeze‐dried DCL and HP‐β‐CD/Euc inclusion complex show a higher retention of eucalyptol than CL delivery system. Both carrier systems HP‐β‐CD/Euc and Euc‐loaded DCL decrease Euc evaporation and improve its retention. Practical Applications: Eucalyptol is a natural insecticide. It is highly volatile and poorly soluble in water. To enlarge its application, its encapsulation in three delivery systems (conventional liposomes, cyclodextrin/drug inclusion complex, combined system composed of cyclodextrin inclusion complex and liposome) is studied. In this paper it is proved that cyclodextrin/eucalyptol inclusion complex and eucalyptol‐in‐cyclodextrin‐in‐liposome are effective delivery systems for encalyptol encapsulation, retention, and release.     

ITER CODAC interface for the visible and infra-red wide angle viewing cameras

The amount of data generated by the infra-red and visible cameras at ITER is expected to be considerably larger than most diagnostics. ITER will have 12 infra-red cameras plus 12 visible cameras in four different equatorial port plugs. Each of the ports will have a Plant System Host (PSH) that will provide a standard image of the plant system to the ITER's Control and Data Access and Communication (CODAC) system.The two key functions of these cameras will be the scientific exploitation with the detection of interesting physics events and the operational protection of the machine, namely the first wall. Already assuming high bandwidth requirements for both audio and video, ITER will provide a separate network for this kind of communication, which will be used to transmit both the experimental and informational data provided by the cameras.This paper presents the current camera plant system design and its interaction with ITER CODAC system and networks. Starting from the camera specifications several alternatives for data collection and compression are discussed. The required inputs from CODAC and a first specification for the internal finite state machine are also presented. Finally, a possible hardware straw man design solution for the plant system hardware is proposed.     

Morphological and adhesive properties of polypyrrole films synthesized by sonoelectrochemical technique

This paper presents electrochemical synthesis of polypyrrole films under high frequency sonication. The films are characterized in terms of adhesion and surface morphology. Comparison to classical electrosynthesized polypyrrole films is made. In particular, the use of high frequency sonication (500 kHz) during electrodeposition of polypyrrole on Si was reported for the first time. Chronocoulometry was used for polymer films electrosynthesis on Si substrates. Influence of polymer thickness on the rugosity, morphology and adhesion force properties was studied. Scanning Electron Microscopy and mechanical probe profiling showed that sonication led to less rough and more homogeneous surface structure. Adhesion force properties of polypyrrole films were also studied by means of force–distance curves obtained by atomic force microscopy. For polymer films obtained in the absence of sonication, adhesion forces decreases with increase in polymer thickness. On the contrary, for polymer films obtained under sonication, the adhesion of the films decreased when the polymer thickness decreased.     

Macroporous calcium phosphate ceramic implants for sustained drug delivery

Limited blood circulation to the skeletal tissue is a major cause of reduced therapeutic effects seen with drugs administered by conventional systemic ways. To resolve this issue and obtain a sufficiently high local concentration to induce therapeutic effects, several implanted drug delivery systems have been developed for hard tissues using biomaterials.We have designed a drug delivery device based on hydroxylapatite (HA) and tested it in vitro using metronidazol and chloramphenicol as model compounds. Porous HA ceramics were prepared with two different shapes (cylindrical and spherical). Known amounts of drugs were introduced inside a drilled hole and sealed with wax. The ceramics were then suspended in stirred distilled water in closed polypropylene vials. Drug release was observed during several weeks.A mathematical model used to describe drug release from HA was elaborated based on the expressions of Fick's laws. The experimental kinetic results could be related to ceramic constitution and to drug localization.