Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system

Lactic acid bacteria (LAB) are Gram-positive bacteria and are generally regarded as safe (GRAS) organisms. Therefore, LAB could be used for heterologous protein secretion and they are good potential candidates as antigen delivery vehicles. To develop such live vaccines, a better control of protein secretion is required. We developed an efficient secretion system in the model LAB, Lactococcus lactis. Staphylococcal nuclease (Nuc) was used as the reporter protein. We first observed that the quantity of secreted Nuc correlated with the copy number of the cloning vector. The nuc gene was cloned on a high-copy number cloning vector and no perturbation of the metabolism of the secreting strain was observed. Replacement of nuc native promoter by a strong lactococcal one led to a significant increase of nuc expression. Secretion efficiency (SE) of Nuc in L. lactis was low, i.e., only 60% of the synthesized Nuc was secreted. Insertion of a synthetic propeptide between the signal peptide and the mature moiety of Nuc increased the SE of Nuc. On the basis of these results, we developed a secretion system and we applied it to the construction of an L. lactis strain which secretes a bovine coronavirus (BCV) epitope-protein fusion (BCV-Nuc). BCV-Nuc was recognized by both anti-BCV and anti-Nuc antibodies. Secretion of this antigenic fusion is the first step towards the development of a novel antigen delivery system based on LAB-secreting strains.

Design of automatic control system for the precipitation of bromelain from the extract of pineapple wastes

In this work, bromelain was recovered from ground pineapple stem and rind by means of precipitation with alcohol at low temperature. Bromelain is the name of a group of powerful protein-digesting, or proteolytic, enzymes that are particularly useful for reducing muscle and tissue inflammation and as a digestive aid. Temperature control is crucial to avoid irreversible protein denaturation and consequently to improve the quality of the enzyme recovered. The process was carried out alternatively in two fed-batch pilot tanks: a glass tank and a stainless steel tank. Aliquots containing 100 mL of pineapple aqueous extract were fed into the tank. Inside the jacketed tank, the protein was exposed to unsteady operating conditions during the addition of the precipitating agent (ethanol 99.5%) because the dilution ratio "aqueous extract to ethanol" and heat transfer area changed. The coolant flow rate was manipulated through a variable speed pump. Fine tuned conventional and adaptive PID controllers were on-line implemented using a fieldbus digital control system. The processing performance efficiency was enhanced and so was the quality (enzyme activity) of the product.

Chitosan as a wall material for a microencapsulated delivery system for Macrobrachium rosenbergii(de Man) larvae

Chitosan has beenwidely accepted as awall material for preparing microcapsules of various purposes in human medicine. The possibility of using chitosan as a wall material for microencapsulating nutrients and drugs for aquaculture purposes, speci¢cally to Macrobrachium rosenbergii larvae was evaluated in this study. Two types of chitosan-coated microcapsules were prepared using either acetone (MEC-A) or NaOH (MEC-N) as the cross-linking agents. They were compared with a microbound diet relative to total leaching of nutrients and free amino acids (FAA). Among the microcapsules, MEC-N showed the lowest level of total leaching of nutrients (23.3%) during 5 h of immersion in seawater and released 65% FAA after 60min. During laboratory trials,75% larvae had accepted the MEC-N capsule. The results of the study suggest that chitosan can be used as a wall material for preparing microcapsules to deliver drugs and nutrients to M. rosenbergii larvae.

Logistics Network Design with Differentiated Delivery Lead-Time: Benefits and Insights

Most logistics network design models assume exogenous customer demand that is independent of the service time or level. This paper examines the benefits of segmenting demand according to lead-time sensitivity of customers. To capture lead-time sensitivity in the network design model, we use a facility grouping method to ensure that the different demand classes are satisfied on time. In addition, we perform a series of computational experiments to develop a set of managerial insights for the network design decision making process.

A nipple shield delivery system for oral drug delivery to breastfeeding infants : Microbicide delivery to inactivate HIV

A new drug delivery method for infants is presented which incorporates an active pharmaceutical ingredient (API)-loaded insert into a nipple shield delivery system (NSDS). The API is released directly into milk during breastfeeding. This study investigates the feasibility of using the NSDS to deliver the microbicide sodium dodecyl sulfate (SDS), with the goal of preventing mother-to-child transmission (MTCT) of HIV during breastfeeding in low-resource settings, when there is no safer alternative for the infant but to breastfeed. SDS has been previously shown to effectively inactivate HIV in human milk. An apparatus was developed to simulate milk flow through and drug release from a NSDS. Using this apparatus milk was pulsed through a prototype device containing a non-woven fiber insert impregnated with SDS and the microbicide was rapidly released. The total SDS release from inserts ranged from 70 to 100% of the average 0.07 g load within 50 ml (the volume of a typical breastfeed). Human milk spiked with H9/HIVIIIB cells was also passed through the same set-up. Greater than 99% reduction of cell-associated HIV infectivity was achieved in the first 10 ml of milk. This proof of concept study demonstrates efficient drug delivery to breastfeeding infants is achievable using the NSDS.

Design of a forecasting system for ECMWF

As laid out in its convention there are 8 different objectives for ECMWF. One of the major objectives will consist of the preparation, on a regular basis, of the data necessary for the preparation of medium-range weather forecasts. The interpretation of this item is that the Centre will make forecasts once a day for a prediction period of up to 10 days. It is also evident that the Centre should not carry out any real weather forecasting but merely disseminate to the member countries the basic forecasting parameters with an appropriate resolution in space and time. It follows from this that the forecasting system at the Centre must from the operational point of view be functionally integrated with the Weather Services of the Member Countries. The operational interface between ECMWF and the Member Countries must be properly specified in order to get a reasonable flexibility for both systems. The problem of making numerical atmospheric predictions for periods beyond 4-5 days differs substantially from 2-3 days forecasting. From the physical point we can define a medium range forecast as a forecast where the initial disturbances have lost their individual structure. However we are still interested to predict the atmosphere in a similar way as in short range forecasting which means that the model must be able to predict the dissipation and decay of the initial phenomena and the creation of new ones. With this definition, medium range forecasting is indeed very difficult and generally regarded as more difficult than extended forecasts, where we usually only predict time and space mean values. The predictability of atmospheric flow has been extensively studied during the last years in theoretical investigations and by numerical experiments. As has been discussed elsewhere in this publication (see pp 338 and 431) a 10-day forecast is apparently on the fringe of predictability.

AC Biosusceptometry as a Tool for Monitoring the Gastrointestinal Transit of Multiparticulate Drug Delivery System

Drug delivery systems based on natural polysaccharides, such as chitosan (CS) and pectin (PC), rather than on synthetic polymers, have been widely studied. Some reasons for that are low toxicity and costs and high biodegradability of the formers. A multiparticulate system based on CS and PC was developed in our laboratories, including the addition of an enteric polymer, cellulose acetate phtalate (CAP). Such improvement promoted stronger gastric and enteric resistances, as assessed in vitro, making the systems more selective to enzymatic degradation in the colon. Although in vitro dissolution tests can simulate some properties concerning the gastrointestinal transit (GT), collaborating to characterize the systems behavior in the biological fluids, frequently they do not result in satisfactory in vitro/in vivo correlations. The objective of this work was to follow in vivo the GT of the particles developed by means of AC biosusceptometry (ACB), a non-invasive and of low cost methodology. The particles containing ferrite in powder form were prepared by complex coacervation using an ideal 3:1:1 mass ratio for PC:CS:CAP. The magnetic particles were administered to healthy volunteers by oral route. The GT was monitored by using multi-sensor ACB system and the signal acquisition was performed every IS min until the colonic region was reached. By means of ACB technique, it was possible to acquiring images generated by the magnetic particles within the whole gastrointestinal tract including the colonic region. Variable particles transit times were observed among the volunteers, but without interference on the mapping of the particles until the colonic region. The particles were able to produce magnetic field strong enough to generate signals adequate for mapping the particles. The results suggest that integral particles reached the colon, after they resisted against gastric and enteric media. Studies associating transit time and in vivo drug release are in development in order to confirm the efficiency of the systems.

Natural rubber latex used as drug delivery system in guided bone regeneration (GBR)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Design of a control system using linear matrix inequalities for the active vibration control of a plate

The study of algorithms for active vibration control in smart structures is an area of interest, mainly due to the demand for better performance of mechanical systems, such as aircraft and aerospace structures. Smart structures, formed using actuators and sensors, can improve the dynamic performance with the application of several kinds of controllers. This article describes the application of a technique based on linear matrix inequalities (LMI) to design an active control system. The positioning of the actuators, the design of a robust state feedback controller and the design of an observer are all achieved using LMI. The following are considered in the controller design: limited actuator input, bounded output (energy) and robustness to parametric uncertainties. Active vibration control of a flat plate is chosen as an application example. The model is identified using experimental data by an eigensystem realization algorithm (ERA) and the placement of the two piezoelectric actuators and single sensor is determined using a finite element model (FEM) and an optimization procedure. A robust controller for active damping is designed using an LMI framework, and a reduced model with observation and control spillover effects is implemented using a computer. The simulation results demonstrate the efficacy of the approach, and show that the control system increases the damping in some of the modes.

Oil-in-water lecithin-based microemulsions as a potential delivery system for amphotericin B

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Software architecture for web-based project management system

This article presents software architecture for a web-based system to aid project managing, conceptually founded on guidelines of the Project Management Body of Knowledge (PMBoK) and on ISO/IEC 9126, as well as on the result of an empiric study done in Brazil. Based on these guidelines, this study focused on two different points of view about project management: the view of those who develop software systems to aid management and the view of those who use these systems. The designed software architecture is capable of guiding an incremental development of a quality system that will satisfy today's marketing necessities, principally those of small and medium size enterprises.