•LAB have been successfully used in the last years as safe mucosal delivery vectors.•L. lactis is considered as the model LAB and an excellent vector of medical proteins.•We report on the development ...of a Stress-Inducible Controlled Expression (SICE) system.•We validated SICE system in a model of therapy against IBD.•We validated SICE system in a model of mucosal vaccination against HPV-16.
In recent years, recombinant lactic acid bacteria (LAB) have been successfully used as safe mucosal delivery vectors. Herein, we report on the development of a Stress-Inducible Controlled Expression (SICE) system in L. lactis for the production and delivery of proteins of health interest (both therapeutic and vaccine related) at mucosal surfaces. This system is episomal in nature and is composed of a vector carrying an expression cassette under the transcriptional control of a stress-inducible promoter. The functionality of the SICE system was validated in vivo using two different routes of administration: oral and intranasal, and in two different murine models of human pathologies: (i) a model of therapy against inflammatory bowel diseases (IBD) and (ii) a model of vaccination against human papillomavirus type-16 (HPV-16).
Human papillomavirus (HPV) is the causative agent of cervical cancer (CxCa) and the most commonly sexually transmitted pathogen worldwide. HPV type 16 (HPV-16) E7 oncoprotein is constitutively ...produced in CxCa and considered as a good antigen candidate for the development of new therapeutic CxCa vaccines. Here, we report the use of non-genetically modified, E7-expressing lactic acid bacteria (LAB) by using the cell-binding domain from
Lactobacillus casei
A2 phage lysin as a cell wall anchor. The versatility of this system was validated by investigating E7 stability at the surface of
Lactococcus lactis
and
L. casei
, two major species of LAB. Moreover, we demonstrated the successful use of these LAB displaying E7 antigen as a mucosal live vaccine in mice. Altogether, these results show the feasibility of using non-genetically modified LAB for low-cost mucosal immunotherapy against HPV-related CxCa in humans.
Celotno besedilo
Dostopno za:
CEKLJ, DOBA, EMUNI, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK
The lysin gene (
lysA2
) of the
Lactobacillus casei
bacteriophage A2 was cloned and expressed in
Escherichia coli
. LysA2 is an endopeptidase that hydrolyzes the bond between the terminal
d
-alanine ...of the peptidoglycan tetrapeptide and the aspartic acid residue that forms the bridge with the
l
-lysine of a neighboring peptidoglycan chain, characteristic of Gram-positive bacteria included into the A4 peptidoglycan subgroup. This includes most lactobacilli,
Lactococcus lactis
,
Pediococcus acidilactici
, and
Pediococcus pentosaceus
, the walls of all of which were substrates for the enzyme. Specific binding of LysA2 to the wall of these bacteria is mediated by its C-terminal moiety, does not need the N-terminal catalytic domain for recognition, and is stable: at least 88% of the molecules were still bound to
L. casei
after 3 days in phosphate buffer at 4°C. The enzyme acts as a monomer, is active at pH values between 4 and 6, and at temperatures ranging between 18°C and 50°C while being independent of divalent cation addition. The enzyme showed strong resistance to incubation at high and low pH values but became progressively inactivated at 50°C and above. LysA2 is bactericidal, the viability of
L. casei
cultures dropping to 1% in 10 min, under the standard conditions used for the enzymatic assay.
This work reports on the nucleation of the β-phase of poly(vinylidene fluoride) (PVDF) by incorporating CoFe2O4 and NiFe2O4 nanoparticles, leading in this way to the preparation of magnetoelectric ...composites. The fraction of filler nanoparticles needed to produce the same β- to α-phase ratio in crystallized PVDF is 1 order of magnitude lower in the cobalt ferrite nanoparticles. The interaction between nanoparticles and PVDF chains induce the all-trans conformation in PVDF segments, and this structure then propagates in crystal growth. The nucleation kinetics is enhanced by the presence of nanoparticles, as corroborated by the increasing number of spherulites with increasing nanoparticle content and by the variations of the Avrami’s exponent. Further, the decrease of the crystalline fraction of PVDF with increasing nanoparticle content indicates that an important fraction of polymer chains are confined in interphases with the filler particle.
AbstractThe problem presented in this edition of the Battle of the Water Networks is to define district metering areas (DMAs) in a large network. The problem is addressed in two phases. First, the ...complexity of the network is simplified by dividing it into three operational areas. Second, an optimization algorithm defines DMAs, looking for the best feasible solution. A preliminary simulation of the network is made. From this, engineering judgment allows for defining an initial set of elements suitable to change. In the second stage, a heuristic algorithm is used to search for the best DMA definition by selecting the locations and settings of the pressure-reducing valves and isolation valves. The network is then divided into two categories: the main pipes and the distribution pipes. Only the distribution pipes can be closed. With these restrictions and those described in the problem, the algorithm looks for the best DMA definition based on both the pressure and demand distribution among all the DMAs.
Abstract Objectives Missing data imputation is an important task in cases where it is crucial to use all available data and not discard records with missing values. This work evaluates the ...performance of several statistical and machine learning imputation methods that were used to predict recurrence in patients in an extensive real breast cancer data set. Materials and methods Imputation methods based on statistical techniques, e.g., mean, hot-deck and multiple imputation, and machine learning techniques, e.g., multi-layer perceptron (MLP), self-organisation maps (SOM) and k-nearest neighbour (KNN), were applied to data collected through the “El Álamo-I” project, and the results were then compared to those obtained from the listwise deletion (LD) imputation method. The database includes demographic, therapeutic and recurrence-survival information from 3679 women with operable invasive breast cancer diagnosed in 32 different hospitals belonging to the Spanish Breast Cancer Research Group (GEICAM). The accuracies of predictions on early cancer relapse were measured using artificial neural networks (ANNs), in which different ANNs were estimated using the data sets with imputed missing values. Results The imputation methods based on machine learning algorithms outperformed imputation statistical methods in the prediction of patient outcome. Friedman’s test revealed a significant difference ( p = 0.0091 ) in the observed area under the ROC curve (AUC) values, and the pairwise comparison test showed that the AUCs for MLP, KNN and SOM were significantly higher ( p = 0.0053 , p = 0.0048 and p = 0.0071 , respectively) than the AUC from the LD-based prognosis model. Conclusion The methods based on machine learning techniques were the most suited for the imputation of missing values and led to a significant enhancement of prognosis accuracy compared to imputation methods based on statistical procedures.
Capecitabine is an active drug in metastatic breast cancer (BC). GEICAM/2003-10 is an adjuvant trial to investigate the integration of capecitabine into a regimen of epirubicin and docetaxel for ...node-positive early BC.
Patients with operable node-positive BC (T1-3/N1-3) were eligible. After surgery, 1,384 patients were randomly assigned to receive epirubicin plus cyclophosphamide (EC; 90 and 600 mg/m(2), respectively, × four cycles), followed by docetaxel (100 mg/m(2) × four cycles; EC-T) or epirubicin plus docetaxel (ET; 90 and 75 mg/m(2), respectively, × four cycles), followed by capecitabine (1,250 mg/m(2) twice a day on days 1 to 14, × four cycles; ET-X); all regimens were given every 3 weeks. The primary end point was invasive disease-free survival. Secondary end points included safety (with an alopecia-specific study) and overall survival (OS).
After a median follow-up of 6.6 years and 297 events, 86% of patients who received EC-T and 82% of those who received ET-X were invasive disease free at 5 years (hazard ratio, 1.30; 95% CI, 1.03 to 1.64; log-rank P = .03). The OS difference between arms was not statistically significant (hazard ratio, 1.13; 95% CI, 0.82 to 1.55; log-rank P = .46). The most frequent grade 3 to 4 adverse events in the EC-T versus ET-X arms were neutropenia (19% v 10%), with 7% febrile neutropenia across arms; fatigue (13% v 11%); diarrhea (3% v 11%); hand-foot syndrome (2% v 20%); mucositis (6% v 5%); vomiting (both, 5%); and myalgia (4.5% v 1%). Incomplete scalp hair recovery was more frequent in the EC-T than ET-X arm (30% v 14%), and patients who received EC-T wore wigs significantly longer than those who received ET-X (8.35 v 6.03 months).
Invasive disease-free survival, but not OS, was significantly superior for patients with node-positive early BC who received the adjuvant standard schedule EC-T than for those who received the experimental ET-X regimen. Toxicity profiles differed substantially across arms.
Battle of the Water Networks II Marchi, Angela; Salomons, Elad; Ostfeld, Avi ...
Journal of water resources planning and management,
07/2014, Letnik:
140, Številka:
7
Journal Article
Recenzirano
Odprti dostop
AbstractThe Battle of the Water Networks II (BWN-II) is the latest of a series of competitions related to the design and operation of water distribution systems (WDSs) undertaken within the Water ...Distribution Systems Analysis (WDSA) Symposium series. The BWN-II problem specification involved a broadly defined design and operation problem for an existing network that has to be upgraded for increased future demands, and the addition of a new development area. The design decisions involved addition of new and parallel pipes, storage, operational controls for pumps and valves, and sizing of backup power supply. Design criteria involved hydraulic, water quality, reliability, and environmental performance measures. Fourteen teams participated in the Battle and presented their results at the 14th Water Distribution Systems Analysis conference in Adelaide, Australia, September 2012. This paper summarizes the approaches used by the participants and the results they obtained. Given the complexity of the BWN-II problem and the innovative methods required to deal with the multiobjective, high dimensional and computationally demanding nature of the problem, this paper represents a snap-shot of state of the art methods for the design and operation of water distribution systems. A general finding of this paper is that there is benefit in using a combination of heuristic engineering experience and sophisticated optimization algorithms when tackling complex real-world water distribution system design problems.
Several types of C4‐functionalized 4‐alkyl‐2‐aryl‐1,2,3,4‐tetrahydroquinolines underwent rearrangement of their functional groups to C3, with concomitant aromatization, by simple reflux in ...1,2‐dichlorobenzene. The functional groups that were shown to undergo the C4 to C3 migration were –CH=CH‐Z (where Z = CO2Et, CN, NO2, COCH3, CH2OH) and –CH=C(Y)–Z (where Y = CN and Z = CO2Et or Y = Z = CN). On the other hand, the dimethylhydrazono group failed to migrate under thermal conditions but was shown to undergo a smooth dehydrogenation/C4 to C3 rearrangement/ dehydrogenation sequence at room temperature in the presence of DDQ, with a broad scope that includes 4‐alkyl‐2‐aryl‐ and 2‐acyl‐1,2,3,4‐tetrahydroquinolines. We also report a computational and experimental study of the mechanism of both reactions, which supports an unusual intramolecular aza‐ene pathway. The ready availability by this method of 2,3‐difunctionalized quinolines allowed the simple preparation of fused heterocyclic systems derived from the pyrrolo3,4‐bquinoline framework, using both reductive and non‐reductive domino processes.
Highly substituted and functionalized quinolines were obtained by thermal or DDQ‐induced C4 to C3 migration of functionalized vinyl substituents in racemic 4,4‐disubstituted 1,2,3,4‐tetrahydroquinolines.
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