A new family of sugar-inducible expression vectors for Escherichia coli

A set of 11 expression vectors was constructed, each of themharbouring a cloning cassette under the control of the promoter.Some of these vectors enable expression of foreign proteinsin the cytoplasm, while others include a synthetic sequencecoding for a very efficient secretion signal sequence. Otherfeatures are an fl origin of replication (in plus or minus orientation)and a promoter^up mutation that enhances the already very highlevel of expression from these vectors. With such a versatilevector family, cloning, sequencing and sitedirected mutagenesiscan be performed on the same vector, and the level of expressioncan be defined according to the specific constraints of a givenprotein.     

Fibroblast growth factor,estrogen, and prolactin receptor features in different grades of prostatic adenocarcinoma in elderly men

The objective was to characterize and associate the receptor reactivities of fibroblastic growth factor (FGF)‐2, FGF‐7, FGF‐8, epidermal growth factor (EGF), α‐actin and vimentin in relation to the androgen receptor (AR), α and β estrogen receptors (ERα and ERβ), and prolactin receptor in the prostate of elderly men showing low‐ and high‐grade adenocarcinoma. Thirty prostatic samples were taken from 60‐ to 90‐year‐old patients without prostatic lesions and with low‐grade cancer and high‐grade cancer, from the University Hospital, School of Medicine, the State University of Campinas. The results showed that increased FGF‐2, FGF‐7, and FGF‐8 receptor reactivities and decreased AR reactivity were verified in both high‐ and low‐grade cancer. However, the FGF‐8 receptor showed greater involvement at the beginning of the malignancy alterations. Increased EGF receptor (EGFR) reactivity and diminished α‐actin immunohistochemistry were identified in both cancer groups. Also, increased ERα, PR, and vimentin receptors were verified in both cancer groups. To conclude, the ERα involvement in the reactive stroma activation led to a microenvironment, which was favorable to cancer progression, due to maximizing stromal imbalance. The prolactin could be related to cancer progression due to its interaction with ERα action, indicating that this hormone could be a relevant target to prevent the estrogenic effects in the prostatic lesions. Both FGF receptor (FGFR)‐2 and FGFR‐8 play a fundamental role in the early stages of prostate cancer, suggesting that these molecules could be a promising therapeutic target. The differential localization of the fibroblastic factors between the prostatic epithelium and stroma of elderly men, who presented prostate cancer, could indicate a favorable distinction for tumoral progression. Microsc. Res. Tech. 76:321–330, 2013. © 2013 Wiley Periodicals, Inc.     

Photo-Fenton-peroxide process using FE (II)-embedded composites based on activated carbon: Characterization of catalytic tests

Cost-efficient Fenton-like catalysts for the removal of organic molecules in aqueous solutions were elaborated by a simple process. Porous activated carbons (ACs) were directly impregnated with a precursor solution using the wet impregnation method. Their efficiency as Fenton-like catalysts was studied. Photo-Fenton tests were performed to establish the performance of the prepared Fe-impregnated activated carbons in relation to the degradation of an organic pollutant (Indigo Carmine) model in aqueous solution, under different conditions. Photo-catalytic tests were carried out by means of a laboratory photo-reactor (UV-Consulting Peschl). The influence of several parameters such as solution pH value, initial concentration of the model pollutant, and hydrogen peroxide dose on the process performance was investigated. The ACs and prepared catalysts were characterized by nitrogen adsorption-desorption isotherms at 77 K, SEM, and thermogravimetric analyses. The total Fe content of the synthesized composites was estimated by the phenanthroline method using UV-vis spectrophotometry. The results show an increase in the degradation rate when the heterogeneous photo-Fenton process is conducted with an ozone generating UV lamp.     

Site-directed mutagenesis of active site contact residues in a hydrolytic abzyme: evidence for an essential histidine involved in transition state stabilization

Specific molecular interactions involved in catalysis by antibody 6D9 were investigated by site-directed mutagenesis. The catalytic antibody 6D9, which was generated against a transition state analog (III), hydrolyzes a non-bioactive chloramphenicol monoester derivative (I) to produce chloramphenicol (II). Construction of a three-dimensional molecular model of 6D9 and sequence comparison within a panel of related antibodies suggested candidates for catalytic residues, His (L27d), Tyr (L32), Tyr (H58) and Arg (H100b); these were targeted for the site-directed mutagenesis study. The Y-H58-F and R-H100b-A mutants possessed catalytic activities comparable to that of the wild-type, and the Y-H58-H and Y-L32-F mutant displayed an approximately fivefold decrease in k(cat)/Km. In the transition state analysis, the plots of logK(TSA) versus log(k(cat)/Km) for the mutants are linear, with a slope of approximately 1.0, indicating that the entire hapten-binding energy in the mutants is also utilized to bind the transition state and to accelerate the catalysis. In addition, a dramatic change in the catalytic activity was observed when the histidine residue (27d) in the CDR1 light chain was replaced with alanine. The H-L27d-A mutant had no detectable catalytic activity. This mutation led to a large, 40-fold reduction in transition state binding, with no change in substrate binding. Coupled with the previous kinetic studies and chemical modifications of the intact 6D9 antibody, this mutagenesis study has demonstrated that His L27d plays an essential role in stabilization of the transition state, the mechanism of catalysis by the 6D9 antibody.     

Removal of dark coloured and polyphenolic compounds of sugar beet vinasse by adsorption onto activated carbons: Application to a crosscurrent adsorption process

Sugar beet distillery vinasse is rich in betaine but the recovery of betaine by ionic exchange first requires removing ethanol‐insoluble compounds as well as dark coloured and polyphenolic compounds. The adsorption kinetics and isotherms of the latter compounds were studied on two commercial activated carbons. The results show that the pseudo‐second order kinetic model provided the best correlation, and the Langmuir model was selected for describing the isotherms. The data can be used to conduct experiments with a view to applying the crosscurrent process methodology to determine the number of stages necessary for the discolouration of the solution. © 2011 Canadian Society for Chemical Engineering     

Morphological alterations in the prostate stroma of rats submitted to chronic nicotine treatment

The stroma plays a fundamental role in the function of different glandular systems. In the prostate, the stroma is responsible for the development and maintenance of the differentiated state of the epithelium. Nicotine induces tobacco dependence and promotes morphological alterations in the epithelial compartment. However, its effects on the prostate stroma are unclear. The objective of this study was to evaluate the morphology of the stromal microenvironment in the ventral prostate lobe of rats submitted to chronic nicotine administration. Twenty rats (Rattus norvegicus) were divided into two groups: 10 animals received subcutaneous nicotine and 10 animals received physiological saline by the same route. After treatment, samples were collected from the ventral prostate lobe, processed and submitted to histology, histochemistry, and ultrastructural analysis by transmission and scanning electron microscopy. The level of circulating testosterone was also analyzed. The results showed a significant increase in the density of type I collagen (56.3% to 85.9%, P < 0.01) and a decrease in the density of type III collagen (43.7% to 14.1%, P < 0.01). In addition, there was a qualitative increase in elastic fibers and in the number of smooth muscle cells with a secretory phenotype. Circulating testosterone levels were significantly reduced (898.3 to 363.1 ng/mL, P < 0.01). The results showed that nicotine modifies different components of the prostate stroma, suggesting that this drug is a risk factor for morphofunctional alterations in the prostate gland.     

Activated carbon porosity tailoring by cyclic sorption/decomposition of molecular oxygen

Activated-carbon pore size tailoring is usually achieved by pore size reduction, pore size widening or more rarely by direct activation. Successive widening cycles composed of an initial molecular oxygen sorption step, followed by a carbonization step under nitrogen, have been applied to three different materials in order to study their gradual pore size modifications. For years, the whole microporosity obtained after several cycles is known to present better pore size distribution than those inherited from conventional activations. Moreover, as shown in the present paper, the gradual obtained microporosities cover a wide and valuable range of porous textures highly dependent on the initial material origin and initial activation. Gradual mean pore size evolutions of 1 or 2 Å per cycle were observed linked to a simultaneous increase in microporous specific volume in the case of pitch-based and coconut activated carbons, respectively. Comparatively, the microporous specific volume of a commercial coconut carbon molecular sieve was increased by 26% without modification of its mean pore size and therefore of its sieving effect. Those results have been used to test a simple textural model of activated carbon porosity proposed in the recent literature.     

Importance of the structure of paper support in gas transfer properties of protein‐coated paper

Composite materials made from fibrous support coated with agro‐polymers are widely commercialized for different applications. But, their transfer properties have only been sparingly studied despite their importance in membrane technologies or food packaging. Furthermore, most of past studies on the topic have been focused on the impact of surface properties of the support on the final coated material structure and its properties; leaving unexplored the potential impact of the in‐bulk structure of the support. This study demonstrated the influence of in‐bulk structure and especially fibers refining degree of 4 kraft papers (SP28, 36, 60, and 80) on the structure, and surface and gas transfer properties of their respective wheat gluten (WG) coated papers (WGP28, 36, 60, and 80). Paper presenting a high fibers refining degree (SP28) exhibited very tight and narrow in bulk fibers network which prevented most of the WG‐coated layer penetration, maintaining an important WG apparent layer on top of the paper and a small fibers/wheat gluten composite zone inside (WGP28). Such structure gave strong “WG‐like” properties to the final coated material with moderate oxygen permeation and high permselectivity (1.50 × 10^?11 mol m^?2 s^?1 Pa^?1 and 8.09, respectively for WGP28) whereas the highly impregnated structure of coated papers built on lowly refined papers (SP80, with wide and loose structure) gave coated materials presenting weak “WG‐like” properties, supposedly due to a thick composite zone presenting interfacial defects, with higher oxygen permeation and very limited permselectivity (11.90 × 10^?11 mol m^?2 s^?1 Pa^?1 and 1.06, respectively for WGP80). © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 2848–2858, 2013     

Pore structure modification of pitch-based activated carbon by NaOCl and air oxidation/pyrolysis cycles

Two complementary wearing off cycling methods based on an initial wet oxidation in a sodium hypochlorite solution or an initial dry oxidation under air, both followed by a thermal pyrolysis under nitrogen, have been applied to a same carbon molecular sieve to study its gradual pore structure modifications. The changes in microporous properties resulting from these cycles were studied by N₂ physisorption at 77 K and analyzed by using the Dubinin–Radushkevich equation. The observed textural behaviour is different from those usually observed using conventional activation processes. The surface complexes created on the surface of carbon by each NaOCl oxidation were characterized by temperature-programmed desorption (TG-TPD-MS), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). Corresponding surface functional species have been identified and quantified.

Each of the two methods leads to the formation of two distinct microporous domains developed on the obtained modified CMS series. The gradual mean pore size values of one of those domains are common to the two series while the others are much more enlarged by the oxygen than by the NaOCl treatments. Therefore, the two methods lead to complementary data allowing experimental differentiation of the different porosities encountered in activated carbons. Such series of materials of common origin and surface chemistry but different gradual textural properties are of great interest for activated carbons structural studies as well as for characterization technique improvements.     

Steroid hormone receptors, matrix metalloproteinases, insulin-like growth factor, and dystroglycans interactions in prostatic diseases in the elderly men

OBJECTIVES: The aim of this study was to evaluate the reactivity of steroid hormone receptors (SHRs), dystroglycans (DGs), matrix metalloproteinases (MMPs), insulin‐like growth factor receptor (IGFR‐1), and laminin (Lam) in both prostatic stromal and epithelial compartments showing different diseases in elderly men. METHODS: Sixty prostatic samples were obtained from 60‐ to 90‐year‐old patients (mean 63 years) with and without prostatic lesions from Hospital of the School of Medicine, State University of Campinas (UNICAMP). The Samples were divided into standard (no lesions); high grade prostatic intraepithelial neoplasia (HGPIN); prostatic cancer (PC); and benign prostatic hyperplasia (BPH) groups. The samples were submitted to immunohistochemistry and Western blotting analyses. Research Ethics Committee of the School of Medicine, University of Campinas/UNICAMP (number 0094.0.146.000‐08). RESULTS: The results showed increased IGFR‐1 and MMPs protein levels in the PC and HGPIN groups. Decreased αDG and βDG protein levels were verified in the PC and HGPIN groups. Androgen receptor (AR) reactivity was similar among all groups. Estrogen receptor α (Erα) immunoreactivity was more intense in the epithelium in the PC and HGPIN groups. Estrogen receptor β (ERβ) immunoreactivity was weak in the epithelium of the HGPIN and PC groups. CONCLUSIONS: To conclude, there was an association among IGFR‐1, MMPs, and SHRs, indicating IGFR‐1 as a target molecule in prostate therapy, considering the IGF proliferative properties. Also, the distinct SHRs reactivities in the lesions in both prostatic compartments indicated different paracrine signals and pointed out the importance of estrogenic pathways in the activation of these disorders. Microsc. Res. Tech. 75:1197–1205, 2012. © 2012 Wiley Periodicals, Inc.