Chemical modification of botryosphaeran: structural characterization and anticoagulant activity of a water-soluble sulfonated (1-->3)(1-->6)-β-D-glucan

The exopolysaccharide botryosphaeran (EPS(GLC); a (1--> 3)(1-->6)-β-D-glucan from Botryosphaeria rhodina MAMB- 05) was sulfonated to produce a water-soluble fraction (EPS(GLC)-S) using pyridine and chlorosulfonic acid in formamid. This procedure was then repeated twice to produce another fraction (EPSGLC-RS) with a higher degree of substitution (DS, 1.64). The purity of each botryosphaeran sample (unsulfonated and sulfonated) was assessed by gel filtration chromatography (Sepharose CL-4B), where each polysaccharide was eluted as a single symmetrical peak. The structures of the sulfonated and re-sulfonated botryosphaerans were investigated using ultraviolet-visible (UV-Vis), Fourier-transform infrared (FT-IR), and (13)C nuclear magnetic resonance ((13)C NMR) spectroscopies. EPS(GLC) and EPS(GLC)-RS were also assayed for anticoagulation activity, and EPS(GLC)-RS was identified as an anticoagulant.     

A novel mechanism of modulation of 5-HT₃A receptors by hydrocortisone

Modulation of Cys-loop receptors by steroids is of physiological and therapeutical relevance. Nonetheless, its molecular mechanism has not been elucidated for serotonin (5-HT) type 3 receptors. We deciphered the mechanism of action of hydrocortisone (HC) at 5-HT type 3A receptors. Single-channel currents from the high-conductance form (∼4.7 pA, −70 mV) appear as a series of long opening events forming bursts, which group into long clusters. Although they are very infrequent, subconductance events (∼2.4 pA) are detected within clusters. HC produces a significant concentration-dependent reduction in open and burst durations, demonstrating open-channel block. In addition, it increases the appearance of subconductance levels in a concentration- and slightly voltage-dependent manner. The amplitude of the subconductance level does not change with HC concentration and its open duration is briefer than that of full amplitude events, indicating lower open-channel stability. Dual effects are distinguished from macroscopic responses: HC reduces amplitude by acting from either open or closed states, and it increases decay rates from the open state. Thus, HC acts as a negative modulator of 5-HT type 3A receptors by different mechanisms: It acts as an open-channel blocker and it favors opening to a preexisting subconductance level. The latter constitutes a novel, to our knowledge, mechanism of channel modulation, which might be applicable to other steroids and channels.     

Four members of the HSP101 gene family are differently regulated in Triticum durum Desf

Heat shock proteins play an essential role in preventing deleterious effects of high temperatures. In many plants, HSP101 has a central role in heat stress survival. We report the isolation and characterization of four cDNAs corresponding to different members of the durum wheat HSP101 gene family. Expression analysis revealed differences in their induction. Accordingly, durum wheat HSP101 genes are differently regulated, therefore having distinct roles in stress response and thermotolerance acquisition. These findings are important for further dissection of the molecular mechanisms underlying the stress response and for understanding the functions of the HSP101 family members. This information could be important for the exploitation of specific alleles in marker assisted selection for abiotic stress resistance.     

Experimental endocrine therapies promote epithelial cytodifferentiation and ciliogenesis in the gerbil female prostate

The incidence of ciliated cells in the prostate gland of the female gerbil (Meriones unguiculatus) is uncommon and apparently becomes more frequent during androgen (testosterone cypionate) and anti-estrogen (letrozole) endocrine therapies. To evaluate the effects of such drug therapies on the induction of ciliogenesis in the glandular epithelium of female prostate glands, adult female gerbils aged 90 days were treated for 14 days with testosterone and letrozole after which their prostate glands were removed for histological, ultrastructural, and serological analyses. The cytodifferentiation of the ciliated phenotype in the alveolar epithelium became more frequent after both the testosterone and the letrozole treatments. The ciliogenesis phenomenon of the epithelial cells in the prostate gland of female gerbils thus appears to be induced by variations in the increase of androgen levels.     

Recycling of EOL CRT glass into ceramic glaze formulations and its environmental impact by LCA approach

Background, Aims and Scope The interest in recycling materials at the end of their life is growing in the industry in general. As regards the Wastes of Electrical and Electronic Equipment (WEEE), an appreciable increase of these materials has been noticed in the last decades, 117 · 10³ tons of WEEE have been produced in Italy in 2002 according to Ecohitech [1] and the increase in this kind of waste is three times higher than that of the municipal waste according to the FISE ASSOAMBIENTE report [2]. Within WEEE, End-of-Life Cathode Ray Tube (EOL CRT) glass, the main part of TV sets and PC monitors, is here analysed using both a technical approach to establish a possible reuse of the glass in a open-loop recycling field (ceramic industry) and a methodology (LCA) capable of providing environmental evaluations. Methods The technological characterization was performed by chemical resistance tests (UNI EN ISO 10545-13), staining tests (UNI EN ISO 10545-14) with blue methylene and potassium permanganate (KMnO₄), and surface abrasion tests (UNI EN ISO 10545-7). The LCA study was conducted using the SimaPro 5.0 software and Eco-Indicator 99 as an evaluation method. Results and Discussion The good technical results, reached by using cleaned EOL CRT panel glass inside a ceramic glaze formulation instead of a commercial frit, are supported by the environmental impact evaluation, which shows a decrease of the overall potential damage (measured in Points) of 36% and, in particular, a reduction of 53% in ‘Human health’, 31% in ‘Eco-system quality’ and 24% in ‘Resources’. Conclusions This study has demonstrated that this new, open-loop recycling strategy for the CRT glass significantly reduces the environmental impact of the ceramic glaze production process. In fact, in all damage categories examined in this study, there is a minor impact. An improvement is evident in the respiratory inorganics sub-category related to the lowering of dusts mainly and to a lesser amount with NO_x and SO_x in the climate change sub-category, due mainly to the reduction of CO₂ emission correlated to the avoided combustion of the mixture which feeds melting furnaces in the frit production. Thus, the damage decrease in ‘Ecosystem quality’ is prevalently due to the lower NO_x emissions by the kilns in the frit production that is evident in the acidification/eutrophication sub-category. Finally, the significant saving in the ‘Resource’ category is principally linked to the fossil fuels sub-category, thanks to the methane saving which stokes the melting furnaces. Perspectives Furthermore, the decrease in CO₂ emission (94.4%) evident in the climate change sub-category is a very important topic because it is in line with the Kyoto protocol (1997), where significant efforts have been exerted for the reduction of the green house gases emission, notably CO₂. The CO₂ emission is correlated to the combustion of the mixture which feeds melting kilns in the frit production, therefore the recycling of secondary raw materials, already in a glass state, can reduce the emissions of this gas. This reduction can be termed as environmental credit and it is an example of an allocation of environmental loads in a open-loop recycling, where waste from one industrial system are used as raw materials in another product system.     

The Effect of Aging on the Specialized Conducting System: A Telemetry ECG Study in Rats over a 6 Month Period

Advanced age alone appears to be a risk factor for increased susceptibility to cardiac arrhythmias. We previously observed in the aged rat heart that sinus rhythm ventricular activation is delayed and characterized by abnormal epicardial patterns although conduction velocity is normal. While these findings relate to an advanced stage of aging, it is not yet known when and how ventricular electrical impairment originates and which is the underlying substrate. To address these points, we performed continuous telemetry ECG recordings in freely moving rats over a six-month period to monitor ECG waveform changes, heart rate variability and the incidence of cardiac arrhythmias. At the end of the study, we performed in-vivo multiple lead epicardial recordings and histopathology of cardiac tissue. We found that the duration of ECG waves and intervals gradually increased and heart rate variability gradually decreased with age. Moreover, the incidence of cardiac arrhythmias gradually increased, with atrial arrhythmias exceeding ventricular arrhythmias. Epicardial multiple lead recordings confirmed abnormalities in ventricular activation patterns, likely attributable to distal conducting system dysfunctions. Microscopic analysis of aged heart specimens revealed multifocal connective tissue deposition and perinuclear myocytolysis in the atria. Our results demonstrate that aging gradually modifies the terminal part of the specialized cardiac conducting system, creating a substrate for increased arrhythmogenesis. These findings may open new therapeutic options in the management of cardiac arrhythmias in the elderly population.     

Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design

Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors.