Beneficial Effect of H2S-Releasing Molecules in an In Vitro Model of Sarcopenia: Relevance of Glucoraphanin

Sarcopenia is a gradual and generalized skeletal muscle (SKM) syndrome, characterized by the impairment of muscle components and functionality. Hydrogen sulfide (H₂S), endogenously formed within the body from the activity of cystathionine-γ-lyase (CSE), cystathionine- β-synthase (CBS), and mercaptopyruvate sulfurtransferase, is involved in SKM function. Here, in an in vitro model of sarcopenia based on damage induced by dexamethasone (DEX, 1 μM, 48 h treatment) in C2C12-derived myotubes, we investigated the protective potential of exogenous and endogenous sources of H₂S, i.e., glucoraphanin (30 μM), L-cysteine (150 μM), and 3-mercaptopyruvate (150 μM). DEX impaired the H₂S signalling in terms of a reduction in CBS and CSE expression and H₂S biosynthesis. Glucoraphanin and 3-mercaptopyruvate but not L-cysteine prevented the apoptotic process induced by DEX. In parallel, the H₂S-releasing molecules reduced the oxidative unbalance evoked by DEX, reducing catalase activity, O₂⁻ levels, and protein carbonylation. Glucoraphanin, 3-mercaptopyruvate, and L-cysteine avoided the changes in myotubes morphology and morphometrics after DEX treatment. In conclusion, in an in vitro model of sarcopenia, an impairment in CBS/CSE/H₂S signalling occurs, whereas glucoraphanin, a natural H₂S-releasing molecule, appears more effective for preventing the SKM damage. Therefore, glucoraphanin supplementation could be an innovative therapeutic approach in the management of sarcopenia.     

Development and Biochemical Characterization of Self-Immolative Linker Containing GnRH-III-Drug Conjugates

The human gonadotropin releasing hormone (GnRH-I) and its sea lamprey analogue GnRH-III specifically bind to GnRH receptors on cancer cells and can be used as targeting moieties for targeted tumor therapy. Considering that the selective release of drugs in cancer cells is of high relevance, we were encouraged to develop cleavable, self-immolative GnRH-III-drug conjugates which consist of a p-aminobenzyloxycarbonlyl (PABC) spacer between a cathepsin B-cleavable dipeptide (Val-Ala, Val-Cit) and the classical anticancer drugs daunorubicin (Dau) and paclitaxel (PTX). Alongside these compounds, non-cleavable GnRH-III-drug conjugates were also synthesized, and all compounds were analyzed for their antiproliferative activity. The cleavable GnRH-III bioconjugates revealed a growth inhibitory effect on GnRH receptor-expressing A2780 ovarian cancer cells, while their activity was reduced on Panc-1 pancreatic cancer cells exhibiting a lower GnRH receptor level. Moreover, the antiproliferative activity of the non-cleavable counterparts was strongly reduced. Additionally, the efficient cleavage of the Val-Ala linker and the subsequent release of the drugs could be verified by lysosomal degradation studies, while radioligand binding studies ensured that the GnRH-III-drug conjugates bound to the GnRH receptor with high affinity. Our results underline the high value of GnRH-III-based homing devices and the application of cathepsin B-cleavable linker systems for the development of small molecule drug conjugates (SMDCs).     

Progress Towards an Acoustic Measurement of the Molar Gas Constant at INRIM

Progress in developing an experiment for the determination of the molar gas constant R and the Boltzmann constant k at INRIM is reported. The experiment involves simultaneous measurements of the acoustic and microwave resonance frequencies of a stainless steel spherical resonator for which its hemispheres were deliberately misaligned. For the present work, these frequencies were measured in helium near 273.16 K, in the pressure range from 100 to 800 kPa. From microwave data, the radius of the resonator was determined as a function of pressure with an estimated uncertainty of 6.0 ppm. Using acoustic data and the microwave determination of the resonator radius, the speed of sound in helium was deduced, and these values were compared with those predicted by recent accurate ab initio calculations. Over most of the pressure range, the present values agreed with the ab initio values within the uncertainty of the measurements (standard uncertainty of approximately 7.0 ppm). Many suggestions for reducing the uncertainty are provided.     

Supporting design via the System Operational Dependency Analysis methodology

In this paper, we introduce the system operational dependency analysis methodology. Its purpose is to assess the effect of dependencies between components in a monolithic complex system, or between systems in a system-of-systems, and to support design decision making. We propose a parametric model of the behavior of the system. This approach results in a simple, intuitive model, whose parameters give a direct insight into the causes of observed, and possibly emergent, behavior. Using the proposed method, designers, and decision makers can quickly analyze and explore the behavior of complex systems and evaluate different architecture under various working conditions. Thus, the system operational dependency analysis method supports educated decision making both in the design and in the update process of systems architecture, without the need to execute extensive simulations. In particular, in the phase of concept generation and selection, the information given by the method can be used to identify promising architectures to be further tested and improved, while discarding architectures that do not show the required level of global features. Application of the proposed method to a small example is used to demonstrate both the validation of the parametric model, and the capabilities of the method for system analysis, design and architecture.     

Cystic disease of the breast. Ten-year experience

Cystic breast disease is a relatively widespread pathological condition in the female sex, it has an incidence of around 7% and predominantly affects women aged between 40 and 50 years old. The authors report their experience based on the observation of 1046 cases taken from a series consisting of over 30,000 examinations. Patients were studied following a standardised diagnostic protocol including breast examination, breast scan and, depending on the patient's age or the presence of pathological findings, mammography. The protocol examines the distribution according to age, the number of cysts, their localization and diameter, as well as parameters concerning reproductive or sexual life, age at first pregnancy, the use of oral contraceptives, the number of abortions and menopausal status. Moreover, the efficacy of diagnostic tests, such as ecography, mammography, cytology and the assay of electrolytes using fine needle aspiration, was evaluated. The authors conclude that, when the patient follows a complete and valid diagnostic iter, breast cystic disease is an easily dominable pathology that can be simply controlled periodically without the need for therapeutic surgery.     

Mutation analysis of the RSK2 gene in Coffin-Lowry patients: extensive allelic heterogeneity and a high rate of de novo mutations

Coffin-Lowry syndrome (CLS) is an X-linked disorder characterized by severe psychomotor retardation, facial and digital dysmorphisms, and progressive skeletal deformations. By using a positional cloning approach, we have recently shown that mutations in the gene coding for the RSK2 serine-threonine protein kinase are responsible for this syndrome. To facilitate mutational analysis, we have now determined the genomic structure of the human RSK2 gene. The open reading frame of the RSK2 coding region is split into 22 exons. Primers were designed for PCR amplification of single exons from genomic DNA and subsequent single-strand conformation polymorphism analysis. We screened 37 patients with clinical features suggestive of CLS. Twenty-five nucleotide changes predicted to be disease-causing mutations were identified, including eight splice-site alterations, seven nonsense mutations, five frameshift mutations, and five missense mutations. Twenty-three of them were novel mutations. Coupled with previously reported mutations, these findings bring the total of different RSK2 mutations to 34. These are distributed throughout the RSK2 gene, with no clustering, and all but two, which have been found in two independent patients, are unique. A very high (68%) rate of de novo mutations was observed. It is noteworthy also that three mutations were found in female probands, with no affected male relatives, ascertained through learning disability and mild but suggestive facial and digital dysmorphisms. No obvious correlation was observed between the position or type of the RSK2 mutations and the severity or particular clinical features of CLS.     

Material Waste in Building Industry: Main Causes and Prevention

Material waste has been recognized as a major problem in the construction industry that has important implications both for the efficiency industry and for the environmental impact of construction projects. Moreover, waste measurement plays an important role in the management of production systems since it is an effective way to assess their performance, allowing areas of potential improvement to be pointed out. This paper describes the main results of two research studies carried out in Brazil that investigated the occurrence of material waste at 74 building sites located in different regions of that country. Some typical figures for the waste of some key construction materials are provided, and the main causes of waste in the sector are discussed. The results indicate that the waste of materials in the Brazilian building industry is fairly high and that a large variability in waste incidence is found across different projects. Most of this waste can be avoided by implementing inexpensive preventive measures, mostly related to managerial improvements.     

Heteroepitaxial growth of vertical GaAs nanowires on Si(111) substrates by metal-organic chemical vapor deposition

Epitaxial growth of vertical GaAs nanowires on Si(111) substrates is demonstrated by metal-organic chemical vapor deposition via a vapor-liquid-solid growth mechanism. Systematic experiments indicate that substrate pretreatment, pregrowth alloying temperature, and growth temperature are all crucial to vertical epitaxial growth. Nanowire growth rate and morphology can be well controlled by the growth temperature, the metal-organic precursor molar fraction, and the molar V/III ratio. The as-grown GaAs nanowires have a predominantly zinc-blende crystal structure along a <111> direction. Crystallographic {111} stacking faults found perpendicular to the growth axis could be almost eliminated via growth at high V/III ratio and low temperature. Single nanowire field effect transistors based on unintentionally doped GaAs nanowires were fabricated and found to display a strong effect of surface states on their transport properties.