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Background: Increasing evidence suggests that diabetes increases the risk of developing different types of cancer. Hyperinsulinemia, hyperglycemia and chronic inflammation, characteristic of diabetes, could represent possible mechanisms involved in cancer development in diabetic patients. At the same time, cancer increases the risk of developing new-onset diabetes, mainly caused by the use of specific anticancer therapies. Of note, diabetes has been associated with a ∼10% increase in mortality for all cancers in comparison with subjects who did not have diabetes. Diabetes is associated with a worse prognosis in patients with cancer, and more recent findings suggest a key role for poor glycemic control in this regard. Nevertheless, the association between glycemic control and cancer outcomes in oncologic patients with diabetes remains unsettled and poorly debated. Purpose: The current review seeks to summarize the available evidence on the effect of glycemic control on cancer outcomes, as well as on the possibility that timely treatment of hyperglycemia and improved glycemic control in patients with cancer and diabetes may favorably affect cancer outcomes.

Analog In-Memory Computing (AiMC) based on Non-Volatile Memories (NVM) is a promising candidate to reduce latency and power consumption of neural network (NN) inference in edge-computing applications. This kind of computational accelerators allows both storing weights and performing in-situ analog computation inside the array. This tutorial explores trade-offs and strategies in the design of DACs and ADCs for this kind of systems, highlighting the strong interdependence between the two converters. Starting from an analysis of input and weights encoding techniques this tutorial will then propose a discussion aiming at exploring critical aspects that constrain the design of D-A and A-D converters drawing some co-design considerations.

AbstractThe Italian National Pig Breeders Association (ANAS) manages the breeding programs of the Italian Large White (ILW), Landrace (IL), and Duroc (ID) breeds, mainly oriented to the production of PDO hams. ANAS evaluates the inclusion of genetic resistance in its breeding scheme. This study aimed to estimate variance components and breeding values (EBV) using threshold (TM) and linear (LM) models. During the sib test performed at the genetic station of ANAS from 1997 to 2021, 9,595 (respiratory diseases) and 12,046 (enteritis) diagnoses were collected by the veterinary. The trait was recorded as a dichotomous variable: affected animals with 1, whereas healthy with 2. A multi-breed model was applied with the breed, sex, and farm sector as fixed effects and litter, animal, and residual as random effects. The same model was also applied within each breed, removing the breed effect. The limited data size within single breed did not allow to estimate accurate variance components. With the multi-breed model, low heritabilities were estimated for respiratory (0.09) and enteritis (0.15), with TM and LM leading to the same values. The multibreed model led to more precise variance components estimation and to EBV very close to the ones from the single breed analyses. Pearson and Spearman rank correlations between EBV estimated with TM and LM in the multibreed scenario were ≥0.97. Results of this study demonstrated the feasibility of including disease resistance among the breeding goals of ANAS and that both TM and LM could be used to reach this objective.HighlightsGenetic selection for disease resistance in heavy pigs is possibleLow heritabilities estimated for enteritis and respiratory disordersThreshold and linear models gave similar results

The aim of the present experiments was to study the effect of nonselective electrolytic lesions of the rat dorsal hippocampus on 2 learning phenomena: the L. J. Kamin (1969) blocking effect and latent inhibition of taste aversion learning. Bilateral dorsal hippocampal lesions selectively impaired blocking induced by 1 saccharin-lithium chloride pairing previous to 1 serial compound (saccharin-cider vinegar)-lithium pairing, but lesions had no effect on latent inhibition of a saline aversion, induced by 6 saline preexposures, in the same group of animals. Moreover, dorsal hippocampal lesions did not affect latent inhibition of saccharin-conditioned aversion induced by 1 or 6 preexposures. It is argued that blocking and latent inhibition of taste aversion learning do not share a common neural mechanism.

With the proliferation of ultrahigh-speed mobile networks and internet-connected devices, along with the rise of artificial intelligence (AI)1, the world is generating exponentially increasing amounts of data that need to be processed in a fast and efficient way. Highly parallelized, fast and scalable hardware is therefore becoming progressively more important2. Here we demonstrate a computationally specific integrated photonic hardware accelerator (tensor core) that is capable of operating at speeds of trillions of multiply-accumulate operations per second (1012MAC operations per second or tera-MACs per second). The tensor core can be considered as the optical analogue of an application-specific integrated circuit (ASIC). It achieves parallelized photonic in-memory computing using phase-change-material memory arrays and photonic chip-based optical frequency combs (soliton microcombs3). The computation is reduced to measuring the optical transmission of reconfigurable and non-resonant passive components and can operate at a bandwidth exceeding 14 gigahertz, limited only by the speed of the modulators and photodetectors. Given recent advances in hybrid integration of soliton microcombs at microwave line rates3–5, ultralow-loss silicon nitride waveguides6,7, and high-speed on-chip detectors and modulators, our approach provides a path towards full complementary metal–oxide–semiconductor (CMOS) wafer-scale integration of the photonic tensor core. Although we focus on convolutional processing, more generally our results indicate the potential of integrated photonics for parallel, fast, and efficient computational hardware in data-heavy AI applications such as autonomous driving, live video processing, and next-generation cloud computing services.

The present work is a survey on aircraft hybrid electric propulsion (HEP) that aims to present state-of-the-art technologies and future tendencies in the following areas: air transport market, hybrid demonstrators, HEP topologies applications, aircraft design, electrical systems for aircraft, energy storage, aircraft internal combustion engines, and management and control strategies. Several changes on aircraft propulsion will occur in the next 30 years, following the aircraft market demand and environmental regulations. Two commercial areas are in evolution, electrical urban air mobility (UAM) and hybrid-electric regional aircraft. The first one is expected to come into service in the next 10 years with small devices. The last one will gradually come into service, starting with small aircraft according to developments in energy storage, fuel cells, aircraft design and hybrid architectures integration. All-electric architecture seems to be more adapted to UAM. Turbo-electric hybrid architecture combined with distributed propulsion and boundary layer ingestion seems to have more success for regional aircraft, attaining environmental goals for 2030 and 2050. Computational models supported by powerful simulation tools will be a key to support research and aircraft HEP design in the coming years. Brazilian research in these challenging areas is in the beginning, and a multidisciplinary collaboration will be critical for success in the next few years.

AbstractSpring-fed streams in Tokyo are important habitats for various aquatic species, whereas urbanization as well as introduction of invasive species is threatening the sustainability of such aquatic ecosystems. This study applies the System for Environmental Flow Analysis (SEFA) in a small urban river in Tokyo to assess the dynamics of the suitable habitats for the endangered freshwater fish Lefua echigonia(Jordan and Richardson 1907). A set of Habitat Suitability Curves (HSCs) for water depth, velocity and substrate was developed to evaluate the suitable habitats. The habitat assessment indicated that the Area Weighted Suitability (AWS) reached the maximum at 0.02 m3/s, which is close to the base flow of the target river; a gradual decrease in AWS was observed for higher flows. The temporal distribution of AWS, during forty-one consecutive months, showed that, on average, the best habitat conditions for adult L. echigoniaoccur during the period between January and July, whereas the worst situation occurs during the period between August and December. This work presents information and tools for instream habitat analysis that should help managers to conserve this aquatic species and prioritize actions to further rehabilitate urban rivers, using L. echigoniaas a case study.

In many modern data, the number of variables is much higher than the number of observations and the within-group scatter matrix is singular. Then, the Fisher's linear discriminant analysis (LDA) cannot be applied. The work considers a way to circumvent this problem by doing principal component analysis (PCA) enhanced with additional discriminating features. Two approaches are proposed: the original PCs are rotated to maximize the Fisher's LDA criterion, and second, penalized PCs are produced to achieve simultaneous dimension reduction and maximization of the Fisher's LDA criterion. Both approaches are illustrated and compared to other existing methods on several well known data sets. [ABSTRACT FROM AUTHOR]

Extensive density functional theory (DFT) calculations dedicated to analyze the stability, thermal behavior, as well as the infrared (IR) and Raman spectra of low hydroxylated C60(OH)12fullerenols are presented. Adsorbed configurations in which OH groups form various types of molecular islands on the carbon surface are the preferred atomic arrays, while random distributions of hydroxyl species are the highest energy molecular structures. It is found that the formation of local networks of hydrogen-bonded OH groups plays a fundamental role in the stability of these complexes. The calculated dipole moments, polarizability values, optical gap, and Fukui functions of C60(OH)12isomers strongly depend on the structure of the hydroxyl overlayer, thus being an important parameter to tune the material properties. DFT Born–Oppenheimer molecular dynamics calculations at T= 300 K reveal that aggregated forms of OH groups on the fullerene surface show an interesting dynamical behavior, characterized by a continuous proton-exchange process between neighboring hydroxyl molecules that modifies the structure and chemical nature of the molecular coating. From nudged elastic band studies analyzing OH diffusion on the C60surface, energy barriers opposing OH migration of ∼1 eV are found. However, in the presence of surrounding H2O species, a water-assisted diffusion process is obtained which can reduce the energy barriers to values as low as 0.25 eV. The comparison between experimental and calculated IR spectra of various C60(OH)12isomers shows well-defined spectral features which can be very helpful to identify the structure of these fullerene complexes. Finally, simulations of the wavelength-dependent Raman spectra of hydroxylated fullerenes reveals (i) that the intensities of the Raman active modes strongly depend on the excitation laser and (ii) the importance of the wavelength-dependent calculations to reveal precise features of different regions of the spectra. The combination of IR and Raman spectroscopies is an efficient approach to reveal the atomic structure of these sub-nanometer-sized carbon nanostructures.

La etapa final de la vida de personas con discapacidad intelectual es un aspecto poco conocido en nuestro entorno.

•Emphysematous gastritis is an entity with aspecific clinical manifestations : diagnosis is directed by a suspicion index.•Endoscopy is primordial to direct treatment strategy by determining the extent of necrosis and by revealing a potential microbial causal factor•Surgical treatment is necessary in case of medical treatment failure, extended gastric necrosis, severe sepsis or gastric perforation.•Clostridium ventriculiis a rare pathogen that can occur commensally but can also lead to emphysematous gastritis and gastric perforation when mucosal injury is present.•Chemotherapy agents are associated with gastrointestinal toxicity that should be identified and treated properly.

Measurements on vibrating structures has been a topic of interest for decades. Vibrating structures are however generally assumed to behave linearly and in a noise-free environment, which is not the case in practice. This paper provides a methodology that allows for the autonomous estimation of nonlinearities and assessment of uncertainties by bootstrap on a given vibrating structure. Nonlinearities are estimated by means of a block-oriented nonlinear model approach based on parallel Hammerstein models and on exponential sine sweeps. Estimation uncertainties are simultaneously assessed using repetitions of the input signal (multi-sine sweeps) as the input of a bootstrap procedure. Mathematical foundations and a practical implementation of the method are discussed using an experimental example. The experiment chosen here consists in exciting a steel plate under various boundary conditions with exponential sine sweeps and at different levels in order to assess the evolution of nonlinearities and uncertainties over a wide range of frequencies and input amplitudes.

We carried out a case–control study to investigate the possible role of occupational and environmental exposure to endocrine disruptors in the onset of testicular cancer (TC). We evaluated 125 TC patients and 103 controls. Seminal fluid examination and organochlorine analysis were performed in all subjects. Cases and controls were also interviewed using a structured questionnaire to collect demographic information, residence, andrological medical history and dietary information. We found that a higher level of reproductive tract birth defects was associated with a higher risk of TC. With regard to diet, cases reported a higher consumption of milk and dairy products than controls. Overall, there was a statistically significant increase in TC risk in cases with detectable values of total polychlorinated organic compounds against controls (14.4 vs. 1.0 %; p< 0.001). TC patients with detectable levels of organochlorines had lower mean semen parameters than those with undetectable levels, although this difference was not statistically significant. The International Agency for Research on Cancer recently included dioxin-like polychlorinated biphenyls (PCBs) in Group 1 of known human carcinogens. Our study confirmed and identified various risk factors for testicular cancer: cryptorchidism, consumption of milk and dairy products, parents’ occupation and serum concentration of hexachlorobenzene and PCBs and, for the first time, we showed the correlation between semen quality and the serum concentration of these pollutants.

The article presents a study on how varying animal housing and rearing conditions affect the Time to Integrate to Nest Test (TINT). It discusses the meaning of TINT and indicates the hypothesis stating that all mice, regardless of their lack of exposure to nesting material during fetal development and rearing will achieve positive TINT outcomes. Experimental conditions, measurement of outcomes and statistical analysis of data are explained.

The issue of chiral drug is now a major theme in the design, discovery and development of new drugs. It has been shown for many pharmaceuticals that only one enantiomer contains the desired activity, and the synthesis of such drug molecules in their optically pure form is becoming increasingly important. Mitsunobu reaction was carried out between (R)- and (S)-3,4-dihydro-2H-1,5-benzoxathiepin-3-ol and purines under microwave irradiation. A contraction into a six-membered ring takes place with concomitant inversion at the stereocentre with excellent enatiomeric excesses giving rise to the homochiral 9-(2,3-dihydro-1,4-benzoxathiin-3-ylmethyl)-9H-purines. The anti-tumour activity of all enantiomers is reported against the caspase-3-deficient MCF-7 and the wild type SKBR-3 human breast cancer cells. The most active homochiral compound displays an IC50 of 1.85 μM and induces inhibition of the translation initiation factor eIF2. All homochiral compounds included in this study show different apoptotic effects between both enantiomers with levels up to 99%. We have analyzed caspase-mediated apoptotic pathways on enantiomers and racemates. We have found a homochiral derivative that activates the canonical intrinsic caspase-8/caspase-3 apoptotic pathway on the MCF-7 cells, and a racemic compound that induces caspase-2 activation. Moreover, we demonstrate the involvement of caspase activation during cell death induced by these compounds in SKBR-3 cells.

During the last years, various novel anti-diabetic drugs have considerably enriched the therapeutic armamentarium for subjects with Type 2 diabetes. In the meantime, much interest has recently been focused on the potential cardiovascular and oncological adverse effects of these new therapies. As to glucagon-like peptide 1 (GLP-1) analogs, medullary thyroid tumors were reported to be more common in rodent toxicology studies with liraglutide, although the relevance of this finding in humans has been questioned. Analyses of sequential changes in calcitonin levels in several thousands of subjects did not reveal a relationship between liraglutide therapy and plasma calcitonin. Furthermore, no medullary thyroid cancer has been detected in humans taking liraglutide. Nevertheless, the long-term consequences of sustained GLP-1 receptor activation in the human thyroid remain unknown and deserve further investigation.

The control of cytosolic calcium is a major determinant not only of cardiac function, but also of the capability of myocardial tissue to survive damage. Increase of diastolic calcium leads rapidly to cell injury, and may be induced by a wide range of causes. In this review we describe the major points of calcium control in cardiac myocytes, mainly in mammalian ventricle, focusing on mechanisms of intracellular calcium influx during excitation, voltage gated channels of the sarcolemma and ryanodine receptors of the sarcoplasmic reticulum (SR), and efflux during relaxation, principally the sodium/calcium exchanger in membrane and the SR calcium complex. Mitochondria also depend on calcium concentration while also participating in its control. Moreover, we will outline receptor check points and their roles in physiology and pathology. We will focus on some new aspects of potential protective mechanisms that have been recently described and that involve peptide ligands and that in the case of the Neuregulin1beta/ErbB pathway are already reaching the clinical trial relevance.

Background:Correct histone/protamine replacement is an important stage in chromatin condensation during spermiogenesis in humans. There are two types of protamines: protamine 1 (P1) and the protamine 2 family (P2, P3, and P4), coded by the genes PRM1 and PRM2. Aim:We analyze the sequences and gene expression of PRM1 and PRM2 and their relationship with defective spermatogenesis. Materials and methods:Sequence analysis was carried out on 163 patients attending our laboratory for analysis of seminal fluid. Patients were divided into three groups: normozoospermic (53), teratozoospermic (60), and azoospermic (50). Gene expression was analyzed in seven patients with azoospermia and one with cryptozoospermia. Results:Seven single nuclotide polymorphisms (SNP) were identified: G54A, G102T and C230A for PRM1, and C246T, G288C, G298C and C373A for PRM2. For C230A, the CA genotype was present in 38% of teratozoospermic vs 55% of normozoospermic and 64% of azoospermic patients; for C373A, CA was found in 37% of teratozoospermic vs 47% of normozoospermic and 64% of azoospermic patients. In contrast, for G298C, GC was more common in the teratozoospermic (63%) than in the normozoospermic (49%) or azoospermic (48%) groups. These differences could suggest a greater susceptibility of these patients to abnormal sperm morphology. In five patients the levels of transcripts were reduced with respect to the control. Conclusion:These data suggest that premeiotic arrest is associated with extremely reduced protamine expression. New studies of both PRM1 and PRM2 and their mRNA expression could help us better understand the molecular mechanisms underlying the protamine transcription and translation processes.

The involvement of Chromogranin A (CgA) in the cardiovascular function regulation is attributed to its function as a prohormone. Several studies indicated that CgA-derived peptides, particularly Vasostatin-1 (VS-1) and Catestatin (CST), exert signaling effects in numerous organs/systems, including the cardiovascular system. This review focuses on the recently described signaling pathways activated by VS-1 and CST, giving insights into the mechanisms at the basis of their cardiac negative inotropic action, their vasodilator effects and their cardioprotective role observed in different experimental conditions. Accumulated evidences provided convincing support for VS-1 and CST as vasoactive peptides indirectly acting on cardiomyocytes through a Ca2-independent/PI3-K-dependent NO release from endothelial cells. This pathway is supposed to be triggered by the interaction of these peptides with the plasma membrane. The premise of these studies grounds on the biochemical features of VS-1 and CST, which are structurally characterized by amphipathic properties and the ability to interact with mammalian and microbial membranes. On the other hand, recent data obtained in both isolated heart and isolated cardiomyocytes suggest that the VS-1 and CSTmediated cardioprotective effects are primarily direct on the myocardium, rather than endothelium-dependent. Anyway, both direct and indirect pathways seem to be characterized by the absence of specific membrane receptors on target cells, highlighting intriguing novelties in the topic of cell signaling, in particular respect to an hypothetical receptor-independent eNOS activation.

Nitric oxide (NO), which is produced by oxidation of L-arginine to L-citrulline in a process catalyzed by different isoforms of nitric oxide synthase (NOS), exhibits diverse roles in several physiological processes, including neurotransmission, blood pressure regulation and immunological defense mechanisms. On the other hand, an overproduction of NO is related with several disorders as Alzheimer’s disease, Huntington´s disease and the amyotrophic lateral sclerosis. Taking melatonin as a model, our research group has designed and synthesized several families of compounds that act as NOS inhibitors, and their effects on the excitability of N-methyl-D-aspartate (NMDA)-dependent neurons in rat striatum, and on the activity on both nNOS and iNOS were evaluated. Structural comparison between the three most representative families of compounds (kynurenines, kynurenamines and 4,5-dihydro-1H-pyrazole derivatives) allows the establishment of structure-activity relationships for the inhibition of nNOS, and a pharmacophore model that fulfills all of the observed SARs were developed. This model could serve as a template for the design of other potential nNOS inhibitors. The last family of compounds, pyrrole derivatives, shows moderate in vitro NOS inhibition, but some of these compounds show good iNOS/nNOS selectivity. Two of these compounds, 5-(2-aminophenyl)-1H-pyrrole-2-carboxylic acid methylamide and cyclopentylamide, have been tested as regulators of the in vivo nNOS and iNOS activity. Both compounds prevented the increment of the inducible NOS activity in both cytosol (iNOS) and mitochondria (i-mtNOS) observed in a MPTP model of Parkinson´s disease.

Polychlorinated biphenyl (PCB) congeners differentially reduce serum thyroxine (T4) in rats, but little is known about their ability to affect biliary excretion of T4. Thus, male Sprague-Dawley rats were orally administered Aroclor-1254, Aroclor-1242 (32 mg/kg per day), PCB-95, PCB-99, PCB-118 (16 mg/kg per day), PCB-126 (40 μg/kg per day), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (3.9 μg/kg per day), or corn oil for 7 days. Twenty-four hours after the last dose, [125I]T4was administered intravenously, and blood, bile, and urine samples were collected for quantifying [125I]T4and in bile [125I]T4metabolites. Serum T4concentrations were reduced by all treatments, but dramatic reductions occurred in response to Aroclor-1254, PCB-99 [phenobarbital (PB)-type congener], and PCB-118 (mixed-type congener). None of the treatments increased urinary excretion of [125I]T4. Aroclor-1254, PCB-118, TCDD, and PCB-126 (TCDD-type congener) increased biliary excretion of T4-glucuronide by 850, 756, 710, and 573%, respectively, corresponding to marked induction of hepatic UDP-glucuronosyltransferase (UGT) activity toward T4. PCB-95 and PCB-99 did not induce UGT activity; therefore, the increased biliary excretion of T4-glucuronide was related to the affinity of congeners for the aryl hydrocarbon receptor. The disappearance of [125I]T4from serum was rapid (within 15-min) and was increased by Aroclor-1254, PCB-99 and PCB-118. Thus, reductions in serum T4in response to PCBs did not always correspond with UGT activity toward T4or with increased biliary excretion of T4-glucuronide. The rapid disappearance of [125I]T4from the serum of rats treated with PB-like PCBs suggests that increased tissue uptake of T4is an additional mechanism by which PCBs may reduce serum T4.

Large kindreds segregating familial Alzheimer disease (FAD) offer the opportunity of studying clinical variability as observed for presenilin 1 (PSEN1) mutations. Two early-onset FAD (EOFAD) Calabrian families with PSEN1Met146Leu (ATG/CTG) mutation constitute a unique population descending from a remote common ancestor. Recently, several other EOFAD families with the same mutation have been described worldwide.

The purine ring system is undoubtedly among the most ubiquitous of all the heterocyclic compounds. In recent years modified purine structures both of natural and synthetic origin have been a rich source of biologically active materials. The halogen at 6 position of the purine moiety of the (RS)-9 or 7-(2,3-dihydro-5H-1,4-benzodioxepin-3-yl)-9H-or 7Hpurines shows an interesting reactivity which is presented and discussed. The anticarcinogenic potential of the target molecules is reported against the MCF-7 cancer cell line.

There are some concerns about the use of Pseudomonas-based products as biocontrol agents because of the hemolytic activity shown by their metabolites. The effects of Pseudomonaslipodepsipeptides (LDPs) on mammals via ingestion and the LDP degradation during the digestion and intestinal permeability have not been evaluated. In this research, the susceptibility of different LDPs to degradation was assayed with enzymatic gastrointestinal digestion, and intestinal permeability to LDPs was investigated in an in vitro system based on an intestinal cell layer system. Results demonstrated that trypsin and chymotrypsin hydrolyze up to 50% of the various LDPs, and that proteolysis was further increased by pronase E treatment. A decrease in LDP hemolytic activity matched LDP degradation during the various steps of the digestion process. Moreover, it was shown that syringomycin E (SRE), the main known LDP, was not able to cross the intestinal cell layer, suggesting that SRE does not reach the bloodstream in vivo. It was concluded that the Pseudomonas-based biocontrol products do not represent a serious risk for consumer health. In fact, LDPs possibly present on biocontrol-treated agricultural commodities would likely be partially digested by gastrointestinal enzymes and would not be absorbed at the intestinal level.

Abstract: A 3-year-old male chamois (Rupicapra rupicapra) shot during a harvest plan in Piedmont (Italy) presented periocular alopecic and thickened crusty lesions, some of which slightly red in colour. Hair still present was broken and easily removed. Direct microscopic examination of the pathological material collected by skin scraping led to the diagnosis of dermatophytosis, as the hair shafts appeared invaded by unstained spherical spores (arthroconidia). Fungal growth was obtained by culturing hair and crusts on thiamine/inositol enriched Sabouraud’s medium at 37°C. The macro- and microscopic characteristics of the organism were typical of the dermatophyte Trichophyton verrucosum. Wild ruminants are rarely affected by dermatophytosis, whereas in cattle, sheep and goats, infection because of this dermatophyte is quite common. This seems to be the first case of infection by T. verrucosum in chamois.

The identification of the molecular components involved in the aberrant processes that control proliferation, differentiation and apoptosis, is necessary for the development of chemotherapeutic interventions to restore or to destroy selectively the transformed cells. The discovery of new chemotherapeutic agents is probably one of the most reliable ways to improve our success against cancer, and intelligent drug design is a key factor to achieve this goal. Thus, the identification of novel targets for anticancer drug discovery is needed. Here we provide evidence that choline kinase (ChoK) is a novel target for the design of antitumor drugs. In this review we present the evolution of ChoK inhibitors by using the Hansch approach, starting from hemicholinium-3 (HC-3) as a lead compound. To start with we synthesized and evaluated ten bis-quaternary derivatives, in which the modifications affect both the spacer and the two cationic heads of the prototype. In the second phase 56 biscationic dibromides with distinct polar heads [bis(4-substituted)pyridinium, bis(4-substituted)quinolinium, and bisisoquinolinium moieties] and several spacers were synthesized and assayed for biological activity. This oriented synthesis produced 45 inhibitors of ChoK with antitumor activity against the HT-29 cell line. Finally, 40 bisquinolinium compounds were prepared and the corresponding QSAR equation was obtained for the whole set of compounds for the antiproliferative activity, the electronic parameter σR of R4, the molar refractivity of R8, and the lipophilic parameters clog P and linker. The most potent antiproliferative agent so far described shows IC50 0.20 M, while its theoretical value is 0.45 M.

Frontotemporal dementia (FTD) in several 17q21-linked families was recently explained by truncating mutations in the progranulin gene (GRN).

Most of the signal transduction pathways are mediated by protein kinases regulating every aspect of cell function. Mutations which deregulate their expression or their function or both result in cancers. Therefore, protein kinase inhibitors have become the focus of development of new therapies for cancer. A comprehensive review of Choline kinase (ChoK) was published by us in 2003. Since then, molecular information of ChoK inhibitors has been accumulated. In this review, we intend to summarize the new lines of evidence that will include the design of the most active antiproliferative agents so far described against ChoK. Studies have been aimed at the establishment of structure-activity relationships and the structural parameters that define ChoK inhibitory and antiproliferative activities of a set of twenty-five acyclic biscationic pyridophane and forty acyclic biscationic quinolinephane compounds. The corresponding QSAR equation was obtained for the whole set of bisquinolinium compounds for the antiproliferative activity, taking into consideration the electronic parameter R of R4, the molar refractivity (MR) of R8, and the lipophilic parameters clog P and linker. The most potent antiproliferative agent shows an IC50 0.45 M, predicted by the QSAR equation, whilst its experimental value is IC50 0.20 M. Finally, toxicity assays were performed for the most promising compounds because of their interesting antiproliferative activities [IC50 HT-29 0.70, 0.80, 1.50 and 1.90 M] and low toxicity [LD50 16.7, 12.5, > 25 and > 20 g/kg of mouse]. These biological activities justify further analysis for antitumoral assays under in vivo conditions.

Objectivesand study design: Increasing evidences support an inflammatory origin for gestational hyperglycemia. This paper aims at investigating, cross-sectionally and prospectively, the relationships between tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) levels in normoglycemic and hyperglycemic pregnancies of women with and without conventional risk factors for gestational diabetes (GDM). Results: Both at simple and multiple correlations TNF-α levels are associated to fasting insulin, homeostasis model assessment-insulin resistance (HOMA-IR) values and gestational hyperglycemia, while high sensitivity CRP (hsCRP) levels to body mass index (BMI). Furthermore, the TNF-α levels of the second trimester and their increments in the third trimester are significant predictors of insulin levels measured at 32–36 weeks in the subgroup of hyperglycemic women with ≤35 yr, BMI <25 kg/m2and the absence of a first-degree relative with Type 2 diabetes (respectively, β=1.1; 95%CI 0.66–1.48; p=0.002 and β=1.0; 95%CI 0.36–1.66; p=0.02), in a multiple regression model, after multiple adjustments. In a second cohort of women at low risk for GDM (<25 yr, BMI <25 kg/m2and absence of a first-degree relative with Type 2 diabetes), 24–28 weeks TNF-α levels are highly associated with corresponding insulin and HOMA values in the same model (respectively, β=0.27; 95%CI 0.11–0.43; p=0.001 and β=0.30; 95%CI 0.14–0.46; p<0.001). Conclusions: the data support the developing hypothesis that low-grade systemic inflammation is associated to GDM, in particular for pregnant women without conventional risk factors for gestational hyperglycemia, whose insulin resistance seems less explainable.

Studies have been aimed at the establishment of structure−activity relationships that define choline kinase inhibitory and antiproliferative activities of 40 bisquinolinium compounds. These derivatives have electron-releasing groups at position 4 of the quinolinium ring. It is found that the enzymatic inhibition is closely related to the size of the linker, the 3,3‘-biphenyl moiety being the most suitable. On the other hand, the antiproliferative activity against the HT-29 cancer cell line is less influenced by the linker type and by substituent R4. The corresponding QSAR equation was obtained for the whole set of compounds for the antiproliferative activity, the electronic parameter σR of R4, the molar refractivity of R8, and the lipophilic parameters clog P and πlinker. The most potent antiproliferative agent so far described is 40 for which an IC50 = 0.45 μM was predicted by the QSAR equation, while its experimental value is IC50 = 0.20 μM.

Peptides corresponding to three alpha helices present in the C-terminal region of the human prion protein have been synthesized and their structural autonomy analyzed by circular dichroism (CD) and NMR spectroscopy. The results obtained indicate that the protein fragment corresponding to the α3-helix, in contrast to α1 and α2 peptides, shows a complete structural autonomy. The chemical shifts values found for NH and CHa resonances of the isolated α3 peptide, formed by 30 aminoacid residues, were markedly and surprisingly similar to the corresponding values of the α3-helix in the protein. The structural autonomy of the α3-helix is profoundly determined by the presence of the conserved capping box and, in part, by the ionic bond formed between Glu200 and Lys204. On the basis of these observations a novel PrP consensus pattern, centered on the α3-helix region, has been defined. The data indicate that this autonomous and highly conserved region of the PrPclikely plays a critical role in folding and stability. This gives an explanation of why many of pathogenic mutations occur in this part of the molecule, sharing relevant effects on the overall protein conformation. In particular the D202N capping mutation almost completely destabilizes the isolated α3 peptide. While it is well known that the D202N substitution is associated with a GSS disease, the possible structural basis of this fatal pathology has never been investigated. We propose that a lower α3-helical propensity leading to a major destabilization of the PrPcmolecule initiates the pathogenic process associated with D202N capping mutation.

In an attempt to find new compounds with neuroprotective activity, we have designed, synthesized and characterized 19 new nNOS inhibitors with a 4,5-dihydro-1H-pyrazole structure. Compounds 11r [1-cyclopropanecarbonyl-3-(2-amino-5-chlorophenyl)-4,5-dihydro-1H-pyrazole] and 11e [1-cyclopropanecarbonyl-3-(2-amino-5-methoxyphenyl)- 4,5-dihydro-1H-pyrazole] show the highest activities with inhibition percentages of 70% and 62%, respectively. A structure−activity relationship for the nNOS inhibition can be established from the structural comparison of these new pyrazole derivatives and the described synthetic kynurenines 10.

Studies have been aimed to establish the structure−activity relationship that define choline kinase (ChoK) inhibitory potency and antiproliferative activity of a set of 25 bispyridinium compounds with electron-releasing groups at position 4. Here we report that, according to their inhibitory activities against human ChoK, the enzymatic inhibitory potency is closely related to the size of the linker, the 3,3‘-biphenyl moiety being the most suitable. The N-methylanilino and its derivatives, 4-chloro-N-methylanilino and 3,5-dichloro-N-methylanilino, render higher ChoK inhibitory and antiproliferative activities against the HT-29 human colon cancer cell line.

Several investigators have suggested that certain hydroxylated metabolites of 17β-estradiol (E2) are the proximate carcinogens that induce mammary carcinomas in estrogen-sensitive rodent models. The studies reported here were designed to examine the carcinogenic potential of different levels of E2and the effects of genotoxic metabolites of E2in an in vivomodel sensitive to E2-induced mammary cancer. The potential induction of mammary tumors was determined in female ACI rats subcutaneously implanted with cholesterol pellets containing E2(1, 2, or 3 mg), or 2-hydroxyestradiol (2-OH E2), 4-hydroxyestradiol (4-OH E2), 16α-hydroxyestradiol (16α-OH E2), or 4-hydoxyestrone (4-OH E1) (equimolar to 2 mg E2). Treatment with 1, 2, or 3 mg E2resulted in the first appearance of a mammary tumor between 12 and 17 weeks, and a 50% incidence of mammary tumors was observed at 36, 19, and 18 weeks respectively. The final cumulative mammary tumor incidence in rats treated with 1, 2, or 3 mg E2for 36 weeks was 50%, 73%, and 100% respectively. Treatment of rats with pellets containing 2-OH E2, 4-OH E2, 16α-OH E2, or 4-OH E1did not induce any detectable mammary tumors. The serum levels of E2in rats treated with a 1 or 3 mg E2pellet for 12 weeks was increased 2- to 6-fold above control values (~30 pg/ml). Treatment of rats with E2enhanced the hepatic microsomal metabolism of E2to E1, but did not influence the 2- or 4-hydroxylation of E2. In summary, we observed a dose-dependent induction of mammary tumors in female ACI rats treated continuously with E2; however, under these conditions 2-OH E2, 4-OH E2, 16α-OH E2, and 4-OH E1were inactive in inducing mammary tumors.

A series of quinoline derivatives as aza analogues of the naphthalene chromophore and a series of “nonfused” tricyclic aromatic systems, in particular 5-arylquinolines and 5- or 6-aryl and heteroaryl naphthalene systems, were synthesized and evaluated for growth-inhibitory properties in several human cell lines. The analysis of quantitative structure−antitumor activity relationships for the growth-inhibitory properties is also reported. Findings suggest that these compounds may not express their cytotoxicity via interaction on DNA.

The synthesis and biological activities of four novel bispyridinium cyclophanes as choline kinase (ChoK) inhibitors are presented. Their synthetic methodology has been optimized according to dilution, temperature, and reaction time and provides pure bispyridinium cyclophanes in high yields very easily. One of these cyclophanes (6, 4,8-diaza-3(1,4),9(4,1)-dipyridina-1(1,4),6(1,3)-dibenzenacyclodecaphan-3¹,9¹-bis(ilium) dibromide) has an IC50(ChoK) of 0.3 μM and is the most potent human ChoK inhibitor described to date.

The design, synthesis, characterization, DNA binding properties, and cytotoxic activity of a novel series of hybrids, namely, a molecular combination of the natural antibiotic distamycin A and the antineoplastic agent uramustine, are reported, and the structure−activity relationships are discussed. This homologous series 29−34 consisted of the minor groove binder distamycin A joined to uramustine (uracil mustard) by suitable aliphatic carboxylic acid moieties containing a flexible polymethylene chain that is variable in length [(CH2)n, where n = 1−6). All the hybrid compounds in this series exhibit enhanced activity compared to both distamycin A and uramustine derivatives 22−27 used for conjugation, giving IC50 values in the range 7.26−0.07 μM following a 1 h exposure of human leukemic K562 cells, with maximal activity shown when n = 6. The distance between the uramustine and distamycin frame is crucial for the cytotoxicity, with compounds having linker lengths of four to six being at least 20-fold more cytotoxic than liker lengths one to three. Taq polymerase stop experiments demonstrated selective covalent binding of uramustine−distamycin hybrids to A/T rich DNA sequences, which was again more efficent with compounds 32−34 with a longer linker length. Two consequences can be derived from our study:  (a) the distamycin moiety directs binding to the minor groove of A/T rich DNA sequences and, consequently, is responsible for the alkylation regioselectivity found in footprinting studies; (b) the higher flexibility due to a longer linker between the distamycin and uracil moieties allows the formation of complexes with the mustard moiety situated more deeply in the minor groove and, hence, with better alkylating properties.

Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS®can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a control cell line not expressing the antigen. Antibodies binding only to the transfected cell line are then selected for further analysis. This approach can be challenging if a large number of antibodies need to be screened and the antibody quantities are limited. Here we describe a novel method that combines the above two steps of FACS screening into a single step, based on the use of two fluorochromes, CMFDA and CMTMR, to stain transfected and control cell lines, respectively. Antibodies conjugated to a third fluorochrome are then added to the combined cells, followed by three-color FACS analysis. The newly modified FACS method is simple, sensitive, and high throughput. It can be used for antibody screening in multiple cell lines simultaneously.

In the present study we have investigated whether pharmacophore models may account for the activity and selectivity of the known cyclooxygenase-2 (COX-2) selective inhibitors of the phenylsulfonyl tricyclic series, i.e., Celecoxib (1) and Rofecoxib (3), and whether transferring this structural information onto the frame of a nonsteroidal antiinflammatory drug (NSAID), known to tightly bind the enzyme active site, may be useful for designing novel COX-2 selective inhibitors. With this aim we have developed a pharmacophore based on the geometric disposition of chemical features in the most favorable conformation of the COX-2 selective inhibitors SC-558 (2; analogue of Celecoxib (1)) and Rofecoxib (3) and the more restrained compounds 4 (DFU) and 5. The pharmacophore model contains a sulfonyl S atom, an aromatic ring (ring plane A) with a fixed position of the normal to the plane, and an additional aromatic ring (ring plane B), both rings forming a dihedral angle of 290° ± 10°. The final disposition of the pharmacophoric groups parallels the geometry of the ligand SC-558 (2) in the known crystal structure of the COX-2 complex. Moreover, the nonconserved residue 523 is known to be important for COX-2 selective inhibition; thus, the crystallographic information was used to position an excluded volume in the pharmacophore, accounting for the space limits imposed by this nonconserved residue. The geometry of the final five-feature pharmacophore was found to be consistent with the crystal structure of the nonselective NSAID indomethacin (6) in the COX-2 complex. This result was used to design indomethacin analogues 8 and 9 that exhibited consistent structure−activity relationships leading to the potent and selective COX-2 inhibitor 8a. Compound 8a (LM-1685) was selected as a promising candidate for further pharmacological evaluation.

The overstimulation of the N-methyl-d-aspartate (NMDA) subtype of glutamate receptors is involved in excitotoxicity, a process participating in neurodegeneration that characterizes some neurological disorders and acute cerebral insults. In looking for compounds with neuroprotective properties, a series of kynurenine derivatives were synthesized, and their effects on both the NMDA and nNOS activity in rat striatum were evaluated. Two compounds, 15a (2-acetamido-4-(2-amino-5-methoxyphenyl)-4-oxobutyric acid) and 15c (2-butyramido-4-(2-amino-5-methoxyphenyl)-4-oxobutyric acid), displayed more potent activities than the other synthetic compounds tested for the inhibition of NMDA excitability and nNOS activity. Two other compounds, 18a (2-acetamido-4-(3-methoxyphenyl)-4-oxobutyric acid) and 18c (2-butyramido-4-(3-methoxyphenyl)-4-oxobutyric acid), that have the same structure as 15a and 15c, except the amino group in R1, showed different effects. Whereas compound 18a showed lower electrophysiological potency than compounds 15a and 15c in the inhibition of the NMDA-dependent excitability, compound 18c showed the opposite effect. Moreover, compounds 18a and 18c were unable to modify nNOS activity. The remaining kynurenines tested behave like compound 18a. These results suggest that a structure-related activity of these synthetic kynurenines and a N-H bond in a specific direction is necessary for some kynurenine analogues to inhibit nNOS activity.