Monthly Archives: August 2017

Previous studies have also focused

Previous studies have also focused on the effect of a generalized shutoff, or when specific, virus-induced transcription shutoff on induction of IFN-α/β responses (Fros et al., 2013; Frolov et al., 2012), while the role of translation shutoff in antiviral state antagonism has not been emphasized. For SINV, both transcription and translation shutoff are induced by the same protein (Gorchakov et al., 2005), and the relative contribution of these functions in resisting the antiviral state is difficult to explore. Similarly, most previous work with VEEV or EEEV has implicated pgc-1α inhibitor induced transcription shutoff to play a major role in suppression of IFN-α/β induction, despite the temporally deferred synthesis of this viral protein during infection (Atasheva et al., 2010; Atasheva et al., 2015). Induction of host translation shutoff by VEEV has been localized to the nonstructural protein region of the genome, which is translated before the capsid region during infection (Yin et al., 2009), suggesting a role for this activity in the antiviral state resistance of VEEV.
Here we have examined possible mechanisms underlying resistance of VEEV to an IFN-α/β induced, pre-established antiviral state and identified/confirmed the proteins that mediate host transcription and translation shutoff with CHIKV, SINV, VEEV and EEEV through individual protein expression. In in vitro testing, VEEV was more resistant than SINV, CHIKV and EEEV to the global antiviral state in mouse and human cells, and this resistance became evident at a point after initial translation of the incoming virus genome. Furthermore, a panel of mutant viruses deficient in host macromolecular synthesis shutoff demonstrated that sensitivity to the antiviral state was correlated with slower rates of this activity. Using a plasmid expression system to study host macromolecular synthesis shutoff independent of virus replication rates, we found that expression of VEEV, CHIKV or SINV nsP2, or VEEV or EEEV capsid expression, but not control nsPs or GFP was sufficient to block host translation, with VEEV and EEEV capsid translation blockade likely secondary to transcription shutoff. VEEV and EEEV nsP2 did not inhibit transcription. VEEV or EEEV capsid and CHIKV or SINV nsP2 expression directly inhibited host transcription. EEEV nsP2 also failed to block host translation revealing a stark difference between VEEV and EEEV. Importantly, in the absence of transcription shutoff, host translation in IFN-primed cells was inhibited more efficiently by VEEV nsP2 than that of SINV nsP2. Furthermore, when VEEV nsP2 was expressed in IFN-primed cells, levels of ISG\’s were lower, and replication of an unrelated IFN-sensitive virus (yellow fever virus 17−D) was enhanced over IFN-primed control cells. Overall, we conclude that VEEV nsP2-induced host translation shutoff early after infection downregulates the antiviral state by decreasing levels of ISG\’s and creating an environment more permissive for viral replication.

Materials and methods




Viruses must maintain particle stability in order to survive in the environment and to carry out their replication cycles within hosts. Thermal fluctuations are some of the main environmental perturbations faced by viruses, and are particularly relevant for mammalian viruses, which are subject to periodic increases in temperature during febrile episodes. Similarly, thermostability is important for phages that infect thermophilic microbes (Dellas et al., 2014). Experimental data also suggest that thermal adaptation plays an important role in the evolution of arboviruses, which alternate between vectors and hosts whose temperatures can be quite disparate (Novella et al., 1999). More recently, climate change may be increasing the extent of selection for thermostability in viruses and their hosts (Cox et al., 2010). The ability of a virus to develop stability under thermal selection depends on its evolutionary history (Alto and Turner, 2010). From an applied point of view, thermostability is highly desirable in vaccines and viral vectors that may be used therapeutically (Ashcroft et al., 2005; Hwang and Schaffer, 2013; Porta et al., 2013; Rincon et al., 2014).

The first finding revealed by

The first finding revealed by my empirical inquiry was that the share of ethnic Muslims in the overall buy tranylcypromine of Russia is often overestimated. According to the 2002 national census, they constituted 9.93 percent of the population. Table 1 lists the ethnic Muslim groups of Russia in the descending order of their numeric sizes. An additional piece of information provided in the table refers to the shares of ethnic Muslim populations residing in their ethnic homelands, republics. This information is useful for the assessment of the territorial dispersion of ethnic Muslins, as well as of some political aspects to be discussed below. Twelve groups (Crimean Tatars, Dunguns, Karakalpaks, Khemshils, Meskhetin Turks, Middle-Asian Arabs, Middle-Asian Gipsies, Persians, Shapsugs, Talysh, Tats, Uyghurs), each of them constituting less then 0.01 percent of the overall population, are joined together under the rubric of Others. It is important to note that for all other calculations reported in this study, they were counted separately. The republic of Dagestan is officially a homeland for 10 groups (Aguls, Avars, Dargins, Kumyks, Laks, Lezghins, Nogai, Rutuls, Tabasarans, and Tsakhurs), none of which constituting a majority. Kabardino-Balkaria and Karachaevo-Cherkesia are homelands for two groups each, Balkarians and Kabardinians, and Cirkassians and Karachai, respectively. Thus of the 31 groups for which the data are given in the table, 19 do have their ethnic homelands in the Russian Federation, even though some of them are shared, while the remaining 12, with the only exception of Abazins, have their majorities residing in foreign countries. This means that the Soviet-time nationality politics were quite efficient in supplying every significant ethnic Muslim group with its own ‘statehood’.
As follows from the data, the vast majority of ethnic Muslim populations of Russia reside mostly in those regions where they are ‘titular nationalities’. The most salient exception is, of course, provided by Tatars. While Tatars are by far the largest ethnic Muslim group in Russia, almost two thirds of them live outside of their ethnic homeland, Tatarstan. Then saprophytes is not surprising that Tatars constitute the largest ethnic Muslim minorities in the vas majority of Russia\’s regions. Yet it is also important to note that a large part of Tatars live in a neighboring republic, Bashkortostan. The data on the ethnic compositions of the Russian regions are provided in Table 2. The regions are listed in the descending order of the percentage shares of their ethnic Muslim populations. Individual ethnic groups are listed only if they constitute more than 1 percent of the overall population. However, the largest ethnic Muslim minorities are listed irrespective of their relative size. The words ‘autonomous district’ are abbreviated to AD, and ‘autonomous province’, to AP.

How many ethnic Muslims are there in regional legislative assemblies?
The reason for taking December 2003 as a starting point for my inquiry was institutional. Starting with that time, the federal law made it imperative for the regions to elect no less than a half of their assembly deputies by a system of proportional representation. This makes the whole set of cases internally homogeneous, buy tranylcypromine and the cases themselves, comparable (Golosov, 2006), which will have especially important consequences for the next section of this analysis. At this point, however, certain clarifications regarding the institutional framework of the observed elections are in order. Each of the 95 elections included a proportional component, and ten of them were held exclusively by proportional formulas. Sverdlovsk province uses a system of staggered elections, with a half of its assembly being elected each two years. I counted each of these elections separately. The average size of the elected assembly was 45.08, while the average size of the proportional part was slightly more than a half of that, 24.18. To understand this, it is important to take into account that mixed-system assemblies tended to be larger than those elected by pure proportional systems.

Between and Russia received more than million people of which

Between 1990 and 2003, Russia received more than 10 million people, of which more than half were ethnic Russians (60%) or belonged to peoples with national autonomous status in Russia (Tatars, Bashkirs, etc.). Among Central Asian republics Tajikistan became the first to emigrate the ethnic Russians because of the difficult conditions caused by the Civil War there; followed by Kyrgyzstan where the transition to a market economy initiated by Bishkek impoverished rural areas resulted in a considerable loss of the ethnic Russians. Disintegration of the Union was the major reason itself for UNC2025 of people from one area to other.
Magnitude of migration
Alone in 1990s more than half of the Russians out migrated from Central Asia to Russia and by now more than 4.5 million people have out migrated from Central Asia and most of them were Russians (70%). From 1989 to 2002 the number of those Central Asian nationals who settled down in Russia legally and permanently rose from 882,000 to 963,000.
Numerically Kazakhstan ranks highest in emigration of non-natives in the entire Commonwealth of Independent States. From Kazakhstan 728,000 Slavs left between 1989 and 1996 and by 2006 there were less than 4 million Russians living there and many more, about 47.4%, were willing to leave Kazakhstan. This out migration in Uzbekistan was mostly of Slavs rather than of Russians as 300,000 Germans, left Kazakhstan in 1993.
Tajikistan and Kyrgyzstan experienced large scale out-migration of Russians as compared to the remaining states of Central Asia. From Tajikistan almost 100,000 Russians left soon after February 1990 when in Dushanbe violent confrontations occurred. By the end of 1992 some 150,000 Russian and other Russian speaking people (Ukrainian, Korean, German) left Tajikistan. Before the eruption of Civil War in Tajikistan, there were about half a million Russian and Russian speakers in Tajikistan, but by 1996 only half of them remained. Tajik Civil War compelled many Russians and other Europeans to emigrate out of its fear and as a result Russians who contributed 7.6% (388,000) in 1989 in Tajikistan were mere 1.1% (68,200) by 2000.
The number of Russians in Kyrgyzstan fell from 916,500 in 1989 to 720,000 in 1995. This number further decreased to 603,000 in 1999 and to approximately 500,000 in 2006. Between 1990 and 1994, 200,000 Russians out-migrated from Kyrgyzstan. Here the primary reason for the out-migration was economic because a significant number of Russians worked there in agriculture. Following table expresses the situation that arose in Central Asia from 1990 onwards.
Uzbekistan alone lost 170,000 Russians in 1992–1993 and 200,000 in 1993–1994. Between 1992 and 1996, Central Asia accounted 59% of its net migrations to Russia, of which 25% was from Uzbekistan alone.
Turkmenistan in 1989 contained 9.5% Russians (334,000) which in 1995 had shrank to 6.7% and in 2006 it had further decreased to just 2% of the total population.

Disintegration of USSR
There was internal conflict or rather inherent conflict in the policies of USSR. Since USSR was framed as an ideological state based on non-religiosity of communism yet in spite of vigorous campaign of the state many times it failed. Similarly Socialism based on Marxism could not develop the way it should have as political order remained in few hands during the 70 years of the life of the Union. Accordingly the common people were not part of those who were the “decision-makers” and social developments, therefore, were found to be suffering as compared to the earlier years of the history of the Union. It was also because the economic depression was found in the Union after 1970s when economic growth was not at par with that of markets outside. This slide saw people finding it difficult to stay in jobs they were engaged earlier. Accordingly many industries started to become sick forcing professionals and labourers to shift from one place to other giving rise to migration within the state.

The role of petro diplomacy was glaringly

The role of ‘petro-diplomacy’ was glaringly obvious in the negotiations on the new treaty. In fact, the treaty talks had begun when Yanukovych sent his Prime Minister and Energy Minister to Moscow to ask for a cut in gas prices (Big, 2010). Moscow decided to reward Yanukovych for his loyalty by cutting the price the Ukraine would pay for Russian natural gas by thirty percent for the next 10 years (a benefit which could save the Ukraine about $40 billion) (Barry, 2010). Including this provision openly in the new agreement was galling to the Orange forces, who felt that their country was being sold out for the proverbial ‘thirty pieces of silver.’ They protested vehemently in the Ukrainian Rada (Parliament), pelting the Speaker with eggs and smoke bombs. Despite this, Yanukovych\’s party was able to rush through ratification of the Kharkov Agreement only six days after it was signed (Protests, 2010). Russia\’s decision to directly link the NIBR189 lease and gas prices, while perhaps crass, fits the classic theory of economic sanctions: if the target state concedes, sanctions can quickly become economic incentives, to reward the target\’s ‘improved’ behavior.
All in all, the result of the 2010 elections and the concessions made by the new government make Russia\’s policy of energy sanctions look quite successful in this case. Of course, many in the West—and in Ukraine—are critical. Even in Russia, some opposition figures have questioned the policy. For example, Boris Nemtsov of the Union of Right Forces criticized the Kharkov Agreement for costing Russia an estimated $40 billion over 10 years. The main reason for maintaining the Sevastopol base, he argued, was the war with Georgia. Was keeping Georgia in check, he asked, really worth $40 billion? (Boris, 2010). However, it could be argued that Nemtsov\’s view was too narrow: the Kremlin was not paying that amount just to maintain its naval base. It was paying to maintain a pro-Russian President in Ukraine, which resulted (in the Kremlin\’s view) in a fundamental improvement in Russia\’s geopolitical position in the world.

The relevance of this conclusion for real-world politics is clear. The U.S. and others are constantly using various forms of economic leverage. Often, as for example with the sanctions imposed against Iran, the state targeted has some sort of democratic or quasi-democratic system. In Iran there have often been important contested elections between anti-Western fundamentalists and reform candidates. This paper has shown that sanctions do not have to force the country to collapse: they just have to cause enough economic ‘pain’ to force the Islamic hard-liners to lose elections. This is an important point: a major criticism of sanctions has been that if they are designed to target a country\’s entire economy, they can cause a humanitarian disaster, as took place in Iraq from 1991 to 2003, when harsh sanctions targeted Saddam Hussein\’s regime. Yet this paper argues that sanctions do not have to bring economic collapse to be effective; they may only have to NIBR189 lower a target state\’s GNP by a few percent, enough to cause a leader to lose an election.
Alternatively, as the EU has shown in its relations with countries like Serbia, states can follow the more positive tactic now being used by the Russians in Ukraine: rewarding favored leaders by boosting their states\’ economies to try to help them win elections. Again, this could potentially be a relatively ‘cost-effective’ tactic. One does not have to totally rebuild a country or flood Telomerase with billions of dollars in aid–just boost the target state\’s economy by a crucial few percent when an election is near. In either case, as this paper has attempted to show, such tactics can potentially be effective. Ideally, future research will be able to show that they can be effective in measurable and predictable ways.

tubocurarine cost br From China to Rome from Rome to China a

From China to Rome – from Rome to China: a short focus on Silk and Silk Roads
We cannot conceive describing the trade relationship between Rome and the Far East without considering silk, one of the most legendary goods of the Eurasian networks (Hildebrand, 2016: non vidi). If it tubocurarine cost is true that the Romans tended to identify goods with their supposed places of origin (for example pepper is associated with India, etc.), this identification is particularly evident in the case of China: the Latin word Seres used to designate the Chinese people. The same word was used as an adjective to identify the silk: sericum ‘silk’, serica meant ‘silk garments’ or also ‘from the land of the Seres’. Finally the word sericum in the plural form serici was also a noun for ‘merchants of silk’.
It is only at the beginning of the Roman Empire that this material becomes a widespread luxury good (Thorley, 1971). Hardly a coincidence then to find silk mentioned in the poems of Martial –the poet of the Roman ‘daily life’– who, at the end of the 1st century CE, speaks of silk products in Rome, as such pillows or clothes, to convey an image of wealth and sophisticated lifestyle.
As far as the economic evaluation is concerned, it is worthwhile to mention the famous tax on the imported luxury goods from Alexandria (dated to the beginning of the 3rd century CE, see Appendix text 3) that concerned the precious textiles from the land of the Seres as an important source of revenue. On the list of the “articles subject to duty” it is possible to find the words “raw silk”, “garments made completely or partly from silk”, “silk yarn”, together with other luxury goods coming from the Eurasian trade networks (Parker, 2002).
Another reason why the Palmyra silk fragments are incredibly important is the fact that, thanks to archeometric analysis, it is possible to reconstruct the different ways of processing silk material when arrived in the West. The textile products made of Chinese silk reached the West as finished goods with typical original decorations and embroidered Chinese letters. But we know from ancient Classical sources and from the Chinese ones that once arrived in the Western cities, the Chinese silk finished products were completely unravelled and rewoven (Schmidt-Colinet & Stauffer, 2000).
The new textile was extremely thin, shiny and transparent: this new creation was something that much more suited the Roman taste. It is highly significant that the beauty of these new ‘Chinese’ textiles, (re)manufactured in the Roman cities (Appendix text 4 a), could generate great interest and attraction in the very places from which that silk was originally being made, i.e. China. According to the brilliant suggestion of Thorley: “It was this that the Romans knew as silk, not the brocade with which we usually associate Chinese silk. This is what the Chinese were buying, totally unaware that they were simply buying back their own silk” (Thorley, 1971: 77–78; Leslie & Gardiner, 1996: 227).
This is the case of the remains of a silk funerary veil found together with Sri Lanka sapphires. Approximately 26 km south-east far from modern Rome, at the town of Colonna, a monumental grave of the middle 3rd century CE was accurately excavated in 2005: the sarcophagus was found with remains of a rich Roman lady, dressed with a silk veil decorated with a gold-strip and wearing a beautiful gold necklace or diadem decorated with sapphires and probably originally in combination with pearls. The style of the jewel and the presence of the very rare Ceylon sapphires (and probably of Indian pearls) recall examples attested in Palmyra and the Syrian jewellery of the 2nd–3rd century CE (Altamura, Angle, Cerino, De Angelis, & Tomei, 2013). We can consider this finding as one of the rare pieces of evidence of the trade and cultural Eurasian connections beyond the frontiers of Rome.

The D potential energy surfaces are presented

The 1D potential BQ-788 sodium salt surfaces are presented in Fig. 3. The 2D potential energy surfaces were constructed using, and, coordinates. Coordinate values were determined with an increment of 0.1 Å within the range −0.5Å to –1.2Å. Energy values were calculated in the nodes of the 2D mesh on the square with the side 1.7Å and with the step of 0.1Å. Energy values between the mesh nodes for the 1D and 2D PESs were found using the interpolation with the third order splines in the Mathematica package [58]. The 2D PESs of FAD obtained within the second approach are presented in Fig. 4.
Taking into account the absence of a kinematic interaction between and as well as between and coordinates, we can write down the following Schrodinger equations for three types of PESs:where
Coefficients of the expansion of potential energy into one-dimensional or two-dimensional Fourier series with the given number of harmonic components were found using the results of the interpolation [58]. The wave function was expressed as one-dimensional or two-dimensional Fourier series with a larger (in comparison with the case of the potential energy) number of harmonic components. After substituting the potential energy and the wave functions with such Fourier series in Eqs. (2)–(4), a Hamiltonian matrix was formed. Its diagonalization allows the energy values and the wave functions of the most deeply lying vibrational levels to be found. The frequency values of the and vibrations found in such a way were specified by taking into account the anharmonic interaction of these modes with other normal modes within the PT2 approximation. Such a hybrid approach to the estimation of an anharmonicity influence, that takes into account all vibrational degrees of freedom, was successfully employed earlier for intermolecular H-bonds [53,59,60] and for intramolecular H-bond in malonaldehyde molecule [61]. In frame of the PT2 approximation [54], the frequency value of the fundamental vibration can be found if its harmonic frequency is known and the matrix of the anharmonicity constants is determined:Since the intrinsic anharmonicity of the analyzed mode i is taken into account with the help of 1D PES calculation more precisely, then the first two members in the right part of equation (7) are substituted for the frequency value of the ith mode obtained by numerically solving the 1D Schrodinger equation:
Within the framework of 2D PES calculation for the i-th and k-th normal modes their anharmonic interaction together with the intrinsic anharmonicity are fully taken into account. In this case the frequency values obtained during the numerical solution of 2D Schrodinger equation substitute not only for the first two members in the right part of the equation (7) – the member is also substituted. Such changes allow us to rewrite equation (7) as:
Eqs. (8)–(9) and the data from Table 2 were used for calculating the frequencies of the analyzed vibrations in a hybrid approach. The full dimensionality of the vibrational problem is taken into account in Chromosome approach. The results of the calculations of the frequencies , , , and using the mentioned methods are listed in Table 3.

Discussion of the results
The analysis of the results of the first approach to the PES construction in coordinates and , is carried out. The dynamic coefficients (6) are used when solving Eq. (4). It is notable that the value of the splitting of the ground vibrational state is 292.8сm−1. The values of the corresponding splitting of symmetric and antisymmetric stretching vibrations of ОН group are equal to 1345.4 and 825.0сm−1. The frequency values in the doublets of symmetric and antisymmetric vibrations are equal to 1582.3; 2908.4 and 2134.3; 2959.3сm−1, respectively. The frequency values of the low-frequency components in both doublets can hardly be considered as acceptable. However, the more obvious fact, which contradicts to the experimental data, is the calculated value of the ground state splitting. Really, according to [62] the experimental value of this frequency is 0.0029сm−1. We explain the discrepancy between the calculated and experimental values by the motion of heavy atoms accompanying the tunneling process. The reduced masses in this case are essentially larger, and the kinematic coefficients are essentially smaller than those predicted by formula (6). If vibrating hydrogen atoms of the ОН group are affected by the same potential surface as in the case of tunneling and, hence, the vibrations of ОН groups are really accompanied by essential shifting of heavy atoms, then we can estimate the frequencies of symmetric and antisymmetric vibrations. Such estimations can be performed by reducing the values of kinematic coefficients to the ones at which the value of the ground vibrational state splitting will be equal to the experimental one. Indeed, having the value of reduced from 17.79 to 1.429сm−1, we obtained the value of 0.00292сm−1 for the ground state splitting, which is in excellent agreement with the experimental ones. However, in this case for the doublets of the frequencies of the symmetric and antisymmetric vibrations the values are equal to 698.89; 699.33 and 823.63 and 823.66сm−1 respectively. Since the values of these frequency is not acceptable, later on we will use the results of PES calculation within the second approach.

br Materials and methods PerkinElmer Spectrum One instrument in

Materials and methods
PerkinElmer Spectrum One instrument in ATR (attenuated total reflection) mode was used to record the infrared spectra of the samples. Because of the non-destructiveness of the employed set-up, every single specimen was directly placed onto the ATR plate with no further preparation and an optimal contact between the surface of the sample and the diamond crystal was ensured by means of a piston [8]. For all samples, thirty-two scans were collected with 4cm−1 Recent data from in the range between 4000 and 600cm−1. Samples were also analyzed by a Horiba XploRA micro-Raman spectrometer coupled to an Olympus BX-41 confocal microscope. The excitation source was a 785nm (90–100mW) laser. To prevent any sample alteration, laser power at the sample surface was reduced by neutral density filters to a value between 0.25 and 0.001mW, on an estimated spot size of 1064μm. Raman signal was collected using a 100× microscope objective. An acquisition time of 30s and 20 scans were carried out for each spectrum (spectral range 100–3000cm−1). One thousand lire (1000£) samples were taken from a private collection which includes specimens from the first issue for the Italian Republic in 1947 until the last issue in 2002 (see Table 1). The analyses have been focused on watermarks, security marks and serial numbers, as reported in Fig. 1. Furthermore, fibers, fillers, glues and coating, distributed on the surface, were investigated.

Results and discussion
Nowadays banknote recognition is realized by programmed softwares that check print quality; but, while this process ponders the print quality, it is interesting that the majority of features remain to be authenticated through manual inspection [9].

Raman and infrared spectroscopy has been used to collect vibrational spectra of inks, papers, glues and fillers used in banknotes belonging to 1000£ series, issued in Italy until January 31st, 2001. Five different typologies of banknotes were issued during 55 years.
Even though such paper-moneys are not currently valid, they are growing in interest for collectors. For example, differences in red pigment composition have been revealed between the exemplars used in 1947 and those one of 1962 by means of Raman spectroscopy and variations between the paper pulp used in the banknotes. For each type of exemplars, 3 samples were characterized, and two certified fakes were analyzed, highlighting important shift on the main peak of chrome orange and azo-pigment. Furthermore, the spectrum of paper banknote evidences changes in the papermaking an can be seen, as suggested for example by the band at 1727cm−1 in the infrared spectrum of the fake specimen.

The authors thank Studio Filatelico Salentino in Lecce for the kind loan of the 1000£ Busto d’Italia specimen. This research was supported by the Projects PON 254/Ric. Potenziamento del CENTRO RICERCHE PER LA SALUTE DELL’UOMO DELL’AMBIENTE Cod. PONa3_00334, PRIN 2012 Nanostrutture gerarchiche fotosintetiche per la produzione di energia and the Project PON S.I.Mi.S.A. Cod. PON02_00186_3417512, Project PON Pro.Ali.Fun Cod. PON02_00186_2937475, by Regione Puglia, Costituzione di Reti di Laboratori Pubblici di Ricerca, Progetto Esecutivo 09, WAFITECH.

At the microscopic level hydrogen bonding can lead to a wealth of complex suprastructures that constantly break and reform. This thermally driven property governs the functionality of many bio-macromolecules. Relatively small hydrogen bonded molecules are currently under scrutiny as well, like for example sugars [1,2], pharmaceuticals [3], alcohols [4,5], and other substances of interest for the life sciences. The detailed nature of their supramolecular organization depends on the hydrogen bond strength and on molecular structure. Therefore, the study of isomeric species has been exploited for a long time using vibrational spectroscopy [6]. By focusing on the OH stretching vibration, the hydrogen-bonding partners can be accessed directly. This allows one to study the consequences of intra- and intermolecular hydrogen bonding [7,8] as well as that of hydrogen bond cooperativity [9]. The latter term refers to the observation that the addition of molecules to a hydrogen bonded network enhances the mutual bonding strength of its constituents and thus shifts the corresponding vibrational frequencies. The impact of hydrogen bond cooperativity is interesting to study in the condensed [10] as well as in the gas phase [11].

br Conclusions This study described the vibrational properties

This study described the vibrational properties of diterpene C20H28O4 based on first principle calculations as well as effects of low temperature onto the Raman active vibrational modes. The ab initio calculations reproduced the characteristics of the pak1 inhibitor in good agreement with the experimental spectra recorded by Raman and infrared spectroscopy. In particular, in the region void of bands between 2800 and 1750cm−1, the experiments confirmed the theoretical result. Beyond this, it was possible to associate most of the normal modes (observed between 1750 and 20cm−1 and between 3100 and 2800cm−1) of the substance with defined vibrations by calculation of the potential energy distributions. In the entire spectrum, bands appearing in the Raman spectra practically do not change their wavenumbers when the substance is exposed to temperature variations between 20 and 300K. As a consequence of these observations, the statement is allowed that the diterpene C20H28O4 appears to be structurally very stable within the temperature range investigated.

The authors would like to acknowledge the financial support from the Brazilian agencies FUNCAP and CNPq. We also thank the CENAPAD-SP for the use of the GAUSSIAN 98 software package and the computational facilities through the project reference proj373. We also acknowledge Dr. D. Anderson for reading the manuscript.

Thin films of metal nitrides such TiN and AlN are well-known for their thermal, chemical and mechanical stability. TiN also has a dielectric function that can produce a surface plasmon resonance and high localized electric fields under suitable conditions [1–4]. In this respect it offers similar functionality to the conventional materials used to support the surface plasmon oscillations responsible for the unusual optical properties of plasmonic devices [5,6]. Silver (Ag) and gold (Au) have the lowest optical losses compared to most other elements or compounds [5] but their dielectric permittivity is generally fixed and cannot be adjusted easily. The imaginary part of the dielectric permittivity of metals is relatively large in magnitude in the optical range which is limiting the realization of devices in this region of the spectrum. The microstructure of metals (grain boundaries, defects) on the nanoscale also introduces additional losses. Finally, noble metals are quite expensive [7]. Therefore, we need to find new materials (compounds or alloys) that can support surface plasmons instead of Ag and Au. These materials should be easy to fabricate and cheap. Titanium nitride (TiN) – a gold-colored semiconductor – is a potential plasmonic material which is inexpensive, extremely hard and biocompatible (an important attribute for some applications in sensing). It is also compatible with complementary metal oxide semiconductor (CMOS) technology and has a very high melting temperature which makes it highly suitable for applications in thermal radiation engineering and thermophotovoltaics. In contrast, aluminum nitride (AlN) thin films also have remarkable hardness, and excellent thermal and chemical stability but are insulating [8,9].
Raman spectroscopy is a useful technique for providing information on the structure of molecules and solid surfaces [10]. The Raman signal can be enhanced if the analyte is located within a locally amplified electromagnetic field generated by a plasmon resonance. This amplification effect is known as Surface Enhanced Raman Spectroscopy (SERS) [11]. TiN has the necessary dielectric function to support localised plasmon resonances, with the absorption peak falling in the vicinity of 600nm [1]. For this reason, there has been some recent interest in its use for SERS [12–14]. The present work extends this possibility to more complex thin films of mixed Ti/Al nitrides.

In order to control the thickness of the thin films, the deposition parameters for pure TiN and AlN were calibrated first, then metal nitride thin films were deposited on glass substrates using a high vacuum, cryo-pumped chamber sputtering system. The pressure of nitrogen (N2) and argon (Ar) were 0.3mTorr and 1mTorr respectively. A Zeiss Supra 55VP scanning electron microscope (SEM) and Bruker D8 Discover X-ray diffraction (XRD) were used to characterize these thin films. Finally, Raman spectra of Rhodamine 6G (R6G) dye on the thin films were obtained using a Raman spectrometer (Renishaw In Via Raman Spectrometer (Gloucestershire, UK)) equipped with a Leica DMLB microscope (Wetzlar, Germany) and silicon reference standard. The Wire 3.4 spectroscopic suite software was used to process the analytical outputs.

A different study design was developed by

A different study design was developed by Gradoni et al. (2005) to test the efficacy of a multi-subunit recombinant Leishmania antigen (MML) in combination with two different adjuvants. Forty-five naïve young Beagles of both sexes were exposed to natural sand fly bites in a dedicated kennel sited in an endemic area of southern Italy. The study duration was two years, including two transmission seasons. Follow-up examinations were performed every two months from the end of the first season and consisted of clinical and clinicopathological evaluation, serology (IFAT), culture and nested-PCR from tissue aspirates (bone marrow and interleukins nodes). While the study failed to demonstrate a prophylactic efficacy of the MML vaccine, however both the investigation design and the elevated 2-year cumulative incidence of infections recorded in the study location (about 80%) was considered suitable for testing novel promising antigens.

The regular assessments performed for each dog through a standard set of diagnostic markers for infection and clinical scoring proposed by Gradoni et al. (2005) have resulted in a highly reproducible model for CanL staging confirmed by subsequent studies and found useful for the interpretation of Leishmania vaccine efficacy (Oliva et al., 2006; Foglia Manzillo et al., 2013). Basically, in a context of a cumulative incidence of 80% of Leishmania infection, three conditions were invariably identified during 2 years from initial exposure of untreated naïve dogs:
Following the MML recombinant vaccine study, further four Leishmania vaccines have been tested in the frame of this standardized field model. Three of them were found ineffective to protect dogs against both infection and disease (confidentiality-protected information, unpublished data). The fourth vaccine consisted of purified excreted-secreted proteins (ESP) of L. infantum (LiESP) produced by means of a patented cell-free, serum-free culture system and with QA-21 saponin as adjuvant. This vaccine had been found highly immunogenic in dogs (Moreno et al., 2011) and showed 57% efficacy against L. infantum active infection following artificial challenge administered one year after the primary vaccine course (Martin et al., 2014). In the field model, the vaccine efficacy was assessed by exposing 90 naïve dogs (46 vaccinated and 44 controls). The vaccine provided a significant reduction in the risk of progressing to active infection (p=0.025) or overt disease (p=0.046), with a clinical efficacy of 68%. The probability of a dog becoming PCR positive in tissue aspirates was similar between groups, but the probability of returning to a PCR negative condition was higher in the vaccinated group (p=0.04) (Oliva et al., 2014). In a subsample of vaccinated dogs that developed disease despite vaccination and that were exposed to the bites of reared sand flies, the reduction in parasite transmission was found significant when compared to matched controls (Bongiorno et al., 2013). LiESP/QA-21 has been the first commercially licensed vaccine for CanL in Europe (CaniLeish®, Virbac).

Despite advances in Leishmania genomics and proteomics (Myler and Fasel, 2008), modern biotechnology for antigen expression, purification and delivery, and the large availability of murine models in the field of experimental immunology, Leishmania vaccinology still suffers from several bottlenecks that limit the progress towards effective and universal vaccines. Among them, the great genetic diversity of parasites, the difficulty of identifying surrogate markers of resistance in naturally immunized hosts, and the lack of practical infectious challenge systems mimicking the natural one. Research in canine vaccines is further hampered by the polymorphic nature of the infection (“resistant” versus “susceptible” patterns) and the chronic clinical course of the disease. This implies very high costs for vaccine investigations: on one hand, a high number of study dogs is required as many of them, in a totally unpredictable way, will naturally resist to the infection and hence will be “useless” to the data analysis; on the other hand, it takes years before assessments on clinical efficacy can reasonably be concluded. Both these aspects conflict with international rules for animal welfare, making difficult the approval of long-term protocols including numerous dogs that require periodical examinations by invasive techniques such as tissue aspirates. The only two field trials leading to commercially available canine vaccines have enrolled less than 100 dogs each and, because the vaccines conferred only partial protection, efficacy parameters were close to the threshold of significance. Furthermore, only a small pilot study was performed to assess the infectiousness to phlebotomine vectors of vaccinated dogs in comparison with appropriate controls. Post-marketing data on vaccine efficacy and its impact on parasite transmission would therefore be needed from a much higher number of dogs from the “real life”.

There is only limited information available about the

There is only limited information available about the immune response of chickens or turkeys following experimental infection with H. meleagridis. Measuring mRNA of IL-1ß, CXLi2 and IL-6 in tissue samples taken from experimentally infected chicken caeca revealed an increase expression of such genes, which was absent in turkeys (Powell et al., 2009). It was concluded that this first defence reaction prevents heavy invasion of the chicken liver, a process different in turkeys. In livers of turkeys an uncontrolled immune reaction could be noticed with an upregulation of certain cytokines and an influx of different T-lympohcytes. In another study IFN-γ was upregulated indicating a Th1 immune response similar to other protozoan infections (Schwarz et al., 2011). Furthermore, a lower percentage of CD4+ cells was noticed in the spleen whereas the caecal lamina propria of infected chickens displayed a heavy infiltration with certain T-cells. However, the aforementioned studies used H. gallinarum bearing histomonads to infect the imidazoline receptors and a possible influence of the nematode needs to be considered. In vitro grown histomonads applied orally to chickens resulted in an elevated level of different immunoglobulins, most prevalent in the caeca (Windisch and Hess, 2010).

Treatment and prophylactic strategies

Conclusions and perspectives
Recent intensification of research on H. meleagridis and histomonosis in turkeys and chickens has substantially improved our existing knowledge. Despite the progress noticed in various fields great efforts are still needed in order to improve the understanding of the parasite\’s life cycle, its epidemiology, the introduction into and transmission within a flock. Numerous additional questions remain to be addressed in future studies as agreed upon amongst specialists during a technical meeting hosted by the European Food Safety Authority (EFSA, 2013).

Conflict of interest statement

Infection of livestock with Fasciola hepatica is a worldwide, economic problem with increasing importance (Fairweather, 2009). In the north of Germany for instance, the prevalence was previously reported to be 0.6% (Epe et al., 2004), while more recent studies have found mean herd prevalences of between 32.2% and 69.1% (Koch, 2005; Rabeler, 2011; Kuerpick et al., 2012).
Despite strategic treatments with different flukicides and sophisticated forecast models, high and even increasing prevalences have been reported elsewhere (Bennema et al., 2009; Rapsch et al., 2006). In addition to likely increases in disease prevalence, future fluke control will also have to deal with the threat of drug (i.e. triclabendazole) resistance (Fairweather, 2011). Economic losses due to bovine fasciolosis can be considerable. In Switzerland for example, such losses have been estimated at several thousand Euros per affected farm (Schweizer et al., 2005). Consequently, control of this parasite is important for economic reasons in addition to reasons of animal welfare. Of the several strategies for the control of bovine fasciolosis described, in practice usually strategic anthelmintic control is used with flukicide treatment time and interval depending on the climatic zone (Armour, 1975; Boray, 1971, 1972; Harris and Charleston, 1971; Schneider et al., 1975; Torgerson and Claxton, 1999; Whitehead, 1976).
A variety of fasciolicides is available for the treatment of bovine fasiolosis: triclabendazole, albendazole, oxyclozanide, clorsulon, nitroxynil, rafoxanide or oxfendazole (Fairweather, 2011; Gomez-Puerta et al., 2012; Martínez-Valladares et al., 2010). Of all these agents, triclabendazole is the most effective against immature stages of the parasite, which makes uterus the drug of choice for the treatment of acute fasciolosis, which is more commonly seen in sheep than in cattle. Due to increasing resistance against triclabendazole (Fairweather, 2011), there are efforts to develop new drugs which are efficacious against immature parasites, but results to date have been disappointing (Fairweather, 2011). Combining compounds can help to increase efficacy against immature stages. For example the combination of clorsulon (2.0mg/kg) and nitroxynil (10.2mg/kg) reached an efficiency of 95% against 2-week-old flukes and 99% against 4-week-old flukes (Hutchinson et al., 2009).