Regulation of Growth, Cell Shape, Cell Division, and Gene Expression by Second Messengers (p)ppGpp and Cyclic Di-GMP in Mycobacterium smegmatis

The alarmone (p)ppGpp regulates transcription, translation, replication, virulence, lipid synthesis, antibiotic sensitivity, biofilm formation, and other functions in bacteria. Signaling nucleotide cyclic di-GMP (c-di-GMP) regulates biofilm formation, motility, virulence, the cell cycle, and other functions. In Mycobacterium smegmatis, both (p) ppGpp and c-di-GMP are synthesized and degraded by bifunctional proteins Rel(Msm) and DcpA, encoded by rel(Msm) and dcpA genes, respectively. We have previously shown that the Delta rel(Msm) and Delta dcpA knockout strains are antibiotic resistant and defective in biofilm formation, show altered cell surface properties, and have reduced levels of glycopeptidolipids and polar lipids in their cell wall (K. R. Gupta, S. Kasetty, and D. Chatterji, Appl Environ Microbiol 81:2571-2578, 2015, http://dx.doi.org/10.1128/AEM.03999-14). In this work, we have explored the phenotypes that are affected by both (p) ppGpp and c-di-GMP in mycobacteria. We have shown that both (p) ppGpp and c-di-GMP are needed to maintain the proper growth rate under stress conditions such as carbon deprivation and cold shock. Scanning electron microscopy showed that low levels of these second messengers result in elongated cells, while high levels reduce the cell length and embed the cells in a biofilm-like matrix. Fluorescence microscopy revealed that the elongated Delta rel(Msm) and Delta dcpA cells are multinucleate, while transmission electron microscopy showed that the elongated cells are multiseptate. Gene expression analysis also showed that genes belonging to functional categories such as virulence, detoxification, lipid metabolism, and cell-wall-related processes were differentially expressed. Our results suggests that both (p) ppGpp and c-di-GMP affect some common phenotypes in M. smegmatis, thus raising a possibility of cross talk between these two second messengers in mycobacteria. IMPORTANCE Our work has expanded the horizon of (p) ppGpp and c-di-GMP signaling in Gram-positive bacteria. We have come across a novel observation that M. smegmatis needs (p) ppGpp and c-di-GMP for cold tolerance. We had previously shown that the Delta rel(Msm) and Delta dcpA strains are defective in biofilm formation. In this work, the overproduction of (p) ppGpp and c-di-GMP encased M. smegmatis in a biofilm-like matrix, which shows that both (p) ppGpp and c-di-GMP are needed for biofilm formation. The regulation of cell length and cell division by (p) ppGpp was known in mycobacteria, but our work shows that c-di-GMP also affects the cell size and cell division in mycobacteria. This is perhaps the first report of c-di-GMP regulating cell division in mycobacteria.

Flow Characteristics of Liquids in Microtubes Driven by a High Pressure

The flow characteristics of liquids in microtubes driven by a high pressure ranging from 1 MPa to 30 MPa are studied in this paper. The diameter of the microtube is from 3 μm to 10 μm and liquids composed of simple small molecules are chosen as the working fluids. The Reynolds number ranges from 0. 1 to 24. The behavior of isopropanol and carbon tetrachloride under high pressure is found different from the prediction from conventional Hagen-Poiseuille (HP) equation. The normalized friction coefficient C* increases significantly with the pressure. From an analysis of the microtube deformation, liquid compressibility, viscous heating and wall slip, it may be seen that the viscosity at high pressure plays an important role here. An exponential function of viscosity vs pressure is introduced into the HP equation to counteract the difference between experimental and theoretical values. However, this difference is not so marked for di-water.

Impossibilità di pensare Dio non esistente : il Proslogion di Anselmo d'Aosta come applicazione implicita dei moderni procedimenti di logica aletica

Resumen: La vera intenzione teologica di Anselmo d’Aosta nello scrivere il Proslogion e il vero significato del suo celebre unum argumentum vanno visti nella funzione che la ragione svolge necessariamente all’interno della vita di fede del cristiano. Anselmo, come Tommaso d’Aquino, non sostiene che l’esistenza di Dio sia un “articulus fidei” ma piuttosto uno dei “praeambula fidei”. La ragione naturale ha la certezza che Dio esiste, ancora prima della dimostrazione metafisica, e questo non fa che confermare l’assurdità di pensare che non esista il fondamento reale di tutte le cose esistenti.

La distruzione delle statue di Pausania nella Sparta tardoantica (Lib. Ep. 1518)

Abstract: The late antique destruction of two bronze statues of Pausanias - the Spartan general responsible for the Greek victory at the Battle of Plataea (479 BC) - housed in the temple of Athena Chalkioikos in Sparta (Lib. Ep. 1518), has been interpreted as one of the few cases of a violent conflict between pagan and Christian population in Greece. Nevertheless the sources suggest that late antique Sparta was a bastion of Hellenic paganism and give a picture of a small and quiet town ruled by a pagan educated élite, where pagans like Libanius wanted to live. Since there is no evidence of a violent conflict between pagans and Christians in Sparta, and Libanius confirms that in 365 AD all the temples and cult statues were still in place, this paper addresses the issue from a different point of view and offers a new contribution to the history of Sparta in Late Antiquity. By using literary, archaeological and epigraphic evidence the paper explores: 1) the relationship between Roman administration and Spartan élite in the IVth century AD; 2) the historical memory of Pausanias in Late Antiquity. It will be emphasized that the obscure burning of the two statues helped to remove from Sparta the memory of Pausanias - a controversial figure, misrepresented in Late Antiquity and connected to the ancient staseis in Laconia - in order to promote a positive image of Sparta as a city without conflicts and ruled by the political system of Lycurgus (eunomia). As documented by local inscriptions in praise of late Roman governors, the mythical lawgiver Lycurgus was the paradigm of the imperial governors who rebuilded the town in the IVth cent. AD. It can be assumed that while Rome, Constantinople, Antioch and Athens were troubled by political and religious violence or by seditions between different factions, Sparta aimed to revive its traditional model of civic order in the new historical context of Late Antiquity.

Evaluación de la eficacia de la técnica de floating system para la producción de radicchio rosso var. di Verona en condiciones controladas

La técnica de producción en Floating system constituye una alternativa económica y veloz de producción en hidroponia. Este sistema permite obtener hojas de vegetales de pequeño tamaño “Baby leaf” listas para consumir (IV Gama), cuyo consumo a nivel mundial muestra una tendencia creciente. Dicha técnica no se encuentra aún muy difundida para la producción de radicchio rosso. El objetivo de este trabajo fue evaluar el efecto del tamaño de celda, del substrato y de la técnica de floating system sobre la producción de radicchio. Los ensayos se realizaron en cámara a 25 ºC con un fotoperíodo de 16 hs de luz y 8 de oscuridad y con una intensidad de luz de 10 000 Lux. Se utilizó un diseño experimental factorial totalmente aleatorizado, con 5 repeticiones y 3 factores, cuyos niveles fueron: 1) Tamaño de celda, (T), con 3 niveles: grande, mediano y pequeño, 2) Técnica de producción, (F), con 2 niveles: floating system y sistema convencional, 3) Sustrato, (S), con 2 niveles: vermiculita y mezcla. Se analizaron las siguientes variables: 1) peso fresco, 2) peso seco, 3) largo de planta, 4) número de hojas y 5) área foliar. Los resultados obtenidos resultan alentadores respecto del empleo de la técnica de floating system. Para la obtención de plántulas con calidad de IV Gama resulta más conveniente el empleo de celdas de 80 cm3 y sustrato mezcla.

Vita, et fatti dell'invitissimo imperatore Carlo Qvinto, et historie universali del mondo, de' suoi tempi... opera vitilissima a prencipi, signori, & capitani, & qual si voglia persona che d'historie si diletta composta dal Sig. Alfonso Ulloa, con la Pace, & Capitolationi seguite, tra li due potentissimi Prencipi Di Francia, & Spagna : & Con Toaule Copiosissime : & In questa ultima qditione in molti moughi mccresciuta, & migliorata

Afonso de Ulloa, escritor espanhol, nasceu em Zamora na primeira metade do século XVl e morreu em Veneza, Itália, em 1580. Filho de Francisco de Ulloa, oficial que esteve na África com Carlos V, seguiu também a carreira das armas, estabelecendo-se depois em Veneza. Desempenhou várias missões diplomáticas, escreveu muitas obras e traduziu para o italiano, língua que conhecia tanto quanto a materna, autores espanhóis e portugueses. Quanto à obra Vita, et fatti dell' invitissimo imperatore Carlo Qvinto.. . sabe-se que foi pela primeira vez impressa em Veneza em 1560 por Vincenzo Valgrisio e que foi muitas vezes reeditada. Salvo pela época, a edição supra referenciada não oferece grande interesse artístico, mas há uma interessante gravura com a efígie de Carlos V.

Palmerino d'Inghilterra : figliuolo del Re Don Duardo, que si contano gli fran fatti in arme che egli fece per amore della infanta Polinarda, figliuola di Primalcone, doue acquisto mortal fama, et di Floriano dal Deserto suo fratello, con alcuni gloriosi fatti del Prencipe Florendo, figliuolo di Primaleone

Nuouamente tradotto di spagnuolo in italiano

Istoria delle guerre del Regno del Brasile : accadute tra la corona di Portogallo e la Republica di Olanda, composta, ed offerta alla sagra reale maesda di Pietro Secondo, Re di Portogallo & C. dal P. F. Gio. Gioseppe di S. Teresa Carmelitano Scalzo : Parte prima e Parte seconda

As negociações entre Portugal e Holanda a respeito da devolução dos territórios portugueses ocupados se desenvolveram ao longo de um decênio. As propostas iniciais de Portugal referiam-se aos territórios ocupados pelos holandeses na África e no Maranhão. Estão no raríssimo folheto.

Le Historie delle Indie Orientali : ... con una scielta di lettere scritte delle Indie, fra lequali ve ne fono molte non più stampate, tradotte dal medesimo : con um indice copiosissimi

Tradotte di latino in lingua toscana, da M. Francesco Serdonati Fiorentino.

Historia della disunione del Regno di Portogallo dalla Corona di Castiglia

Essa é a segunda edição de uma obra clássica sobre a restauração da monarquia portuguesa, com referências sobre o Brasil e adesão a D.João IV. O Séc. XVII foi, para Portugal, um tempo de lutas, tanto na Europa como no Brasil e na África. Foi, ainda, um período de reorganização jurídica e econômica da sociedade.

Synthesis and characterization of 1,1-di-tert-butyldiazene

The synthesis and direct observation of 1,1-di-tert-butyldiazene (16) at -127°C is described. The absorption spectrum of a red solution of 1,1-diazene 16 reveals a structured absorption band with λ max at 506 run (Me_2O, -125°C). The vibrational spacing in S_1 is about 1200 cm^(-1). The excited state of 16 emits weakly with a single maximum at 715 run observed in the fluorescence spectrum (Me_2O:CD_2Cl_2, -196°C). The proton NMR spectrum of 16 occurs as a singlet at 1.41 ppm. Monitoring this NMR absorption at -94^0 ± 2°C shows that 1,1-diazene 16 decomposes with a first-order rate of 1.8 x 10^(-3) sec(-1) to form isobutane, isobutylene and hexarnethylethane. This rate is 10^8 and 10^(34) times faster than the thermal decomposition of the corresponding cis and trans 1,2-di-tert-butyldiazene isomers. The free energy of activation for decomposition of 1,1-diazene 16 is found to be 12.5 ± 0.2 kcal/mol at -94°C which is much lower than the values of 19.1 and 19.4 kcal/lmole calculated at -94°C for N-(2,2,6,6- tetramethylpiperidyl)nitrene (3) and N-(2,2,5,5- tetrarnethylpyrrolidyl)nitrene (4), respectively. This difference between 16 and the cyclic-1,1-diazenes 3 and 4 can be attributed to a large steric interaction between the tert-butyl groups in 1,1-diazene 16.

In order to investigate the nature of the singlet-triplet gap in 1,1-diazenes, 2,5-di-tert-butyl-N-pyrrolynitrene (22) was generated but was found to be too reactive towards dimerization to be persistent. In the presence of dimethylsulfoxide, however, N-pyrrolynitrene (22) can be trapped as N-(2,5-di-tert-butyl- N'-pyrrolyl)dimethylsulfoxirnine (38). N-(2,5-di-tert-butyl-N'-pyrrolyl)dimethylsulfoximine (38-d^6) exchanges with free dimethylsulfoxide at 50°C in solution, presumably by generation and retrapping of pyrrolynitrene 22.

Megacylops viridis Jurine, abyssal form in Lake Maggiore. [Translation of: Memorie dell'Istituto Italiano di Idrobiologia Dott.Marco de Marchi 17 57-79, 1964]

Megacyclops viridis Jurine, noticed in Lago Maggiore in 1912 by De Marchi from the littoral vegetation of Pallanza, is a normal member of the littoral plankton of the lake. The subgenus Megacyclops, created by Kiefer in his revision of the viridis-vernalis group, contains european and american species some of which are today considered as varieties of the species viridis. This paper examines morphology and of the distribution of the Italian viridis in Lake Maggiore.

Study on the dynamics of the population of a copepod (Eudiaptomus vulgaris Schmeil). [Translation from: Memorie dell'Istituto Italiano di Idrobiologia Dott.Marco de Marchi 13 179-202, 1961. ]

Eudiaptomus vulgaris Schmeil is the most abundant copepod in Lake Maggiore and forms also, in respect to other entomostraca, the most important element, through its average biomass and because it is fairly numerous throughout the year. Plankton samples collected in a systematic and quantitative way, gave the opportunity to study some aspects of the dynamics of the population of this copepod, in safety in view of the uncertainty which in this kind of study can ensue when samples are taken only at a single station - in consequence of the changes in size of population between different water masses. The results of the biometrical observations are of the population of Eudiaptomus vulgaris is presented.

Ecological significance of seasonal variation in clutch-size of Arctodiaptomus bacillifer in alpine lakes of high altitudes (Val Bognanco) [Translation from: Bollettino di Zoologia 37 491-492, 1970]

This brief paper describes the significance of seasonal variation in clutch-size of the copepod Arctodiaptomus bacillifer in alpine lakes of high altitudes (Val Bognanco). Seasonal dynamics of the zooplankton of these lakes was studied during summer and autumn of 1968 and 1969and results are summarised.