Prolonged fluconazole exposure leads to a shift to Candida species with decreased susceptibility in breakthrough candidemia: prospective survey of the Fungal Infection Network of Switzerland

Objectives: To compare the clinical characteristics, species distribution and antifungal susceptibility of Candida bloodstream isolates (BSI) in breakthrough (BTC) vs. non-breakthrough candidemia (NBTC) and to study the effect of prolonged vs. short fluconazole (F) exposure in BTC.Methods: Candida BSI were prospectively collected during 2004- 2006 from 27 hospitals (seven university, 20 affiliated) of the FUNGINOS network. Susceptibility to F, voriconazole (V) and caspofungin (C) was tested in the FUNGINOS mycology reference laboratory by microtitre broth dilution method with the Sensititre YeastOneTM test panel. Clinical data were collected using standardized CRFs. BTC was defined as occurring during antifungal treatment/prophylaxis of at least three days duration prior to the candidemia. Susceptibility of BSI was defined according to 2010/2011 CLSI clinical breakpoints.Results: Out of 567 candidemia episodes, 550 Candida BSI were available. Of these, 43 (7.6%) were from BTC (37/43, 86% were isolated after F exposure). 38 BTC (88.4%) and 315 NBTC (55.6%) occurred in university hospitals (P < 0.001). The majority of patients developing BTC were immunocompromised: higher proportions of haematological malignancies (62.8% in BTC vs. 47.1% in NBTC, P < 0.001), neutropenia (37.2% vs. 11.8%, P < 0.001), acute GvHD (14% vs. 0.2%, P < 0.001), immunosuppressive drugs (74.4% vs. 7.8%, P < 0.001), and mucositis (32.6% vs. 2.3%, P < 0.001) were observed. Other differences between BTC and NBTC were higher proportions of patients with central venous catheters in the 2 weeks preceding candidemia (95.3% vs. 83.4%, P = 0.047) and receiving total parenteral nutrition (62.8% vs. 35.9%, P < 0.001), but a lower proportion of patients treated with gastric proton pump inhibitors (23.3% vs. 72.1%, P < 0.001). Overall mortality of BTC and NBTC was not different (34.9% vs. 31.7%, P = 0.73), while a trend to higher attributable mortality in BTC was found (13.9% vs. 6.9%, P = 0.12). Species identification showed a majority of C. albicans in both groups (51.2% in BTC vs. 62.9% in NBTC, P = 0.26), followed by C. glabrata (18.6% vs. 18.5%), C. tropicalis (2.3% vs. 6.3%) and C. parapsilosis (7.0% vs. 4.7%). Significantly more C. krusei were detected in BTC versus NBTC (11.6% vs. 1.6%, P = 0.002). The geometric mean MIC for F, V and C between BTC and NBTC isolates was not significantly different. However, in BTC there was a significant association between duration of F exposure and the Candida spp.: >10 days of F was associated with a significant shift from susceptible Candida spp. (C. albicans, C. parapsilosis, C. tropicalis, C. famata) to non-susceptible species (C. glabrata, C. krusei, C. norvegensis). Among 21 BTC episodes occurring after £10 days of F, 19% of the isolates were non-susceptible, in contrast to 68.7% in 16 BTC episodes occurring after >10 days of F (P = 0.003).Conclusions: Breakthrough candidemia occurred more often in immunocompromised hosts. Fluconazole administered for >10 days was associated with a shift to non-susceptible Candida spp.. Length of fluconazole exposure should be taken into consideration for the choice of empirical antifungal treatment.

Cell death and autophagy after severe hypoxic-ischemic encephalopathy in term newborns

Introduction: Various studies from hypoxic-ischemic animals haveinvestigated neuroprotection by targeting necrosis and apoptosis with inconclusive results. Three types of cell death have been described: apoptosis, necrosis and more recently, autophagic cell death. While autophagy is a physiological process of degradation of cellular components, excessive autophagy may be involved in cell death. Recent studies showed that inhibition of autophagy is neuroprotective in rodent neonatal models of cerebral ischemia. Furthermore, neonatal hypoxia-ischemia strongly increased neuronal autophagic flux which is linked to cell death in a rat model of perinatal asphyxia. Following our observations in animals, the aim of the present study was to characterize the different neuronal death phenotypes and to clarify whether autophagic cell death could be also involved in neuronal death in the human newborns after perinatal asphyxia. Methods: we selected retrospectively and anonymously all newborns who died in our unit of neonatology between 2004 and 2009, with the following criteria: gestational age >36 weeks, diagnosis of perinatal asphyxia (Apgar <5 at 5 minutes, arterial pH <7.0 at 1 hour of life and encephalopathy Sarnat III) and performed autopsy. The brain of 6 cases in asphyxia group and 6 control cases matching gestational age who died of pulmonary or other malformations were selected. On histological sections of thalamus, frontal cortex and hippocampus, different markers of apoptosis (caspase 3, TUNEL), autophagosomes (LC3-II) and lysosomes (LAMP1, Cathepsin D) were tested by immunohistochemistry. Results: Preliminary studies on markers of apoptosis (TUNEL, caspase 3) and of autophagy (Cathepsin D, LC3II, LAMP1) showed an expected increase of apoptosis, but also an increase of neuronal autophagic flux in the selected areas. The distribution seems to be region specific. Conclusion: This is the first time that autophagic flux linked with cell death is shown in brain of human babies, in association with hypoxicischemic encephalopathy. This work leads to a better understanding of the mechanisms associated with neuronal death following perinatal asphyxia and determines whether autophagy could be a promising therapeutic target.

Novel FGF8 mutations associated with recessive holoprosencephaly, craniofacial defects, and hypothalamo-pituitary dysfunction.

Context: Fibroblast growth factor (FGF) 8 is important for GnRH neuronal development with human mutations resulting in Kallmann syndrome. Murine data suggest a role for Fgf8 in hypothalamo-pituitary development; however, its role in the etiology of wider hypothalamo-pituitary dysfunction in humans is unknown.Objective: The objective of this study was to screen for FGF8 mutations in patients with septo-optic dysplasia (n = 374) or holoprosencephaly (HPE)/midline clefts (n = 47).Methods: FGF8 was analyzed by PCR and direct sequencing. Ethnically matched controls were then screened for mutated alleles (n = 480-686). Localization of Fgf8/FGF8 expression was analyzed by in situ hybridization in developing murine and human embryos. Finally, Fgf8 hypomorphic mice (Fgf8(loxPNeo/-)) were analyzed for the presence of forebrain and hypothalamo-pituitary defects.Results: A homozygous p.R189H mutation was identified in a female patient of consanguineous parentage with semilobar HPE, diabetes insipidus, and TSH and ACTH insufficiency. Second, a heterozygous p.Q216E mutation was identified in a female patient with an absent corpus callosum, hypoplastic optic nerves, and Moebius syndrome. FGF8 was expressed in the ventral diencephalon and anterior commissural plate but not in Rathke's pouch, strongly suggesting early onset hypothalamic and corpus callosal defects in these patients. This was consolidated by significantly reduced vasopressin and oxytocin staining neurons in the hypothalamus of Fgf8 hypomorphic mice compared with controls along with variable hypothalamo-pituitary defects and HPE.Conclusion: We implicate FGF8 in the etiology of recessive HPE and potentially septo-optic dysplasia/Moebius syndrome for the first time to our knowledge. Furthermore, FGF8 is important for the development of the ventral diencephalon, hypothalamus, and pituitary. (J Clin Endocrinol Metab 96: E1709-E1718, 2011)

An overview of L-2-hydroxyglutarate dehydrogenase gene (L2HGDH) variants: a genotype-phenotype study.

L-2-Hydroxyglutaric aciduria (L2HGA) is a rare, neurometabolic disorder with an autosomal recessive mode of inheritance. Affected individuals only have neurological manifestations, including psychomotor retardation, cerebellar ataxia, and more variably macrocephaly, or epilepsy. The diagnosis of L2HGA can be made based on magnetic resonance imaging (MRI), biochemical analysis, and mutational analysis of L2HGDH. About 200 patients with elevated concentrations of 2-hydroxyglutarate (2HG) in the urine were referred for chiral determination of 2HG and L2HGDH mutational analysis. All patients with increased L2HG (n=106; 83 families) were included. Clinical information on 61 patients was obtained via questionnaires. In 82 families the mutations were detected by direct sequence analysis and/or multiplex ligation dependent probe amplification (MLPA), including one case where MLPA was essential to detect the second allele. In another case RT-PCR followed by deep intronic sequencing was needed to detect the mutation. Thirty-five novel mutations as well as 35 reported mutations and 14 nondisease-related variants are reviewed and included in a novel Leiden Open source Variation Database (LOVD) for L2HGDH variants (http://www.LOVD.nl/L2HGDH). Every user can access the database and submit variants/patients. Furthermore, we report on the phenotype, including neurological manifestations and urinary levels of L2HG, and we evaluate the phenotype-genotype relationship.

Lack of Mycobacterium tuberculosis-specific interleukin-17A-producing CD4(+) TÂ cells in active disease.

Protective immunity to Mycobacterium tuberculosis (Mtb) is commonly ascribed to a Th1 profile; however, the involvement of Th17 cells remains to be clarified. Here, we characterized Mtb-specific CD4(+) T cells in blood and bronchoalveolar lavages (BALs) from untreated subjects with either active tuberculosis disease (TB) or latent Mtb infection (LTBI), considered as prototypic models of uncontrolled or controlled infection, respectively. The production of IL-17A, IFN-γ, TNF-α, and IL-2 by Mtb-specific CD4(+) T cells was assessed both directly ex vivo and following in vitro antigen-specific T-cell expansion. Unlike for extracellular bacteria, Mtb-specific CD4(+) T-cell responses lacked immediate ex vivo IL-17A effector function in both LTBI and TB individuals. Furthermore, Mtb-specific Th17 cells were absent in BALs, while extracellular bacteria-specific Th17 cells were identified in gut biopsies of healthy individuals. Interestingly, only Mtb-specific CD4(+) T cells from 50% of LTBI but not from TB subjects acquired the ability to produce IL-17A following Mtb-specific T-cell expansion. Finally, IL-17A acquisition by Mtb-specific CD4(+) T cells correlated with the coexpression of CXCR3 and CCR6, currently associated to Th1 or Th17 profiles, respectively. Our data demonstrate that Mtb-specific Th17 cells are selectively undetectable in peripheral blood and BALs from TB patients.

Sporadic late-onset nemaline myopathy with MGUS: Long-term follow-up after melphalan and SCT.

OBJECTIVE: Sporadic late-onset nemaline myopathy (SLONM) is a rare, late-onset myopathy that progresses subacutely. If associated with a monoclonal gammopathy of unknown significance (MGUS), the outcome is unfavorable: the majority of these patients die within 1 to 5 years of respiratory failure. This study aims to qualitatively assess the long-term treatment effect of high-dose melphalan (HDM) followed by autologous stem cell transplantation (SCT) in a series of 8 patients with SLONM-MGUS. METHODS: We performed a retrospective case series study (n = 8) on the long-term (1-8 years) treatment effect of HDM followed by autologous SCT (HDM-SCT) on survival, muscle strength, and functional capacities. RESULTS: Seven patients showed a lasting moderate-good clinical response, 2 of them after the second HDM-SCT. All of them had a complete, a very good partial, or a partial hematologic response. One patient showed no clinical or hematologic response and died. CONCLUSIONS: This case series shows the positive effect of HDM-SCT in this rare disorder. Factors that may portend an unfavorable outcome are a long disease course before the hematologic treatment and a poor hematologic response. Age at onset, level and type of M protein (κ vs λ), and severity of muscle weakness were not associated with a specific outcome. CLASSIFICATION OF EVIDENCE: This study provides Class IV evidence that for patients with SLONM-MGUS, HDM-SCT increases the probability of survival and functional improvement.

Tumor targeting with newly designed biparatopic antibodies directed against two different epitopes of the carcinoembryonic antigen (CEA).

In an attempt to improve tumor targeting and tumor retention time of monoclonal antibodies (MAbs), we prepared biparatopic antibodies (BpAbs) having the capability of binding 2 different non-overlapping epitopes on the same target antigen molecule, namely, the carcinoembryonic antigen (CEA). Six BpAbs were constructed by coupling 2 different Fab' fragments from 4 different specific anti-CEA MAbs recognizing 4 CEA epitopes (Gold 1-4). Demonstration of the double paratopic binding of these antibodies for CEA was confirmed in vitro by inhibition radioimmunoassay and cross-inhibition analysis by surface plasmon resonance (SPR; BIACORE) technology. Using the latter technique, the affinity constants for CEA immobilized onto the sensor chip were found to range from 0.37 to 1.54 x 10(9) M(-1) for the 4 parental F(ab')2 fragments and from 1.88 to 10.14 x 10(9) M(-1) for the BpAbs, demonstrating the advantage of biparatopic binding over conventional F(ab')2 binding. The Ka improvement was particularly high for BpAb F6/35A7 and BpAb F6/B17 with a 9.5- and 8.1-fold increase, respectively, as compared with the parental F(ab')2. In vivo, the 6 BpAbs were compared with their 2 respective parental F(ab')2 by injection of 131I-BpAb/125I-F(ab')2 parental fragments into nude mice xenografted with the human colon carcinoma T380. Dissection 72 hr post-injection demonstrated that BpAb B17/CE25 and BpAb F6/B17 gave higher tumor uptake than that of their parental F(ab')2. This finding is particularly interesting for BpAb F6/B17, which compared favorably with the F6 F(ab')2, one of the best parental F(ab')2 fragments used in our study.

Anà lisis antropològica de les restes neolítiques de la Caserna de Sant Pau (Biometria, Dentició, ADN i Microestriació Dentà ria).

Les restes esquelètiques analitzades corresponen tant al nivell neolític (n=26) com a l"època del bronze (n=2), encara que aquí es presenten tan sols els resultats de la població neolítica (taula 1). En tots els casos les sepultures són individuals, amb l"única excepció de la sepultura 20, on s"han recuperat les restes corresponents a un individu femení adult (CSP201) i un fetus (CSP202) (taula 1). Per tal de caracteritzar biomètricament la població s"ha emprat la metodologia proposada per Martin i Saller (1959), Brothwell (1981) i Bass (1971). La determinació del sexe s"ha realitzat a través dels caràcters sexuals secundaris del crani i de la pelvis, i se"n pot trobar una descripció més detallada a Estebaranz et alii (2007). En el cas específic de les paleopatologies es van seguir les recomanacions de Campillo (Campillo, 1977).

Diferentes substratos e recipientes na formação de mudas de mamoeiro 'sunrise solo'

Objetivando avaliar a viabilidade de diferentes substratos e recipientes na formação de mudas de mamoeiro 'Sunrise Solo', realizou-se o presente experimento em condições de casa de vegetação, no Pomar da Universidade Federal de Lavras-UFLA. O delineamento experimental foi o inteiramente casualizado, em esquema fatorial 4x3, com quatro repetições e cinco plantas por parcela. Os tratamentos constituíram-se dos seguintes substratos: Plantimaxâ; substrato A (esterco de curral + carvão vegetal + solo e areia na proporção de 2:1:1:1 v/v; substrato B (vermicomposto + carvão vegetal + solo e areia na proporção de 1:1:1:1 v/v ) e substrato C (Plantimaxâ + carvão vegetal + solo e areia na proporção de 1:1:1:1 v/v); e dos seguintes recipientes: saco de polietileno com de 750 ml, bandeja de isopor com capacidade de 70 ml/célula de capacidade e tubetes de 50 ml. Foram avaliados altura das mudas, número de folhas, matéria fresca e seca da parte aérea e raiz. Pelos resultados, concluiu-se que o recipiente saco de polietileno, juntamente com o substrato A, foram os que apresentaram resultados favoráveis para todas as características avaliadas, e o substrato C proporcionou resultados desfavoráveis ao desenvolvimento das mudas.

Theta-duality on Prym varieties and a Torelli Theorem

Let $\pi : \widetilde C \to C$ be an unramified double covering of irreducible smooth curves and let $P$ be the attached Prym variety. We prove the scheme-theoretic theta-dual equalities in the Prym variety $T(\widetilde C)=V^2$ and $T(V^2)=\widetilde C$, where $V^2$ is the Brill-Noether locus of $P$ associated to $\pi$ considered by Welters. As an application we prove a Torelli theorem analogous to the fact that the symmetric product $D^{(g)}$ of a curve $D$ of genus $g$ determines the curve.

Transcranial Magnetic Stimulation Combined with EEG Reveals Covert States of Elevated Excitability in the Human Epileptic Brain.

BACKGROUND: Transcranial magnetic stimulation combined with electroencephalogram (TMS-EEG) can be used to explore the dynamical state of neuronal networks. In patients with epilepsy, TMS can induce epileptiform discharges (EDs) with a stochastic occurrence despite constant stimulation parameters. This observation raises the possibility that the pre-stimulation period contains multiple covert states of brain excitability some of which are associated with the generation of EDs. OBJECTIVE: To investigate whether the interictal period contains "high excitability" states that upon brain stimulation produce EDs and can be differentiated from "low excitability" states producing normal appearing TMS-EEG responses. METHODS: In a cohort of 25 patients with Genetic Generalized Epilepsies (GGE) we identified two subjects characterized by the intermittent development of TMS-induced EDs. The high-excitability in the pre-stimulation period was assessed using multiple measures of univariate time series analysis. Measures providing optimal discrimination were identified by feature selection techniques. The "high excitability" states emerged in multiple loci (indicating diffuse cortical hyperexcitability) and were clearly differentiated on the basis of 14 measures from "low excitability" states (accuracy = 0.7). CONCLUSION: In GGE, the interictal period contains multiple, quasi-stable covert states of excitability a class of which is associated with the generation of TMS-induced EDs. The relevance of these findings to theoretical models of ictogenesis is discussed.

Impacte tecnològic en el nou món colonial. Aculturació en arqueologia i arqueometria cerà mica (TECNOLONIAL)

El projecte se centra en l"expansió colonial atlàntica espanyola, especialment durant el segle XVI, estudiada a partir del coneixement arqueològic i arqueomètric de la ceràmica de pisa o majòlica, de les ceràmiques vidrades i dels contenidors de transport, però també de les ceràmiques indígenes d"estil europeu. Amb això, l"objectiu bàsic del projecte és aprofundir en els aspectes d"aculturació o de transculturació, de relacions i d"influències que es generen en un procés colonitzador, entre els colonitzadors i els grups humans colonitzats, però també l"aparició de noves identitats que sorgeixen en els espais colonitzats, amb grups humans generalment mixtos i complexos. Aquest objectiu general es concreta en quatre objectius concrets: la caracterització arqueològica i arqueomètrica dels centres productors i de difusió de Barcelona, Sevilla i el País Basc; la caracterització d"aquests materials, i dels materials indígenes d"influència europea, dels centres receptors de Gran Canària, Perú, Colombia i Canadà; la interpretació d"aquests materials ceràmics tant en termes de comerç, intercanvi i difusió, com des de la perspectiva del seu consum, com a artefactes que marquen o confereixen identitat social o estatus, i la seva participació en aspectes com les tradicions culturals, l"etnicitat o la creació de noves identitats; i la derivació de models teòrics aplicables a contextos colonials, basats en els estudis arqueològics i arqueomètrics de les ceràmiques implicades.

Impacte tecnològic en el nou món colonial. Aculturació en arqueologia i arqueometria cerà mica (TECNOLONIAL)

El projecte TECNOLONIAL té com a principal objectiu aprofundir en aspectes relacionats amb la interacció, influència i aculturació durant el procés de colonització a Amèrica. En aquest sentit, cal destacar que s"utilitza el terme"aculturació" com a nom genèric per denominar un procés complex de contactes entre societats diferents i diverses; procés que donarà com a resultat diverses respostes a la integració i/o resistència d"acord amb diferents situacions i estratègies, com el desenvolupament de noves identitats en les àrees colonitzades on grups humans mixtos i complexos es troben ubicats. El projecte se centra en l"expansió atlàntica espanyola, especialment durant el segle XVI, com a punt de partida del comerç global que es practica avui dia entre societats completament interconnectades.La investigació es basa essencialment en l"estudi de la ceràmica (majòlica, vidriada i contenidors de transport, com també ceràmiques indígenes), que és un bé o un bé comercial de valor intrínsec, d"ús intensiu i relacionat amb activitats quotidianes i simbòliques. A més, la seva ubiqüitat i abundància en el registre arqueològic converteixen la ceràmica en l"objecte ideal per contestar aquest tipus de qüestions. En aquest sentit, la caracterització de la tecnologia ceràmica, que inclou les seves decoracions i vidriats les propietats mecàniques, com també l"avaluació dels seus dissenys formals, ens ha de permetre aconseguir un profund coneixement de la tecnologia europea i del seu impacte transformador sobre la producció ceràmica indígena

La cerà mica de Barcelona. Organització i producció entre els segles XIII i XVIII a través de la seva caracterització arqueomètrica

L"estudi sobre la producció ceràmica de Barcelona en època medieval i postmedieval s'està fent des del Museu d"Història de Barcelona i des de l"ARQUB en aquest darrers anys. Aquest estudi s"ha reforçat amb els treballdesenvolupats en el marc del projecte de recerca Tecnolonial, cosa que ha permès ampliar les classes ceràmiques i el marc cronològic d"estudi. Els resultats aconseguits fins ara mostren que per a un mateix moment cronològic es preparen diverses pastes segons el producte final que es vol obtenir. Finalment, el canvi d"ubicació dels tallers que segurament es va produïr cap a finals del segle XIII / inicis del segle XIV podria estar relacionat amb l"explotació d"un nou llit d"argiles, la qual cosa coincidiria amb el fet que la producció del segle XIII és completament diferent químicament a la del segle XIV en endavant.