Detection of <i>aac(6’)-Ib-cr</i> in Avian Pathogenic <i>Escherichia coli</i> Isolates in Japan

We investigated the prevalence of plasmid-mediated quinolone resistance (PMQR) genes in avian pathogenic Escherichia coli (APEC) strains in Japan. A total of 117 APEC strains collected between 2004 and 2007 were examined for PMQR genes (qnrA, qnrB, qnrC, qnrD, qnrS, aac(6')-Ib-cr, qepA and oqxAB) by polymerase chain reaction. None of the APEC strains carried qnrA, qnrB, qnrC, qnrD, qnrS, qepA or oqxAB, but one of the isolates was identified as an AAC (6')-Ib-cr producer. Phylogenetic grouping, multi-locus sequence typing and serotyping showed that this isolate belonged to phylogenetic group A, sequence type 167 and untypable serogroup. To our knowledge, this is the first report of the aac (6')-Ib-cr gene in bacteria from food-producing animals in Japan.

O acometimento renal na hipertensão arterial e diabetes mellitus tipo 2: como identificar e prevenir? – A visão do cardiologista

Revista HUPE, Rio de Janeiro, 2013;12(Supl 1):44-52 doi:10.12957/rhupe.2013.7082 A hipertensao arterial (HA) e o diabetes mellitus tipo 2 (DM2) sao fatores de risco cardiovascular que frequentemente coexistem. Estas condicoes clinicas tem grande impacto epidemiologico, com elevados custos medicosociais e economicos. Muito comumente estas duas condicoes cursam com algum grau de lesao renal e representam as principais causas de doenca renal terminal. Por tudo isto o controle da pressao arterial (PA), da glicemia e dos lipides sericos sao fundamentais para prevenir ou postergar o desenvolvimento da nefropatia diabetica. O beneficio do controle glicemico mais intenso na prevencao da nefropatia diabetica e de complicacoes micro e macrovasculares nao esta completamente estabelecido. De forma semelhante, evidencias de literatura tem demonstrado beneficio da reducao da PA para valores de 130-135/80 mmHg em diabeticos. No entanto, reducoes mais intensas da PA apresentam evidencias limitadas. O tratamento da HA e do DM deve envolver mudancas de habitos de vida e tratamento medicamentoso concomitantemente. Todas as classes de anti-hipertensivos sao capazes de reduzir a PA e, portanto contribuir para a prevencao da nefropatia diabetica; entretanto, destacam-se os beneficios dos bloqueadores do sistema renina angiotensina aldosterona. A escolha do esquema hipoglicemiante sera baseada nos valores da glicemia de jejum, pos-prandial e da HbA1c, no peso e idade do paciente, na presenca de complicacoes associadas e na interacao com outras drogas. Os autores discutem neste artigo as metas de tratamento da PA e da glicemia, as linhas gerais de tratamento nao medicamentoso e medicamentoso da HA e do DM, bem como as principais estrategias para protecao renal, na visao do cardiologista. Descritores: Hipertensao; Diabetes mellitus; Nefropatias diabeticas.

Analysis of Lumbar Movement under Different Muscle Activation Strategies During Active Leg Raise in Normal Subjects

Analysis of the root morphology of European anterior teeth.

Aim of this study was to investigate the gross anatomy of the root of European anterior teeth. A review of the dental literature shows that in the past the root morphology was investigated from the inner pulp chamber for endodontic therapies. In order to be admitted to the study, the teeth had to be undamaged. Each tooth was identified by a serial number and gauged by a millimeter tape (for the root length), a goniometer (for the root angle), and a millimeter gauge (for the root diameter). Furthermore, a statistical elaboration of the data was performed to underline the shape variations of the surface around the different sides of the root. At the end of the analysis, 12 parameters for each single-root tooth were described. The study highlights significant differences (p<0.01) only in two teeth of the maxillary arch (central incisor and canine) and in one tooth of the mandibular arch (central incisor). In both cases, the observed differences may be due to the sinuosity of the cement-enamel line. The Tables for each measured parameter were obtained for all examined classes of teeth, but a comparison with literature data was possible only for the "root length" parameter. This study can be considered innovative for the absence, in the scientific literature, of a statistical analysis of all parameters with the exception of the "root length". Moreover, it gives a detailed updating of the data relative to the European population creating a useful tool as well for surgical interventions during periodontal therapy (for example in the choice of the right ultrasonic handpiece) as for new CAD/CAM assisted implant manufacturing techniques.

Secondhand tobacco smoke in public venues in three Indonesian cities

Background: The aim of this study was to measure secondhand tobacco (including kretek ) smoke (SHS) concentrations in public places in Jakarta, Bogor, and Palembang before laws banning smoking in public spaces went into effect. Methods: Particulate matter (PM 2.5 ) was measured in 15 hospitals, 15 government offices, 30 restaurants, and 26 entertainment venues throughout the three cities. Also, in Jakarta, vapor-phase nicotine was measured in 5 schools, 5 hospitals, 5 government offices, 9 restaurants, and 10 entertainment venues. Data were analyzed descriptively. Differences by city and venue characteristics were analyzed by Student’s t-test, ANOVA, and Bonferroni pairwise statistical tests. Results: Geometric mean PM 2.5 levels were highest in entertainment venues (96 μg/m 3 ), followed by restaurants (78 μg/m 3 ), government offices (57 μg/m 3 ), and hospitals (46 μg/m 3 ). Air nicotine levels in Jakarta were highest in designated smoking areas (4.71 μg/m 3 ) and designated non-smoking areas (1.55 μg/m 3 ) of entertainment venues. These were followed by government offices (0.30 μg/m 3 ), designated smoking areas (0.24 μg/m 3 ) and designated non-smoking areas (0.19 μg/m 3 ) of restaurants, hospitals (0.01 μg/m 3 ), and schools (0.01 μg/m 3 ). Conclusion: SHS was detected in all venues in the three cities in Indonesia. High levels of air nicotine were found in non-smoking areas of restaurants and entertainment venues, indicating that designated smoking areas are not an effective solution to eliminate SHS. There is no safe level of SHS exposure and thus SHS in these venues increases the risk of adverse health effects among children and adults. These findings support the need for 100% smoke-free laws covering all public venues in these and other Indonesian cities. (Med J Indones. 2013;22:232-7. doi: 10.13181/mji.v22i4.606) Keywords: Cigarettes, kretek, protection, protect, secondhand smoke, smoke-free policy, tobacco

Soulamarin Isolated from Calophyllum brasiliense (Clusiaceae) Induces Plasma Membrane Permeabilization of Trypanosoma cruzi and Mytochondrial Dysfunction

Chagas disease is caused by the parasitic protozoan Trypanosoma cruzi . It has high mortality as well as morbidity rates and usually affects the poorer sections of the population. The development of new, less harmful and more effective drugs is a promising research target, since current standard treatments are highly toxic and administered for long periods. Fractioning of methanol (MeOH) extract of the stem bark of Calophyllum brasiliense (Clusiaceae) resulted in the isolation of the coumarin soulamarin, which was characterized by one- and two-dimensional 1 H- and 13 C NMR spectroscopy as well as ESI mass spectrometry. All data obtained were consistent with a structure of 6-hydroxy-4-propyl-5-(3-hydroxy-2-methyl-1-oxobutyl)-6″,6″-dimethylpyrane-[2″,3″:8,7]-benzopyran-2-one for soulamarin. Colorimetric MTT assays showed that soulamarin induces trypanocidal effects, and is also active against trypomastigotes. Hemolytic activity tests showed that soulamarin is unable to induce any observable damage to erythrocytes (c max.  = 1,300 µM). The lethal action of soulamarin against T. cruzi was investigated by using amino(4-(6-(amino(iminio)methyl)-1H-indol-2-yl)phenyl)methaniminium chloride (SYTOX Green and 1H,5H,11H,15H-Xantheno[2,3,4-ij:5,6,7-i′j′]diquinolizin-18-ium, 9-[4-(chloromethyl)phenyl]-2,3,6,7,12,13,16,17-octahydro-chloride (MitoTracker Red) as fluorimetric probes. With the former, soulamarin showed dose-dependent permeability of the plasma membrane, relative to fully permeable Triton X-100-treated parasites. Spectrofluorimetric and fluorescence microscopy with the latter revealed that soulamarin also induced a strong depolarization (ca. 97%) of the mitochondrial membrane potential. These data demonstrate that the lethal action of soulamarin towards T. cruzi involves damages to the plasma membrane of the parasite and mitochondrial dysfunction without the additional generation of reactive oxygen species, which may have also contributed to the death of the parasites. Considering the unique mitochondrion of T. cruzi , secondary metabolites of plants affecting the bioenergetic system as soulamarin may contribute as scaffolds for the design of novel and selective drug candidates for neglected diseases, mainly Chagas disease.

Differentially Expressed Genes in Hirudo medicinalis Ganglia after Acetyl-L-Carnitine Treatment

Acetyl- l -carnitine (ALC) is a naturally occurring substance that, when administered at supra-physiological concentration, is neuroprotective. It is involved in membrane stabilization and in enhancement of mitochondrial functions. It is a molecule of considerable interest for its clinical application in various neural disorders, including Alzheimer’s disease and painful neuropathies. ALC is known to improve the cognitive capability of aged animals chronically treated with the drug and, recently, it has been reported that it impairs forms of non-associative learning in the leech. In the present study the effects of ALC on gene expression have been analyzed in the leech Hirudo medicinalis . The suppression subtractive hybridisation methodology was used for the generation of subtracted cDNA libraries and the subsequent identification of differentially expressed transcripts in the leech nervous system after ALC treatment. The method detects differentially but also little expressed transcripts of genes whose sequence or identity is still unknown. We report that a single administration of ALC is able to modulate positively the expression of genes coding for functions that reveal a lasting effect of ALC on the invertebrate, and confirm the neuroprotective and neuromodulative role of the substance. In addition an important finding is the modulation of genes of vegetal origin. This might be considered an instance of ectosymbiotic mutualism.

bis-Dehydroxy-Curcumin Triggers Mitochondrial-Associated Cell Death in Human Colon Cancer Cells through ER-Stress Induced Autophagy

Background The activation of autophagy has been extensively described as a pro-survival strategy, which helps to keep cells alive following deprivation of nutrients/growth factors and other stressful cellular conditions. In addition to cytoprotective effects, autophagy can accompany cell death. Autophagic vacuoles can be observed before or during cell death, but the role of autophagy in the death process is still controversial. A complex interplay between autophagy and apoptosis has come to light, taking into account that numerous genes, such as p53 and Bcl-2 family members, are shared between these two pathways. Methodology/Principal Findings In this study we showed a potent and irreversible cytotoxic activity of the stable Curcumin derivative bis -DeHydroxyCurcumin (bDHC) on human colon cancer cells, but not on human normal cells. Autophagy is elicited by bDHC before cell death as demonstrated by increased autophagosome formation -measured by electron microscopy, fluorescent LC3 puncta and LC3 lipidation- and autophagic flux -measured by interfering LC3-II turnover. The accumulation of poly-ubiquitinated proteins and ER-stress occurred upstream of autophagy induction and resulted in cell death. Cell cycle and Western blot analyses highlighted the activation of a mitochondrial-dependent apoptosis, which involves caspase 7, 8, 9 and Cytochrome C release. Using pharmacological inhibitions and RNAi experiments, we showed that ER-stress induced autophagy has a major role in triggering bDHC-cell death. Conclusion/Significance Our findings describe the mechanism through which bDHC promotes tumor selective inhibition of proliferation, providing unequivocal evidence of the role of autophagy in contrasting the proliferation of colon cancer cells.

The Cancer Stem Cell Marker CD133 Interacts with Plakoglobin and Controls Desmoglein-2 Protein Levels

The pentaspan membrane glycoprotein CD133 (also known as prominin-1) has been widely used as a marker for both cancer and normal stem cells. However, the function of CD133 has not been elucidated. Here we describe a cancer stem cell line established from clear cell carcinoma of the ovary (CCC) and show that CD133 interacts with plakoglobin (also known as γ-catenin), a desmosomal linker protein. We further demonstrate that knockdown of CD133 by RNA interference (RNAi) results in the downregulation of desmoglein-2, a desmosomal cadherin, and abrogates cell-cell adhesion and tumorigenicity of CCC stem cells. We speculate that CD133 may be a promising target for cancer chemotherapy.

Inflammation Subverts Hippocampal Synaptic Plasticity in Experimental Multiple Sclerosis

Abnormal use-dependent synaptic plasticity is universally accepted as the main physiological correlate of memory deficits in neurodegenerative disorders. It is unclear whether synaptic plasticity deficits take place during neuroinflammatory diseases, such as multiple sclerosis (MS) and its mouse model, experimental autoimmune encephalomyelitis (EAE). In EAE mice, we found significant alterations of synaptic plasticity rules in the hippocampus. When compared to control mice, in fact, hippocampal long-term potentiation (LTP) induction was favored over long-term depression (LTD) in EAE, as shown by a significant rightward shift in the frequency–synaptic response function. Notably, LTP induction was also enhanced in hippocampal slices from control mice following interleukin-1β (IL-1β) perfusion, and both EAE and IL-1β inhibited GABAergic spontaneous inhibitory postsynaptic currents (sIPSC) without affecting glutamatergic transmission and AMPA/NMDA ratio. EAE was also associated with selective loss of GABAergic interneurons and with reduced gamma-frequency oscillations in the CA1 region of the hippocampus. Finally, we provided evidence that microglial activation in the EAE hippocampus was associated with IL-1β expression, and hippocampal slices from control mice incubated with activated microglia displayed alterations of GABAergic transmission similar to those seen in EAE brains, through a mechanism dependent on enhanced IL-1β signaling. These data may yield novel insights into the basis of cognitive deficits in EAE and possibly of MS.