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Anti-TNF treatment with adalimumab induces changes in the microbiota of Crohn's disease. - Journal of Crohn's & colitis
The composition of the intestinal microbiota is altered in Crohn's disease (CD) patients. The objective of this study was to evaluate the qualitative and quantitative changes in the microbiota of CD patients in three months of treatment with adalimumab (ADA), and determine whether or not these changes are produced towards the recovery of the normal, healthy-like, microbiota.The microbiota composition and the Faecalibacterium prausnitzii / Escherichia coli quantitative relationship as dysbiosis indicator, were studied at baseline (T0), one month (T1) and three months (T3) after starting treatment using a polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rRNA gene fragments and quantitative PCR, respectively in colonic mucosal biopsies from fifteen CD patients and four healthy subjects.T0 and T3 fingerprints were different in all patients; whereas T1 and T3 presented similar patterns. Recovered phylogroups were Firmicutes (79.1%), Bacteroides (12.5%), and Actinobacteria (6.25%). The prevalence of E. coli decreased along the treatment. Relative E. coli loads in CD samples were significantly reduced at every analysed step (T1 and T3) (P<0.005) whereas no significant changes were observed in relative F. prausnitzii counts.Treatment with ADA induces short-term changes in the microbiota composition, which seems to be parallel to the partial recovery of the gut bacterial ecology, with recovery parameters tending to eubiosis recovery. The quantitative determination of dysbiosis representative bacteria, such as E. coli, may provide a fast and reliable indicator of the healing state of the colonic mucosa.Copyright Â© 2015 European Crohnâ€™s and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: firstname.lastname@example.org.
[Ecodialysis: first strategies to limit damages and reduce costs]. - Giornale italiano di nefrologia : organo ufficiale della SocietÃ italiana di nefrologia
In the medical field, the attention to the environmental impact of industrial processes and products is still limited. In recent years there has been an increased sensitivity towards the environment; meanwhile, the economic burden of hazardous waste disposal is becoming evident. Dialysis is a "big producer" of waste and it has been estimated that disposal costs can be up to 10-40% of the cost of disposables. So there are several reasons of interest on "ecodialysis": the high amount of waste defined as "potentially hazardous", which requires a very expensive management and the recyclability potential of the non-contaminated waste, that has not yet been fully explored in dialysis. This primary study has been performed in collaboration with the Politecnico di Torino. Its aim has been to define a schedule of activities by a few brainstorming sessions. This schedule is to be readily performed or it should be developed in detail to optimize, by reducing and recycling, the waste production during the dialysis session. The discussion identified seven basic points for the eco-sustainability of haemodialysis to:  reduce packaging;  facilitate separation of materials, and  their discharge;  differentiate materials;  clearly highlight the potentially hazardous materials;  improve the recyclability of plastic products;  propose a path of recovery and reuse. Although a full optimization requires a close cooperation with the manufacturers and is achievable only in the long term, the reduction of one pound of potentially contaminated materials could presently lead, on a national scale, to a saving of several million euros, which can be better employed in investments to improve our treatments.
A versatile complementation assay for cell-to-cell and long distance movements by cucumber mosaic virus based agro-infiltration. - Virus research
Microinjection, bombardment or tobamovirus and potexvirus based assay has been developed to identify the putative movement protein (MP) or to characterize plasmodesma-mediated macromolecular transport. In this study, we developed a versatile complementation assay for the cell-to-cell and long distance movements of macromolecules by agro-infiltration based on the infectious clones of cucumber mosaic virus (CMV). The movement-deficient CMV reporter was constructed by replacing the MP on RNA 3 with ER targeted GFP. The ectopic expression of CMV MP was able to efficiently move the RNA3-MP::erGFP reporter from the original cell to neighboring cells, whereas CMV MP-M5 mutant was unable to initiate the movement. Importantly, the presence of CMV RNA1 and RNA2 can dramatically amplify the movement signals once the RNA3-MP::erGFP reporter moves out of the original cell. The appropriate observation time for this movement complementation assay was at 48-72 hours post infiltration (hpi), whereas the optimal incubation temperature was between 25 and 28 Â°C. The ectopic co-expression of MPs from other virus genera, NSm from tomato spotted wilt tospovirus (TSWV) or NSvc4 from rice stripe tenuivirus (RSV), could also facilitate the movement of the RNA3::erGFP reporter from the original cell into other cells. The chimeric mutant virus created by substituting the MP of CMV RNA3 with NSm from TSWV or NSvc4 from RSV move systemically in Nicotiana benthamiana plants by agro-infiltration. This agro-infiltration complementation assay is simple, efficient and reliable. Our approach provides an alternative and powerful tool with great potentials in identifying putative movement protein and characterizing macromolecular trafficking.Copyright Â© 2014 Elsevier B.V. All rights reserved.
The 2a protein of Cucumber mosaic virus induces a hypersensitive response in cowpea independently of its replicase activity. - Virus research
Resistance in cowpea to infection with strains of Cucumber mosaic virus (CMV) involves a local hypersensitive response (HR), and previous studies indicated that the 2a replicase of CMV is involved in HR induction. In this study, we confirmed and extended this observation by demonstrating that the nonviral expression of the 2a protein encoded by CMV is able to induce a cell death response in cowpea plants, whereas no other CMV-encoded proteins elicits such response. The 2a single-amino acid mutant, F631Y, no longer induces the necrosis response, yet the A641S mutant still induces cell death. The 2a double mutant, F631Y and A641S, does not induce HR. However, the three 2a mutants have comparable replicase activities in a fluorescence reporter assay. The 2a(D610A) mutant that alters the highly conserved GDD motif abolishes the replicase activity, however it does not affect HR induction in cowpea. The 2a(301-778aa) fragment introduced with the same D610A mutation in the GDD motif is also capable of inducing HR in cowpea. Collectively, these findings suggest that the 2a protein of CMV is sufficient to induce HR in cowpea independently of its replicase activity.Copyright Â© 2012 Elsevier B.V. All rights reserved.
One-step detection of Bean pod mottle virus in soybean seeds by the reverse-transcription loop-mediated isothermal amplification. - Virology journal
Bean pod mottle virus (BPMV) is a wide-spread and destructive virus that causes huge economic losses in many countries every year. A sensitive, reliable and specific method for rapid surveillance is urgently needed to prevent further spread of BPMV.A degenerate reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primer set was designed on the conserved region of BPMV CP gene. The reaction conditions of RT-LAMP were optimized and the feasibility, specificity and sensitivity of this method to detect BPMV were evaluated using the crude RNA rapidly extracted from soybean seeds.The optimized RT-LAMP parameters including 6â€‰mM MgCl2, 0.8â€‰M betaine and temperature at 62.5-65Â°C could successfully amplify the ladder-like bands from BPMV infected soybean seeds. The amplification was very specific to BPMV that no cross-reaction was observed with other soybean viruses. Inclusion of a fluorescent dye makes it easily be detected in-tube by naked eye. The sensitivity of RT-LAMP assay is higher than the conventional RT-PCR under the conditions tested, and the conventional RT-PCR couldn't be used for detection of BPMV using crude RNA extract from soybean seeds.A highly efficient and practical method was developed for the detection of BPMV in soybean seeds by the combination of rapid RNA extraction and RT-LAMP. This RT-LAMP method has great potential for rapid BPMV surveillance and will assist in preventing further spread of this devastating virus.
OS048 Mitochondrial content and function in placental cells and tissuesof preeclampsia and IUGR. - Pregnancy hypertension
Early onset placenta Preeclampsia (ePE) with Intrauterine Growth Restriction (IUGR) is associated with insufficient placental function, leading to decreased nutrient and oxygen (O2) availability for the fetus . Mitochondria (mt) are the cell energy producers. Mt dysfunctions could be involved in altered placental metabolism leading to ePE and IUGR. We previously demonstrated higher levels of mtDNA in human IUGR placentas .Here we investigate mtDNA levels in ePE and PE without IUGR placentas, and we present an innovative technique, High Resolution Respirometry (HRR), on cytotrophoblast cells (CTC) from PE, IUGR and control placentas (C), measuring cell O2 consumption which represents respiratory chain efficiency.mtDNA was measured by Real-Time PCR in 20 PE placentas, with (n=14) or without (n=6) IUGR, and 45 C. CTC were isolated from 4 PE, 4 IUGR and 6 C and characterized by flow cytometry, staining samples with anti-cytokeratin-7 and anti-vimentin antibodies. Cells were located in chambers with atmospheric O2levels; 2 different protocols were used, with or without digitonin permeabilization, allowing to measure the O2 consumption of the respiratory chain complexes singularly or all together. Substrates and inhibitors of different respiratory chain complexes were sequentially administered (succinate, ADP, oligomycin, FCCP, rotenone, antimycin A, glutamate, malate, myxothiazol, TMPD, ascorbate, pyruvate, cytochrome C, differently combined depending on the protocol) and O2 consumption levels were recorded. Data were normalized by Citrate Synthase (CS) activity and CTC mtDNA content.PE placentas: mtDNA content was significantly increased in ePE+IUGR (p=0.02) vs C; opposite to this, mtDNAÂ was decreased in PE without IUGR (p=0.03). CTC: single mt O2 consumption (obtained by normalizing data both by CS activity and mtDNA) was slightly increased both in PE and IUGR. The global cell respiration was increased, though not significantly. The trend towards higher O2 consumption studied on permeabilized cells Â was confirmed for all the respiratory chain complexes.Our study showed that mtDNA is increased also in ePE with IUGR and added the novel observation that mtDNA is decreased in PE without IUGR. In both conditions placental mitochondria present an altered respiratory chain activity, with a trend to a higher respiratory capacity. This could lead to higher ATP production likely as an attempt to compensate for other aspects of placental disease due to small or inefficient exchange capabilities. Further data are needed to confirm these preliminary results, together with specific enzymatic assays to asses the respiratory chain complexes functionality. Supported by Fondazione Giorgio Pardi, Associazione Studio Malformazion(ASM) and by a Grant COFIN (Italian Ministry of Research) on: New markers for preterm deliveries.Copyright Â© 2012. Published by Elsevier B.V.
Determinants of ante-partum depression: a multicenter study. - Social psychiatry and psychiatric epidemiology
Ante-partum depression (APD) is usually defined as a non-psychotic depressive episode of mild to moderate severity, beginning in or extending into pregnancy. APD has received less attention than postpartum depression. This is a cross-sectional study carried out in the Obstetrics and Gynaecology (OG) departments of four different general hospitals in Italy.Women attending consecutively the OG departments for their first ultrasound examination were asked to fill in the Edinburgh Postnatal Depression Scale (EPDS) in its Italian validated version. We used the total scores of the EPDS as a continuous variable for univariate and linear regression analyses; in accordance with the literature, the item analysis of EPDS was carried out by classifying the sample as women with "no depression" (scores 0-9), "possible depression" (scores 10-12), "probable depression" (scores 13+) and "probable APD" (scores 15+).The number of women recruited was 1,608. The EPDS assessment classified 10.9 % of the women as possibly depressed, 8.3 % as probably depressed and 4.7 % probably affected from an APD. EPDS score distribution was associated with nationality (higher scores for foreigners), cohabitation (higher scores for women living with friends or in a community), occupation (higher scores for housewives), past episodes of depression and use of herbal drugs. Non-depressed women had significantly lower values on all ten items as compared with depressed women, however, the pattern of item distribution on the EPDS scale remained similar across depression severity groups. In all four groups item 4 (anxious depression) attained the highest scores, while item 10 (suicidality) attained the lowest scores.
Effects of aging and hypertension on the participation of endothelium-derived constricting factor (EDCF) in norepinephrine-induced contraction of rat femoral artery. - European journal of pharmacology
Endothelium-dependent contraction elicited by high concentrations of acetylcholine was described in hypertensive as well as in aged normotensive rats. The contribution of endothelium-derived constricting factor (EDCF) to norepinephrine-induced contraction is still unknown. We aimed to compare EDCF participation to norepinephrine-induced arterial contraction in spontaneously hypertensive rats (SHR) and aged normotensive Wistar-Kyoto (WKY) rats. Femoral arteries from either adult (7-months-old) or aged (14-months-old) animals were placed in myograph and norepinephrine-induced concentration-response curves were recorded under control conditions and in the presence of indomethacin (cyclooxygenase inhibitor, 10(-5) mol/l) or L-NNA (NO synthase inhibitor, 10(-4) mol/l) or both. Norepinephrine-induced concentration-response curve was enhanced in SHR compared to WKY rats, but concentration-response curve of aged WKY rats was similar to those of adult SHR. Cyclooxygenase inhibition largely attenuated concentration-response curves in all groups. However, this effect was greater in aged WKY rats and adult SHR compared to adult WKY rats. NO synthase inhibition augmented norepinephrine-induced contraction in arteries of adult WKY rats, but not in arteries from aged WKY rats or adult SHR. The combined administration of L-NNA and indomethacin had no additive effects on concentration-response curves. EDCF contribution to norepinephrine-induced contractions of arteries was considerably greater in adult SHR (80Â±3%) and aged WKY rats (86Â±2%) compared to adult WKY rats (35Â±10%). The inhibition of NO synthase augmented EDCF contribution to norepinephrine-induced contraction only in arteries from adult WKY rats (76Â±9%). We conclude that EDCF contribution to norepinephrine-induced contraction of conduit arteries is similarly enhanced in adult hypertensive and aged normotensive rats.Copyright Â© 2011 Elsevier B.V. All rights reserved.
Urinary tract infections in the era of newer immunosuppressant agents: a tertiary care center study. - Saudi journal of kidney diseases and transplantation : an official publication of the Saudi Center for Organ Transplantation, Saudi Arabia
We studied the incidence and the risk factors predisposing to post transplantation urinary tract infection (UTI) and the association with use of different immunosuppressive regimens. We performed a retrospective analysis of 152 recipients of renal transplantation over a period of two years. Seventy one (46.71%) patients had culture positive UTI, Escherichia coli (45.1%) being the commonest. Thirty four (22.39%) patients had acute rejection and 14.4% of those had suffered UTI in the early post transplant period. Immunosuppression included induction with various antibodies and maintenance on antirejection medications. Trimethoprim-sulphamethoxazole was given as prophylaxis throughout the period. The UTI was treated according to microbiological sensitivity. 2.8% died due to urosepsis. In our retrospective analysis renal transplant recipients under the age of 45, female gender and diabetics suffered more UTI. Combination therapy with micro-emulsion form of cyclosporine A, prednisolone and azathioprine developed more UTI (P= 0.0418).
Gender differences in delusional disorder: Evidence from an outpatient sample. - Psychiatry research
Our objective was to study gender differences in delusional disorder (DD), by comparing potential risk factors, clinical correlates, illness course characteristics, and functionality. The sample was composed of 86 outpatients with DD (according to the SCID-I for DSM-IV criteria). The following assessment instruments were used service use and demographic questionnaires, Standardized Assessment of Personality (SAP), the Positive and Negative Symptom Scale (PANSS), Montgomery-Asberg Depression Rating Scale (MADRS), Mini-Mental State Examination (MMSE), Mini International Neuropsychiatry Interview (MINI), Sheehan Disability Inventory (SDI), and the Global Assessment of Functioning (GAF) scale. The female-to-male ratio was 1.6:1. Men were more likely to be single, while women were more likely to be widows. Men had a greater frequency of schizoid and schizotypal premorbid personality disorders and of premorbid substance abuse. There were no differences for other risk factors (immigration, deafness, late onset, other personality disorders, and family history). Men were younger at onset and more frequently had acute onset of the disorder. Men had more severe symptoms (higher score on the global or separate PANSS scales). There were no gender differences for the remaining symptomatological variables (types of DD, presence and severity of depression, presence of hallucinations, severity of global cognitive functioning and presence of axis I comorbidity). Global and partial (work, family, and social) functioning was significantly poorer among men. Course type and consumption of resources appeared to be similar. We conclude that men with DD had significantly more severe symptoms and worse functionality. They also had a higher frequency of schizoid and schizotypal premorbid personality disorders and premorbid substance abuse.Copyright 2010 Elsevier Ireland Ltd. All rights reserved.
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