95 research outputs found
An unusual Erdheim-Chester disease with orbital involvement: A case report
Erdheim-Chester disease is a rare non-Langerhans cell histiocytosis with multiorgan involvement and a specific tropism for perivascular and fatty connective tissue, of unclear origin, with poor response to therapy. Its identification is difficult because of the variable clinical presentation and its lack of knowledge. We report the case of a 63-years-old woman, with a history of bilateral orbital pseudotumor, who comes to our attention because of progressively worsening asthenia, vomiting and systemic inflammation. Total body computerized tomography scan showed a volumetric increase of choroid plexus of the temporal horn of the left lateral ventricle, presence of solid retrobulbar tissue at the level of both maxillary sinuses, lung fibrosis, and retroperitoneal and peri-aortic infiltration. The association of these signs addressed to a diagnosis of Erdheim-Chester disease. Thus, although extremely rare, the diagnosis of Erdheim-Chester disease must be considered in the case of bilateral retro-orbital tumors and multisystemic involvement
A phenobarbital overdose: a case report
Background: Phenobarbital is a long-acting barbiturate,
responsible for many cases of poisoning, from unintentional
overdose or attempted suicide. We report a case of phenobarbital
overdose in a patient with history of depression.
Patients and Methods: A 60 year old woman was admitted to
our Internal Medicine Unit for drowsiness, irritability, difficulties in
the maintenance of an upright position, dysphasia and weakness.
She was suffering from depression and epilepsy and treated with
phenobarbital 150 mg/die.
Results: At the admittance, she had high fever and neck stiffness;
phenobarbital serum levels were 71.2 mcg/ml (3 times u.n.l.);
aminotransferases were 12-17u.n.l. Arterial blood pressure was
80/50 mmHg. An inflammatory meningeal process was excluded
by lumbar puncture; a brain and spinal cord CT scan excluded
spine bone lesions and ischemic stroke. In the suspect of an
overdose, a protocol of urine alkalinization was applied resulting
in a reduction of phenobarbital levels below the therapeutic range
in about 6 days, with state of consciousness, cognitive and
behavioral functions improvement. A rapid normalization in
aminotransferases levels was noted and serology for hepatitis
viruses (HAV, HBV, CMV, EBV, HSV) resulted negative.
Conclusions: In our patient phenobarbital was responsible for
stupor, hypotension, hypertonicity and aminotransferases
elevation, whereas fever was due to a concomitant pulmonary
inflammatory process resolved after antibiotic therapy. Despite the
use of these drugs has been progressively reduced, the number
overdose reports remains still hig
Clinical course and management of acute and chronic viral hepatitis during pregnancy.
Pregnancy is a para-physiologic condition, which usually evolves without any complications in the majority of women, even if in some circumstances moderate or severe clinical problems can also occur. Among complications occurring during the second and the third trimester very important are those considered as concurrent to pregnancy such as hyperemesis gravidarum, intrahepatic cholestasis of pregnancy, HELLP syndrome and acute fatty liver of pregnancy. The liver diseases concurrent to pregnancy typically occur at specific times during the gestation and they may lead to significant maternal and foetal morbidity and mortality. Commonly, delivery of the foetus, even preterm, usually terminates the progression of these disorders. All chronic liver diseases, such as chronic viral hepatitis, autoimmune hepatitis, Wilson's disease, and cirrhosis of different aetiologies may cause liver damage, independently from pregnancy. In this review we will also comment the clinical implications of pregnancies occurring in women who received a orthotopic liver transplantation (OLT) Therefore, the management of immunosuppressive therapy before and after the delivery in women who received liver transplant is becoming a relevant clinical issue. Finally, we will focus on acute and chronic viral hepatitis occurring during pregnancy, on management of advanced liver disease and we will review the literature on the challenging issue regarding pregnancy and OLT
Prevalence and phase variable expression status of two autotransporters, NalP and MspA, in carriage and disease isolates of Neisseria meningitidis.
Neisseria meningitidis is a human nasopharyngeal commensal capable of causing life-threatening septicemia and meningitis. Many meningococcal surface structures, including the autotransporter proteins NalP and MspA, are subject to phase variation (PV) due to the presence of homopolymeric tracts within their coding sequences. The functions of MspA are unknown. NalP proteolytically cleaves several surface-located virulence factors including the 4CMenB antigen NhbA. Therefore, NalP is a phase-variable regulator of the meningococcal outer membrane and secretome whose expression may reduce isolate susceptibility to 4CMenB-induced immune responses. To improve our understanding of the contributions of MspA and NalP to meningococcal-host interactions, their distribution and phase-variable expression status was studied in epidemiologically relevant samples, including 127 carriage and 514 invasive isolates representative of multiple clonal complexes and serogroups. Prevalence estimates of >98% and >88% were obtained for mspA and nalP, respectively, with no significant differences in their frequencies in disease versus carriage isolates. 16% of serogroup B (MenB) invasive isolates, predominately from clonal complexes ST-269 and ST-461, lacked nalP. Deletion of nalP often resulted from recombination events between flanking repetitive elements. PolyC tract lengths ranged from 6-15 bp in nalP and 6-14 bp in mspA. In an examination of PV status, 58.8% of carriage, and 40.1% of invasive nalP-positive MenB isolates were nalP phase ON. The frequency of this phenotype was not significantly different in serogroup Y (MenY) carriage strains, but was significantly higher in invasive MenY strains (86.3%; p<0.0001). Approximately 90% of MenB carriage and invasive isolates were mspA phase ON; significantly more than MenY carriage (32.7%) or invasive (13.7%) isolates. This differential expression resulted from different mode mspA tract lengths between the serogroups. Our data indicates a differential requirement for NalP and MspA expression in MenB and MenY strains and is a step towards understanding the contributions of phase-variable loci to meningococcal biology
Identification of a novel zinc metalloprotease through a global analysis of clostridium difficile extracellular proteins
Clostridium difficile is a major cause of infectious diarrhea worldwide. Although the cell surface proteins are recognized to be important in clostridial pathogenesis, biological functions of only a few are known. Also, apart from the toxins, proteins exported by C. difficile into the extracellular milieu have been poorly studied. In order to identify novel extracellular factors of C. difficile, we analyzed bacterial culture supernatants prepared from clinical isolates, 630 and R20291, using liquid chromatography-tandem mass spectrometry. The majority of the proteins identified were non-canonical extracellular proteins. These could be largely classified into proteins associated to the cell wall (including CWPs and extracellular hydrolases), transporters and flagellar proteins. Seven unknown hypothetical proteins were also identified. One of these proteins, CD630_28300, shared sequence similarity with the anthrax lethal factor, a known zinc metallopeptidase. We demonstrated that CD630_28300 (named Zmp1) binds zinc and is able to cleave fibronectin and fibrinogen in vitro in a zinc-dependent manner. Using site-directed mutagenesis, we identified residues important in zinc binding and enzymatic activity. Furthermore, we demonstrated that Zmp1 destabilizes the fibronectin network produced by human fibroblasts. Thus, by analyzing the exoproteome of C. difficile, we identified a novel extracellular metalloprotease that may be important in key steps of clostridial pathogenesis
A Full-Genomic Sequence-Verified Protein-Coding Gene Collection for Francisella tularensis
The rapid development of new technologies for the high throughput (HT) study of proteins has increased the demand for comprehensive plasmid clone resources that support protein expression. These clones must be full-length, sequence-verified and in a flexible format. The generation of these resources requires automated pipelines supported by software management systems. Although the availability of clone resources is growing, current collections are either not complete or not fully sequence-verified. We report an automated pipeline, supported by several software applications that enabled the construction of the first comprehensive sequence-verified plasmid clone resource for more than 96% of protein coding sequences of the genome of F. tularensis, a highly virulent human pathogen and the causative agent of tularemia. This clone resource was applied to a HT protein purification pipeline successfully producing recombinant proteins for 72% of the genes. These methods and resources represent significant technological steps towards exploiting the genomic information of F. tularensis in discovery applications
BPGA- an ultra-fast pan-genome analysis pipeline
Recent advances in ultra-high-throughput sequencing technology and metagenomics have led to a
paradigm shift in microbial genomics from few genome comparisons to large-scale pan-genome studies
at different scales of phylogenetic resolution. Pan-genome studies provide a framework for estimating
the genomic diversity of the dataset, determining core (conserved), accessory (dispensable) and
unique (strain-specific) gene pool of a species, tracing horizontal gene-flux across strains and providing
insight into species evolution. The existing pan genome software tools suffer from various limitations
like limited datasets, difficult installation/requirements, inadequate functional features etc. Here we
present an ultra-fast computational pipeline BPGA (Bacterial Pan Genome Analysis tool) with seven
functional modules. In addition to the routine pan genome analyses, BPGA introduces a number of
novel features for downstream analyses like core/pan/MLST (Multi Locus Sequence Typing) phylogeny,
exclusive presence/absence of genes in specific strains, subset analysis, atypical G + C content analysis
and KEGG & COG mapping of core, accessory and unique genes. Other notable features include
minimum running prerequisites, freedom to select the gene clustering method, ultra-fast execution,
user friendly command line interface and high-quality graphics outputs. The performance of BPGA has
been evaluated using a dataset of complete genome sequences of 28 Streptococcus pyogenes strains
Bordetella pertussis Autotransporter Vag8 Binds Human C1 Esterase Inhibitor and Confers Serum Resistance
Bordetella pertussis employs numerous strategies to evade the immune system, including the ability to resist killing via complement. Previously we have shown that B. pertussis binds a complement regulatory protein, C1 esterase inhibitor (C1inh) to its surface in a Bvg-regulated manner (i.e. during its virulence phase), but the B. pertussis factor was not identified. Here we set out to identify the B. pertussis C1inh-binding factor. Using a serum overlay assay, we found that this factor migrates at approximately 100 kDa on an SDS-PAGE gel. To identify this factor, we isolated proteins of approximately 100 kDa from wild type strain BP338 and from BP347, an isogenic Bvg mutant that does not bind C1inh. Using mass spectrometry and bioinformatics, we identified the autotransporter protein Vag8 as the putative C1inh binding protein. To prove that Vag8 binds C1inh, vag8 was disrupted in two different B. pertussis strains, namely BP338 and 18–323, and the mutants were tested for their ability to bind C1inh in a surface-binding assay. Neither mutant strain was capable of binding C1inh, whereas a complemented strain successfully bound C1inh. In addition, the passenger domain of Vag8 was expressed and purified as a histidine-tagged fusion protein and tested for C1inh-binding in an ELISA assay. Whereas the purified Vag8 passenger bound C1inh, the passenger domain of BrkA (a related autotransporter protein) failed to do so. Finally, serum assays were conducted to compare wild type and vag8 mutants. We determined that vag8 mutants from both strains were more susceptible to killing compared to their isogenic wild type counterparts. In conclusion, we have discovered a novel role for the previously uncharacterized protein Vag8 in the immune evasion of B. pertussis. Vag8 binds C1inh to the surface of the bacterium and confers serum resistance
Transcriptome Analysis of Neisseria meningitidis in Human Whole Blood and Mutagenesis Studies Identify Virulence Factors Involved in Blood Survival
During infection Neisseria meningitidis (Nm) encounters multiple
environments within the host, which makes rapid adaptation a crucial factor for
meningococcal survival. Despite the importance of invasion into the bloodstream
in the meningococcal disease process, little is known about how Nm adapts to
permit survival and growth in blood. To address this, we performed a time-course
transcriptome analysis using an ex vivo model of human whole
blood infection. We observed that Nm alters the expression of ≈30% of
ORFs of the genome and major dynamic changes were observed in the expression of
transcriptional regulators, transport and binding proteins, energy metabolism,
and surface-exposed virulence factors. In particular, we found that the gene
encoding the regulator Fur, as well as all genes encoding iron uptake systems,
were significantly up-regulated. Analysis of regulated genes encoding for
surface-exposed proteins involved in Nm pathogenesis allowed us to better
understand mechanisms used to circumvent host defenses. During blood infection,
Nm activates genes encoding for the factor H binding proteins, fHbp and NspA,
genes encoding for detoxifying enzymes such as SodC, Kat and AniA, as well as
several less characterized surface-exposed proteins that might have a role in
blood survival. Through mutagenesis studies of a subset of up-regulated genes we
were able to identify new proteins important for survival in human blood and
also to identify additional roles of previously known virulence factors in
aiding survival in blood. Nm mutant strains lacking the genes encoding the
hypothetical protein NMB1483 and the surface-exposed proteins NalP, Mip and
NspA, the Fur regulator, the transferrin binding protein TbpB, and the L-lactate
permease LctP were sensitive to killing by human blood. This increased knowledge
of how Nm responds to adaptation in blood could also be helpful to develop
diagnostic and therapeutic strategies to control the devastating disease cause
by this microorganism
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