Orientia tsutsugamushi, is an endemic disease in Taiwan and may be potentially fatal if diagnosis is delayed.Scrub typhus, a mite-transmitted zoonosis caused by We encountered a 23-year-old previously healthy Taiwanese male soldier presenting with the right ear pain after training in the jungle and an eleven-day history of intermittent high fever up to 39°C. Amoxicillin/clavulanate was prescribed for otitis media at a local clinic. Skin rash over whole body and abdominal cramping pain with watery diarrhea appeared on the sixth day of fever. He was referred due to progressive dyspnea and cough for 4 days prior to admission in our institution. On physical examination, there were cardiopulmonary distress, icteric sclera, an eschar in the right external auditory canal and bilateral basal rales. Laboratory evaluation revealed thrombocytopenia, elevation of liver function and acute renal failure. Chest x-ray revealed bilateral diffuse infiltration. Doxycycline was prescribed for scrub typhus with acute respiratory distress syndrome and multiple organ failure. Fever subsided dramatically the next day and he was discharged on day 7 with oral tetracycline for 7 days.Scrub typhus should be considered in acutely febrile patients with multiple organ involvement, particularly if there is an eschar or a history of environmental exposure in endemic areas. Rapid and accurate diagnosis, timely administration of antibiotics and intensive supportive care are necessary to decrease mortality of serious complications of scrub typhus. Orientia tsutsugamushi, is an endemic disease in Taiwan and may be potentially fatal if diagnosis is delayed. It is an acute febrile illness characterized by a typical necrotic primary lesion (eschar), generalized lymphadenopathy, maculopapular skin rash, and nonspecific symptoms and signs. The incidence of eschar reported in patients with scrub typhus is low, < 25%, and only 5% are found over head, face and neck. An eschar in the external auditory canal has not been reported previously, so the diagnosis of scrub typhus was delayed with serious complications, including acute respiratory distress syndrome (ARDS) and multiple organ failure (MOF). In patients with scrub typhus, the incidence and mortality rate of ARDS are 11.1%-15.2% and 20%-25%, respectively. Reported here is a case of an eschar in the external auditory canal in scrub typhus complicated by ARDS and MOF in a 23-year-old previously healthy Taiwanese male soldier.Scrub typhus, a mite-transmitted zoonosis caused by 3 (reference range [RR]: 4000-11000/mm3) with 87.9% segmented neutrophils, hemoglobin 14.6 g/dL (RR: 14-16 g/dL), platelet counts 24 × 103/mm3 (RR: 140-400 × 103/mm3), blood urea nitrogen 25 mg/dL, creatinine 1.5 mg/dl (RR: 0.7-1.4 mg/dL), sodium 140 mEq/L, potassium 3.9 mEq/L, chloride 108 mEq/L, calcium 8.2 mg/dL, total protein 5.8 g/dL (RR: 6.0-8.0 g/dL), albumin 3 g/dL (RR: 3.5-5.0 g/dL), total bilirubin 4.8 mg/dL (RR: 0.1-1.2 mg/dL), direct bilirubin 2.7 mg/dL (RR: 0.0-0.2 mg/dL), C-reactive protein 16.23 mg/dL (RR <0.3 mg/dL), aspartate aminotransferase (AST) 368 IU/L (RR: 8-38 IU/L), alanine aminotransferase (ALT) 271 IU/L (RR: 4-44 IU/L), alkaline phosphatase (ALK) 324 IU/L (RR: 50-190 IU/L), lactate dehydrogenase 783 IU/L (RR: 120-240 IU/L), glucose 94 mg/dL, creatine phosphokinase 229 IU/L (RR: 10-160 IU/L), and a positive for Weil-Felix reaction with a Proteus OX-K titer of 1:1280 on day 11 of fever. Arterial blood gas analysis was pH 7.501, PaCO2 38.1 mmHg, PaO2 76 mmHg, HCO3- 30.1 mmol/l, and BEB 7.2 with a FiO2 of 60%. A central venous line was setup for monitoring his central venous pressure and fluid replacement because of hypotension. Chest x-ray , is a disease distributed throughout the Asia Pacific rim and endemic in South Korea, China, Japan, Taiwan, Pakistan, India, Thailand, Malaysia and northern Australia. It is an acute febrile illness characterized by a typical necrotic primary lesion (eschar), generalized lymphadenopathy, maculopapular skin rash, and nonspecific symptoms and signs such as fever, chills, headache, alerted sensorium, seizure, sorethroat, otalgia, cough, dysponea, vomiting, abdominal pain, jaundice, diarrhea, hepatosplenomegaly, abnormal bleeding, arthralgia and myalgia[Orientia tsutsugamushi [Proteus OX-K titer of 1:1280 was obtained on day 11 after exposure in our case. While all current scrub typhus tests have limitations and the Weil-Felix and immunofluorescent antibody (IFA) tests have low sensitivity and specificity[Scrub typhus, a mite-transmitted zoonosis caused by the intracellular Gram-negative bacteria, d myalgia-12. Unfod myalgia,12. Serid myalgia. In our d myalgia. Initiald myalgia. Acute rugamushi ,12. Abnougamushi ,8-10,12.ecificity,12. The ecificity,12. ScruWe recommend that scrub typhus should be taken into consideration in acutely febrile patients with varying degree of respiratory distress, impaired liver function, acute renal failure and hematologic involvement, particularly if there is an eschar or a history of environmental exposure in an endemic area-9,11,12.The authors declare that they have no competing interests.BJL took care of this patient in the intensive care units. CYC drew up the first draft of the report. SYH made a substantial contribution to draft the manuscript and revised the draft all over the course of submission. All authors read and approved the final manuscript.Written informed consent was obtained from the patient for publication of this study. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2334/11/79/prepub


Anopheles gambiae, Aedes aegypti, Culex quinquefasciatus, Ixodes scapularis, Pediculus humanus and Rhodnius prolixus. We observed that (1) disease vectors encode a lower percentage of ubiquitin-related genes when compared to Drosophila melanogaster, Mus musculus and Homo sapiens but not Saccharomyces cerevisiae; (2) overall, there are more proteins categorized as E3 ubiquitin ligases when compared to E2-conjugating or E1-activating enzymes; (3) the ubiquitin machinery within the three mosquito genomes is highly similar; (4) ubiquitin genes are more than doubled in the Chagas disease vector (R. prolixus) when compared to other arthropod vectors; (5) the deer tick I. scapularis and the body louse (P. humanus) genomes carry low numbers of E1-activating enzymes and HECT-type E3 ubiquitin ligases; (6) R. prolixus have low numbers of RING-type E3 ubiquitin ligases; and (7) C. quinquefasciatus present elevated numbers of predicted F-box E3 ubiquitin ligases, JAB and UCH deubiquitinases. Taken together, these findings provide novel opportunities to study the interaction between a pathogen and an arthropod vector. Protein regulation by ubiquitin has been extensively described in model organisms. However, characterization of the ubiquitin machinery in disease vectors remains mostly unknown. This fundamental gap in knowledge presents a concern because new therapeutics are needed to control vector-borne diseases, and targeting the ubiquitin machinery as a means for disease intervention has been already adopted in the clinic. In this study, we employed a bioinformatics approach to uncover the ubiquitin-mediated pathway in the genomes of  Vector-borne diseases are some of the most prevalent infectious illnesses worldwide. According to the World Health Organization, there are approximately 216 million cases of malaria alone, with over 1.2 million estimated deaths . OverallUbiquitin consists of a 76 amino acid protein that carries seven lysine (K) residues  . UbiquitDrosophila, it is established that developmental regulation by the evolutionarily conserved Notch receptor depends on ubiquitin [Drosophila because ubiquitination also regulates these pathways in disease vectors, such as: Anopheles gambiae, Aedes aegypti, Culex quinquefasciatus and Ixodes scapularis [In biquitin , and sombiquitin . Additiobiquitin . This imapularis .An. gambiae, Ae. aegypti, C. quinquefasciatus, I. scapularis, Pediculus humanus and Rhodnius prolixus. We compared our findings to datasets available for Saccharomyces cerevisiae, Drosophila melanogaster, Mus musculus and Homo sapiens [The lack of information regarding the role of ubiquitin in disease vectors is problematic since molecular interactions depend on this posttranslational modification. These biochemical signatures may lead to novel factors to control pathogen transmission and/or acquisition . For ins sapiens -13. Whil sapiens ,15, tran sapiens ,16-19 an sapiens ,20,21, iI. scapularis, An. gambiae, Ae. aegypti, C. quinquefasciatus, P. humanus and R. prolixus were downloaded from VectorBase [S. cerevisiae, D. melanogaster, H. sapiens and M. musculus were downloaded from http://www.yeastgenome.org [Complete protein datasets encoded within the genomes of ctorBase . Proteinnome.org , http://nome.org , and httnome.org , respectnome.org , and downome.org . This in<0.5. Importantly, observed discrepancies between our results and findings previously published for S. cerevisiae, D. melanogaster, H. sapiens and M. musculus [Using the selected Pfam domains, the hmmsearch program of the HMMER v3.0 package  was emplmusculus -13 were I. scapularis, An.gambiae, Ae. aegypti, and C. quinquefasciatus were aligned using the Multiple Sequence Alignment tool MUSCLE [ The resulting protein sequences from l MUSCLE  availabll MUSCLE . The ful For each aligned dataset, protein phylogenies were estimated with the maximum likelihood method, employing the PhyML program  availabl-like modifier (SUMO), neural precursor cell expressed developmentally downregulated gene 8 (NEDD8), autophagy-related protein 8 (ATG8), ubiquitin-related modifier-1 (URM1), homologous to ubiquitin 1 (HUB1), E1s (ubiquitin-activating enzyme 1 (UBA1), UBA2, UBA3, UBA1-like, autophagy-related E1-like enzyme (ATG7)), really interesting new gene (RING) , other E3s (homology to E6AP C-terminus (HECT), Cullin, U-box), and DUBs (ovarian tumor (OTU), Josephin, jun activation domain-binding protein (JAB), domain of unknown function (DUF) 862, Wss1p‐like metalloproteases (WLM), ubiquitin carboxyl-terminal hydrolase (UCH), Peptidase_C12, Peptidase_C48, and Peptidase_C54). For direct classification of ubiquitin-like proteins, we manually analyzed and categorized each protein encoded in the genomes of An. gambiae, Ae. aegypti, C. quinquefasciatus, and I. scapularis. Ubiquitin-like proteins analyzed were SUMO, URM1, NEDD8, HUB1 and ATG8. Proteins containing two or more Pfam domains were observed and recorded in Protein sequences belonging to specific subcategories of the five ubiquitination-components , ubiquitin-conjugating enzymes (E2), ubiquitin ligases (E3) and deubiquitinases) were identified based on the Pfam domain matches obtained from hmmsearch. Whenever possible, proteins were cataloged according to their major branching patterns in their respective phylogenies: ubiquitin/ubiquitin-like proteins , while P. humanus had the highest ratio at 3.13. R. prolixus had a %U of 2.25. When compared to four model species, all disease vectors possessed a much lower %U, with the model species ranging from 4.38 to 5.02. The exception was S. cerevisiae (%U = 2.24). Interestingly, D. melanogaster carried a much higher %U at 4.50.  The ubiquitin machinery regulates fundamental biological processes within eukaryotic cells . Thus, wI. scapularis (31), An.gambiae (40), Ae. aegypti (48), C. quinquefasciatus (40), P. humanus (35) and R. prolixus (129) (S. cerevisiae (7), D. melanogaster (30), H. sapiens (45) and M. musculus (41). Apart from S. cerevisiae (7) and R. prolixus (129), our analysis revealed a similar number of ubiquitin proteins across species. We further analyzed sequence similarities among ubiquitin family members  for An. gambiae, Ae. aegypti, C. quinquefasciatus, and I. scapularis from their multiple alignments  . We alsoignments . An. gambiae, Ae. aegypti, C. quinquefasciatus, and I. scapularis ( Ubiquitin-like proteins were first discovered in 1979 and named after the interferon-stimulated gene 15 (ISG15) . Since iapularis . UbiquitAn. gambiae (2), Ae. aegypti (2), C. quinquefasciatus (1), and I. scapularis (1)  . NEDD8 ih vector .An. gambiae, Ae. aegypti, C. quinquefasciatus, and I. scapularis , Ae. aegypti (2), C. quinquefasciatus (3), and I. scapularis (3) (URM1 is a ubiquitin-like protein that shares very little homology with ubiquitin . Four prapularis . HUB1 dih vector . ATG8 isaris (3) . I. scapularis (16), An. gambiae (29), Ae. aegypti (37), C. quinquefasciatus (30), P. humanus (19) and R. prolixus (26)  have a very similar number of E1 proteins, while I. scapularis and P. humanus have noticeably less E1s than the other disease vectors. We also analyzed the E1 number in S. cerevisiae (8), D. melanogaster (30), H. sapiens (42), and M. musculus (25)  domain. The E1 associated to ubiquitin is known as UBA1. UBA2 and UBA3 serve as activators for SUMO and NEDD8 respectively . The E1 xus (26) . The thrlus (25) . We furttimation . I. scapularis (32), An. gambiae (32), Ae. aegypti (36), C. quinquefasciatus (28), P. humanus (29) and R. prolixus (31)  . The numapularis . I. scapularis (151), An.gambiae (147), Ae. aegypti (161), C. quinquefasciatus (156), P. humanus (127) and R. prolixus (86) (3HisCys4. The zf-C3HC4 subcategory was the most common RING-type ligase within selected vectors: I. scapularis (100), An. gambiae (82), Ae. aegypti (84), C. quinquefasciatus (87), P. humanus (80) and R. prolixus (56)  . The numxus (56) -7. I. scapularis (28), An.gambiae (42), Ae. aegypti (43), C. quinquefasciatus (47), P. humanus (31) and R. prolixus (19) (I. scapularis (17), An. gambiae (12), Ae. aegypti (22), C. quinquefasciatus (12), P. humanus (16) and R. prolixus (11) (2HC2 motif and plays a role in DNA replication [I. scapularis (6), An. gambiae (11), Ae. aegypti (12), C. quinquefasciatus (10), P. humanus (6) and R. prolixus (8) (We report the number of zf-Apc11 proteins in xus (19) -7. RINGvxus (11) -7. Rtf2 lication . We analixus (8) -7. It isixus (8) .An. gambiae, Ae. aegypti, C. quinquefasciatus, and I. scapularis by phylogeny estimation , An. gambiae (17), Ae. aegypti (17), C. quinquefasciatus (23), P. humanus (14) and R. prolixus (16)  . The numtimation . I. scapularis (5), An.gambiae (6), Ae. aegypti (9), C. quinquefasciatus (4), P. humanus (7) and R. prolixus (8) (S. cerevisiae (4), D. melanogaster (14), H. sapiens (16), and M. musculus (11)  . The numlus (11) . We then ligases . SimilarI. scapularis (8), An.gambiae (10), Ae. aegypti (10), C. quinquefasciatus (12), P. humanus (7) and R. prolixus (8)  . Much liixus (8) . I. scapularis (43), An. gambiae (39), Ae. aegypti (49), C. quinquefasciatus (151), P. humanus (32) and R. prolixus (39) , D. melanogaster (46), H. sapiens (111), and M. musculus (104)  . We then species . I. scapularis (7), An. gambiae (9), Ae. aegypti (9), C. quinquefasciatus (9), P. humanus (7), R. prolixus (6), S. cerevisiae (2), D. melanogaster (12), H. sapiens (21), and M. musculus (18)  . In mostapularis , S10. I. scapularis (2), An. gambiae (1), Ae. aegypti (1), C. quinquefasciatus (2), P. humanus (2), R. prolixus (2), S. cerevisiae (0), D. melanogaster (1), H. sapiens (13), and M. musculus (10) , a neurodegenerative condition . We repolus (10) . All selapularis . I. scapularis (12), An. gambiae (10), Ae. aegypti (11), C. quinquefasciatus (18), P. humanus (10) and R. prolixus (8)  DUBs . This spixus (8) . Proteinapularis . I. scapularis (3), An.gambiae (1), Ae. aegypti (2), C. quinquefasciatus (3), P. humanus (2) and R. prolixus (11)  . The numlogenies . I. scapularis (2), An. gambiae (2), Ae. aegypti (0), C. quinquefasciatus (1), P. humanus (0), and R. prolixus (0)  . I. scapularis (25), An. gambiae (29), Ae. aegypti (28), C. quinquefasciatus (34), P. humanus (26) and R. prolixus (29) , Ae. aegypti (6), C. quinquefasciatus (8), and I. scapularis (5). ( DUBs containing the UCH domain have been known to release ubiquitin in polyubiquitin chains by hydrolyzing its C-terminus . We perfxus (29) . UCH-assris (5). . I. scapularis , An. gambiae , Ae. aegypti , C. quinquefasciatus , P. humanus , R. prolixus , S. cerevisiae , D. melanogaster , H. sapiens , and M. musculus   protease family ,52. Pept_C54: 5) . In mosth vector . Vector-borne illnesses threaten public health in the tropics. Environmental changes due to globalization and the lack of effective vaccines are also contributing to the spread of these maladies to temperate climates. Thus, novel treatments are necessary in the clinic. One effective therapeutic strategy recently used to treat cancer, neurodegenerative disorders and some infectious diseases is pharmacological intervention targeting (de)ubiquitination. However, the use of (de)ubiquitination as a therapeutic target to combat arthropod-borne diseases has not been established. A possible reason for this gap in translational research is the inherent complexity of vector-borne diseases. Another possibility is the lack of communication among scientists. Pharmacologists and chemists do not typically interact with vector biologists, and vector biologists have little incentive to interact with the drug development community. An. gambiae, Ae. aegypti, C. quinquefasciatus, I. scapularis, P. humanus and R. prolixus. Although independent work will be necessary to validate our bioinformatics analysis, empirical evidence suggests that the ubiquitin machinery is present in disease vectors. Recently, Severo et al., 2013 characterized a RING-type E3 ubiquitin ligase named x-linked inhibitor of apoptosis protein (XIAP) and showed the importance of ubiquitination for microbial pathogenesis in ticks [Here we took advantage of publicly available arthropod genomes and described the ubiquitin machinery of in ticks . Similarin ticks . Corroboin ticks . In summTable S1Source and release date of protein datasets.(PDF)Click here for additional data file.Table S2An.List of proteins in gambiae identified by the HMM search.(XLSX)Click here for additional data file.Table S3Ae. aegypti identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S4C. quinquefasciatus identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S5I. scapularis identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S6P. humanus identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S7R. prolixus identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S8D. melanogaster identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S9H. sapiens identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S10M. musculus identified by the HMM search.List of proteins in (XLSX)Click here for additional data file.Table S11List of protein identifications containing sequences matching two or more Pfam domains.(XLSX)Click here for additional data file.Figure S1I. scapularis, An.Ubiquitin and ubiquitin-like proteins in gambiae, Ae. aegypti, and C. quinquefasciatus. Protein sequences matched with ubiquitin and ubiquitin-like proteins were aligned using MUSCLE and a phylogeny was estimated with the PhyML software. Sequences with a bootstrap value lower than 50 were manually removed. Bootstrap values ranged from 0.64 - 0.86 for highlighted clustered categories . (TIF)Click here for additional data file.Figure S2I. scapularis, An.Ubiquitin and ubiquitin-like activating enzymes in gambiae, Ae. aegypti, and C. quinquefasciatus. Phylogeny of ubiquitin and ubiquitin-like activating enzymes. Matched sequences for ubiquitin and ubiquitin-like activating enzymes were aligned using MUSCLE and a phylogeny was estimated with the PhyML software. Bootstrap values ranged from 0.43 - 0.83 for highlighted clustered categories .(TIF)Click here for additional data file.Figure S3I. scapularis, An.Ubiquitin and ubiquitin-like conjugating enzymes in gambiae, Ae. aegypti, and C. quinquefasciatus. Protein sequences matched with ubiquitin and ubiquitin-like conjugating enzymes were aligned using MUSCLE and a phylogeny was estimated with the PhyML software. (TIF)Click here for additional data file.Figure S4An.Phylogeny of RING and RING-like ubiquitin ligases in gambiae and Ae. aegypti. Protein sequences matched with ubiquitin and ubiquitin-like ligases were aligned using MUSCLE and a phylogeny was estimated with the PhyML software. Proteins not listed under a specific subset were categorized as zf-C3HC4. Phylogeny of RING and RING-like ubiquitin ligases in (A) An. gambiae and (B) Ae. aegypti. Bootstrap values ranged from (A) 0.41 - 0.80 and (B) 0.41 - 0.83 for highlighted clustered categories .(TIF)Click here for additional data file.Figure S5C. quinquefasciatus and I. scapularis.Phylogeny of RING and RING-like ubiquitin ligases in  Protein sequences matched with ubiquitin and ubiquitin-like ligases were aligned using MUSCLE and a phylogeny was estimated with the PhyML software. Proteins not listed under a specific subset were categorized into the zf-C3HC4 classification. Phylogeny of RING and RING-like ubiquitin ligases in (A) C. quinquefasciatus and (B) I. scapularis. Bootstrap values ranged from (A) 0.43 - 0.83 and (B) 0.46 - 0.83 for highlighted clustered categories .(TIF)Click here for additional data file.Figure S6I. scapularis, An.HECT, Cullin and U-box ligases in gambiae, Ae. aegypti, and C. quinquefasciatus. Protein sequences matched with ubiquitin and ubiquitin-like ligases were aligned using MUSCLE and the phylogeny was estimated with the PhyML software. Sequences with bootstrap values lower than 50 were manually removed. Bootstrap values ranged from 0.35 - 0.88 for highlighted clustered categories .(TIF)Click here for additional data file.Figure S7An.Phylogenetic trees of F-box ubiquitin ligases in gambiae and Ae. aegypti. Protein sequences matched with F-box ligases were aligned using MUSCLE and the phylogeny was estimated with the PhyML software. Sequences with a bootstrap values lower than 50 were manually removed. Phylogenetic trees of F-box ubiquitin ligases in (A) An. gambiae and (B) Ae. aegypti. (TIF)Click here for additional data file.Figure S8C. quinquefasciatus and I. scapularis.Phylogenetic trees of F-box ubiquitin ligases in  Protein sequences matched with F-box ligases were aligned using MUSCLE and the phylogeny was estimated with the PhyML software. Sequences with a bootstrap values lower than 50 were manually removed. Phylogenetic trees of F-box ubiquitin ligases in (A) C. quinquefasciatus and (B) I. scapularis. (TIF)Click here for additional data file.Figure S9An.Phylogenetic trees of deubiquitinases in gambiae and Ae. aegypti. Protein sequences were aligned using MUSCLE and a phylogeny was estimated using the maximum likelihood method. Phylogeny of deubiquitinases in (A) An. gambiae and (B) Ae. aegypti. Bootstrap values ranged from (A) 0.43 - 0.87 and (B) 0.37 - 0.84 for highlighted clustered categories .(TIF)Click here for additional data file.Figure S10C. quinquefasciatus and I. scapularis.Phylogenetic trees of deubiquitinases in  Protein sequences were aligned using MUSCLE and a phylogeny was estimated using the maximum likelihood method. Phylogeny of deubiquitinases in (A) C. quinquefasciatus and (B) I. scapularis. Bootstrap values ranged from (A) 0.39 - 0.83 and (B) 0.37 - 0.78 for highlighted clustered categories .(TIF)Click here for additional data file.


Candida species play an important complications in severely-ill hospitalized patients which may lead to therapeutic failure. During a 5-year period from 2006-2010, 7 Candida species and 14 unidentified strains were identified from 887 candidaemia of patients admitted at Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.Bloodstream infections due to Candida species were identified according to characteristics of chlamydoconidia production and carbohydrate assimilation/fermentation profile using 13/6 carbon substrates. Only a limited number of Candida were tested for antifungal susceptibility by agar gradient diffusion (Etest®)to amphotericin B and fluconazole.Candida albicans remains a major pathogen accounted for 50.3%, followed by C. tropicalis(27.4%), C. parapsilosis (12.5%), C.guilliermondii (6%), C. glabrata (1.8%), C. krusei (0.23%), C. rugosa (0.23%), and unidentified species(1.6%). From a total of 103 C. albicans, amphotericin B MIC ranged from 0.016-1ug/ml whereas fluconazole MIC ranged were between 0.023 to > 256ug/ml. Other species tested were C. tropicalis (73 strains) which yielded MIC range of 0.19-2ug/ml for amphotericin B and 0.5->256 ug/ml for fluconazole. 34 strains of C. parapsilosis gave amphotericin B ranged from 0.023-16ug/ml and 0.094->256 for fluconazole.From a total of 887 isolates, C. albicans remains the most prevalence Candida species causing candidaemia followed by C. tropicalis, C. parapsilosis and C. guilliermondii. Those non-albicans Candida seemed to express decrease susceptibility to both major antifungal agents like amphotericin B and fluconazole whereas a small number of C. albicans gave high MIC to fluconazole.None declared.


Stress perfusion (SP) abnormalities predict prognosis in specific subsets of patients with coronary artery disease (CAD). However, results are more controversial in populations with stable CAD. We tried to assess prognostic value of SP cardiac magnetic resonance (CMR) in unselected patients with chronic CAD, beyond conventional risk factors such as left ventricle ejection fraction (LVEF).We considered consecutive patients undergoing CMR for definite/suspected chronic CAD. CMR included first-pass myocardial perfusion assessment after dipyridamole stress (0.56 mg/kg) and late gadolinium enhancement (LGE). CMR images were semi-quantitatively evaluated by two blinded operators. SP-CMR was judged positive if a perfusion defect ≥ 75% of myocardium wall thickness was detected in areas of myocardium without LGE. We also collected clinical and pharmacological history, atherosclerotic risk factors, ECG end Echocardiography data. Events in the follow-up , were recorded with outpatient/inpatient visits or with structured telephonic interviews. Univariate and multivariate Cox hazard analyses were performed, with continuous variables categorized according to cut-offs from the literature.We recruited 374 patients , between January 2002 and December 2006. During a follow-up period of 38±21 months there were 30 deaths (8%). Moreover, 79 patients (21%) underwent revascularization procedures. Univariate analysis showed an association between SP-CMR and mortality . Stronger predictors of death were LVEF (19.1 [6.5-55.7]), LVWMSI (13.2 [4.6-38.2]), LVESV (13.1 [4.9-35.3]), LVEDV (9.2 [3.5-24.6]), LGE (8.3 [3.5-19.5]), anticoagulant (6.3 [2.7-14.3]), QTc interval (5.9 [2.9-12.2]) and loop diuretics . A significant univariate association was also found with LV diastolic function (4.4 [1.9-9.9]), aldosterone antagonist (4.2 [1.9-9.0]) and QRS duration (3.8 [1.8-7.9]). At multivariate analysis SP-CMR was found to be no more significant and independent predictors of death were only LVEF , QTc interval , non-sinusal rhythm  and LGE .A positive SP-CMR was the strongest predictor of non-CMR related revascularization procedures, at univariate as well as at multivariate analysis . Other independent predictors of revascularization need in the follow-up were only diabetes  and smoking habit .Our study showed lack of independent association between stress induced perfusion defects at CMR and mortality in a large group of unselected patients with stable CAD. Conversely, stress CMR was the strongest predictor of subsequent revascularization procedures unrelated to CMR itself.


This study aimed at assessing left ventricular (LV), left atrial (LA) and right ventricular (RV) volumes in patients before and after MitraClip™ implantation by cardiac magnetic resonance imaging (CMR).The MitraClip™ is a novel device for percutaneous mitral valve repair. Recent studies demonstrated a reduction of LV volumes after MitraClip™ implantation using echocardiography. CMR is currently the reference method to assess cardiac volumes but has not been used to assess LV remodeling after MitraClip™ implantation so far.Twelve patients with functional (n=7) or degenerative (n=5) mitral valve regurgitation grade 3 to 4 underwent CMR at baseline (BL) before and at 6 month follow-up (FU) after successful MitraClip™ implantation. CMR protocol consisted of short- and long-axis slices using a steady-state-free-precession cine sequence for the assessment of LV, LA and RV volumes.2; p=0.03) as well as LVESV (82 (54-91) vs. 69 (48-99) ml/m2; p=0.03) indices significantly decreased from BL to FU. No significant difference was found for RVEDV (94 (75-103) vs. 99 (77-123) ml/m2; p=0.91), RVESV (48 (42-80) vs. 51 (40-81) ml/m2; p=0.48) and LAV (87 (55-124) vs. 92 (48-137) ml/m2; p=0.20) indices between BL and FU.Minor device-related artifacts were observed, enabling reliable delineation of endocardial borders in all patients at FU. Figure CMR enables reproducible measurements of cardiac volumes in patients with implanted MitraClip™ devices. Significantly decreased LV but unchanged LA and RV volumes were found at 6 month after successful MitraClip™ implantation.No external funding.


There was an error in the second author's name. The correct spelling is Sudhakar Agnihothram. The correct citation is: Varshney B, Agnihothram S, Tan Y-J, Baric R, Lal SK (2012) SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b. PLoS ONE 7(1): e29542.


DAI are a serious public health problem worldwide. The PC and optimization of the HHA have proven to be excellent tools for DAI minimization or elimination.Assess the impact of PC and HHA on DAI in our AICU.A quasi-experimental study. We calculated the average rates of DAI using NHSN,CDC methodology in two periods:pre (Pa) and post intervention (Pb). Average rates wereexpressed as number of events per 1000 device days (DD). Study periods: ventilator associated pneumonia (VAP) Pa: January 2004-January 2010, Pb: February 2010-February 2013. Catheter-associated bacteremia (BACT) Pa: January 2004-June 2010, Pb: July 2010-February 2013. Catheter-associated urinary tract infection (UTI) Pa: January 2007-October 2011Pb: November 2011-February 2013. The Institute of Healthcare Improvement (IHI) proposed PC were implemented. PC adherence was assessed periodically. In October 2009, WHO campaign to improve HHA was implemented. Periodic measurements of HHA were performed. Statistical analysis: we compared average rates of HAI in both periods with Mann-Whitney test. We calculated the Incidence Rate Ratio (IRR) as Pb average incidence rate/Pa average incidence rate.NEU rate decreased from 9.88 (Pa) to 2.60 (Pb), P <0.001. IRR 0.26, 74% rate reduction. Attributable risk: 7.28/ 1000 DD. Cases avoided in Pb: 33.4 (4589 DD in three years). BACT rate decreased from 5.35 (Pa) to 2.34 (Pb), p 0.007. IRR: 0.44, 56% rate reduction. Attributable risk: 3.01/ 1000 DD. Cases avoided in Pb: 14.99 (4983 DD in 32 months). ITU rate decreased from 2.45 (Pa) to 1.30 (Pb), P 0.32. IRR: 0.53, 47% rate reduction. Attributable risk: 1.15/ 1000 DD. Cases avoided in Pb: 3.34 (2908 DD in 16 months). Adherence to PC and HHA ranged between 80 and 95%.PC implementation and HHA optimization in our AICU was associated with statistically significant decreases in VAP and BACT rates, and similar tendency of UTI rate. The significant number of cases averted, fully justifies the implementation of these tools.None declared.


Once daily ritonavir-boosted darunavir (DRV/RTV) is a preferred antiretroviral regimen for treatment-naïve patients. The pharmacokinetics (PK) of DRV/RTV may be influenced by patient demographics and co-medications. Also with an increased aging HIV population, investigations into the impact of older age on PK are important.0-24) and baseline CD4 cell count were 39yr (21-63), 74kg (57-105), 24kg/m2 (18-31), 4.35mg.h/L (2.27-10.99) and 500cells/mm3 (227-1129), respectively; 49 patients had undetectable viral load at time of study. PK sampling was performed at steady-state and between 1-3 PK curves were available per patient. Nonlinear mixed effects modelling (NONMEM v. VI 2.0) was applied to determine DRV PK parameters, interindividual and interoccasion variability and residual error. The following covariates were evaluated: age, weight, BMI, sex, ethnicity, RTV AUC0-24 and raltegravir co-medication (400mg twice daily). The model was validated by means of simulation and visual predictive check.Data were pooled from 3 DRV/RTV PK studies. In total 51 HIV-infected patients  stable on DRV/RTV  were included. Median age, weight, BMI, RTV area under the curve over 24h (AUCa 0.914h-1) and lag-time (0.358h) best described the data. Inclusion of a different apparent oral clearance (CL/F) and volume of distribution (V2/F) for one of the studies improved the fit . RTV AUC0-24 and age were significantly associated with DRV CL/F. An increase in age of 1yr produced a fractional decrease in DRV CL/F of 0.014, equivalent to a 14% reduction in CL/F with every 10yr increase in age. Based on the visual predictive check 94% of observed DRV concentrations were within the 95% prediction interval, indicative of an adequate model.A 2-compartment model with first-order absorption (k0-24 and age were significantly related to DRV CL/F. The impact of age requires further investigation and clarification over a wide age range, particularly in an elderly population.A population model describing the PK of once daily RTV-boosted DRV has been developed and validated. RTV AUC


Tocilizumab (TCZ), an IL-6 receptor inhibitor, improves arthritis and systemic symptoms associated with systemic JIA. It may be a valuable option in patients with JIA who show inadequate response to anti-TNF agents.To analyze the short-term effectiveness and safety of TCZ in patients with JIA refractory to anti-TNF agents.All patients who received TCZ due to refractory disease were included. All patients had exhibited anti-TNF primary or secondary failure. Data were retrieved from the Rheumatology Section data base. TCZ was administered intravenously at a dose of 8 mg/kg for patients > 30 Kg, 12 mg/Kg for patients < 30 Kg every 2 weeks. Efficacy endpoints included improvement (ACR Pedi30), disappearance of systemic symptoms, and reduction in corticosteroid dose.9 patients with JIA  were included. Median age at study entry was 10 years, disease duration 6 years. They received TCZ for 3 to 9 months. Patients had been refractory to etanercept (9), adalimumab (7) or infliximab (2). At baseline (medians): active joints: 11, joints with limited motion: 4, wellbeing (0-3): 0.1, disease activity (0-3): 0.45, ESR: 35 mm/h; CHAQ > 0.5 3 patients, fever/rash 1 patient. Six patients were receiving corticosteroids. All patients met improvement criteria, 7 before the 3rd TCZ infusion. At 1 month after TCZ therapy began: active joints: 4, joints with limited motion: 3, wellbeing: 0.05, disease activity: 0.24, ESR: 3 mm/h; CHAQ > 0.5 1 patient, fever/rash 0 patient. During the treatment, 4 patients reduced dose (< 50%) of corticosteroids. No side effects were recorded.Tocilizumab seems to be effective and safe with a rapid onset of action in children with anti-TNF refractory JIA.


KRAS genotype in L-L and other or multiple metastatic (O/MM) MCRC patients treated with the FIr-B/FOx regimen was retrospectively evaluated.Bevacizumab (BEV) plus triplet chemotherapy can increase efficacy of first-line treatment of metastatic colorectal cancer (MCRC), particularly integrated with secondary liver surgery in liver-limited (L-L) patients. The prognostic value of the KRAS codon 12 and 13 and BRAF mutations by SNaPshot and/or direct sequencing. Fit MCRC patients <75 years were consecutively treated with FIr-B/FOx regimen: weekly 12-h timed flat-infusion/5-fluorouracil (TFI 5-FU) 900 mg/m2, days 1, 2, 8, 9, 15, 16, 22 and 23; irinotecan (CPT-11) 160 mg/m2 plus BEV 5 mg/kg, days 1, 15; oxaliplatin (OXP) 80 mg/m2, days 8, 22; every 4 weeks. MCRC patients were classified as L-L and O/MM. Activity and efficacy were evaluated and compared using log-rank test.Tumoral and metastatic samples were screened for KRAS wild-type (53%), 28 KRAS mutant (47%). At 21.5 months median follow-up, objective response rate (ORR), progression-free survival (PFS) and overall survival (OS) were, respectively: KRAS wild-type 90%, 14 months, 38 months; KRAS mutant 67%, 11 months, 20 months. PFS and OS were not significantly different. PFS and OS were significantly different in L-L compared to O/MM evaluable patients. In KRAS wild-type patients, clinical outcome of 12 L-L compared to 18 O/MM was significantly different: PFS 21 versus 12 months and OS 47 versus 28 months, respectively. In KRAS mutant patients, the clinical outcome of 13 L-L compared to 14 O/MM was not significantly different: PFS 11 months equivalently and OS 39 versus 19 months, respectively.In all, 59 patients were evaluated: 31 KRAS genotype wild-type and mutant does not significantly affect different clinical outcomes for MCRC patients treated with the first-line FIr-B/FOx intensive regimen. KRAS wild-type patients with L-L disease may achieve a significantly prolonged clinical outcome due to integration with secondary liver surgery, with respect to KRAS mutant patients.The  KRAS wild-type metastatic colorectal cancer (MCRC), reported overlapping activity and efficacy in phase III trials, ranging between objective response rate (ORR) 39% to 68%, progression-free survival (PFS) 7.2 to 10.6 months, overall survival (OS) 19.9 to 26.1 months [et al. [Triplet regimens consisting of chemotherapeutic drugs, or doublets plus bevacizumab (BEV)  or cetuximab  monoclonal antibody) in EGFR-overexpressing and 1 months . Phase I [et al. , proposeRAS, BRAF, PIK3CA genes, or loss of tumor suppressor function of PTEN, resulting in continuous activation of the RAS-mitogen-activated protein kinase (MAPK) or phosphoinositide 3-kinase (PI3K) pathways, characterize most colorectal cancers (CRC) [KRAS mutations represent an early event in colorectal tumorigenesis [BRAF mutations, prevalently c.1799 T>A (V600E) mutation, characterize 4.7% to 8.7% of CRC [Gain-of-function mutations of rs (CRC) -9. KRAS igenesis ,11 and oigenesis ,13, c.35igenesis , and codigenesis . They imigenesis . BRAF mu% of CRC -20.KRAS wild-type and mutant MCRC patients treated with irinotecan, 5-fluorouracil and leucovorin (IFL) plus BEV were 27.7 and 19.9 months, respectively [KRAS or BRAF wild-type compared to KRAS or BRAF mutant genotype was not significantly different, even though the hazard ratio (HR) was 0.64 and 0.38, respectively. A significantly better prognosis was reported only when KRAS/BRAF wild-type patients were compared with patients harboring mutations in the KRAS or BRAF genes (HR 0.51) [KRAS wild-type genotype significantly predicts a favorable clinical outcome of anti-EGFR or anti-vascular endothelial growth factor (VEGF) drugs added to doublet chemotherapy [the KRAS mutant genotype, BEV addition to IFL significantly prolonged PFS up to 9.3 months, without increasing OS and activity, compared to IFL [Clinical outcome  according to wild-type and mutant genotype assesses the prognostic relevance of a specific biomarker, potentially including the predictive role of effectiveness of treatment strategies. In randomized studies, the predictive relevance of wild-type or mutant genotype can also be specifically assessed by comparing experimental and control arms. The reported median OS values of ectively ,21. The HR 0.51) . KRAS wiotherapy ,21-23. Id to IFL ,21.KRAS genotype in MCRC patients enrolled in a previously reported phase II study [Here, we report a retrospective exploratory analysis evaluating the prognostic value of the II study  and in aII study .MCRC patients were enrolled in a previously reported phase II study  and in tL-oxaliplatin (OXP) [2/day, over 12 h (from 10:00 pm to 10:00 am), days 1, 2, 8, 9, 15, 16, 22 and 23; CPT-11 , 160 mg/m2, days 1, 15; BEV , 5 mg/kg, days 1, 15; l-OXP , 80 mg/m2, days 8, 22; cycles every 4 weeks.The FIr-B/FOx regimen was developed from previously reported doublet and triplet chemotherapy schedules ,25, consin (OXP) : TFI 5-FKRAS and BRAF genetic analyses were performed on paraffin-embedded tissue blocks from the primary tumor and/or metastatic sites. Genotype status was assessed for KRAS codon 12 and 13 mutations and BRAF c.1799 T>A (V600E) mutation by SNaPshot® multiplex screening for KRAS mutations and KRAS/BRAF mutations in 36 and 32 samples, respectively [KRAS mutations in 23 samples and to confirm detected mutations. After treatment with xylene thyocyanate and selection of tumoral cell clusters, DNA was isolated using the RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE Tissues  according to manufacturer's instructions. When considering the contamination of tumoral samples by non-malignant cells, a KRAS mutation in the tumor was defined as appearance of a mutant peak with a height of at least one-third compared to the wild-type.ectively ,27; direKRAS exon 2 and BRAF exon 15 were simultaneously amplified by polymerase chain reaction (PCR) using specific primers and purified using NucleoSpin® Extract II kit . PCR-amplified DNA was analyzed using the ABI PRISM SNaPshot Multiplex kit  and five primers including an additional tail at their 5' end allowing their simultaneous detection. Sense primers allowing the extension at nucleotides KRAS c.34G, c.35G, c.37G, c.38G and BRAF c.1799T were used and a multiplex SNaPshot reaction was performed as reported [KRAS exon 2 sequencing was performed from PCR-amplified tumor DNA using the Big Dye V3.1 Terminator Kit . Labeled products were separated using an ABI Prism 3130xl Genetic Analyzer . Data were analyzed using the GeneMapper Analysis Software version 4.0 .SNaPshot multiplex assay was performed as elsewhere reported ,27. Briereported . KRAS exKRAS genotype on clinical outcome of MCRC patients treated with FIr-B/FOx as first-line treatment. Moreover, patients were classified according to involved metastatic sites, L-L and O/MM [KRAS wild-type and mutant MCRC patients. Patients with L-L metastases were evaluated at baseline and every three cycles of treatment by a multidisciplinary team, consisting of a medical oncologist, liver surgeon and radiologist, to dynamically evaluate resectability defined according to resectability categories previously reported [A retrospective analysis was planned to evaluate prognostic relevance of and O/MM , to evalreported . ResectiKRAS wild-type versus mutant, L-L versus O/MM, and KRAS wild-type L-L versus O/MM, and KRAS mutant L-L versus O/MM MCRC patients [Clinical criteria of activity and efficacy were ORR, PFS and OS. ORR was evaluated according to Response Evaluation Criteria In Solid Tumors (RECIST) criteria ; patholopatients .KRAS wild-type and mutant genotypes was 31 (53%) and 28 (47%), respectively  and 19 (61%); KRAS mutant, 13 (46%) and 15 (54%). Table KRAS mutations detected in 28 patients: codon 12, 24 patients (85.7%), specifically c.35 G>A 15 patients (53.5%), c.35 G>T 7 patients (25%), c.34 G>A and c.35 G>C, 1 patient each; codon 13, 4 patients (14.2%), c.38 G>A 3 patients (10.7%) and c.37_39 dupl, 1 patient. A total of 32 tumoral samples (54%) were also analyzed for BRAF and no BRAF mutation was detected; 18 out of 31 KRAS wild-type MCRC patients were KRAS and BRAF wild-type; 14 out of 28 KRAS mutant MCRC patients were BRAF wild-type. EGFR protein expression was positive in 35 patients (59%) and negative in 24 patients (41%): among KRAS wild-type patients, positive in 23 patients (74%) and negative in 8 patients (26%); among KRAS mutant patients, positive in 13 patients (40%) and negative in 15 patients (60%).A total of 59 tumoral samples of 64 enrolled MCRC patients 92%) were available: 46 primary tumors and 13 metastases . Demographic and baseline features of patients were representative of the overall phase II study population . We observed 27 objective responses: 23 partial responses (77%) and 4 complete responses (CRs) (13%); 2 stable diseases (7%); 1 progressive disease (3%). Disease control rate was 97% (95% CI 90 to 100). Liver metastasectomies were performed in 11 patients (35%), 10 out of 12 L-L patients (83%). Median PFS was 14 months (1+ to 69+ months), 25 events occurred. Median OS was 38 months (1+ to 69+ months), 17 events occurred. Among the 18 KRAS/BRAF wild-type patients, ORR was 83% (95% CI 69 to 97), median PFS was 13 months (4 to 44 months), median OS was 31 months (8 to 66+ months). Among 27 evaluable KRAS mutant patients, ORR was 67% (95% CI 49 to 85). We observed 18 objective responses: 17 partial responses (63%) and 1 CR (4%); 4 progressive diseases (16%). Disease control rate was 85% (95% CI 71 to 99). Liver metastasectomies were performed in 7 patients (25%) out of 13 L-L patients (54%). Median PFS was 11 months (1+ to 60+ months), 20 events occurred. Median OS was 20 months (1+ to 60+ months), 17 events occurred. Overall, R0 liver resections made up 13 out of 18 liver metastasectomies (72%). Pathologic CRs were obtained in 2 patients (11%), both KRAS mutant patients, harboring codon 12 mutations, c.35 G>T and c.34 G>A, with multiple liver-only metastases. In one KRAS wild-type patient with single liver associated with lung metastases, double metastatic resections were performed. KRAS wild-type compared with mutant patients did not show significantly different PFS nor OS, even if OS seems to be favorable in KRAS wild-type patients  with FIr-B/FOx according to extension of metastatic disease  in KRAS ts Table . Among 2ts Table .KRAS wild-type patients, ORR in 12 L-L and 18 O/MM patients were 100% and 80%, respectively. Overall activity was 100% (ten liver metastasectomies and two cCRs) in L-L and 17% (one liver plus lung metastasectomy and two cCRs) in O/MM patients, respectively. Significantly different clinical outcome was confirmed in L-L compared to O/MM, respectively  while no liver metastasectomy nor cCR was obtained in O/MM patients. The comparison of PFS and OS in KRAS mutant L-L and O/MM patients was not significantly different: median PFS 11 months (3 to 60+ months) versus 11 months (1+ to 37 months), respectively; median OS 39 months (8 to 60+ months) versus 19 months (1+ to 59+ months), respectively  significantly increased median PFS up to 9.3 months, while ORR was equivalent to doublet arm , and median OS increased up to 19.9 months, even if not significantly [In ectively ,21,31,32ectively ,23,31-33patients . In KRASficantly .KRAS wild-type and mutant MCRC patients, BEV addition to triplet chemotherapy, according to FIr-B/FOx schedule, reported high activity and efficacy: ORR 90% and 67%, median PFS 14 and 11 months, median OS 38 and 20 months, respectively. A similar clinical outcome was also obtained in KRAS/BRAF wild-type patients. Equivalent efficacy was reported with FOLFOXIRI/BEV regimen: ORR 82% and 71%, median PFS 13.6 and 12.6 months, respectively [In ectively .KRAS wild-type and mutant patients, even if not significantly, while they were equivalent in the FOLFOXIRI plus BEV study [KRAS wild-type and mutant patients, respectively [KRAS/BRAF wild-type patients compared with patients harboring mutations in the KRAS or BRAF genes (HR 0.51) [KRAS wild-type and mutant MCRC patients treated with BEV-containing chemotherapy failed to significantly differentiate prognosis, as in the present study. Thus, intensive regimens adding BEV to triplet chemotherapy can further increase activity and efficacy in KRAS wild-type and mutant patients. Randomized studies would be able to properly evaluate this.Median PFS and OS values of MCRC patients treated with FIr-B/FOx were different in EV study . BEV addectively ,21. SignHR 0.51) . Direct KRAS wild-type L-L patients, accounting for 20% of fit MCRC patients, could gain 100% overall activity with an integrated medical and surgical approach, due to performed liver metastasectomies and long-lasting cCRs; median PFS 21 months and OS 47 months. A significantly favorable prognosis was demonstrated in KRAS wild-type L-L compared to O/MM patients, even if this represents a retrospective, exploratory analysis in a small cohort of MCRC patients. Using neoadjuvant cetuximab with either FOLFOX6 or FOLFIRI for unresectable colorectal liver metastases, metastasectomies were performed in 38% and 30% patients, respectively [KRAS wild-type L-L MCRC patients. In KRAS mutant patients, prevalently harboring c.35 G>A transversion (53.5%), integrated medical and surgical treatment failed to significantly increase PFS and OS in L-L compared to O/MM patients: median PFS was equivalent (11 months), in spite of 54% performed liver metastasectomies in L-L patients; median OS was 39 and 19 months, respectively. These data should be further evaluated in a larger cohort of MCRC patients. A proper multidisciplinary treatment strategy for KRAS mutant patients, showing different aggressiveness [The high activity of triplet chemotherapy plus BEV regimens correlated with increased resection rate of liver metastases and pathologic CR, particularly in L-L MCRC patients ,3,4,6. Wectively . Chrono-ectively . Furthersiveness , sensitiKRAS wild-type and mutant genotypes do not significantly affect the clinical outcomes of MCRC patients treated with the first-line FIr-B/FOx intensive regimen. KRAS wild-type patients with L-L disease may achieve significantly greater benefit from integration with liver metastasectomies compared to O/MM metastatic extension, with respect to KRAS mutant patients. The present findings should be verified in prospective trials of multidisciplinary strategies comparing clinical outcome according to KRAS genotype in patients with L-L and O/MM disease.The authors declare that they have no competing interests.Conception and design: GB, ER. Provision of study materials of patients: GB, AD, GC. Collection and/or assembly of data: all authors. Data analysis and interpretation: GB, KC, ER. Manuscript writing: GB, ER. Final approval of manuscript: all authors.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1741-7015/10/135/prepub


The study investigated cardiovascular changes in adult-onset growth hormone deficiency (GHD) and showed that patients with adult-onset GHD have a left ventricular mass index (LVMi) at or below the lower limit of normal, which improves with one year of growth hormone replacement.GHD causes cardiovascular problems, with loss of cardiac response to exercise and increased cardiac mortality, however the underlying processes are poorly understood.Ten patients with adult-onset GHD and age- and sex-matched controls (n=10) underwent CMR. Patients underwent scans before disease treatment and at twelve months after treatment. Cardiac parameters were calculated and indexed to body surface area (BSA). The comparison between groups was done using Mann-U-Whitney test and within the group using Wilcoxon test. The data are presented as median values.2, p=0.315; end diastolic volume index (EDVi) 69.2 vs. 76.3 ml/m2, p=0.1655; end systolic volume index (ESVi) 24.8 vs. 28.3 ml/m2, p=0.3527 and ejection fraction (EF) 64 vs. 63 %, p=0.8197. However, patients had an LVMi beneath the lower limit of normal when compared to published normal ranges (55.4 - 74.0 g/m2).Patients with GHD did not have significantly different left ventricle (LV) mass or volumetric parameters from controls: LV mass index (LVMi) 55.0 vs. 56.6 g/m2, p=0.393), ESVi  and EF  in the GHD group and controls.There were no differences between right ventricular (RV) EDVi  achieving statistical significance compared to pre-treatment values (p=0.0156).Patients with GHD on GH treatment for 1 year showed an increase in median insulin-like growth factor I (IGF-I) SDS from -1.83 to +0.40 (p=0.0068). There was no correlation between LVMi and IGF-I SDS . At one year the median LVMi moved into the previously published normal range (55.0 to 63.0 g/mPatients with adult-onset GHD have an LVMi at or below the lower limit of normal, which improves with one year of growth hormone replacement.Supported by departmental grants by Pfizer and Novartis.


However, sudden cardiac death may be the first clinical manifestation. Hence early detection of myocardial abnormalities before overt left ventricular (LV) dysfunction is warranted for a better risk stratification.Seventeen paucisymptomatic LM carries , 14 paucisymptomatic DCM patients  with mild LV dysfunction (ejection-fraction 40%-55%) and 12 healthy controls  underwent complete clinical, echocardiographic, biohumoral and contrast-enhanced CMR assessment. Cine CMR was used to derive LV volumes, mass and function, and post-contrast CMR to detect late gadolinium enhancement (LGE) as an index of gross fibrosis. Eight LM patients, all DCM patients and all normal controls underwent pre- and post-contrast myocardial T1 mapping with gadolinium partition coefficient calculation, as an index of interstitial fibrosis.LM carriers presented a similar LV end-diastolic volume  and slightly reduced ejection-fraction (61±6% p<0.001) than controls , while in DCM patients LV end-diastolic volume (102±17 ml/m2) and ejection-fraction (49±5%) were worse than both controls and LM carriers (p<0.001). Moreover, DCM patients presented worse diastolic dysfunction , NT-proBNP plasma levels  and Doppler-estimated pulmonary pressure  than LM carriers. Six (43%) DCM patients presented DE, representing 9±3% of LV mass; similarly, seven (42%) LM carriers had DE, but with a larger extension . In LM carries DE distribution was patchy (2 pts) or midwall (5 pt), similarly to DCM patients . GPC reached a steady state 5 minutes after gadolinium injection. In DCM patients GPC was slightly higher than controls , while in LM carriers it was much higher (0.45±0.07) than controls (p<0.01). Even excluding hyperenhanced myocardium, in LM carriers GPC remained higher than controls, while in DCM patients only slightly higher than controls.Myocardial fibrosis occurs early in LM carrier before LV dilatation and dysfunction and may represent an early phenotypic expression of cardiac involvement.


Bloodstream infections (BSI) are important causes of morbidity and mortality. Most of all, when are caused by drug-resistant organisms (DR).To investigate the epidemiology, etiology, systemic response and treatment of DR-BSI.EPINE-EPPS project. Comparisons between groups were performed by means of the X2 test for categorical variables or analysis of variances (ANOVA) for continuous variables.A retrospective study was conducted about all BSI diagnosed in a secondary hospital during one year. The pattern resistant pathogen study was We included 60 patients  with 63 DR-BSI of which 71.5% were nosocomial and healthcare-associated BSI.Unknown and intravascular catheter-related DR-BSI accounted for 49.2% of cases. Among secundary infections, the source was 37.5% urinary track, 31.2% intra-abdominal and 15.6% respiratory track infections.S. aureus. Among DR-Gram-negative bacilli, 12.2% of enterobacteracea family produced extended-spectrum B-lactamasas. We found 5 DR-BSI caused by Acitetobacter carbapenem resistant and 3 DR-BSI by P. aeruginosa carbapenem resistant.Overall DR-BSI, DR-Gram-positive cocci were 55.6%. The most common isolated pathogens were staphylococcus coagulase-negative and Median time to diagnosis for DR-Nosocomial BSI was 14 days (IQR), 7-35 days after hospital admission. For Gram-negative was 11 days (7.5-31.5) and for Gram-positive 19 days (7-29).Only 31.7% of DR-BSI received appropriate initial empirical antimicrobial therapy versus 73.5% of non DR-BSI (p<0.001). More than one third (36.5%) of the episodes occur with significant systemic response (severe sepsis or septic shock). The crude mortality rate was 25.4 % (p<0.001). If the patient developed severe sepsis or septic shock crude mortality rose to 52.2%.Information about local epidemiology is important to develop prevention and control strategies in drug–resistant microorganism and to improve the management of BSI.None declared


Remarkably, while glucose potently stimulated GIP release, fructose was without effect. Similar patterns were found in the mouse and rat, with both fructose and glucose stimulating GLP-1 secretion, whereas only glucose caused GIP secretion. In GLUTag cells, a murine cell line used as model for L cells, fructose was metabolized and stimulated GLP-1 secretion dose-dependently (EC50 = 0.155 mM) by ATP-sensitive potassium channel closure and cell depolarization. Because fructose elicits GLP-1 secretion without simultaneous release of glucagonotropic GIP, the pathways underlying fructose-stimulated GLP-1 release might be useful targets for type 2 diabetes mellitus and obesity drug development.Nutrients often stimulate gut hormone secretion, but the effects of fructose are incompletely understood. We studied the effects of fructose on a number of gut hormones with particular focus on glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). In healthy humans, fructose intake caused a rise in blood glucose and plasma insulin and GLP-1, albeit to a lower degree than isocaloric glucose. Cholecystokinin secretion was stimulated similarly by both carbohydrates, but neither peptide YY While it has long been known that the presence of nutrients such as glucose, fat, and protein in the intestinal lumen stimulates secretion of many gut hormones, the effects of fructose are poorly understood. While some studies have shown that fructose does not stimulate insulin secretion in: 1) humans when perfused intraduodenally, 2) in rat pancreatic preparations, or 3) in isolated human and rat pancreatic islets  participated in the study, which was approved by the local ethical committee and conducted according to the principles of The Helsinki Declaration. All subjects had normal fasting blood glucose levels , and none had parents or siblings diagnosed with any type of diabetes. No subjects received medication known to interfere with glucose homeostasis. Each subject was studied on two occasions within 3 wk after the first day of study. Subjects were instructed to refrain from vigorous exercise and alcohol for at least 24 h before each study. Study days began at 0830 preceded by a 10-h overnight fast. Venous blood samples were collected at time −10, 0, 15, 30, 45, 60, 90, 120 min in prechilled EDTA (10.8 mg) tubes  through a polyethylene cannula placed in a cubital vein. At time 0 min, the seated, single-blinded, subjects drank a sugar solution containing 75 g fructose or glucose dissolved in 300 ml water (RT) within 2 min. For palatability, solutions were refreshed with lemon juice. Upon collection, blood samples were instantly chilled on ice and centrifuged  within 30 min. Plasma were stored at −20°C until analysis.Nine healthy volunteers  (data not shown). Rats were divided into weight-matched groups (n = 4/occasion) and given a bolus of either glucose or fructose  diluted in milli-Q water to a final concentration of 50% (wt/vol) or a matched volume vehicle (milli-Q water). Rats from the same cage received different treatments. Blood (200 μl) was collected into prechilled EDTA-coated capillary tubes  by sublingual vein puncture and instantly transferred onto ice. The zero sample was collected 10 min before bolus administration. At time 0 min rats were stimulated with either vehicle, glucose, or fructose, and blood was collected at time 15, 30, and 60 min. Blood glucose was measured instantly after collection, and samples were centrifuged  to obtain plasma, which was transferred to fresh Eppendorf tubes and immediately frozen on dry ice. Samples were stored at −20°C until analysis. At the end of the experiment rats were anesthetized by subcutaneous injection of hypnorm/midazolam  and killed by diaphragm perforation.Studies were approved by the Danish Animal Experiments Inspectorate (2013-15-2934-00833). Male Wistar rats were obtained from Taconic  and allowed at least 1 wk of acclimatization before the day of experiment. Rats followed a 12:12-h light-dark cycle with ad libitum access to standard chow and drinking water. Every day for a week up to the experiment, rats were handled to minimize stress-related responses on the day of study. Handling included restraint and gavage feeding with drinking water. Experiments were carried out on two occasions on nonfasted rats (294.9 ± 3.8 g) just before their nocturnal feeding period (1700). Weight did not differ between groups or study occasions (time −30 min and again at time 0 min with fructose or glucose (3 g/kg) diluted in drinking water to a final concentration of 20% (wt/vol). At time 0 and 6 min, blood (75 μl) for hormone measurement was collected into prechilled EDTA-coated capillary tubes  supplemented with dipeptidyl peptidase 4 inhibitor  . After collection, blood samples were immediately transferred to Eppendorf tubes supplemented with 7.5 μl diprotinin A (1 mM) and centrifuged  within 30 min. Plasma was transferred to fresh Eppendorf tubes, snap-frozen on dry ice, and stored at −20°C until analysis. Blood for glucose measurements (3 μl) was obtained by tail puncture immediately after blood sampling and directly processed for blood glucose measurements as described below. After the first day of study (13 wk), the procedure was repeated. At both occasions, mice were randomly divided into glucose- and fructose-receiving groups. Weights did not differ between fructose and glucose groups at the respective days of study . At the end of the experiment, mice were killed by cervical dislocation.Studies were carried out with permission from the Danish Animal Experiments Inspectorate (2012-15-2934-00207). Female C57BL/6 mice were obtained from Charles River  and allowed 1 wk of acclimatization before the day of experiment. Mice  were fasted overnight but allowed free access to water. Mice were prestimulated with a gavage of 100 μl drinking water at 2 until 70–80% confluent, then trypsinized and plated  on a 24-well plate precoated with matrigel (1:100)  as described previously  supplemented with 0.1% (wt/vol) fatty acid-free BSA  and incubated for 2 h  with 250 μl test reagents  dissolved in saline buffer. All reagents were supplied by Sigma Aldrich. Supernatants were collected and centrifuged  to remove floating cells and debris and then either snap-frozen on dry ice and stored at −80°C or directly processed for GLP-1active measurement.GLUTag cells were cultured in T75 cell culture flasks in low-glucose (1.0 g glucose/l) DMEM 5564 medium  supplemented with 10% (vol/vol) FCS, 1% (vol/vol) penicillin/streptomycin, and 1% (vol/vol) glutamine. Cells were incubated at 37°C, 5% COeviously . The folIntracellular NAD(P)H concentration levels were measured using NAD(P)H's autofluorescence properties , 23. GLUtotal (GIP1–42 and GIP3–42) was measured with GIP-ELISA , whereas GLP-1 levels were measured by the same Millipore GLP-1 (active) ELISA kit as for the GLUTag cells. In both cases, data were acquired using a Biotek EL800 plate reader controlled by Gene5 software . For the mouse, GIPtotal concentrations were measured as described above, and GLP-1 levels were determined by the Meso Scale assay system . Data were obtained with a Sector Imager 2400 device controlled by MSD Discovery workbench software. Human plasma concentrations of total GLP-1, GIP, insulin (INS), glucagon (GCG), and intact neutrotensin1–13 (NTS1–13) were determined using in-house radioimmunoassays  with 125I-labeled tracers and hormone-specific antiserum, recognizing COOH-terminal epitopes, as described previously . Free and bound moieties were separated by addition of (bovine) plasma-coated charcoal . Supernatants were collected , and the radioactivity was counted in a Packard Cobra II Gamma Counter (GMI). Plasma polypeptide YY3–36 (PYY3–36) levels were measured by a commercially available RIA kit . Plasma cholecystokinin (CCK) concentrations were measured by RIA measuring all the tyrosyl-sulfated forms of CCK as described  read on a SpectraMax M5 plate reader controlled by SoftMax Pro 5.4.1 software . For both human, rat and mouse, blood glucose levels were measured by the glucose oxidase method immediately after sampling by a handheld Accu-chek Compact plus device . For the rat, plasma GIPescribed . For allt-test, paired Student's t-test, or one-sample t-test, testing values against a hypothetical value of zero, as indicated. P < 0.05 was considered significant. For the human study, baseline values are presented as the mean of the −15 and 0 min levels. EC50 value for fructose-stimulated GLP-1 secretion from GLUTag cells  (P < 0.001) with significantly higher area under the curve (AUC) values after oral glucose vs. fructose  . For both groups, blood glucose concentrations returned to basal levels by the end of the study period (2 h)  A. Both t, n = 9) B. Maximuod (2 h) A.P > 0.05, n = 9). There was a rise in insulin after both glucose and fructose ingestion , albeit glucose elicited a higher maximal response  (P < 0.05) (P < 0.001)  C. Insuli < 0.05) D. Insuli< 0.001) C.P > 0.05) (P = 0.17) (Glucagon levels did not change significantly at any time point after glucose or fructose intake ( > 0.05) E. Glucag = 0.17) F.n = 9)  with progressively increasing concentrations from baseline until 45 min (P < 0.001). Levels then tended to fall, reaching an elevated plateau phase from 60 to 120 min. GLP-1 also rose after oral glucose intake , reaching significantly elevated concentrations at 15 min (P < 0.01). Thereafter, levels remained elevated until 90 min (P < 0.05)  A. GLP-1 l 90 min A. Over t < 0.05) B.P > 0.05, n = 9) , reaching at plateau from 15 to 120 min , fructose did not significantly elevate GIP concentrations , also compared with basal levels (P > 0.05) (P < 0.0001)  C. While  > 0.05) C. GIP AU 0.0001) D.P > 0.05, n = 9) , but levels were slightly elevated at 60 and 90 min after fructose intake  (P > 0.05)  E. PYY co, n = 9) E. Plasma > 0.05) F.P > 0.05, n = 9)  with peak values occurring 15 min after ingestion for both treatments . Thereafter, CCK concentrations decreased for both treatments, albeit with a tendency toward higher values in the glucose treatment group at later time points (P > 0.05)  G. Glucose points G. CCK AU > 0.05) H.P > 0.05, n = 9) (P < 0.001), with similar peak values observed at 15 min for both treatments . NTS AUCs did not differ between treatments (P > 0.05)  I. Glucos > 0.05) J.P < 0.01, n = 7). At 60 min, blood glucose levels had normalized and were not different from basal levels (P > 0.05)  but not in the fructose-treated group   . In contrast, fructose stimulation did not elevate GIPtotal levels at any time points  (P < 0.001), but not after fructose , but GLP-1 AUCs were elevated by both glucose and fructose compared with control (P < 0.05)  A. Blood , n = 6) B. Blood , n = 6) A. GIPtot, n = 6) C. GIP AUfructose D. GLP-1  < 0.05) F.P < 0.05, n = 31) albeit glucose stimulation increased levels to a greater extent than fructose (P < 0.0001)  (P > 0.05). GIPtotal concentrations were elevated by glucose treatment , but fructose had no effect  (P > 0.05) (C).Responses to glucose and fructose were studied on two occasions separated by 13 wk. Blood glucose increased significantly for both groups compared with basal levels  A. GLP-1t n = 23) B to a si n = 23) C. Basal  > 0.05) , A–C.P < 0.05, n = 43)  (P < 0.0001) in a dose-dependent manner, reaching a maximal response at 1 mM  . Addition of gliclazide (GLI) to induce ATP-sensitive potassium (KATP) channel closure did not potentiate fructose-induced GLP-1 secretion , but stimulation with GLI alone caused GLP-1 secretion to a similar degree as tolbutamide  (ATP channel opener diazoxide (340 μM) completely abolished fructose-induced GLP-1 secretion , which reached similar levels as during diazoxide exposure alone  (Metabolic responses to fructose were investigated by the NAD(P)H imaging technique. Representative traces from an experiment are shown in  n = 43) B. Stimul n = 43)  confirme n = 27) C. Fructo = 6–16) D. Again,= 14–15) F. Presen= 10–12) E.1–13 to a similar extent as glucose, and also enhanced insulin secretion, albeit to a lower degree than glucose. In contrast, neither fructose nor glucose altered glucagon or PYY3–36 levels. With a view to perform mechanistic studies regarding the differential responses of the incretin hormones, we also investigated GIP and GLP-1 responses to fructose in mice and rats. In both animals, fructose stimulated GLP-1 secretion to the same extent as isocaloric glucose, but, similar to humans, GIP secretion was not stimulated.The increased intake of dietary fructose in general and high-fructose corn syrup in particular has been suggested to play a role in the current obesity epidemic , 20. MorIn mice and humans, fructose intake also increased blood glucose levels, and a similar tendency was seen in the rat. It is generally accepted that fructose can be converted to glucose in the liver by mechanisms involving rapid phosphorylation of fructose to fructose 1-phosphate by fructose kinase, which by liver-type-B aldolase activity, yields the intermediate metabolites dihydroxyacetone phosphate and glyceraldehyde, with the latter being a substrate for glucose production by further metabolic activity. However, it has also been shown that the small intestine has capacity to transform fructose into glucose by the same pathway . AlthougATP channel closure, causing cell membrane depolarization  and a grant to J. J Holst from the Novo Nordisk Centre for Basic Metabolic Metabolism .The study was supported by a The authors declare no financial or other conflicts of interest.Author contributions: R.E.K., F.M.G., B.H., F.R., J.A.W., and J.J.H. conception and design of research; R.E.K., B.H., J.A.W., and J.F.R. performed experiments; R.E.K., B.H., and J.F.R. analyzed data; R.E.K., F.M.G., B.H., F.R., and J.J.H. interpreted results of experiments; R.E.K. prepared figures; R.E.K. drafted manuscript; R.E.K., F.M.G., B.H., F.R., J.A.W., and J.J.H. edited and revised manuscript; R.E.K., F.M.G., B.H., J.A.W., J.F.R., and J.J.H. approved final version of manuscript.


Objective(s): In the current study, the effects of selected folk medicinal herbs were evaluated in D-galactose-induced aging in male mice.Materials and Methods: Male BALB/c mice were randomly divided into 12 groups composing sham, control, and treated groups. Aging was induced by administration of D-galactose (500 mg/kg/day for 6 weeks). A positive control group was assigned that received vitamin E (200 mg/kg/day). The extract of herbs was prepared, lyophilized, and used in this study. The herbs were administered by gavage for 4 weeks to D-galactose-aged animals at the selected doses (mg/kg/day) as follows: Zingiber officinale (250), Glycyrrhiza glabra (150), Rosmarinus officinalis (300), Peganum harmala (50), Aloe vera (150), Satureja hortensis (200), Teucrium scordium (200), Hypericum perforatum (135) and Silybum marianum (150). One group of animals was assigned as sham and not given D-galactose. Results: At the end of treatment, pro-inflammatory markers including tumor necrosis factor-α (TNF-α), interlukine-1β (IL-β), interlukine-6 (IL-6), NF-kappaB (NF-κb), total antioxidant power (TAP), thiobarbituric acid reactive substances (TBARS) as lipid peroxidation (LPO) marker and male sex hormones i.e. testosterone and dehydroepiandrosterone-sulfate (DHEA-S) were measured in the blood.Conclusion: These data for the first time indicate significant anti-aging potential of examined herbs. Results showed that D-galactose induces a significant oxidative stress and promotes proinflammatory cascade of aging while all herbs more or less recovered these changes. Among 9 herbal extracts, Silybum marianum showed the best effect in restoring aging changes. Aging as a complex of natural circumstance is exhibited by an augmentation in the chance of illness and finally death. Although some theories have been proposed as the mechanisms of aging but the one relating aging and cellular oxidative stress have received more supports. Therefore, it can be said that reduced sex hormones and augmented quantity of oxidative stress parameters or inflammatory cytokines are main biochemical manifestations of aging -2. In acOne of the problems in testing anti-aging compounds is lack of suitable animal models. Although several models have been used so far but among them, typical mouse D-galactose-induced model of aging is the best one that gives closer results to clinical studies. D-galactose is a sugar that at higher levels converts to aldose and hydro-peroxide during the catalysis of galactose oxidase, culminated in the generation of free radicals . These mMany scientists and pharmaceutical companies try to develop a drug to reduce speed of human aging but no effective drug has been discovered yet. In the last decade the importance of folk medicine and herbal medicines have been revisited that resulted in developing many effective drugs for many human diseases. For instance, in the recent years, efficacy of herbal medicines in diseases like inflammatory bowel diseases -10, obesZ. officinale, G. glabra, R. officinalis, P. harmala, A. vera,S. hortensis, T. scor-dium, H. perforatum and S. marianum to test in D-galactose-induced model of mouse aging.We recently proved anti-aging potential of naturally-based drugs like IMOD and Angipars which have strong antioxidant power . On the Chemicals3-6H2O), D-galactose, and vitamin E (Trolox) were purchased from Merck (Germany). Rat specific tumor necrosis factor-α (TNF-α), interlukine-1β (IL-β), interlukine-6 (IL-6), NF-kappa B (NF-κb) ELISA kits were purchased from BenderMed Systems (Austria). Testosterone and dehydroepiandrosterone ELISA kits were purchased from Dia Metra . Thiobarbituric acid (TBA), trichloroacetic acid (TCA), n-butanol, hexadecyltrimethyl ammonium bromide (HETAB), tri (2-pyridyl)-s-triazine (TPTZ), HCl, malondialdehyde (MDA), ferric chloride  were washed in a suitable bactericide (chlorh-exidine). The filets were grounded to a liquid, and the pulp was removed by filtering. The resultant gel was then freeze dried.Herbs were provided from the Research Institute of Medicinal Plants Karaj during June 2009 and were air-dried at room temperature. Samples were authenticated by a botanist (Y. Ajani), and voucher specimens were preserved in the central herbarium of medicinal plants (RIMP). The scientific names and tested parts of the herbal materials are detailed in AnimalsMale BALB/c mice  were provided from Tehran University of Medical Sciences (TUMS) animal house. The animals were housed in standard polypropylene cages with wired-net top in a controlled room  and were allowed free access to standard laboratory pellet diet and water during the experiments. All ethical issues on the use of animals were carefully considered and the study protocol was approved by TUMS review board with code number of 90-03-33-15668.Experimental designth group of animals was the sham group which was not given D-galactose. After 2 weeks, the 11 groups which had been given D-galactose were randomly divided into aging control group , positive control group  and herb-treated groups including 9 groups that each received 500 mg/kg D-galactose per 1 ml drinking water plus Z. officinale (250 mg/kg/day), G. glabra (150 mg/kg/day), R. officinalis (300 mg/kg/day), P. harmala (50 mg/kg/day), A. vera (150 mg/kg/day), S. hortensis (200 mg/kg/day), T. scordium (200 mg/kg/day), H. perforatum (135 mg/kg/day) and S. marianum (150 mg/kg/day), respectively by gavage for 4 weeks , lipid peroxidation (LPO) and male sex hormones including testosterone and dehydroepiandrosterone-sulfate (DHEA-S) were measured in the serum.Measurement of LPO 2SO4 (0.05 M). After addition of TBA (0.2% in sodium sulfate), the sample was heated for 30 min in a boiling water bath. Then, TBA reactive substances (TBARS) as LPO marker adducts were extracted by n-butanol and absorbance was measured at 532 nm as described in details in our previous work (LPO was measured by the reaction of thiobarbituric acid (TBA) with lipid peroxides. Samples were mixed with TCA (20%) and the precipitate was dispersed in Hous work . Data weMeasurement of TNF-α, IL-1β, IL-6 and NF-κbQuantitative detection of TNF-α, IL-1β, IL-6 and NF-κb levels in serum were performed using an enzyme-linked immunosorbent assay rat specific ELISA kit according to each specific brochure. The absorbance of the final colored product was measured in 450 nm as the primary wave length and 620 nm as the reference wave length. TNF-α, IL-1β, IL-6 and NF-κb levels were expressed as pg/mg.Measurement of TAP 3+ to Fe2+. Interaction of TPTZ with Fe2+ results in formation of a blue color with a maximum absorbance at 593. The whole procedure has been described in our previous study . Data were analyzed by one-way ANOVA followed by Tukey post-hoc test for multiple comparisons to ensure the variances of the data are distributed properly. A P< 0.05) and a significant decrease in TAP (P<0.05) were observed when sham group was compared with D-galactose-received aged group. P<0.05; 1.2±0.05 vs. 2.5±0.33, P<0.05; 27±3.9 vs. 49.66±3.4, P<0.05; 45.7±2.4 vs. 97±21.2, P<0.05). As shown in P<0.05; 1.2±0.2 vs. 0.6±0.08, P<0.05) in aged mice was lower than that in the sham.A significant increase in TBARS , and increasing TAP , , respectively. As shown in Z. officinale increased testosterone and DHEA-S  in aged mice.P<0.05), , 2. FiguEffects of G. glabra in aged miceG. glabra .G. glabra recovered D-galactose-induced increase in TNF-α, IL-6, IL-1β and NF-kB , respectively. As shown in G. glabra increased testosterone and DHEA-S levels  in aged mice.D-galactose-induced elevation of TBARS and reduction of TAP , 2 were Effects of R. officinalis in aged miceR. officinalis treatment recovered D-galactose-induced rats by reducing TBARS , and increasing TAP  , respectively. As shown in R. officinalis increased testosterone,  and DHEA-S  in aged mice. P<0.05) , 2. FiguEffects of P. harmala in aged miceP. harmala . P. harmala recovered D-galactose-induced increase in TNF-α, IL-6, IL-1β, and NF-kB , respectively. As shown in P. harmala increased testosterone and DHEA-S  in aged mice.D-galactose-induced elevation of TBARS and reduction of TAP , 2 were Effects of A. vera in aged miceA. vera treatment recovered D-galactose-induced elevation of TBARS (P<0.05), and improved reduction of TAP (P< 0.05). A. vera recovered D-galactose-induced increase in TNF-α, IL-6, IL-1β, and NF-kB , respectively. As shown in A. vera increased testosterone and DHEA-S levels  in aged mice.Effects of S. hortensis in aged miceS. hortensis treatment recovered D-galactose-induced elevation of TBARS (P<0.05), and increased TAP (P<0.05). S. hortensis recovered D-galactose-induced increase in TNF-α, IL-6, IL-1β, and NF-kB , respectively. As shown in S. hortensis increased testosterone and DHEA-S  in aged mice.Effects of T. scordium in aged miceT. scordium . T. scordium recovered D-galactose-induced increase in TNF-α, IL-6, IL-1β, and NF-kB , respectively. As shown in T. scordium increased testosterone and DHEA-S  in aged mice.D-galactose-induced elevation of TBARS and reduction of TAP , 2 were Effects of H. perforatum in aged miceH. perforatum treatment recovered D-galactose-induced rats by reducing TBARS  and increasing TAP  , respectively. As shown in H. perforatum increased testosterone and DHEA-S  in aged mice. P<0.05) , 2. FiguEffects of S. marianum in aged miceS. marianum . S. marianum recovered D-galactose-induced increase in TNF-α, IL-6, IL-1β, and NF-kB , respectively. As shown in S. marianum recovered D-galactose-induced reduction of testosterone and DHEA-S  in aged mice.D-galactose-induced elevation of TBARS and reduction of TAP , 2 were Effects of vitamin E in aged miceP<0.05; 190±13.1 vs. 120±7.5, P<0.05). P<0.05; 1.4±0.26 vs. 2.5±0.33, P<0.05; 30.1±2.2 vs. 49.66±3.4, P<0.05; 57±3.9 vs. 97±21.2, P<0.05), respectively. As shown in P<0.05; 1±0.16 vs. 0.6±0.08, P<0.05) in aged mice.D-galactose-induced elevation of TBARS and reduction in TAP , 2 were In this study, for the first time, we analyzed the anti-aging potentials of nine famous herbs in a well-setup animal aging model using chronic administration of D-galactose. Our results showed that production of free radicals is the principal reason of up-regulation of pro-inflammatory cytokines and the main determinant involved in the D-galactose-induced aging model. Furthermore, these herbs dramatically diminished oxidative stress and proinflammatory cytokines in the aged mice. Supporting the mechanism of action of these herbs and the theory of oxidative stress in aging, vitamin E was used as the standard and showed the similar effects in examined markers of aging.Adapted from corresponding author’s previous paper published in open access source .Interestingly, present results indicated improve-ment of testosterone and DHEA-S by herbs in the aged mice. Decline of steroid hormones with aging is already known and is believed a major contributor to elevation of pro-inflammatory markers .Recent studies have shown the mechanisms of action of anti-aging herbs in reducing aging process that is divided into four categories including anti-oxidant, anti-inflammatory, effect on memory/cog-nition/mood, and the sex hormones . This inGinger [Zingiber officinale Roscoe (Zingiberaceae)] and supplements derived from ginger like zingerone, shogaols and gingerols posse the abilities for the treatment of chronic inflammation. The protective effects of Z. officinale in lessening macromolecular damage in aged mice were shown in this study. Besides, recent study has shown that ginger extracts owns antioxidant activity  or licorice is the root of G. glabra from which a sweet flavor can be extracted. The results of this study showed that G. glabra has the protective effects in declining macromolecular damage in aged mice. It has been shown that G. glabra extract is the safest pigment-lightening agent with the fewest side effects  (Fam. Liliaceace) are the source of aloe vera gel. A. vera gel is greatly used in cosmetics and toiletries for its moisturizing and regenerating action. Also, the leaf of A. vera could assist cellular repairing, imbibition of foods, vitamins, minerals and vital nutrients  comprises 12 species, which possess antioxidant, anti-spasmodic, anti-nociceptive and anti-inflammatory properties -b-farnesene as the major components of T. scordium  Gaertn. . Interestingly, the present findings confirmed that S. marianum causes the best effects in improving antioxidant, anti-inflammatory and male sex hormones in aged mice. This effect is so important and should be considered as an advantage. This can be explained with current knowledge that among many medicinal plants, S. marianum, has been greatly used for centuries as a natural popular complementary medicine for the treatment of several diseases. The main indications for the use of silymarin are related to the hepatoprotection , a flavonoids complex known as ‘milk thistle’ is extracted from the fruit of otection , 63. Alsotection , 65. It otection . Excitinal cells . S. marianum showed the best effect in improving all the D-galactose-induced aging effects. Since all of the selected and examined herbs are already found safe in human and there are good information from traditional medicine, therefore, they can be supplemented into the diet of elderly people to reduce speed of aging. Testing the mixture of these herbs together or with other anti-aging products is among the plans of future.Taking collectively, the present results confirmed our hypothesis that the herbs with highest antioxidant power may reduce speed and rate of aging as evidenced by recovery of proinflammatory cytokines and sex hormones. Among tested herbs,


It has been reported in Mexico that genotypes H and G are the most common, although genotypes prevalence in HIV co-infected patients is unknown.We estimated the prevalence and identified the resistance pattern of HBV/H and G genotypes in HIV co-infected patients and compared them in mono-infected HBV patients.A cross-sectional prevalence and analytic study were realized. Risk factors, HIV or HCV co-infections, antiretroviral therapy (ART) experience, HBsAg, HBeAg, HBV viral load and mutations genetic analysis were collected; CD4+ cells count from HIV co-infected patients and HIV viral load were measured. We calculated the prevalence and exact 95% binomial confidence interval as well the Odds ratios (OR) and 95% confidence intervals to assess the relationship between risks factors and the risk of having HBV/H or G genotype.We enrolled 84 patients, 72 men and 12 women with 41 HIV co-infected patients. The distribution of HBV genotypes was: HBV/H 56 (66%), HBV/G 22 (26.1% [95% CI 17% to 36%]), HBV/F 4 (4.7%) and HBV/A 2 (2.3%). The most frequent mutations presented in 9 HIV co-infected patients and one mono-infected patient with ART experience were rtM204V and seven of them showed genotype G (7/9). Mono-infected HBV patients exposed more probability to HBV/H genotype than co-infected HIV patients OR 13.0 (CI 95% 3.40-49.79), P=0.0001 In contrast co-infected patients presented less possibility to have genotype H, 0.56 (CI 95% 0.42-0.75).Our results suggest that HBV/G genotype predominates in co-infected patients. As well, rtM204V and rtL180M mutations are common in HBV/HIV co-infected patients with genotype G and ART experience.


Urinary tract infections are the main indication for antimicrobials in elderly.Despite the treat for resistance dissemination and therapy failure, clinicians are seldom informed on the per patient evolution of antimicrobial resistance in pathogens.Laboratory results were obtained from 13 voluntary diagnostic laboratories  in Belgium during the year 2005. Susceptibility profiles were done by Kirby Bauer disk diffusion according to CLSI. The first two urine samples from patients older than 65 year were included.E. coli (7188/1654), E. faecalis (1282/403), P. mirabilis (1230/313), K. pneumonia (673/173), P. aeruginosa (293/120), E. aerogenes (375/203), S. aureus (158/54), M. morganii (347/89), Group B streptococci (149/31), C. freundii complex (101/29). When comparing first versus second samples antibiograms for E. coli, a decrease in susceptibility was found for the following antimicrobial agents: cotrimoxazole -6.9%; nitrofurantoin -2.8%, fosfomycin 0.0%; ciprofloxacin -10.8%; cefuroxime -5.6%; amoxicillin-clavulanic acid -5.6%; ampicillin -10.5%. For E. faecalis, marked decreases were found for nitrofurantoin -2.4%; fosfomycin -2.2%; -ciprofloxacin -10.3%; and only mild decreases for amoxicillin-clavulanic acid 0.0%; and ampicillin -1.2%. For K. pneumoniae decreases were in the range of -2.9 to -4.1% for cotrimoxazole, ciprofloxacine, cefuroxime and amoxicillin-clavulanic acid, and was -12.4% for nitrofurantoin. For S. aureus and C. freundii no decrease (<-0.1%) was seen for nitrofurantoin and fosfomycin. For E. aerogenes, decreases of -18 and -12.5% were found for cotrimoxazole and fosfomycin, respectively. M. morganii showed in consecutive samples less susceptibility for cotrimoxazole (-16.2%), fosfomycine (-13.0%) and ciprofloxacin (-10.5%), while only a marginal decrease was found for nitrofurantoin (-0.5%).Following organisms were predominantly isolated (N first samples/N second samples): The resistance selection influence of consecutive samples depends on the antibiotic-bacterium combinations, and thus might be taken into account when empiric therapy guidelines for urinary tract infections in elderly are reviewed.None declared


In patients undergoing atrial fibrillation (AF) procedures, imaging of the left atrium (LA) and pulmonary veins (PV) is important for pre-ablation planning and to identify post ablation complications. Conventional MRA protocols use first-pass, non-gated sequences that require long breath-holds. Quality of non-gated MRA's can be challenging in sick or sedated patients. We developed a novel ECG-gated, respiratory navigated MRA sequence less dependent on patient compliance, which yields better clarity of LA anatomy.Eighty patients with AF underwent either conventional non-gated (n=40) vs. novel ECG-gated (n=40) MRA on a 3T Verio scanner . All MRA's were performed using 0.1 mmol/kg Multihance . A novel ECG-gated, respiratory navigated MRA was developed using 3D saturation recovery prepared, GRE sequence with fast contrast injection  followed by slow infusion . Saturation pulse was applied every heart beat and fat saturation was applied immediately before data acquisition during LA diastole. Additional scan parameters were: axial imaging volume, FOV =400x400x110, voxel size=1.25x1.25x2.5 mm, TR/TE=2.8/1.3ms, flip angle=15 degrees, TI=250ms, phase encoding direction: left to right. Typical scan time was 3-5 minutes.Conventional non-gated MRA were performed with contrast injection rate 2.0 mL/sec and continuous data acquisition during single breath-hold (14 sec.). Scan parameters included: axial imaging volume, FOV=400x263x120 mm, voxel size=1.25x1.25x2.5, TR/TE=2.8/1.1ms, flip angle=27 degrees.All MRA's were randomized and quality scores were determined by two experienced readers for contrast enhancement, border sharpness, and chamber detail of the PV's, LA, and LA appendage (LAA) .  in the non-gated cohort. Border sharpness scored 3.08 vs. 2.35 QS  respectively. Chamber detail was also assessed with specific anatomical positions, which all yielded superiority of ECG-gated MRA ; left atrial appendage 3.11 vs. 2.25 QS ; left atrium 3.08 vs. 2.35 QS .ECG-gated MRA improves image quality with better border sharpness and anatomical detail compared to conventional non-gated MRA. These advantages may translate into improved diagnostics in AF patients including detection of PV stenosis or following LA remodeling post-ablation.No disclosures.


AbstractStenohya Beier, 1967 are described from China: Stenohya pengaesp. n.  and Stenohya huangisp. n. .The presence of Stenohya pengaesp. n. in the tree crown of Castanopsis fabri represents a new habitat for Neobisiidae. A key and a distribution map of the Chinese Stenohya species are also provided.Two new species of the genus  Stenohya Beier, 1967 is a small Asian pseudoscorpion genus of the family Neobisiidae Chamberlin, 1930. At present it includes 12 species , Stenohya curvata Zhao et al., 2011 (Yunnan Province) and Stenohya xiningensis Zhao et al., 2011 (Qinghai Province). species , of whicFagaceae, Styracaceae, Daphniphyllaceae, Lauraceae and Ericaceae  by sweeping vegetation with an entomological net. After examining the specimens in the laboratory, we found them to represent a new species, which is described here under the name Stenohya pengae sp. n. When we examined the pseudoscorpions collected by Prof. Fusheng Huang from Gushan Mountain, Fujian Province, China, we found another new Stenohya species, which is also described and illustrated in this paper as Stenohya huangi sp. n.Damingshan National Nature Reserve is located in the midwest of Guangxi Zhuang Autonomous Region, lying on the Tropic of Cancer, and possesses a rich subtropical primeval forest, which is home to many rare animals and plants. Daming Mountains are densely covered by jungle, including trees of the families ricaceae . In 2011b = basal; sb = sub-basal; st = sub-terminal; t = terminal; ib = interior basal; isb = interior sub-basal; ist = interior sub-terminal; it = interior terminal; eb = exterior basal; esb = exterior sub-basal; est = exterior sub-terminal; et = exterior terminal.The specimens are preserved in 75% alcohol and deposited in the Museum of Hebei University (MHBU). Permanent slide mounts were prepared by removing the chelicerae, pedipalps, leg I and leg IV from specimens with small needles and clearing overnight with lactic acid at room temperature. Drawings were made with the aid of a camera lucida mounted above the eyepiece of a compound microscope. Photographs were taken with a Leica M165 stereomicroscope. Terminology of trichobothria follows urn:lsid:zoobank.org:act:3E8D205C-B127-4BE0-AC87-B029B7F9719Fhttp://species-id.net/wiki/Stenohya_pengae23°08'N, 108°17'E], alt. 1250 m, 21 May 2011, Yan-qiu Peng leg. Tree-crown layer of Castanopsis fabri. Paratypes: 17 males and 25 females, same data as for holotype.Holotype male (Ps.-MHBU-GX110521), China: Guangxi Province, Nanning City, Daming Mountain , 24 February 1975, Fu-sheng Huang. Habitat unknown.PageBreakHolotype female (Ps.-MHBU-FJ750224), China: Fujian Province, Fuzhou City, Gushan Mountain [The specific name is a patronym in honour of Prof. Fu-Sheng Huang, who collected and donated the specimen.it and et at same level.Species with slender pedipalps  and slender legs IV (e.g. femur+patella 7.23 times as long as deep), with low numbers of the teeth (about 30) on movable chelal finger; trichobothria . Colour Carapace  smooth, Abdomen. Pleural membrane strongly granulate. Tergal chaetotaxy: 4: 12: 10: 10: 10: 10: 11: 11: 11: 10: 9, including at least 4 tactile setae on tergites VI–XI; anterior genital sternite  with 22 PageBreakof femur with fine granulation; patella claviform, smooth; chelal fingers long and slender. Trichobothriotaxy: est, et and it grouped together distally; ist situated midway between isb and it, nearer to it than to isb. eb and esb situated on base of the hand, grouped very closely with ib and isb; b and sb closer to each other situated on the basal half, and st andPageBreakt closer to each other situated on the distal half of the movable finger. Fixed chelal finger with 63 pointed teeth of unequal length, movable finger with about 30 teeth which with 20 pointed teeth slightly unequal length in distal half, and 10 rounded teeth in basal half.Pedipalps . Apex ofCheliceral palm  with 7 sLegs . Tibia IDimensions (in mm) and ratios (in parentheses). Body length 4.2. Carapace 1.29/0.89 (1.45); diameter of anterior eye 0.10; diameter of posterior eye 0.09. Pedipalps: trochanter 0.59/0.26 (2.27), femur 1.58/0.25 (6.32), patella 1.38/0.26 (5.31), chela (with pedicel) 2.58/0.53 (4.87), chela (without pedicel) 2.42 (4.57), hand length (without pedicel) 1.04 (1.96), movable finger length 1.44 (1.38 times longer than hand without pedicel). Chelicera 0.67/0.38 (1.76), movable finger length 0.45. Leg I: femur 0.79/0.13 (6.08), patella 0.59/0.13 (4.54), tibia 0.63/0.10 (6.30), basitarsus 0.45/0.10 (4.50), telotarsus 0.43/0.10 (4.30). Leg IV: femur + patella 1.52/0.21 (7.24), tibia 1.22/0.12 (10.17), basitarsus 0.59/0.10 (5.90), telotarsus 0.79/0.10 (7.90).This species is known only from the type locality.Stenohya huangi sp. n. is only known from the female, but it can be easily separated from most other species of this genus by the proportions of pedipalpal femur and patella . Finally, the new species differs from Stenohya vietnamensis in having an epistome. patella . Two speme level  in Stenot and et : fig. 28Neobisiidae the lyrifissures near the trochanteral foramen of the pedipalpal coxa number 3 or 4  has 6 foramenal lyrifissures and 1 dorsal lyrifissure (PageBreakIn most r 3 or 4 . Having dorsally . Males o foramen : fig. 3 ifissure .


AbstractBambusananus cuihuashanensissp. n. , a new bamboo-feeding leafhopper species, is described and illustrated from Shaanxi Province of China. Checklist, host plants and distribution for each species of Bambusananus is given along with a key to all known species. Athysanini, was established by Van Duzee in 1892 and is the largest tribe of Deltocephalinae, including 228 genera and 1123 species  was established by Bambusananus furcatus Li & Xing, 2011, from Guizhou Province of China as its type species, and three new combinations: Bambusananus binotatus , Bambusananus bipunctatus  and Bambusananus maculipennis  were proposed at the same time. Bambusananus bipunctatus , Bambusananus maculipennis , Bambusananus furcatus Li & Xing, 2011, Bambusananus lii  and Bambusananus yangae Xing & Chen, 2013  and are currently known only from southern mainland China and Taiwan .In the present paper, terminology follows 1.Bambusananus bipunctatus  (in 1999) in Indocalamus hirsutissimus Z. P. Wang & P. X. Zhang and Qiongzhuea communis Hsueh & Yi) (Host plant. Bamboo (eh & Yi) .Distribution. China  .PageBreak2.Bambusananus maculipennis Chimonobambusa pachystachys Hsuch & W. P. Zhang and Qiongzhuea communis Hsueh & Yi) (Host plant. Bamboo (eh & Yi) .Distribution. China (Guizhou) .3.Bambusananus cuihuashanensis sp. n.Host plant. Bamboo.Distribution. China (Shaanxi) .4.Bambusananus furcatus Li & Xing, 2011Chimonobambusa angustifolia C. D. Chu & C. S. Chao) (Host plant. Bamboo (S. Chao) .Distribution. China (Guizhou) .5.Bambusananus lii Host plant. Bamboo .Distribution. China (Taiwan) .6.Bambusananus yangae Xing & Chen, 2013Indocalamus sp.) (Host plant. Bamboo (mus sp.) .Distribution. China (Guizhou) .http://zoobank.org/4B853422-EEC6-41EB-AD6B-B1570C4E92E2http://species-id.net/wiki/Bambusananus_cuihuashanensisChina: Shaanxi, Xi’an, Cuihuashan , on bamboo, 37 Aug. 2008, J.-D. Li; paratypes: 2 ♂♂, 4 ♀♀, same data as holotype.Holotype: ♂, The new species is named after its locality, Cuihuashan, Shaanxi Province, China.Body length (from apex of vertex to tip of forewings): male 4.75–4.85 mm (N = 2); female 5.75–5.90 mm (N = 4); forewing length: male 3.95–4.05 mm (N = 2); female 5.00–5.15 mm (N = 4).Crown  pale yelExternal features as in generic description. Crown shorter medially than width between eyes (0.54:1). Pronotum longer medially than crown (2.10:1). Scutellum shorter medially than pronotum (0.89:1). Forewing longer medially than width at widest part (3.38:1).Male pygofer  with basBamboo.PageBreakPageBreakChina (Shaanxi) .Bambusananus bipunctatus  in general appearance, but can be distinguished by: pygofer in lateral view with ventral margin broadly smoothly rounded, without notch at base of ventral process (with notch in Bambusananus bipunctatus); connective with stem slightly shorter than arms (longer in Bambusananus bipunctatus); aedeagal shaft with appendages longer, in lateral view with apex reaching to middle of aedeagus . This new species is also similar to Bambusananus lii , but can be distinguished by: upper area of frontoclypeus with a large kidney-shaped black marking (lacking in Bambusananus lii); ventral margin of pygoger without any lobe (with a lobe near middle in Bambusananus lii); appendages of aedeagal shaft mostly straight basally, apex directed laterally ; ventral margin of aedeagus without any teeth (with a row of teeth at middle in Bambusananus lii).This new species is similar to