bluuebunny/crossref_metadata_2025 huggingface dataset:
Consensus recommendations for the treatment of basal cell carcinomas in Gorlin syndrome with topical methylaminolaevulinate‐photodynamic therapy
Authors: Basset‐Seguin N. , Bissonnette R. , Girard C. , Haedersdal M. , Lear J.T. , Paul C. , Piaserico S.
Published: April 2013

Background

Patients with Gorlin syndrome develop multiple basal cell carcinomas (BCC), for which treatment is often difficult. Methylaminolevulinate‐photodynamic therapy (MAL‐PDT) is approved for the treatment of superficial and nodular BCCs in Canada and several European countries.

Objectives

To establish consensus recommendations for the use of MAL‐PDT in patients with Gorlin syndrome.

Methods

The Gorlin consensus panel was comprised of 7 dermatologists who had treated a total of 83 patients with Gorlin syndrome using MAL‐PDT. Consensus was developed based on the personal experience of the expert and results of literature review (on PUBMED using the keywords ‘MAL’ and ‘PDT’ and ‘Gorlin’ or ‘naevoid basal cell carcinoma syndrome’).

Results

Consensus was reached among the experts and the literature review identified 9 relevant reports. The experts considered MAL‐PDT a generally effective and safe therapy for treatment of BCC in Gorlin syndrome. For superficial BCC (sBCC), all sizes can be treated, and in nodular BCC (nBCC), better efficacy can be achieved in thinner lesions (<2 mm in thickness). MAL‐PDT treatment schedule should be performed according to labelling although in individual cases, it may be adapted and performed on a monthly basis based on clinical assessment. Follow‐up should be related to frequency of recurrence, and severity, number and location of lesions. Multiple lesions and large areas may be treated during the same session; however, adequate pain management should be considered.

Conclusions

MAL‐PDT is safe and effective in patients with Gorlin syndrome. Utilization of these recommendations may improve efficacy and clearance rates in this population.
Treatment of bullous pemphigoid with low‐dose oral cyclophosphamide: a case series of 20 patients
Authors: Gual A. , Iranzo P. , Mascaró Jr J.M.
Published: April 2013

Background

Cyclophosphamide has been commonly used for the treatment of pemphigus vulgaris and mucous membrane pemphigoid with satisfactory results. Published data of this therapeutic approach for bullous pemphigoid are scant and showed significant morbidity and mortality.

Objective

To assess the clinical efficacy and safety of low‐dose oral cyclophosphamide (CFM) (50–100 mg/day) in patients with refractory bullous pemphigoid.

Methods

We conducted a retrospective study including patients with bullous pemphigoid treated with CFM in the department of Dermatology in the Hospital Clínic of Barcelona, Spain.

Results

Complete response was observed in 11 (58%) over 19 evaluable patients. Cyclophosphamide at 50 mg/day was enough to achieve clinical remission in eight of these patients. Partial response was observed in four (21%) more patients. Bone marrow suppression appeared in 12 (60%) patients, but treatment discontinuation was only required in three (15%) cases. Gastrointestinal intolerance occurred in one (5%) patient. One patient died during therapy from heart failure (not attributed to CFM) and another patient developed acute myeloid leukaemia 1.5 years after CFM therapy.

Conclusions

In our series, CFM had a marked therapeutic effect in bullous pemphigoid. These results of efficacy are similar to those described in other autoimmune blistering skin diseases. Only a few patients had to discontinue their treatment due to adverse effects. Therefore, we consider that low‐dose oral CFM can be valuable therapeutic alternative in elderly patients with moderate‐to‐severe bullous pemphigoid.
MUC5AC expression correlates with invasiveness and progression of extramammary Paget's disease
Authors: Hata H. , Abe R. , Hoshina D. , Saito N. , Homma E. , Aoyagi S. , Shimizu H.
Published: April 2013

Background

Patients with in situ extramammary Paget's disease (EMPD) tend to have a good prognosis, although dermal invasion and metastasis are associated with significantly increased morbidity and mortality. Previous studies have addressed mechanisms underlying the EMPD pathogenesis; however, no molecular markers that reflect invasiveness or progression have been established.

Objective

This study aims to identify a reliable marker for predicting the risk of invasion and metastasis in EMPD.

Methods

We performed an initial microarray screening for in situ, invasive or metastatic lymph node lesions of EMPD. We analysed 44 specimens from 38 primary EMPD cases by immunohistochemical staining.

Results

We found that expressions of MUC5AC directly correlate with invasion and prognosis. Labelling rates of tumour cells were scored by staining intensity on a four‐tiered scale (− to 3+) to investigate the correlation between the expression score of these molecular markers and the type of EMPD lesion. All the specimens scored positive (3+) for MUC1 and negative (−) for MUC6. MUC5AC expression was detected in 19 of 44 (43.2%) specimens. Invasive lesions and metastatic lymph nodes tended to express MUC5AC significantly higher than in situ lesions (P < 0.01). MUC2 was positive in 10 specimens (22.7%). There was no significant difference between the degree of MUC2 expression and invasiveness.

Conclusion

The degree of MUC5AC expression may correlate with the invasiveness and progression of EMPD, and may be a useful marker for identifying high‐risk EMPD cases.
Troglitazone, a thiazolidinedione, decreases tau phosphorylation through the inhibition of cyclin‐dependent kinase 5 activity in SH‐SY5Y neuroblastoma cells and primary neurons
Authors: Cho Du‐Hyong , Lee Eun Joo , Kwon Kyoung Ja , Shin Chan Young , Song Kee‐Ho , Park Jung‐Hyun , Jo Inho , Han Seol‐Heui
Published: April 2013

The peroxisome proliferator‐activated receptor gamma (PPARγ) agonists thiazolidinediones (TZDs) are prescribed for the treatment of type 2 diabetes mellitus. Furthermore, it has been reported that TZDs have a beneficial effect on neurodegenerative disorders, such as Alzheimer's disease. However, the molecular mechanisms underlying this effect are not fully understood. Here, we investigated whether and how troglitazone, a parent TZD drug, inhibits tau phosphorylation. Treatment with troglitazone decreased tau‐Thr231 phosphorylation and p35, the specific activator of cyclin‐dependent kinase 5 (CDK5), in a dose‐ and time‐dependent manner. Troglitazone also decreased CDK5 enzymatic activity, and ectopic expression of p25, the cleaved and more active form of p35, restored the troglitazone‐induced decrease in tau‐Thr231 phosphorylation. Treatment with either MG‐132, a reversible proteasome inhibitor, or lactacystin, a specific and irreversible 26S proteasome inhibitor, significantly reversed the observed inhibitory effects of troglitazone. However, GW9662, a specific and irreversible PPARγ antagonist, did not alter the observed inhibitory effects. Similar results were also found when other TZD drugs, pioglitazone and rosiglitazone, were used. Treatment with various inhibitors revealed that troglitazone‐induced inhibitions of tau‐Thr231 phosphorylation and p35 expression were not mediated by glycogen synthase kinase 3β, protein kinase A, and protein phosphatase 2A signaling pathways. Finally, we also found that the same observed inhibitory effects of troglitazone hold true for the use of primary cortical neurons. Taken together, we demonstrated that TZDs repressed tau‐Thr231 phosphorylation via the inhibition of CDK5 activity, which was mediated by the proteasomal degradation of p35 and a PPARγ‐independent signaling pathway.

image

Thiazolidinediones (TZDs) decrease tau‐Thr231 phosphorylation via the inhibition of CDK5 enzymatic activity, which is mediated by the enhanced proteasomal degradation of p35 and a PPARγ‐independent signaling pathway. This molecular mechanism implies a role for TZD drugs in the prevention and treatment of Alzheimer's disease.
Multimodal interventions (chewing gum, early oral hydration and early mobilisation) on the intestinal motility following abdominal gynaecologic surgery
Authors: Terzioglu Fusun , Şimsek Sevgi , Karaca Kubra , Sariince Nilay , Altunsoy Pinar , Salman Mehmet Coskun
Published: April 2013

Aims and objectives

To determine the effects of chewing gum, early oral hydration and early mobilisation on the time of first bowel sounds, first passage of flatus and first defecation following abdominal gynaecologic surgery.

Background

A major complication of abdominal surgical procedures is paralytic ileus which results in patient discomfort, prolonged length of hospital stay and increased cost of treatment.

Design

Prospective randomised case–control study.

Methods

Women who underwent abdominal gynaecological surgery for benign disorders under general anaesthesia were randomised into eight groups according to different combinations of interventions consisting of chewing gum, early oral hydration and early mobilisation. The effects of these interventions on the time of first bowel sounds, first passage of flatus and first defecation following abdominal gynaecologic surgery were investigated. The data were analysed using chi‐square tests, t‐test for independent samples, Tukey's HSD test, pairwise comparison test, one‐way analysis of variance.

Results

It was found that the time when bowel sounds were heard was shorter, the time first passage of flatus was shorter and first defecation occurred earlier in the 1st group of women who chew gum, were hydrated orally and were mobilised early after surgery than the other groups. It was also determined that these periods were longest in the women who did not receive any intervention and received the routine hospital care when compared with other groups. Duration of hospital stay was shorter in the women who chew gum, were hydrated orally and were mobilised early than the other groups.

Conclusions

Early oral feeding, early mobilisation and chewing gum are effective methods in terms of preventing paralytic ileus following abdominal gynaecological surgery, improving patient comfort and shortening the duration of hospitalisation.

Relevance to clinical practice

Nurses may cause early recovery, improve the patient comfort, prevent paralytic ileus and shorten the duration of hospitalisation after gynaecologic abdominal surgery by recommending gum chewing, early mobilisation and early hydration.
Nurses' discharge planning and risk assessment: behaviours, understanding and barriers
Authors: Graham Jane , Gallagher Robyn , Bothe Janine
Published: April 2013

Aims and objectives

To examine nurses' discharge planning understanding, adherence and barriers.

Background

Discharge planning commenced at admission by nurses plays a key role in improving patient outcomes,but policies in place to maintain effective discharge planning are often not followed by nurses.

Design

This is a descriptive study.

Methods

Nurses (n = 64) working in acute wards undertook a self‐report survey of discharge planning understanding, adherence and barriers.

Results

Adherence to discharge planning policy is low (23%), despite a general awareness of the reduced quality of patient outcomes that may result. The most common barriers to discharge planning identified were lack of time and patient factors. Further contradictions occurred in that nurses understood the importance of discharge planning, yet did not comply with discharge planning policies.

Conclusions

Nurses require additional encouragement and support in complying with discharge planning policies, and discharge planning policies should be adapted to better handle unpredictable illness trajectories.

Relevance to clinical practice

Completion of discharge planning is important for the safe transition of patients from one care setting to the next. Before a systematic approach to discharge planning can be implemented, a greater understanding of nurses' discharge planning practice in acute care wards is required. Greater incorporation of discharge planning activities into nurses' daily practice may also occur if nurses are involved in the development and implementation of the discharge processes and then provided with education and regular feedback on monthly audit results.
Utility of the theory of planned behavior to predict nursing staff blood pressure monitoring behaviours
Authors: Nelson Joan M , Cook Paul F , Ingram Jennifer C
Published: April 2013

Aims and objectives

To evaluate constructs from the theory of planned behavior (TPB, Ajzen 2002) – attitudes, sense of control, subjective norms and intentions – as predictors of accuracy in blood pressure monitoring.

Background

Despite numerous initiatives aimed at teaching blood pressure measurement techniques, many healthcare providers measure blood pressures incorrectly.

Design

Descriptive, cohort design.

Methods

Medical assistants and licensed practical nurses were asked to complete a questionnaire on TPB variables. These nursing staff's patients had their blood pressures measured and completed a survey about techniques used to measure their blood pressure. We correlated nursing staff's responses on the TBP questionnaire with their intention to measure an accurate blood pressure and with the difference between their actual blood pressure measurement and a second measurement taken by a researcher immediately after the clinic visit. Patients' perceptions of MAs' and LPNs' blood pressure measurement techniques were examined descriptively.

Results

Perceived control and social norm predicted intention to measure an accurate blood pressure, with a negative relationship between knowledge and intention. Consistent with the TPB, intention was the only significant predictor of blood pressure measurement accuracy.

Conclusions

Theory of planned behavior constructs predicted the healthcare providers' intention to measure blood pressure accurately and intention predicted the actual accuracy of systolic blood pressure measurement. However, participants' knowledge about blood pressure measurement had an unexpected negative relationship with their intentions.

Relevance to clinical practice

These findings have important implications for nursing education departments and organisations which traditionally invest significant time and effort in annual competency training focused on knowledge enhancement by staff. This study suggests that a better strategy might involve efforts to enhance providers' intention to change, particularly by changing social norms or increasing perceived control of the behaviour by nursing staff.
Secondary c‐Kit mutations confer acquired resistance to RTK inhibitors in c‐Kit mutant melanoma cells
Authors: Todd Jason R. , Becker Therese M. , Kefford Richard F. , Rizos Helen
Published: April 2013

Activation of the c‐Kit receptor tyrosine kinase is rare in melanoma, but occurs in 20‐40% of melanoma arising on mucosal membranes, acral skin and skin with chronic sun‐induced damage. Many activating c‐Kit mutations have been shown to be highly sensitive to imatinib mesylate, although the majority of patients with c‐Kit mutant melanoma eventually progress on this inhibitor. We examined acquired resistance to imatinib and the newer generation inhibitor nilotinib in resistant c‐kit mutant melanoma sublines. Four imatinib‐resistant and six nilotinib‐resistant sublines had acquired additional, secondary c‐Kit mutations. The secondary A829P c‐Kit mutation rendered cells resistant to imatinib, but did not suppress the activity of the tyrosine kinase inhibitors nilotinib and dasatinib. Sublines with an additional T670I c‐Kit mutation showed resistance to imatinib, nilotinib and dasatinib, but responded to sunitinib. The concurrent inhibition of the MAPK and PI3K pathways was also effective at promoting apoptosis in the parent and derived resistant sublines. Our data provide a rationale for treating patients with melanoma progressing on imatinib or nilotinib with alternative RTK inhibitors or inhibitors targeting the MAPK and PI3K signalling cascades.
Clinical applicability of dOFM devices for dermal sampling
Authors: Bodenlenz M. , Aigner B. , Dragatin C. , Liebenberger L. , Zahiragic S. , Höfferer C. , Birngruber T. , Priedl J. , Feichtner F. , Schaupp L. , Korsatko S. , Ratzer M. , Magnes C. , Pieber T. R. , Sinner F.
Published: April 2013

Background

Sampling the dermal interstitial fluid (ISF) allows the pharmacokinetics and pharmacodynamics of dermatological drugs to be studied directly at their site of action.

Dermal open‐flow microperfusion (dOFM) is a recently developed technique that can provide minimally invasive, continuous, membrane‐free (thus unfiltered) access to the dermal ISF. Herein, we evaluate the clinical applicability and reliability of novel wearable dOFM devices in a clinical setting.

Methods

Physicians inserted 141 membrane‐free dOFM probes into the dermis of 17 healthy and psoriatic volunteers and sampled dermal ISF for 25 h by using wearable push‐pull pumps. The tolerability, applicability, reproducibility, and reliability of multiple insertions and 25 h continuous sampling was assessed by pain scoring, physician feedback, ultrasound probe depth measurements, and 25 h‐drift and variability of the sodium relative recovery.

Results

Insertion pain was moderate and decreased with each additional probe. Probe insertion was precise, although slightly deeper in lesional skin. The wearable push‐pull pump enabled uninterrupted ISF sampling over 25 h with low variability. The relative recovery was drift‐free and highly reproducible.

Conclusion

dOFM sampling devices are tolerable and reliable for prolonged continuous dermal sampling in a multiprobe clinical setting. These devices should enable the study of a wide range of drugs and their biomarkers in the skin.
Comparison of Genotypes and Antibiotic Resistances of Campylobacter jejuni and Campylobacter coli on Chicken Retail Meat and at Slaughter
Authors: Kittl Sonja , Korczak Bożena M. , Niederer Lilian , Baumgartner Andreas , Buettner Sabina , Overesch Gudrun , Kuhnert Peter
Published: April 2013

Multilocus sequence typing (MLST) and antibiotic resistance patterns of
Campylobacter jejuni
and
Campylobacter coli
from retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection.
Living Side by Side with a Virus: Characterization of Two Novel Plasmids from Thermococcus prieurii, a Host for the Spindle-Shaped Virus TPV1
Authors: Gorlas Aurore , Krupovic Mart , Forterre Patrick , Geslin Claire
Published: April 2013

Microbial cells often serve as an evolutionary battlefield for different types of mobile genetic elements, such as viruses and plasmids. Here, we describe the isolation and characterization of two new archaeal plasmids which share the host with the spindle-shaped
Thermococcus prieurii
virus 1 (TPV1). The two plasmids, pTP1 and pTP2, were isolated from the hyperthermophilic archaeon
Thermococcus prieurii
(phylum
Euryarchaeota
), a resident of a deep-sea hydrothermal vent located at the East Pacific Rise at 2,700-m depth (7ï¿½25′24 S, 107ï¿½47′66 W). pTP1 (3.1 kb) and pTP2 (2.0 kb) are among the smallest known plasmids of hyperthermophilic archaea, and both are predicted to replicate via the rolling-circle mechanism. The two plasmids and the virus TPV1 do not have a single gene in common and stably propagate in infected cells without any apparent antagonistic effect on each other. The compatibility of the three genetic elements and the high copy number of pTP1 and pTP2 plasmids (50 copies/cell) might be useful for developing new genetic tools for studying hyperthermophilic euryarchaea and their viruses.
Functional Screening of a Metagenomic Library Reveals Operons Responsible for Enhanced Intestinal Colonization by Gut Commensal Microbes
Authors: Yoon Mi Young , Lee Kang-Mu , Yoon Yujin , Go Junhyeok , Park Yongjin , Cho Yong-Joon , Tannock Gerald W. , Yoon Sang Sun
Published: April 2013

Evidence suggests that gut microbes colonize the mammalian intestine through propagation as an adhesive microbial community. A bacterial artificial chromosome (BAC) library of murine bowel microbiota DNA in the surrogate host
Escherichia coli
DH10B was screened for enhanced adherence capability. Two out of 5,472 DH10B clones, 10G6 and 25G1, exhibited enhanced capabilities to adhere to inanimate surfaces in functional screens. DNA segments inserted into the 10G6 and 25G1 clones were 52 and 41 kb and included 47 and 41 protein-coding open reading frames (ORFs), respectively. DNA sequence alignments, tetranucleotide frequency, and codon usage analysis strongly suggest that these two DNA fragments are derived from species belonging to the genus
Bacteroides
. Consistent with this finding, a large portion of the predicted gene products were highly homologous to those of
Bacteroides
spp. Transposon mutagenesis and subsequent experiments that involved heterologous expression identified two operons associated with enhanced adherence.
E. coli
strains transformed with the 10a or 25b operon adhered to the surface of intestinal epithelium and colonized the mouse intestine more vigorously than did the control strain. This study has revealed the genetic determinants of unknown commensals (probably resembling
Bacteroides
species) that enhance the ability of the bacteria to colonize the murine bowel.
Butyrate-Producing Bacteria, Including Mucin Degraders, from the Swine Intestinal Tract
Authors: Levine Uri Y. , Looft Torey , Allen Heather K. , Stanton Thad B.
Published: April 2013

To identify bacteria with potential for influencing gut health, 980 anaerobes were cultured from the swine intestinal tract and analyzed for butyrate production. Fifteen isolates in the order
Clostridiales
produced butyrate and had butyryl coenzyme A (CoA):acetate CoA transferase activity. Three of the isolates grew on mucin, suggesting an intimate association with host intestinal mucosa.
Taxonomy and Distribution of Phenazine-Producing Pseudomonas spp. in the Dryland Agroecosystem of the Inland Pacific Northwest, United States
Authors: Parejko James A. , Mavrodi Dmitri V. , Mavrodi Olga V. , Weller David M. , Thomashow Linda S.
Published: April 2013

We investigated the taxonomic placement of phenazine-producing fluorescentPseudomonasspp. in the Inland Pacific Northwest region of the United States. Five distinct species were identified, two of which were provisionally considered to be new. Agroclimatic zone and soil silt content affected the species diversity across the region.
Changes in the Rumen Epimural Bacterial Diversity of Beef Cattle as Affected by Diet and Induced Ruminal Acidosis
Authors: Petri R. M. , Schwaiger T. , Penner G. B. , Beauchemin K. A. , Forster R. J. , McKinnon J. J. , McAllister T. A.
Published: April 2013

Little is known about the nature of the rumen epithelial adherent (epimural) microbiome in cattle fed different diets. Using denaturing gradient gel electrophoresis (DGGE), quantitative real-time PCR (qPCR), and pyrosequencing of the V3 hypervariable coding region of 16S rRNA, epimural bacterial communities of 8 cattle were profiled during the transition from a forage to a high-concentrate diet, during acidosis, and after recovery. A total of 153,621 high-quality gene sequences were obtained, with populations exhibiting less taxonomic variability among individuals than across diets. The bacterial community composition exhibited clustering (
P
< 0.03) by diet, with only 14 genera, representing >1% of the rumen epimural population, differing (
P
≤ 0.05) among diets. During acidosis, levels of
Atopobium
,
Desulfocurvus
,
Fervidicola
,
Lactobacillus
, and
Olsenella
increased, while during the recovery,
Desulfocurvus
,
Lactobacillus
, and
Olsenella
reverted to levels similar to those with the high-grain diet and
Sharpea
and
Succinivibrio
reverted to levels similar to those with the forage diet. The relative abundances of bacterial populations changed during diet transition for all qPCR targets except
Streptococcus
spp. Less than 5% of total operational taxonomic units (OTUs) identified exhibited significant variability across diets. Based on DGGE, the community structures of epithelial populations differed (
P
≤ 0.10); segregation was most prominent for the mixed forage diet versus the grain, acidotic challenge, and recovery diets.
Atopobium
, cc142,
Lactobacillus
,
Olsenella
, RC39,
Sharpea
,
Solobacterium
,
Succiniclasticum
, and
Syntrophococcus
were particularly prevalent during acidosis. Determining the metabolic roles of these key genera in the rumens of cattle fed high-grain diets could define a clinical microbial profile associated with ruminal acidosis.
Mitochondrial Two-Component Signaling Systems in Candida albicans
Authors: Mavrianos John , Berkow Elizabeth L. , Desai Chirayu , Pandey Alok , Batish Mona , Rabadi Marissa J. , Barker Katherine S. , Pain Debkumar , Rogers P. David , Eugenin Eliseo A. , Chauhan Neeraj
Published: April 2013

Two-component signal transduction pathways are one of the primary means by which microorganisms respond to environmental signals. These signaling cascades originated in prokaryotes and were inherited by eukaryotes via endosymbiotic lateral gene transfer from ancestral cyanobacteria. We report here that the nuclear genome of the pathogenic fungus
Candida albicans
contains elements of a two-component signaling pathway that seem to be targeted to the mitochondria. The
C. albicans
two-component response regulator protein Srr1 (
s
tress
r
esponse
r
egulator 1) contains a mitochondrial targeting sequence at the N terminus, and fluorescence microscopy reveals mitochondrial localization of green fluorescent protein-tagged Srr1. Moreover, phylogenetic analysis indicates that
C. albicans
Srr1 is more closely related to histidine kinases and response regulators found in marine bacteria than are other two-component proteins present in the fungi. These data suggest conservation of this protein during the evolutionary transition from endosymbiont to a subcellular organelle. We used microarray analysis to determine whether the phenotypes observed with a
srr1Δ/Δ
mutant could be correlated with gene transcriptional changes. The expression of mitochondrial genes was altered in the
srr1Δ/Δ
null mutant in comparison to their expression in the wild type. Furthermore, apoptosis increased significantly in the
srr1Δ/Δ
mutant strain compared to the level of apoptosis in the wild type, suggesting the activation of a mitochondrion-dependent apoptotic cell death pathway in the
srr1Δ/Δ
mutant. Collectively, this study shows for the first time that a lower eukaryote like
C. albicans
possesses a two-component response regulator protein that has survived in mitochondria and regulates a subset of genes whose functions are associated with the oxidative stress response and programmed cell death (apoptosis).
Poly(ADP-Ribose) Polymerase Is a Substrate Recognized by Two Metacaspases of Podospora anserina
Authors: Strobel Ingmar , Osiewacz Heinz D.
Published: April 2013

The two metacaspases MCA1 and MCA2 of the fungal aging model organismPodospora anserina(PaMCA1 and PaMCA2, respectively) have previously been demonstrated to be involved in the control of programmed cell death (PCD) and life span. In order to identify specific pathways and components which are controlled by the activity of these enzymes, we set out to characterize them further. Heterologous overexpression inEscherichia coliof the two metacaspase genes resulted in the production of proteins which aggregate and form inclusion bodies from which the active protein has been recovered via refolding in appropriate buffers. The renaturated proteins are characterized by an arginine-specific activity and are active in caspase-like self-maturation leading to the generation of characteristic small protein fragments. Both activities are dependent on the presence of calcium. Incubation of the two metacaspases with recombinant poly(ADP-ribose) polymerase (PARP), a known substrate of mammalian caspases, led to the identification of PARP as a substrate of the twoP. anserinaproteases. Using double mutants in whichP. anserina Parp(PaParp) is overexpressed andPaMca1is either overexpressed or deleted, we provide evidence forin vivodegradation of PaPARP by PaMCA1. These results support the idea that the substrate profiles of caspases and metacaspases are at least partially overlapping. Moreover, they link PCD and DNA maintenance in the complex network of molecular pathways involved in aging and life span control.
Regulation of Copper Toxicity by Candida albicans
GPA2
Authors: Schwartz Jennifer A. , Olarte Karen T. , Michalek Jamie L. , Jandu Gurjinder S. , Michel Sarah L. J. , Bruno Vincent M.
Published: April 2013

Copper is an essential nutrient that is toxic to cells when present in excess. The fungal pathogen
Candida albicans
employs several mechanisms to survive in the presence of excess copper, but the molecular pathways that govern these responses are not completely understood. We report that deletion of
GPA2
, which specifies a G-protein α subunit, confers increased resistance to excess copper and propose that the increased resistance is due to a combination of decreased copper uptake and an increase in copper chelation by metallothioneins. This is supported by our observations that a
gpa2
Δ/Δ mutant has reduced expression of the copper uptake genes,
CTR1
and
FRE7
, and a marked decrease in copper accumulation following exposure to high copper levels. Furthermore, deletion of
GPA2
results in an increased expression of the copper metallothionein gene,
CRD2
. Gpa2p functions upstream in the cyclic AMP (cAMP)-protein kinase A (PKA) pathway to govern hyphal morphogenesis. The copper resistance phenotype of the
gpa2
Δ/Δ mutant can be reversed by artificially increasing the intracellular concentration of cAMP. These results provide evidence for a novel role of the PKA pathway in regulation of copper homeostasis. Furthermore, the connection between the PKA pathway and copper homeostasis appears to be conserved in the pathogen
Cryptococcus neoformans
but not in the nonpathogenic
Saccharomyces cerevisiae
.
One Single Basic Amino Acid at the ω-1 or ω-2 Site Is a Signal That Retains Glycosylphosphatidylinositol-Anchored Protein in the Plasma Membrane of Aspergillus fumigatus
Authors: Ouyang Haomiao , Chen Xiaomin , Lü Yang , Wilson Iain B. H. , Tang Guomin , Wang Aoquan , Jin Cheng
Published: April 2013

Although the plasma membrane is the terminal destination for glycosylphosphatidylinositol (GPI) proteins in higher eukaryotes, cell wall-attached GPI proteins (GPI-CWPs) are found in many fungal species. In yeast, some of the
cis
-requirements directing localization of GPI proteins to the plasma membrane or cell wall are now understood. However, it remains to be determined how
Aspergillus fumigatus
, an opportunistic fungal pathogen, signals, and sorts GPI proteins to either the plasma membrane or the cell wall. In this study, chimeric green fluorescent proteins (GFPs) were constructed as fusions with putative C-terminal GPI signal sequences from
A. fumigatus
Mp1p, Gel1p, and Ecm33p, as well as site-directed mutations thereof. By analyzing cellular localization of chimeric GFPs using Western blotting, electron microscopy, and fluorescence microscopy, we showed that, in contrast to yeast, a single Lys residue at the ω-1 or ω-2 site alone could retain GPI-anchored GFP in the plasma membrane. Although the signal for cell wall distribution has not been identified yet, it appeared that the threonine/serine-rich region at the C-terminal half of
Af
Mp1 was not required for cell wall distribution. Based on our results, the
cis
-requirements directing localization of GPI proteins in
A. fumigatus
are different from those in yeast.
Polyphosphate Deficiency in Mycobacterium tuberculosis Is Associated with Enhanced Drug Susceptibility and Impaired Growth in Guinea Pigs
Authors: Singh Ramandeep , Singh Mamta , Arora Garima , Kumar Santosh , Tiwari Prabhakar , Kidwai Saqib
Published: April 2013

Inorganic polyphosphate (polyP), a linear polymer of hundreds of phosphate residues linked by ATP-like phosphoanhydride bonds, is found in all organisms and performs a wide variety of functions. This study shows that polyP accumulation occurs in
Mycobacterium tuberculosis
upon exposure to various stress conditions.
M. tuberculosis
possesses a single homolog of
ppk-1
, and we have disrupted
ppk-1
in the
M. tuberculosis
genome by allelic replacement. The mutant strain exhibited negligible levels of intracellular polyP, decreased expression of
sigF
and
phoP
, and reduced growth in the stationary phase and displayed a survival defect in response to nitrosative stress and in THP-1 macrophages compared to the wild-type strain. We report that reduction in polyP levels is associated with increased susceptibility of
M. tuberculosis
to certain TB drugs and impairs its ability to cause disease in guinea pigs. These results suggest that polyP contributes to persistence of
M. tuberculosis
in vitro
and plays an important role in the physiology of bacteria residing within guinea pigs.
Nonnative Disulfide Bond Formation Activates the σ
32
-Dependent Heat Shock Response in Escherichia coli
Authors: Müller Alexandra , Hoffmann Jörg H. , Meyer Helmut E. , Narberhaus Franz , Jakob Ursula , Leichert Lars I.
Published: April 2013

Formation of nonnative disulfide bonds in the cytoplasm, so-called disulfide stress, is an integral component of oxidative stress. Quantification of the extent of disulfide bond formation in the cytoplasm of
Escherichia coli
revealed that disulfide stress is associated with oxidative stress caused by hydrogen peroxide, paraquat, and cadmium. To separate the impact of disulfide bond formation from unrelated effects of these oxidative stressors in subsequent experiments, we worked with two complementary approaches. We triggered disulfide stress either chemically by diamide treatment of cells or genetically in a mutant strain lacking the major disulfide-reducing systems TrxB and Gor. Studying the proteomic response of
E. coli
exposed to disulfide stress, we found that intracellular disulfide bond formation is a particularly strong inducer of the heat shock response. Real-time quantitative PCR experiments showed that disulfide stress induces the heat shock response in
E. coli
σ
32
dependently. However, unlike heat shock treatment, which induces these genes transiently, transcripts of σ
32
-dependent genes accumulated over time in disulfide stress-treated cells. Analyzing the stability of σ
32
, we found that this constant induction can be attributed to an increase of the half-life of σ
32
upon disulfide stress. This is concomitant with aggregation of
E. coli
proteins treated with diamide. We conclude that oxidative stress triggers the heat shock response in
E. coli
σ
32
dependently. The component of oxidative stress responsible for the induction of heat shock genes is disulfide stress. Nonnative disulfide bond formation in the cytoplasm causes protein unfolding. This stabilizes σ
32
by preventing its DnaK- and FtsH-dependent degradation.
Features of Pro-σ
K
Important for Cleavage by SpoIVFB, an Intramembrane Metalloprotease
Authors: Zhou Ruanbao , Chen Kangming , Xiang Xianling , Gu Liping , Kroos Lee
Published: April 2013

Intramembrane proteases regulate diverse processes by cleaving substrates within a transmembrane segment or near the membrane surface.
Bacillus subtilis
SpoIVFB is an intramembrane metalloprotease that cleaves Pro-σ
K
during sporulation. To elucidate features of Pro-σ
K
important for cleavage by SpoIVFB, coexpression of the two proteins in
Escherichia coli
was used along with cell fractionation. In the absence of SpoIVFB, a portion of the Pro-σ
K
was peripherally membrane associated. This portion was not observed in the presence of SpoIVFB, suggesting that it serves as the substrate. Deletion of Pro-σ
K
residues 2 to 8, addition of residues at its N terminus, or certain single-residue substitutions near the cleavage site impaired cleavage. Certain multiresidue substitutions near the cleavage site changed the position of cleavage, revealing preferences for a small residue preceding the cleavage site N-terminally (i.e., at the P1 position) and a hydrophobic residue at the second position following the cleavage site C-terminally (i.e., P2′). These features appear to be conserved among Pro-σ
K
orthologs. SpoIVFB did not tolerate an aromatic residue at P1 or P2′ of Pro-σ
K
. A Lys residue at P3′ of Pro-σ
K
could not be replaced with Ala unless a Lys was provided farther C-terminally (e.g., at P9′). α-Helix-destabilizing residues near the cleavage site were not crucial for SpoIVFB to cleave Pro-σ
K
. The preferences and tolerances of SpoIVFB are somewhat different from those of other intramembrane metalloproteases, perhaps reflecting differences in the interaction of the substrate with the membrane and the enzyme.
Associations between objectively measured physical activity intensity in childhood and measures of subclinical cardiovascular disease in adolescence: prospective observations from the European Youth Heart Study
Authors: Ried-Larsen Mathias , Grøntved Anders , Møller Niels Christian , Larsen Kristian Traberg , Froberg Karsten , Andersen Lars Bo
Published: April 2013

Background and aim

No prospective studies have investigated the association between physical activity (PA) and carotid subclinical cardiovascular disease across childhood. Therefore, the primary aim was to investigate the association between PA intensity across childhood and carotid intima media thickness (cIMT) and stiffness in adolescence. Second, we included a clustered cardiovascular disease risk score as outcome.

Methods

This was a prospective study of a sample of 254 children (baseline age 8–10 years) with a 6-year follow-up. The mean exposure and the change in minutes of moderate-and-vigorous and vigorous PA intensity were measured using the Actigraph activity monitor. Subclinical cardiovascular disease was expressed as cIMT, carotid arterial stiffness and secondarily as a metabolic risk z-score including the homoeostasis model assessment score of insulin resistance, triglycerides, total cholesterol to high-density lipoprotein ratio, inverse of cardiorespiratory fitness, systolic blood pressure and the sum of four skinfolds.

Results

No associations were observed between PA intensity variables and cIMT or carotid arterial stiffness (p>0.05). Neither change in PA intensity (moderate-and-vigorous nor vigorous) nor mean minutes of moderate-and-vigorous PA intensity was associated to the metabolic risk z-score in adolescence (p>0.05). However, a significant inverse association was observed between mean minutes of vigorous PA and the metabolic risk z-score in adolescence independent of gender and biological maturity (standard β=–0.19 p=0.007).

Conclusions

A high mean exposure to, or changes in, minutes spent at higher PA intensities across childhood was not associated to cIMT or stiffness in the carotid arteries in adolescence. Our observations suggest that a high volume of vigorous PA across childhood independently associated with lower metabolic cardio vascular disease risk in adolescence.
Moderate-and-vigorous physical activity from adolescence to adulthood and subclinical atherosclerosis in adulthood: prospective observations from the European Youth Heart Study
Authors: Ried-Larsen Mathias , Grøntved Anders , Kristensen Peter Lund , Froberg Karsten , Andersen Lars Bo
Published: April 2013

Aim

To investigate the independent associations between mean exposure to or the change in moderate-and-vigorous physical activity (PA) from adolescence to adulthood and subclinical atherosclerosis in adulthood.

Methods

This was a prospective cohort study among Danish boys and girls (N=277) followed for up to 12 years (age 15.7 (0.4) at baseline) enrolled in the European Youth Heart Study. PA intensity was objectively measured at baseline and follow-up, and ultrasonography was performed on the Carotid arteries at follow-up. Data on carotid intima-media thickness (cIMT), Carotid Compliance and Young's Elastic Modules were used as outcome measures.

Results

In the multivariable analyses (adjusted for personal—lifestyle and demographic factors) the mean exposure to moderate-and-vigorous PA from adolescence to adulthood was negatively associated with Young's Elastic Modules (β=−0.001×103 kPa (95% CI −0.0015 to −0.0002), p=0.02) and positively associated with Carotid Compliance (β=0.004 mm2 kPa−1(95% CI 0.002 to 0.008), p=0.003) and cIMT (β=0.0003 mm (95% CI 0.00001 to 0.0007), p=0.013). Increases in moderate-and-vigorous PA from adolescence to adulthood were negatively associated with Young's Elastic Modules in adulthood (β=−0.00007×103 kPa (95% CI −0.0012 to −0.0001), p=0.01). Furthermore, participants with the largest decline in moderate-and-vigorous PA from adolescence to adulthood displayed significantly less compliant arteries compared with the remaining sample (p<0.05).

Conclusions

High mean exposure to moderate-and-vigorous PA levels and increases herein were independently associated with lower levels of carotid arterial stiffness in adulthood.
Assessment of statin-associated muscle toxicity in Japan: a cohort study conducted using claims database and laboratory information
Authors: Chang Chia-Hsien , Kusama Makiko , Ono Shunsuke , Sugiyama Yuichi , Orii Takao , Akazawa Manabu
Published: April 2013

Objective

To estimate the incidence of muscle toxicity in patients receiving statin therapy by examining study populations, drug exposure status and outcome definitions.

Design

A retrospective cohort study.

Setting

16 medical facilities in Japan providing information on laboratory tests performed in and claims received by their facilities between 1 April 2004 and 31 December 2010.

Participants

A database representing a cohort of 35 903 adult statin (atorvastatin, fluvastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin) users was studied. Use of interacting drugs (fibrates, triazoles, macrolides, amiodarone and ciclosporin) by these patients was determined.

Main outcome measure

Statin-associated muscle toxicity (the ‘event’) was identified based on a diagnosis of muscle-related disorders (myopathy or rhabdomyolysis) and/or abnormal elevation of creatine kinase (CK) concentrations. Events were excluded if the patients had CK elevation-related conditions other than muscle toxicity. Incidence rates for muscle toxicity were determined per 1000 person-years, with 95% CI determined by Poisson regression.

Results

A total of 18 036 patients accounted for 42 193 person-years of statin therapy, and 43 events were identified. The incidence of muscle toxicity in the patients treated with statins was 1.02 (95% CI 0.76 to 1.37)/1000 person-years. The estimates varied when outcome definitions were modified from 0.09/1000 person-years, which met both diagnosis and CK 10× greater than the upper limit of normal range (ULN) criteria, to 2.06/1000 person-years, which met diagnosis or CK 5× ULN criterion. The incidence of muscle toxicity was also influenced by the statin therapies selected, but no significant differences were observed. Among 2430 patients (13.5%) received interacting drugs with statins, only three muscle toxicity cases were observed (incidence: 1.69/1000 person-years).

Conclusions

This database study suggested that statin use is generally well tolerated and safe; however, the risk of muscle toxicity related to the use of interacting drugs requires further exploration.
Ω-3 fatty acid supplement use in the 45 and Up Study Cohort
Authors: Adams Jon , Sibbritt David , Lui Chi-Wai , Broom Alex , Wardle Jonathan
Published: April 2013

Objective

There has been a dramatic increase in the use of dietary supplements in Western societies over the past decades. Our understanding of the prevalence of Ω-3 fatty acid supplement consumption is of significance for future nutrition planning, health promotion and care delivery. However, we know little about Ω-3 fatty acid supplement consumption or users. This paper, drawing upon the largest dataset with regard to Ω-3 fatty acid supplement use (n=266 848), examines the use and users of this supplement among a large sample of older Australians living in New South Wales.

Design

A cross-sectional study. Data were analysed from the 45 and Up Study, the largest study of healthy ageing ever undertaken in the Southern Hemisphere.

Setting

New South Wales, Australia.

Participants

266 848 participants of the 45 and Up Study.

Primary and secondary outcome measures

Participants’ use of Ω-3, demographics (geographical location, marital status, education level, income and level of healthcare insurance) and health status (quality of life, history of smoking and alcohol consumption, health conditions) were measured.

Results

Of the 266 848 participants, 32.6% reported having taken Ω-3 in the 4 weeks prior to the survey. Use of Ω-3 fatty acid supplements was higher among men, non-smokers, non-to-mild (alcoholic) drinkers, residing in a major city, having higher income and private health insurance. Osteoarthritis, osteoporosis, high cholesterol and anxiety and/or depression were positively associated with  Ω-3 fatty acid supplement use, while cancer and high blood pressure were negatively associated with use of Ω-3 fatty acid supplements.

Conclusions

This study, analysing data from the 45 and Up Study cohort, suggests that a considerable proportion of older Australians consume Ω-3 fatty acid supplements. There is a need for primary healthcare practitioners to enquire with patients about this supplement use and for work to ensure provision of good-quality information for patients and providers with regard to Ω-3 fatty acid products.
Barriers to recruitment for surgical trials in head and neck oncology: a survey of trial investigators
Authors: Kaur Geetinder , Hutchison Iain , Mehanna Hisham , Williamson Paula , Shaw Richard , Tudur Smith Catrin
Published: April 2013

Objectives

Many randomised trials in surgery suffer from recruitment rates that lag behind projected targets. We aim to identify perceived barriers to recruitment among these pioneering trials in the field of head and neck cancer surgery.

Design

Recruiting centres to all three trials (Selective Elective Neck Dissection (SEND), Positron Emission Tomography (PET)-Neck and Hyperbaric Oxygen in the Prevention of Osteoradionecrosis (HOPON)) were contacted by email by the chief investigators. Responders were asked to complete a web-based survey in order to identify the barriers to recruitment in their centre and grade each by severity.

Setting

Secondary care: 44 head and neck oncology regional referral centres.

Participants

Analysis was based on 85 responses evenly distributed between the three trials.

Results

The most commonly identified perceived barriers to recruitment (more than 50% of responders identified the item as a barrier in all the three trials) in the order of frequency were: patients consent refusal because of expressed treatment preference, patients consent refusal owing to aversion to randomisation, excess complexity/amount of information provided to patients and lack of time in clinic to accommodate research. The most severely rated of these problems was consent refusal because of the expressed treatment preference and lack of time in the clinic.

Conclusions

Our findings confirm others’ work in surgery that the most significant barrier to trial recruitment in head and neck cancer surgery is the patient's preference for one arm of the trial. It may be that additional training for those taking consent may be helpful in this regard. It is also important to adequately resource busy surgical clinics to support clinical trial recruitment.