bluuebunny/crossref_metadata_2025 huggingface dataset:
Bacteriophage F336 Recognizes the Capsular Phosphoramidate Modification of Campylobacter jejuni NCTC11168
Authors: Sørensen Martine C. Holst , van Alphen Lieke B. , Harboe Anne , Li Jianjun , Christensen Bjarke Bak , Szymanski Christine M. , Brøndsted Lone
Published: October 2011

Bacteriophages infecting the food-borne human pathogen
Campylobacter jejuni
could potentially be exploited to reduce bacterial counts in poultry prior to slaughter. This bacterium colonizes the intestinal tract of poultry in high numbers, and contaminated poultry meat is regarded as the major source of human campylobacteriosis. In this study, we used phage F336 belonging to the
Myoviridae
family to select a
C. jejuni
NCTC11168 phage-resistant strain, called 11168R, with the aim of investigating the mechanisms of phage resistance. We found that phage F336 has reduced adsorption to 11168R, thus indicating that the receptor is altered. While proteinase K-treated
C. jejuni
cells did not affect adsorption, periodate treatment resulted in reduced adsorption, suggesting that the phage binds to a carbohydrate moiety. Using high-resolution magic angle spinning nuclear magnetic resonance (NMR) spectroscopy, we found that 11168R lacks an
O
-methyl phosphoramidate (MeO
P
N) moiety attached to the Gal
f
NAc on the capsular polysaccharide (CPS), which was further confirmed by mass spectroscopy. Sequence analysis of 11168R showed that the potentially hypervariable gene
cj1421
, which encodes the Gal
f
NAc MeO
P
N transferase, contains a tract of 10 Gs, resulting in a nonfunctional gene product. However, when 11168R reverted back to phage sensitive,
cj1421
contained 9 Gs, and the Gal
f
NAc MeO
P
N was regained in this strain. In summary, we have identified the phase-variable MeO
P
N moiety, a common component of the diverse capsular polysaccharides of
C. jejuni
, as a novel receptor of phages infecting this bacterium.
Cyclohexane-1,2-Dione Hydrolase from Denitrifying Azoarcus sp. Strain 22Lin, a Novel Member of the Thiamine Diphosphate Enzyme Family
Authors: Steinbach Alma K. , Fraas Sonja , Harder Jens , Tabbert Anja , Brinkmann Henner , Meyer Axel , Ermler Ulrich , Kroneck Peter M. H.
Published: October 2011

Alicyclic compounds with hydroxyl groups represent common structures in numerous natural compounds, such as terpenes and steroids. Their degradation by microorganisms in the absence of dioxygen may involve a C—C bond ring cleavage to form an aliphatic intermediate that can be further oxidized. The cyclohexane-1,2-dione hydrolase (CDH) (EC 3.7.1.11) from denitrifying
Azoarcus
sp. strain 22Lin, grown on cyclohexane-1,2-diol as a sole electron donor and carbon source, is the first thiamine diphosphate (ThDP)-dependent enzyme characterized to date that cleaves a cyclic aliphatic compound. The degradation of cyclohexane-1,2-dione (CDO) to 6-oxohexanoate comprises the cleavage of a C—C bond adjacent to a carbonyl group, a typical feature of reactions catalyzed by ThDP-dependent enzymes. In the subsequent NAD
+
-dependent reaction, 6-oxohexanoate is oxidized to adipate. CDH has been purified to homogeneity by the criteria of gel electrophoresis (a single band at ∼59 kDa; calculated molecular mass, 64.5 kDa); in solution, the enzyme is a homodimer (∼105 kDa; gel filtration). As isolated, CDH contains 0.8 ± 0.05 ThDP, 1.0 ± 0.02 Mg
2+
, and 1.0 ± 0.015 flavin adenine dinucleotide (FAD) per monomer as a second organic cofactor, the role of which remains unclear. Strong reductants, Ti(III)-citrate, Na
+
-dithionite, and the photochemical 5-deazaflavin/oxalate system, led to a partial reduction of the FAD chromophore. The cleavage product of CDO, 6-oxohexanoate, was also a substrate; the corresponding cyclic 1,3- and 1,4-diones did not react with CDH, nor did the
cis
- and
trans
-cyclohexane diols. The enzymes acetohydroxyacid synthase (AHAS) from
Saccharomyces cerevisiae
, pyruvate oxidase (POX) from
Lactobacillus plantarum
, benzoylformate decarboxylase from
Pseudomonas putida
, and pyruvate decarboxylase from
Zymomonas mobilis
were identified as the closest relatives of CDH by comparative amino acid sequence analysis, and a ThDP binding motif and a 2-fold Rossmann fold for FAD binding could be localized at the C-terminal end and central region of CDH, respectively. A first mechanism for the ring cleavage of CDO is presented, and it is suggested that the FAD cofactor in CDH is an evolutionary relict.
Deletion Strains Reveal Metabolic Roles for Key Elemental Sulfur-Responsive Proteins in Pyrococcus furiosus
Authors: Bridger Stephanie L. , Clarkson Sonya M. , Stirrett Karen , DeBarry Megan B. , Lipscomb Gina L. , Schut Gerrit J. , Westpheling Janet , Scott Robert A. , Adams Michael W. W.
Published: October 2011

Transcriptional and enzymatic analyses of
Pyrococcus furiosus
previously indicated that three proteins play key roles in the metabolism of elemental sulfur (S
0
): a membrane-bound oxidoreductase complex (MBX), a cytoplasmic coenzyme A-dependent NADPH sulfur oxidoreductase (NSR), and sulfur-induced protein A (SipA). Deletion strains, referred to as MBX1, NSR1, and SIP1, respectively, have now been constructed by homologous recombination utilizing the uracil auxotrophic COM1 parent strain (Δ
pyrF
). The growth of all three mutants on maltose was comparable without S
0
, but in its presence, the growth of MBX1 was greatly impaired while the growth of NSR1 and SIP1 was largely unaffected. In the presence of S
0
, MBX1 produced little, if any, sulfide but much more acetate (per unit of protein) than the parent strain, demonstrating that MBX plays a critical role in S
0
reduction and energy conservation. In contrast, comparable amounts of sulfide and acetate were produced by NSR1 and the parent strain, indicating that NSR is not essential for energy conservation during S
0
reduction. Differences in transcriptional responses to S
0
in NSR1 suggest that two sulfide dehydrogenase isoenzymes provide a compensatory NADPH-dependent S
0
reduction system. Genes controlled by the S
0
-responsive regulator SurR were not as highly regulated in MBX1 and NSR1. SIP1 produced the same amount of acetate but more sulfide than the parent strain. That SipA is not essential for growth on S
0
indicates that it is not required for detoxification of metal sulfides, as previously suggested. A model is proposed for S
0
reduction by
P. furiosus
with roles for MBX and NSR in bioenergetics and for SipA in iron-sulfur metabolism.
Hopanoid Production Is Required for Low-pH Tolerance, Antimicrobial Resistance, and Motility in Burkholderia cenocepacia
Authors: Schmerk Crystal L. , Bernards Mark A. , Valvano Miguel A.
Published: October 2011

Hopanoids are pentacyclic triterpenoids that are thought to be bacterial surrogates for eukaryotic sterols, such as cholesterol, acting to stabilize membranes and to regulate their fluidity and permeability. To date, very few studies have evaluated the role of hopanoids in bacterial physiology. The synthesis of hopanoids depends on the enzyme squalene-hopene cyclase (Shc), which converts the linear squalene into the basic hopene structure. Deletion of the 2 genes encoding Shc enzymes in
Burkholderia cenocepacia
K56-2,
BCAM2831
and
BCAS0167
, resulted in a strain that was unable to produce hopanoids, as demonstrated by gas chromatography and mass spectrometry. Complementation of the Δ
shc
mutant with only
BCAM2831
was sufficient to restore hopanoid production to wild-type levels, while introducing a copy of
BCAS0167
alone into the Δ
shc
mutant produced only very small amounts of the hopanoid peak. The Δ
shc
mutant grew as well as the wild type in medium buffered to pH 7 and demonstrated no defect in its ability to survive and replicate within macrophages, despite transmission electron microscopy (TEM) revealing defects in the organization of the cell envelope. The Δ
shc
mutant displayed increased sensitivity to low pH, detergent, and various antibiotics, including polymyxin B and erythromycin. Loss of hopanoid production also resulted in severe defects in both swimming and swarming motility. This suggests that hopanoid production plays an important role in the physiology of
B. cenocepacia
.
Regulation of Transcription by SMU.1349, a TetR Family Regulator, in Streptococcus mutans
Authors: Chattoraj Partho , Mohapatra Saswat Sourav , Rao J. L. Uma Maheswar , Biswas Indranil
Published: October 2011

The TetR family of transcriptional regulators is ubiquitous in bacteria, where it plays an important role in bacterial gene expression.Streptococcus mutans, a Gram-positive pathogen considered to be the primary etiological agent in the formation of dental caries, encodes at least 18 TetR regulators. Here we characterized one such TetR regulator, SMU.1349, encoded by the TnSmu2 operon, which appeared to be acquired by the organism via horizontal gene transfer. SMU.1349 is transcribed divergently from the rest of the genes encoded by the operon. By the use of a transcriptional reporter system and semiquantitative reverse transcription-PCR (RT-PCR), we demonstrated that SMU.1349 activates the transcription of several genes that are encoded within the TnSmu2 operon. Gel mobility shift and DNase I footprinting assays with purified SMU.1349 protein demonstrated binding to the intergenic region between SMU.1349 and the TnSmu2 operon; therefore, SMU.1349 is directly involved in gene transcription. Using purifiedS. mutansRpoD andEscherichia coliRNA polymerase, we also demonstrated in anin vitrotranscription assay that SMU.1349 could activate transcription from the TnSmu2 operon promoter. Furthermore, we showed that SMU.1349 could also repress transcription from its own promoter by binding to the intergenic region, suggesting that SMU.1349 acts as both an activator and a repressor. Thus, unlike most of the TetR family proteins, which generally function as transcriptional repressors, SMU.1349 is unique in that it can function as both.
Adeno-Associated Virus Activates an Innate Immune Response in Normal Human Cells but Not in Osteosarcoma Cells
Authors: Laredj Leila N. , Beard Peter
Published: September 2011

Adeno-associated virus (AAV) is a small, DNA-containing
dependovirus
with promising potential as a gene delivery vehicle. Given the variety of applications of AAV-based vectors in the treatment of genetic disorders, numerous studies have focused on the immunogenicity of recombinant AAV. In general, AAV vectors appear not to induce strong inflammatory responses. We have found that AAV2, when it infects the osteosarcoma cells U2OS, can initiate part of its replicative cycle in the absence of helper virus. This does not occur in untransformed cells. We set out to test whether the cellular innate antiviral defenses control this susceptibility and found that, in nonimmune normal human fibroblasts, AAV2 induces type I interferon production and release and the accumulation of nuclear promyelocytic leukemia bodies. AAV fails to mobilize this defense pathway in the U2OS cells. This permissiveness is in large part due to impairment of the viral sensing machinery in these cells. Our investigations point to Toll-like receptor 9 as a potential intracellular sensor that detects AAV2 and triggers the antiviral state in AAV-infected untransformed cells. Efficient sensing of the AAV genome and the ensuing activation of an innate antiviral response are thus crucial cellular events dictating the parvovirus infectivity in host cells.
Viusid, a nutritional supplement, increases survival and reduces disease progression in HCV-related decompensated cirrhosis: a randomised and controlled trial
Authors: Vilar Gomez Eduardo , Sanchez Rodriguez Yoan , Torres Gonzalez Ana , Calzadilla Bertot Luis , Arus Soler Enrique , Martinez Perez Yadina , Yasells Garcia Ali , Abreu Vazquez Maria del Rosario
Published: October 2011

Objectives

Viusid is a nutritional supplement with recognised antioxidant and immunomodulatory properties which could have beneficial effects on cirrhosis-related clinical outcomes such as survival, disease progression and development of hepatocellular carcinoma (HCC). This study evaluated the efficacy and safety of viusid in patients with HCV-related decompensated cirrhosis.

Design

A randomised double-blind and placebo-controlled study was conducted in a tertiary care academic centre (National Institute of Gastroenterology, Havana, Cuba). The authors randomly assigned 100 patients with HCV-related decompensated cirrhosis to receive viusid (three oral sachets daily, n=50) or placebo (n=50) during 96 weeks. The primary outcome of the study was overall survival at 96 weeks, and the secondary outcomes included time to disease progression, time to HCC diagnosis, time to worsening of the prognostic scoring systems Child–Pugh and Model for End-Stage Liver Disease, and time to a new occurrence or relapse for each one of the main clinical complications secondary to portal hypertension at 96 weeks.

Results

Viusid led to a significant improvement in overall survival (90%) versus placebo (74%) (HR 0.27, 95% CI 0.08 to 0.92; p=0.036). A similar improvement in disease progression was seen in viusid-treated patients (28%), compared with placebo-treated patients (48%) (HR 0.47, 95% CI 0.22 to 0.89; p=0.044). However, the beneficial effects of viusid were wholly observed among patients with Child–Pugh classes B or C, but not among patients with Child–Pugh class A. The cumulative incidence of HCC was significantly reduced in patients treated with viusid (2%) as compared with placebo (12%) (HR 0.15, 95% CI 0.019 to 0.90; p=0.046). Viusid was well tolerated.

Conclusions

The results indicate that treatment with viusid leads to a notable improvement in overall clinical outcomes such as survival, disease progression and development of HCC in patients with HCV-related decompensated cirrhosis.

Trial registration number

http://ClinicalTrials.gov(NCT00502086).
Suicide-related discussions with depressed primary care patients in the USA: gender and quality gaps. A mixed methods analysis
Authors: Vannoy Steven D , Robins Lynne S
Published: October 2011

Objective

To characterise suicide-risk discussions in depressed primary-care patients.

Design

Secondary analysis of recordings and self reports by physicians and patients. Descriptive statistics of depression and suicide-related discussion, with qualitative extraction of disclosure, enquiry and physician response.

Setting

12 primary-care clinics between July 2003 and March 2005.

Participants

48 primary-care physicians and 1776 adult patients.

Measures

Presence of depression or suicide-related discussions during the encounter; patient and physician demographics; depression symptom severity and suicide ideation as measured by the Patient Health Questionnaire (PHQ9); physician's decision-making style as measured by the Medical Outcomes Study Participatory Decision-Making Scale; support for autonomy as measured by the Health Care Climate Questionnaire; trust in their physician as measured by the Primary Care Assessment Survey; physician response to suicide-related enquiry or disclosure.

Results

Of the 1776 encounters, 128 involved patients scoring >14 on the PHQ9. These patients were seen by 43 of the 48 physicians. Suicide ideation was endorsed by 59% (n=75). Depression was discussed in 52% of the encounters (n=66). Suicide-related discussion occurred in only 11% (n=13) of encounters. 92% (n=12) of the suicide discussions occurred with patients scoring <2 on PHQ9 item 9. Suicide was discussed in only one encounter with a male. Variation in elicitation and response styles demonstrated preferred and discouraged interviewing strategies.

Conclusions

Suicide ideation is present in a significant proportion of depressed primary care patients but rarely discussed. Men, who carry the highest risk for suicide, are unlikely to disclose their ideation or be asked about it. Patient-centred communication and positive healthcare climate do not appear to increase the likelihood of suicide related discussion. Physicians should be encouraged to ask about suicide ideation in their depressed patients and, when disclosure occurs, facilitate discussion and develop targeted treatment plans.
Is there an association between wheezing and constipation in preschool children? Explanations from a longitudinal birth cohort
Authors: Kiefte-de Jong J C , Lebon A , Jaddoe V W V , Hofman A , de Jongste J C , Moll H A
Published: October 2011

Objective

To assess whether wheezing and atopic dermatitis were associated with constipation in preschool children and to what extent shared risk factors contribute to this relationship.

Methods

A population-based sample of 4651 preschool children was used. At the age of 24, 36 and 48 months, a parental report of functional constipation was available according to the Rome II criteria, and data on atopic dermatitis and wheezing were available using age-adapted questionnaires from the International Study of Asthma and Allergies in Childhood. Stepwise multivariate analyses were performed to assess whether body mass index, infection exposure, food allergy and infant nutrition, and parental stress explained the association between wheezing, atopic dermatitis and constipation.

Results

Out of 4651 children, 12–17% had functional constipation between 24 and 48 months. Symptoms of wheezing decreased from 20% to 12% and atopic dermatitis decreased from 30% to 18% at the age of 24 and 48 months respectively. Between the age of 24 and 48 months, wheezing symptoms were significantly associated with functional constipation (OR 1.17; 1.02 to 1.34) but these results were mainly explained by the child's exposure to infections and use of antibiotics (adjusted odds ratio 1.08; 95% CI 0.95 to 1.24). No significant association was found between symptoms of atopic dermatitis and functional constipation (OR 1.08; 95% CI 0.94 to 1.23).

Conclusions

These findings suggest that functional constipation coexists with wheezing in childhood but is mainly explained by the child's infection exposure and use of antibiotics. Therefore, an independent association between respiratory symptoms and functional bowel disorders as suggested in previous studies is questionable.
A cluster randomised controlled trial of a comprehensive accreditation intervention to reduce alcohol consumption at community sports clubs: study protocol: Figure 1
Authors: Kingsland Melanie , Wolfenden Luke , Rowland Bosco C , Tindall Jennifer , Gillham Karen E , McElduff Patrick , Rogerson John C , Wiggers John H
Published: October 2011

Introduction

Excessive alcohol consumption is responsible for considerable harm from chronic disease and injury. Within most developed countries, members of sporting clubs consume alcohol at levels above that of communities generally. Despite the potential benefits of interventions to address alcohol consumption in sporting clubs, there have been no randomised controlled trials to test the effectiveness of these interventions. The aim of this study is to examine the effectiveness of a comprehensive accreditation intervention with community football clubs (Rugby League, Rugby Union, soccer/association football and Australian Rules football) in reducing excessive alcohol consumption by club members.

Methods and analysis

The study will be conducted in New South Wales, Australia, and employ a cluster randomised controlled trial design. Half of the football clubs recruited to the trial will be randomised to receive an intervention implemented over two and a half winter sporting seasons. The intervention is based on social ecology theory and is comprehensive in nature, containing multiple elements designed to decrease the supply of alcohol to intoxicated members, cease the provision of cheap and free alcohol, increase the availability and cost-attractiveness of non-alcoholic and low-alcoholic beverages, remove high alcohol drinks and cease drinking games. The intervention utilises a three-tiered accreditation framework designed to motivate intervention implementation. Football clubs in the control group will receive printed materials on topics unrelated to alcohol. Outcome data will be collected pre- and postintervention through cross-sectional telephone surveys of club members. The primary outcome measure will be alcohol consumption by club members at the club, assessed using a graduated frequency index and a seven day diary.

Ethics and dissemination

The study was approved by The University of Newcastle Human Research Ethics Committee (reference: H-2008-0432). Study findings will be disseminated widely through peer-reviewed publications and conference presentations.

Trial registration number

Australian New Zealand Clinical Trials Registry: ACTRN12609000224224.
Estimation of cell density to aid in assessment of percentage cells of a particular lineage or of cells expressing a specific antigen in bone marrow trephine biopsies
Authors: Al-Shieban Saeed , Tadrous Paul J , Naresh Kikkeri N
Published: October 2011

The authors aimed to develop a tool to assess total cell numbers in a microscope's field of vision, which would provide the denominator for calculating the percentage of positive cells for a given antigen in bone marrow trephine biopsies (BMTBs) of varying cellularities. Precise estimates of cell densities were made from 179 images of BMTBs of varying cellularities using a cell-counting software. The estimates were then validated on an independent set of 20 BMTBs. Among the 179 images, there was a strong linear association between marrow cellularity and cell numbers (Pearson correlation: 0.788). Then standardised cell densities (cells/mm2of bone marrow) were deduced for BMTBs of varying cellularities. In the validation study, the actual and the estimated cell numbers correlated strongly (Pearson correlation: 0.990). The cell density estimates provided in this study can be adapted for any microscope and the same method can be used for calculation of the percentage-positive cells for any antigen.
Human papillomavirus coinfection in the oral cavity of HIV-infected children
Authors: Pinheiro Raquel Santos , de França Talita Ribeiro , Rocha Barbara , Ferreira Dennis Carvalho , Ribeiro Camila Maria Beder , Cavalcanti Silvia Maria Beata , de Souza Ivete Pomarico Ribeiro , Leão Jair Carneiro , Castro Gloria Fernanda Barbosa
Published: October 2011

Aim

To identify the early presence of human papillomavirus (HPV) DNA in the oral cavity of HIV-infected children.

Methods

The study group comprised 50 HIV-infected children and 50 healthy aged paired controls. Oral mucosa samples were collected by swab, and a 450 bp HPV DNA fragment was amplified by PCR, using degenerate consensus primers directed to the L1 gene. HPV typing was performed using specifics primers for types 6/16 and 11/18.

Results

All HPV co-infected children had vertical transmission of HIV. HPV DNA was detected in 6/50 (12.0%) of HIV children and three control group individuals 3/50 (6.0%) (p>0.01). In the HIV group, one had type 11, in the control group one had co-infection of types 11/16, and the others had type 16 and 18.

Conclusions

Based upon the results, it is concluded that HPV DNA may be present in HIV disease, but no association with immunosuppression, delivery, or medical records (CD4 cell count, viral load and use of highly active antiretroviral therapy) was observed, and no differences could be observed between the groups either.
Histological examination in sudden unexpected death in infancy: evidence base for histological sampling
Authors: Weber M A , Pryce J W , Ashworth M T , Malone M , Sebire N J
Published: October 2011

Aim

Pathologists currently follow the ‘Kennedy guidelines’ when performing autopsies for sudden unexpected death in infancy (SUDI); these include extensive histological sampling. This study establishes the frequency with which histological examination of visceral organs determines cause of death and examines associations between clinical, macroscopic and microscopic findings.

Methods

Retrospective review of 546 SUDI autopsies performed for a 10-year period (1996–2005) at a single centre. The proportion of cases in which non-neuropathological histological examination directly determined the cause of death was identified, and clinical, macroscopic and histological findings at autopsy were compared.

Results

Of 510 SUDIs included, 166 cases were explained SUDI, and of these, 54% (89/166) were identified solely on microscopic examination, based on histology of the lungs in 71 (43%), heart in 13 (8%), liver in 4 (2%) and kidneys in 1 (<1%). The proportions of macroscopically normal organs with significant histological findings were 26% lungs, 2% heart and 1% each of liver and kidneys, but none of spleen, thymus, pancreas or adrenals. Macroscopically abnormal organs were more likely to yield significant histological features. Symptoms preceding death were more common in cases with significant histological findings in lungs, heart, liver and adrenals.

Conclusion

A non-neuropathological cause of death in explained SUDI can be established from histological examination of lungs, heart, liver and kidneys. Significant histological abnormalities may be detected in selected organs with macroscopically normal appearances. Routine histological sampling of other organs in the absence of specific clinical history or macroscopic abnormalities has a low yield for establishing cause of death.
A clinicopathological aspect of primary small-cell carcinoma of the uterine cervix: a single-centre study of 25 cases
Authors: Li Jun-Dong , Zhuang Yuan , Li Yan-Fang , Feng Yan-Ling , Hou Jin-Hui , Chen Liang , Zhu An-Na , Wu Qiu-liang , Yun Jing-Ping
Published: October 2011

Aims

Small-cell carcinoma is a variant of poorly differentiated neuroendocrine carcinoma. Primary small-cell carcinoma of the cervix (SCCC) is recognised as a rare and aggressive malignant tumour with poor prognosis. In this study, the authors report 25 Chinese cases of SCCC, with a particular focus on their clinical and pathological characteristics.

Material and methods

The records of 25 patients from 4075 Chinese patients with cervical cancer were collected and reviewed, including the patients' age, initial symptoms, cervical tumour size, International Federation of Gynaecology and Obstetrics clinical stage, lymph-node metastasis, treatments and follow-up results. Immunohistochemical detection was performed for cytokeratin, epithelial membrane antigen, neuron-specific enolase (NSE), synaptophysin (Syn), chromogranin A (CgA), neuronal cell adhesion molecules (CD56), thyroid transcriptional factor-1 and S100 protein (S100).

Results

The median age of 25 patients with SCCC was 43.7 years. The most common symptom was abnormal vaginal bleeding. Histologically, there were 19 ‘homogenous’ SCCC samples and six samples of SCCC mixed with adenocarcinoma. The proportion of SCCC samples with positive immunoreactivity were 100.0% for NSE, 96.0% for Syn, 68.0% for CD56, 76.0% for CgA, 40.0% for thyroid transcriptional factor-1, 84.0% for epithelial membrane antigen, 68.0% for cytokeratin and 8.0% for S100, respectively. Every patient received one to three types of treatments, including surgery, chemotherapy and radiotherapy. The median survival time of patients was 20.9 months after diagnosis.

Conclusion

The higher proportion of positive labelling of Syn, CD56, CgA, and NSE in SCCC implicated that they are valuably applied in a differential diagnosis of the malignancy. The patients with SCCC receive one to three types of therapies, including surgery, chemotherapy and radiotherapy, and have a poor prognosis.
Multicentric evaluation of a second generation assay to detect antiviral citrullinated peptide antibodies: a collaborative study by the Forum Interdisciplinare per la Ricerca nelle Malattie Autoimmuni
Authors: Bizzaro N , Allegri F , Alpini C , Doria A , Gerli R , Lotzniker M , Mathieu A , Morozzi G , Bellisai F , Riccieri V , Tampoia M , Migliorini P
Published: October 2011

Aims

A novel immunoenzymatic assay using viral citrullinated peptides derived from Epstein–Barr virus-encoded proteins (viral citrullinated peptide 2 (VCP2)) has been developed and evaluated by means of a multicentre collaborative study.

Methods

Three hundred nine sera from patients with established rheumatoid arthritis (RA), 36 with early arthritis, 12 with juvenile arthritis and 453 controls were tested for VCP2 and cyclic citrullinated peptide (CCP) antibodies.

Results

The VCP2 assay showed 78.3% sensitivity and 97.1% specificity. VCP2 and CCP had a high concordance rate in patients with RA (88%) and controls (97%). However, 36 RA sera were positive in the CCP assay but negative on VCP2, and two RA sera reacted only on VCP2.

Conclusions

The new VCP2 assay is endowed with high sensitivity and specificity. VCP2-positive RA sera are mostly but not completely contained in the CCP-positive population. Studies are in progress to establish whether the VCP2 assay can detect clinically distinct subsets of patients with RA.
Handling and reporting of nephrectomy specimens for adult renal tumours: a survey by the European Network of Uropathology
Authors: Algaba Ferran , Delahunt Brett , Berney Daniel M , Camparo Philippe , Compérat Eva , Griffiths David , Kristiansen Glen , Lopez-Beltran Antonio , Martignoni Guido , Moch Holger , Montironi Rodolfo , Varma Murali , Egevad Lars
Published: October 2011

Aim

To collect information on current practices of European pathologists for the handling and reporting of nephrectomy specimens with renal tumours.

Methods and Results

A questionnaire was circulated to the members of the European Network of Uropathology, which consists of 343 pathologists in 15 European countries. Replies were received from 48% of members. These replies indicated that nephrectomy specimens are most often received in formalin. Lymph nodes are found in less than 5% of nephrectomy specimens. All respondents give an objective measure of tumour size, most commonly in three diameters. The most common method to search for capsule penetration is to slice tissue outside the tumour perpendicularly into the tumour. The most common sampling algorithm from tumours greater than 2 cm is one section for every centimetre of maximum tumour diameter. Most respondents use the 2004 WHO renal tumour classification although only slightly over half consider small papillary tumours malignant if the diameter is greater than 5 mm. The Fuhrman grading system is widely used. Almost all use immunohistochemistry for histological typing in some cases, while only 7% always use it. The most utilised special stains are CK7 (95%), CD10 (93%), vimentin (86%), HMB45 (68%), c-kit (61%) and Hale's colloidal iron (52%). Only 18% use other ancillary techniques for diagnosis in difficult cases.

Conclusions

While most pathologists appear to follow published guidelines for reporting renal carcinoma, there is still a need for the development of consensus and further standardisation of practice for contentious areas of specimen handling and reporting.
Endoscopic ultrasound guided transbronchial fine needle aspiration: a French Department of Pathology's 4-year experience
Authors: Delattre Claire , Fournier Clément , Bouchindhomme Brigitte , Renaud Florence , Escande Fabienne , Ramon Philippe , Copin Marie-Christine
Published: October 2011

Background

Endobronchial ultrasound (EBUS) guided transbronchial needle aspiration (TBNA) is an accurate outpatient procedure used to explore mediastinal lymph nodes for lung cancer staging and unexplained mediastinal masses.

Aims and methods

A retrospective study was conducted over four years on EBUS-TBNA sampled lymph nodes investigated for the staging of lung cancer or unexplained mediastinal lymphadenopathies, first using the conventional method (CM) and then a liquid based cytology (LBC).

Results

Of the 628 specimens (355 patients) collected, the overall rate of adequacy was 88% and the diagnosis of malignancy was achieved in 43% of cases. The inadequate rate was 6% with LBC and 21% with CM. A paraffin cytoblock was available in 80% with LBC and 62% with CM. Of the 628 aspirates, 270 (43%) were categorised as negative for malignancy including 26 cases consistent with sarcoidosis, 272 (43%) as malignant, 9 (1.4%) as suspicious for non-small-cell carcinoma and 77 as inadequate samples (12%). Of the 272 cases diagnosed as malignant, 87 (32%) were classified as non-small-cell carcinoma, 106 (39%) as adenocarcinoma, 48 (18%) as squamous cell carcinoma and 20 (7%) as small cell carcinoma. Five lymphomas, four metastatic melanomas and two carcinoids were also diagnosed.

Conclusions

EBUS-TBNA is a reliable method for the staging of lung cancer and for unexplained mediastinal mass exploration. The LBC has a lower rate of inadequate samples, a better yield of cytoblock for immunohistochemistry and a dramatically reduced time requirement for interpretation as compared to CM.
Chloride channel ClC-2 modulates tight junction barrier function via intracellular trafficking of occludin
Authors: Nighot Prashant K. , Blikslager Anthony T.
Published: September 2011

Previously, we have demonstrated that the chloride channel ClC-2 modulates intestinal mucosal barrier function. In the present study, we investigated the role of ClC-2 in epithelial barrier development and maintenance in Caco-2 cells. During early monolayer formation, silencing of ClC-2 with small interfering (si)RNA led to a significant delay in the development of transepithelial resistance (TER) and disruption of occludin localization. Proteomic analysis employing liquid chromatography-mass spectrometry /mass spectrometry revealed association of ClC-2 with key proteins involved in intracellular trafficking, including caveolin-1 and Rab5. In ClC-2 siRNA-treated cells, occludin colocalization with caveolin-1 was diffuse and in the subapical region. Subapically distributed occludin in ClC-2 siRNA-treated cells showed marked colocalization with Rab5. To study the link between ClC-2 and trafficking of occludin in confluent epithelial monolayers, a Caco-2 cell clone expressing ClC-2 short hairpin (sh)RNA was established. Disruption of caveolae with methyl-β-cyclodextrin (MβCD) caused a marked drop in TER and profound redistribution of caveolin-1-occludin coimmunofluorescence in ClC-2 shRNA cells. In ClC-2 shRNA cells, focal aggregations of Rab5-occludin coimmunofluorescence were present within the cytoplasm. Wortmannin caused an acute fall in TER in ClC-2 shRNA cells and subapical, diffuse redistribution of Rab5-occludin coimmunofluorescence in ClC-2 shRNA cells. An endocytosis and recycling assay for occludin revealed higher basal rate of endocytosis of occludin in ClC-2 shRNA cells. Wortmannin significantly reduced the rate of recycling of occludin in ClC-2 shRNA cells. These data clearly indicate that ClC-2 plays an important role in the modulation of tight junctions by influencing caveolar trafficking of the tight junction protein occludin.
Angiotensin-converting enzyme 2 overexpression improves central nitric oxide-mediated sympathetic outflow in chronic heart failure
Authors: Zheng Hong , Liu Xuefei , Patel Kaushik P.
Published: October 2011

Angiotensin (ANG)-converting enzyme (ACE)2 in brain regions such as the paraventricular nucleus (PVN) controlling cardiovascular function may be involved in the regulation of sympathetic outflow in chronic heart failure (CHF). The purpose of this study was to determine if ACE2 plays a role in the central regulation of sympathetic outflow by regulating neuronal nitric oxide (NO) synthase (nNOS) in the PVN. We investigated ACE2 and nNOS expression within the PVN of rats with CHF. We then determined the effects of ACE2 gene transfer in the PVN on the contribution of NO-mediated sympathoinhibition in rats with CHF. The results showed that there were decreased expressions for ACE2, the ANG-(1–7) receptor, and nNOS within the PVN of rats with CHF. After the application of adenovirus vectors encoding ACE2 (AdACE2) into the PVN, the increased expression of ACE2 in the PVN was confirmed by Western blot analysis. AdACE2 transfection significantly increased nNOS protein levels (change of 50 ± 5%) in the PVN of CHF rats. In anesthetized rats, AdACE2 treatment attenuated the responses of renal sympathetic nerve activity (RSNA), mean arterial pressure, and heart rate to the NOS inhibitor N-monomethyl-l-arginine in rats with CHF (RSNA: 28 ± 3% vs. 16 ± 3%, P < 0.05) compared with CHF + AdEGFP group. Furthermore, neuronal NG-108 cells incubated with increasing doses of AdACE2 showed a dose-dependent increase in nNOS protein expression (60% at the highest dose). Taken together, our data highlight the importance of increased expression and subsequent interaction of ACE2 and nNOS within the PVN, leading to a reduction in sympathetic outflow in the CHF condition.
Human microvascular dysfunction and apoptotic injury induced by AL amyloidosis light chain proteins
Authors: Migrino Raymond Q. , Truran Seth , Gutterman David D. , Franco Daniel A. , Bright Megan , Schlundt Brittany , Timmons Mitchell , Motta Angelica , Phillips Shane A. , Hari Parameswaran
Published: October 2011

Light chain amyloidosis (AL) involves overproduction of amyloidogenic light chain proteins (LC) leading to heart failure, yet the mechanisms underlying tissue toxicity remain unknown. We hypothesized that LC induces endothelial dysfunction in non-AL human microvasculature and apoptotic injury in human coronary artery endothelial cells (HCAECs). Adipose arterioles ( n = 34, 50 ± 3 yr) and atrial coronary arterioles ( n = 19, 68 ± 2 yr) from non-AL subjects were cannulated. Adipose arteriole dilator responses to acetylcholine/papaverine were measured at baseline and 1 h exposure to LC (20 μg/ml) from biopsy-proven AL subjects (57 ± 11 yr) without and with antioxidant cotreatment. Coronary arteriole dilation to bradykinin/papaverine was measured post-LC exposure. HCAECs were exposed to 1 or 24 h of LC. LC reduced dilation to acetylcholine (10−4M: 41.6 ± 7 vs. 85.8 ± 2.2% control, P < 0.001) and papaverine (81.4 ± 4.6 vs. 94.8 ± 1.3% control, P < 0.01) in adipose arterioles and to bradykinin (10−6M: 68.6 ± 6.2 vs. 90.9 ± 1.6% control, P < 0.001) but not papaverine in coronary arterioles. There was an increase in superoxide and peroxynitrite in arterioles treated with LC. Adipose arteriole dilation was restored by cotreatment with polyethylene glycol-superoxide dismutase and tetrahydrobiopterin but only partially restored by mitoquinone (mitochondria-targeted antioxidant) and gp91ds-tat (NADPH oxidase inhibitor). HCAECs exposed to LC showed reduced NO and increased superoxide, peroxynitrite, annexin-V, and propidium iodide compared with control. Brief exposure to physiological amounts of LC induced endothelial dysfunction in human adipose and coronary arterioles and increased apoptotic injury in coronary artery endothelial cells likely as a result of oxidative stress, reduced NO bioavailability, and peroxynitrite production. Microvascular dysfunction and injury is a novel mechanism underlying AL pathobiology and is a potential target for therapy.
Cardiac regeneration therapy: connections to cardiac physiology
Authors: Takehara Naofumi , Matsubara Hiroaki
Published: October 2011

Without heart transplantation, a large number of patients with failing hearts worldwide face poor outcomes. By means of cardiomyocyte regeneration, cardiac regeneration therapy is emerging with great promise as a means for restoring loss of cardiac function. However, the limited success of clinical trials using bone marrow-derived cells and myoblasts with heterogeneous constituents, transplanted at a wide range of cell doses, has led to disagreement on the efficacy of cell therapy. It is therefore essential to reevaluate the evidence for the efficacy of cell-based cardiac regeneration therapy, focusing on targets, materials, and methodologies. Meanwhile, the revolutionary innovation of cardiac regeneration therapy is sorely needed to help the millions of people who suffer heart failure from acquired loss of cardiomyocytes. Cardiac regeneration has been used only in limited species or as a developing process in the rodent heart; now, the possibility of cardiomyocyte turnover in the human heart is being revisited. In the pursuit of this concept, the use of cardiac stem/progenitor stem cells in the cardiac niche must be focused to usher in a second era of cardiac regeneration therapy for the severely injured heart. In addition, tissue engineering and cellular reprogramming will advance the next era of treatment that will enable current cell-based therapy to progress to “real” cardiac regeneration therapy. Although many barriers remain, the prevention of refractory heart failure through cardiac regeneration is now becoming a realistic possibility.
Studies on the cell biology of interendothelial cell gaps
Authors: Ochoa Cristhiaan D. , Stevens Troy
Published: October 2011

Pain, redness, heat, and swelling are hallmarks of inflammation that were recognized as early as the first century AD. Despite these early observations, the mechanisms responsible for swelling, in particular, remained an enigma for nearly two millennia. Only in the past century have scientists and physicians gained an appreciation for the role that vascular endothelium plays in controlling the exudation that is responsible for swelling. One of these mechanisms is the formation of transient gaps between adjacent endothelial cell borders. Inflammatory mediators act on endothelium to reorganize the cytoskeleton, decrease the strength of proteins that connect cells together, and induce transient gaps between endothelial cells. These gaps form a paracellular route responsible for exudation. The discovery that interendothelial cell gaps are causally linked to exudation began in the 1960s and was accompanied by significant controversy. Today, the role of gap formation in tissue edema is accepted by many, and significant scientific effort is dedicated toward developing therapeutic strategies that will prevent or reverse the endothelial cell gaps that are present during the course of inflammatory illness. Given the importance of this field in endothelial cell biology and inflammatory disease, this focused review catalogs key historical advances that contributed to our modern-day understanding of the cell biology of interendothelial gap formation.
Airway tissue factor-dependent coagulation activity in response to sulfur mustard analog 2-chloroethyl ethyl sulfide
Authors: Rancourt Raymond C. , Veress Livia A. , Guo XiaoLing , Jones Tara N. , Hendry-Hofer Tara B. , White Carl W.
Published: October 2011

Acute lung injury is a principal cause of morbidity and mortality in response to mustard gas (SM) inhalation. Obstructive, fibrin-containing airway casts have recently been reported in a rat inhalation model employing the SM analog 2-chloroethyl ethyl sulfide (CEES). The present study was designed to identify the mechanism(s) causing activation of the coagulation cascade after CEES-induced airway injury. Here we report that CEES inhalation elevates tissue factor (TF) activity and numbers of detached epithelial cells present in lavage fluid (BALF) from rats after exposure (18 h). In vitro studies using 16HBE cells, or with rat BALF, indicated that detached epithelial cells could convert factor X (FX) to the active form FXa when incubated with factor VII and could elicit rapid clotting of plasma. In addition, immunocytochemical analysis demonstrated elevated cell surface (TF) expression on CEES-exposed 16HBE cells as a function of time. However, total cell TF expression did not increase. Since membrane surfaces bearing TF are important determinants of clot initiation, anticoagulants directed against these entities were tested for ability to limit plasma clotting or FX activation capacity of BALF or culture media. Addition of tifacogin, a TF pathway inhibitor, effectively blocked either activity, demonstrating that the procoagulant actions of CEES were TF pathway dependent. Lactadherin, a protein capable of competing with clotting factors for phospholipid-binding sites, was partially effective in limiting these procoagulant actions. These findings indicate that TF pathway inhibition could be an effective strategy to prevent airway obstruction after SM or CEES inhalation.
Baroreflex control of muscle sympathetic nerve activity as a mechanism for persistent sympathoexcitation following acute hypoxia in humans
Authors: Querido Jordan S. , Wehrwein Erica A. , Hart Emma C. , Charkoudian Nisha , Henderson William R. , Sheel A. William
Published: September 2011

This study tested the hypothesis that acute isocapnic hypoxia results in persistent resetting of the baroreflex to higher levels of muscle sympathetic nerve activity (MSNA), which outlasts the hypoxic stimulus. Cardiorespiratory measures were recorded in humans (26 ± 1 yr; n = 14; 3 women) during baseline, exposure to 20 min of isocapnic hypoxia, and for 5 min following termination of hypoxia. The spontaneous baroreflex threshold technique was used to determine the change in baroreflex function during and following 20 min of isocapnic hypoxia (oxyhemoglobin saturation = 80%). From the spontaneous baroreflex analysis, the linear regression between diastolic blood pressure (DBP) and sympathetic burst occurrence, the T50 (DBP with a 50% likelihood of a burst occurring), and DBP error signal (DBP minus the T50) provide indexes of baroreflex function. MSNA and DBP increased in hypoxia and remained elevated during posthypoxia relative to baseline ( P < 0.05). The DBP error signal became progressively less negative (i.e., smaller difference between DBP and T50) in the hypoxia and posthypoxia periods (baseline: −3.9 ± 0.8 mmHg; hypoxia: −1.4 ± 0.6 mmHg; posthypoxia: 0.2 ± 0.6 mmHg; P < 0.05). Hypoxia caused no change in the slope of the baroreflex stimulus-response curve; however, there was a shift toward higher pressures that favored elevations in MSNA, which persisted posthypoxia. Our results indicate that there is a resetting of the baroreflex in hypoxia that outlasts the stimulus and provide further explanation for the complex control of MSNA following acute hypoxia.
Biodegradation of Aged Residues of Atrazine and Alachlor in a Mix-Load Site Soil by Fungal Enzymes
Authors: Chirnside Anastasia E. M. , Ritter William F. , Radosevich Mark
Published: September 2011

Soils from bulk pesticide mixing and loading (mix-load) sites are often contaminated with a complex mixture of pesticides, herbicides, and other organic compounds used in pesticide formulations that limits the success of remediation efforts. Therefore, there is a need to find remediation strategies that can successfully clean up these mix-load site soils. This paper examined the degradation of atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT) and alachlor (2-chloro-, -diethyl-N-[methoxymethyl]-acetanilide) in contaminated mix-load site soil utilizing an extracellular fungal enzyme solution derived from the white rot fungus,Phanerochaete chrysosporium, grown in a packed bed bioreactor. Thirty-two percent of AT and 54% of AL were transformed in the biometers. The pseudo first-order rate constant for AT and AL biodegradation was 0.0882 d−1and 0.2504 d−1, respectively. The half-life () for AT and AL was 8.0 and 3.0 days, respectively. Compared to AT, the initial disappearance of AL proceeded at a faster rate and resulted in a greater amount of AL transformed. Based on the net evolved from the biometers, about 4% of the AT and AL initially present in the soil was completely mineralized.