bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence of familial malignancy in a prospectively screened cohort of patients with lymphoproliferative disorders
Authors: Brown Jennifer R. , Neuberg Donna , Phillips Kimberly , Reynolds Hazel , Silverstein Jason , Clark Jennifer C. , Ash Megan , Thompson Christina , Fisher David C. , Jacobsen Eric , LaCasce Ann S. , Freedman Arnold S.
Published: August 2008

Increasing evidence points to a heritable contribution in the development of lymphoma. The goal of this study was to determine the rate of familial lymphoproliferative malignancy among consecutive lymphoma patients presenting to a tertiary care center and to enrol families with multiple affected first‐degree relatives on a data and tissue collection study. Beginning in 2004 all new patients presenting to the Dana‐Farber Cancer Institute with non‐Hodgkin (NHL) or Hodgkin lymphoma (HL) or chronic lymphocytic leukaemia (CLL) were asked to complete a one‐page self‐administered family history questionnaire. 55·4% of 1948 evaluable patients reported a first‐degree relative with a malignancy, with the highest rate among CLL probands. Lymphoid malignancies were particularly common, with 9·4% of all probands reporting a first‐degree relative with a related lymphoproliferative disorder (LPD). This frequency was again highest for CLL, at 13·3% of CLL probands, compared to 8·8% of NHL probands and 5·9% of HL probands (P = 0·002). The prevalence of CLL was significantly increased in parents of CLL probands (P < 0·05), and a greater risk of NHL was seen in fathers of NHL probands than in mothers (P = 0·026). We conclude that familial aggregation of LPDs is common among newly diagnosed patients, varies significantly by diagnosis and contributes meaningfully to the population disease burden.
Secondary non‐Hodgkin lymphoma (NHL) in children and adolescents after childhood cancer other than NHL
Authors: Landmann Eva , Oschlies Ilske , Zimmermann Martin , Moser Olga , Graf Norbert , Suttorp Meinolf , Greiner Jeannette , Reiter Alfred ,   on behalf of the Berlin‐Frankfurt‐Münster group
Published: August 2008

The emergence of non‐Hodgkin lymphoma (NHL) during childhood and adolescence as a secondary neoplasm (SN) after previous cancer other than NHL is rare. To describe the characteristics and outcome of NHL following previous cancer other than NHL in children and adolescents, this study analysed the data of patients reported to the NHL‐Berlin‐Frankfurt‐Münster study centre from 1986 to 2005. Out of the total of 2968 NHL‐patients registered, 11 patients were assessed as having suffered from NHL as a proven SN. Four additional children had most likely suffered from NHL as an SN, but a late relapse of the first neoplasm could not be ruled out unequivocally. In the patients with proven SN, median age at diagnosis of the primary malignancy was 3·9 years (range 2–11·7). The median age at diagnosis of NHL was 7·6 years (range 4·7–18). Only lymphoblastic (n = 7) and diffuse large B‐cell (n = 4) lymphomas were diagnosed as SN. The estimated 5‐year event‐free survival from time of diagnosis of NHL was 91% [95% confidence interval (CI) 74–100%] in patients with proven SNs and 84% (95% CI 63–100%) when the patients with probable SNs were included in the analysis. We concluded that secondary NHL in children and adolescents confers a favourable prognosis.
Diagnostic performance of serum free light chain measurement in patients suspected of a monoclonal B‐cell disorder
Authors: Vermeersch Pieter , Van Hoovels Lieve , Delforge Michel , Mariën Godelieve , Bossuyt Xavier
Published: August 2008

The present study aimed to determine the diagnostic performance of different testing strategies to diagnose malignant B‐cell disorder or monoclonal gammopathy of unknown significance (MGUS). Sensitivity and specificity were determined in 833 consecutive patients investigated for a monoclonal gammopathy. Serum protein electrophoresis (PE), serum κ/λ free light chain (FLC) ratio, and serum and urine immunofixation electrophoresis (IFE) were performed in all patients. Twenty‐eight patients were diagnosed with a malignant plasma cell disorder, 25 with B‐cell non‐Hodgkin lymphoma and 156 with MGUS. Serum PE (with follow‐up IFE) plus FLC had a sensitivity of 82·3% and a specificity of 96·8% and missed one plasmacytoma and 23 patients with MGUS. Serum IFE plus urine IFE had a sensitivity of 92·3% and a specificity of 100% and missed two MGUS patients. Serum IFE plus FLC had a sensitivity of 93·8% and a specificity of 96·8% and missed one MGUS patient. Serum PE plus FLC had a significantly lower sensitivity than serum IFE plus FLC or serum IFE plus urine IFE for the diagnosis of MGUS. The sensitivity of serum IFE plus FLC was comparable to the sensitivity of serum IFE plus urine IFE. The specificity of serum IFE plus FLC, however, was lower than the specificity of serum IFE plus urine IFE.
Ghrelin and the differential regulation of des‐acyl (DSG) and oct‐anoyl ghrelin (OTG) in human adipose tissue (AT)
Authors: Kos K. , Harte A. L. , O’Hare P. J. , Kumar S. , McTernan P. G.
Published: August 2008

Objectives  Ghrelin, an important central acting orexigenic hormone, is predominantly secreted in the gastrointestinal tract. However little is known about the action of ghrelin in human adipose tissue (AT).

Aim  To study the expression of ghrelin in AT, the effects of octanoyl–(OTG) and des‐acyl (DSG) ghrelin on lipolysis and lipogenesis, leptin release and potential peripheral signalling through the Y1 receptor.

Methods  Ex vivo human AT was obtained from women undergoing elective surgery (46 (mean ± SD) 6·8 years, body mass index (BMI): 25·6 ± 5·0 kg/m2, n = 20). Abdominal‐subcutaneous (AbdSc) adipocytes were isolated and treated with recombinant human (rh) OTG and DSG to assess lipid metabolism leptin release and the influence of Y1‐receptor blocker.

Results  Ghrelin was expressed in AbdScAT and negatively correlated with BMI (lean: 3·6 ± 0·74 optical‐density‐units (OD), obese: 1·64 ± 0·45OD, *P < 0·05). Only DSG significantly suppressed glycerol release (Control (C): 286 ± 58 µl/l; DSG 1 nm: 224 ± 38 µl/l↓*; DSG 100 nm: 172 ± 13 µl/l↓*,*↓P < 0·05, n = 7) and reduced hormone sensitive lipase expression (C: 1·0 ± 0·3OD; DSG 1 nm: 0·8 ± 0·3OD↓*; DSG 100 nm: 0·6 ± 0·1OD↓*, n = 4). However, both isoforms increased lipoprotein lipase expression (C: 1·0 ± 0·3OD; DSG 100 nm: 0·2 ± 0·4OD↑*; OTG 100 nm: 2·5 ± 0·3OD↑*, n = 4), whilst blockade of Y1 eliminated this effect in both. Leptin was down‐regulated by DSG only (DSG 1 nm: 5·3 ± 0·7 ng/ml; DSG 100 nm: 4·1 ± 0·7 ng/ml*) and was significant after BMI adjustment (P = 0·029).

Conclusion  Ghrelin was expressed in human AbdSc AT. In vitro, both OGT and DSG appear to mediate fat deposition with the lipogenic effects in part mediated by the Y1 receptor, whilst the influence of DSG affected lipolysis, lipogenesis and leptin secretion. Taken together, these studies support a local action for ghrelin isoforms on lipid and adipokine metabolism that further supports a cross talk between organs.
Functional studies of new TSH receptor (TSHr) mutations identified in patients affected by hypothyroidism or isolated hyperthyrotrophinaemia
Authors: De Marco Giuseppina , Agretti Patrizia , Camilot Marta , Teofoli Francesca , Tatò Luciano , Vitti Paolo , Pinchera Aldo , Tonacchera Massimo
Published: August 2008

Objectives  To establish the role of new TSH receptor (TSHr) variants (P27T, E34 K, R46P, D403N, W488R and M527T) recently identified in children with congenital hypothyroidism (CH) or subclinical hypothyroidism (SH) with a thyroid gland of normal size.

Design and measurements  TSHr variants were obtained by mutagenesis. Wild‐type (wt) and TSHr mutants were expressed in COS cells and cAMP assay, 125I‐TSH binding and microchip flow cytometry analyses were performed.

Results  D403N and M527T mutants showed a lower cAMP response to bovine TSH (bTSH) with respect to the wtTSHr. R46P and W488R mutants did not show any response to bTSH stimulation in terms of cAMP production. The E34 K mutant showed a significantly lower cAMP response to stimulation with bTSH, while P27T had a lower cAMP response only to the highest dose of bTSH used. P27T, E34 K, D403N and M527T mutants showed a lower TSH binding capacity with respect to the wtTSHr. R46P and W488R mutants did not show any TSH binding.

Conclusions  E34 K, D403N, M527T, R46P and W488R TSHr variants seem to cause a functional abnormality of the receptor which is responsible for the observed phenotype. The P27T TSHr variant does not seem to play a functional role in the pathogenesis of CH and should be considered as a polymorphism.
Anaemia and marrow fibrosis in patients with primary hyperparathyroidism before and after curative parathyroidectomy
Authors: Bhadada Sanjay K. , Bhansali Anil , Ahluwalia Jasmina , Chanukya G. V. , Behera Arunanshu , Dutta Pinaki
Published: August 2008

Objective To determine the relationship between anaemia and myelofibrosis in patients with symptomatic primary hyperparathyroidism (PHPT) and to assess the effect of curative parathyroidectomy on anaemia and marrow fibrosis.

Design and methods In this prospective follow‐up study of 28 consecutive patients with symptomatic PHPT from January 2005 to June 2006, 15 patients were diagnosed with anaemia (haemoglobin < 130 g/l in males and < 120 g/l in females), eight (53%) of whom were finally recruited for the study. Complete blood cell count, serum calcium, phosphorus, alkaline phosphatase, intact PTH and 25‐hydroxyvitamin D and bone marrow examination were performed both before and after parathyroidectomy in all (n = 8) patients, but bone marrow examination after surgery was performed only in those who had marrow fibrosis at baseline (n = 6).

Results Anaemia was observed in 15 (53·3%) of the 28 patients with symptomatic PHPT. Normocytic normochromic anaemia that is characteristic of PHPT was found in 14 (50%) patients. Eight of the 15 patients with anaemia had a bone marrow examination and marrow fibrosis was observed in six (75%). Both anaemia and marrow fibrosis improved after successful parathyroidectomy, but improvement in anaemia was significant (P = 0·02) only in those with marrow fibrosis at baseline. Marrow fibrosis did not correlate with duration of the disease (P = 0·17), degree of hypercalcaemia (P = 0·53) or serum levels of intact PTH (P = 0·60).

Conclusions Anaemia is common in patients with symptomatic PHPT, and was associated with marrow fibrosis in the majority of the patients who underwent bone biopsy. Both anaemia and marrow fibrosis improved after curative parathyroidectomy, but improvement in anaemia was noticeable only in those who had marrow fibrosis at presentation.
Voice characteristics in women with congenital adrenal hyperplasia due to 21‐hydroxylase deficiency
Authors: Nygren Ulrika , Södersten Maria , Falhammar Henrik , Thorén Marja , Hagenfeldt Kerstin , Nordenskjöld Agneta
Published: August 2008

Objective  Congenital adrenal hyperplasia (CAH) is an autosomal recessive inherited disorder resulting in lack of cortisol and often aldosterone and overproduction of androgens. Raised levels of androgens can lead to increased mass in the laryngeal tissues, which may lower the fundamental frequency (F0) of the voice.

Design/patients  A follow‐up study of voice characteristics in 38 women with CAH and 24 age‐matched controls between 18 and 63 years of age using subjective self‐ratings of voice symptoms and acoustic and perceptual analyses. The results were correlated to mutation and treatment.

Measurements  The subjects rated degree of hoarseness, dark voice, voice problems and vocal fatigue on visual analogue scales. Audio recordings were made of a standard reading passage in a sound treated booth. Acoustic analyses were made of the subjects’ mean, minimum and maximum F0 in the habitual speech range. Three voice clinicians made perceptual evaluations of dark voice in terms of timbre.

Results  Women with CAH rated higher values than controls with regard to the statement ‘my voice is a problem in my daily life’. They spoke with significantly lower mean, lower minimum and lower maximum F0, as well as darker voice quality as compared with the controls. Affected voice symptoms were associated with a late diagnosis or problems with medication.

Conclusion  The voices in women with CAH can be virilized because of late diagnosis or suboptimal treatment with glucocorticoids. These voice problems may need referral for voice assessment and should be considered when treating CAH.
Serum total antioxidant status and lipid peroxidation marker malondialdehyde levels in overt and subclinical hypothyroidism
Authors: Torun Ayse Nur , Kulaksizoglu Sevsen , Kulaksizoglu Mustafa , Pamuk Baris Onder , Isbilen Elif , Tutuncu Neslihan Bascil
Published: August 2008

Background  Mitochondria are the main production site of free oxygen radicals, which can cause organ dysfunction by oxidation of cellular macromolecules such as carbohydrates, lipids and proteins. Oxidative stress may result from either overproduction of these species or from failure of the antioxidant defence systems. Thyroid hormones have well‐known effects on mitochondrial oxygen consumption, but data about how hypothyroidism affects oxidative stress are controversial, and little is known about oxidative stress in subclinical hypothyroidism. Total antioxidant status (TAS) gives information about all of the antioxidants in the organism, while malondialdehyde (MDA) is a lipid peroxidation marker used to assess lipid peroxidation due to increased oxidative stress. We aimed to determine how hypothyroidism and subclinical hypothyroidism affect serum MDA and TAS.

Subjects and methods  Serum TAS, MDA, C‐reactive protein levels and lipid compositions were studied in 20 hypothyroid, 40 subclinical hypothyroid and 40 healthy subjects.

Results  MDA was elevated in both hypothyroid and subclinical hypothyroid patients compared with controls, while TAS levels show no significant differences between groups. Low‐density lipoprotein (LDL) cholesterol levels were significantly high in both hypothyroid and subclinical hypothyroid patients. Triglyceride levels were high only in hypothyroid patients when compared with the controls. MDA showed a correlation with LDL cholesterol, total cholesterol and triglyceride.

Conclusions  These results suggest an increased oxidative stress in both hypothyroid and subclinical hypothyroidism states, which can be explained by both the insufficient increase in the antioxidant status and the altered lipid metabolism in these cases.
Down‐regulation of NR4A1 in follicular thyroid carcinomas is restored following lithium treatment
Authors: Camacho Cléber P. , Latini Flavia R. M. , Oler Gisele , Hojaij Flavio C. , Maciel Rui M. B. , Riggins Gregory J. , Cerutti Janete M.
Published: August 2008

Introduction  The identification of follicular thyroid adenoma‐associated transcripts will lead to a better understanding of the events involved in pathogenesis and progression of follicular tumours. Using Serial Analysis of Gene Expression, we identified five genes that are absent in a malignant follicular thyroid carcinoma (FTC) library, but expressed in follicular adenoma (FTA) and normal thyroid libraries.

Methods  NR4A1, one of the five genes, was validated in a set of 27 normal thyroid tissues, 10 FTAs and 14 FTCs and three thyroid carcinoma cell lines by real time PCR. NR4A1 can be transiently increased by a variety of stimuli, including lithium, which is used as adjuvant therapy of thyroid carcinoma with 131I. We tested if lithium could restore NR4A1 expression. The expression of other genes potentially involved in the same signalling pathway was tested. To this end, lithium was used at different concentration (10 mm or 20 mm) and time (2 h and 24 h) and the level of expression was tested by quantitative PCR. We next tested if Lithium could affect cell growth and apoptosis.

Results  We observed that NR4A1 expression was under‐expressed in most of the FTCs investigated, compared with expression in normal thyroid tissues and FTAs. We also found a positive correlation between NR4A1 and FOSB gene expression. Lithium induced NR4A1 and FOSB expression, reduced CCDN1 expression, inhibited cell growth and triggered apoptosis in a FTC cell line.

Conclusions  NR4A1 is under‐expressed in most of FTCs. The loss of expression of both NR4A1 and the Wnt pathway gene FOSB was correlated with malignancy. This is consistent with the hypothesis that its loss of expression is part of the transformation process of FTCs, either as a direct or indirect consequence of Wnt pathway alterations. Lithium restores NR4A1 expression, induces apoptosis and reduces cell growth. These findings may explain a possible molecular mechanism of lithium's therapeutic action.
Application of ISO 22000 and comparison with HACCP on industrial processing of common octopus (Octopus vulgaris) – Part I
Authors: Arvanitoyannis Ioannis S. , Varzakas Theodoros H.
Published: August 2008

Critical control points (CCPs) were identified in the risk assessment of octopus (Octopus vulgaris) processing and implemented in the hazard analysis critical control point (HACCP) plan. In the hazard analysis worksheet the different hazards were identified at each processing stage, whereas in the HACCP plan each CCP is identified and accompanied with the relevant significant hazard, critical limit, monitoring of the CCP and corrective actions. In this work comparison of ISO 22000 analysis with HACCP is carried out over octopus processing and packaging. ISO 22000 Analysis Worksheet was employed for determination of some prerequisite programmes (PrPs). Comparison between the two systems has been carried out using the hazard analysis worksheet. The PrPs are the main difference between the two systems. The incorporation of PrPs in the ISO 22000 made the system more flexible as a smaller number of CCPs was introduced.
Viscoelastic properties of reconstituted cassava dough
Authors: Rodríguez‐Sandoval Eduardo , Fernández‐Quintero Alejandro , Sandoval‐Aldana Angélica , Cuvelier Gerard , Bello‐Peréz Luis A.
Published: August 2008

The influence of water (60–70%) and salt (1–2%) content on the viscoelastic properties of cassava dough, reconstituted from cooked flour, was studied using a controlled strain rheometer. Reconstituted cassava dough behaved as a solid‐like material with the storage modulus (G′) predominant over the loss modulus (G″). As the water content was increased, G′ decreased and G″ increased; but tan δ was practically independent of the water content. This behaviour suggested that water had plasticising effects, but probably did not change dough structure. The effect of salt content on the dynamic rheological properties of cassava dough was not significant, except for G″ values at water contents close to 60%.
Study of organic acids, volatile fraction and caseins of a new Halloumi‐type cheese during ripening in whey brine
Authors: Stelios Kaminarides , Paraskevi Stamou , Theophilos Massouras , Aikaterini Georgala
Published: August 2008

Organic acids, fat hydrolysis, volatile compounds and sensory characteristics of a new brine cheese which combines characteristics of Halloumi and Feta cheeses during its ripening in whey brine (100g NaCl L−1) were studied. Thermotolerant protease of Mucor miehei as a coagulant enzyme and a mixture of thermotolerant starter cultures Enterococcus faecium 0165 (0.5% w/w) and Lactobacillus casei 80 10D were used. Good quality new Halloumi‐type cheese was produced with higher proteolysis than traditional Halloumi cheese kept in whey brine. The volatile compounds identified comprised alcohols, aldehydes, ketones, acids, esters, hydrocarbons and sulphur compounds. Ethanol was the dominant volatile compound determined. Lactic acid was the dominant acid produced; its concentration increased during ripening, reaching a maximum value of 9929 mg kg−1 at day 30. Acetic acid was also found in high amounts, which increased during cheese ripening. Lipolysis of cheese was not intense. The most abundant acids of the mature cheese were palmitic, oleic and acetic acid. The Halloumi‐type cheese scored higher in the sensory analysis when fresh than did the mature cheese.
The impact on tigers of poaching versus prey depletion
Authors: Chapron Guillaume , Miquelle Dale G. , Lambert Amaury , Goodrich John M. , Legendre Stéphane , Clobert Jean
Published: August 2008

There exists a continuing dilemma in prioritizing conservation actions for large carnivores. Habitat loss, poaching, and prey depletion have often been cited as the three primary threats, but there is debate over the relative importance of each.

We assess the relative importance of poaching and prey depletion rates, and use existing information in the literature and multi‐type branching process and deterministic felid population models to address four lines of evidence used to infer that tiger populations are inherently resilient to high mortality rates.

Our results suggest that tigers, more so than leopards or cougars, require large populations to persist, are quite susceptible to modest increases in mortality, and less likely to recover quickly after population declines. Demographic responses that would ensure population persistence with mortality rates that are sustainable for cougars or leopards are biologically unrealistic for tigers.

We propose alternative interpretations of evidence used to suggest that tigers are inherently resilient to high mortality rates. In contrast to other solitary felids, tigers breed later and their inter‐birth interval is larger, making them less resilient to poaching. A model used to support the contention that prey depletion has greater impact on population persistence than poaching appears to be based on false premises. Camera‐trapping data that suggest positive population growth despite low survival rate cannot differentiate mortality from emigration, and does not differentiate the impact of varying survival rate on different sex‐age classes; for example, low survival rate of dispersers is tolerable if survival rate of adult breeding females is high.

Synthesis and applications. While high prey numbers are essential to sustain tiger populations, our results suggest prey recovery efforts will not be sufficient if mortality rates reach 15%. Extrapolating demographic responses from other, even closely related species to develop conservation strategies can be misleading. Reduction of human‐caused mortality, especially of resident breeding females, appears to be the most essential short‐term conservation effort that must be made. Since mortality rates are usually unknown and generally stochastic in nature, any management policy that might reduce survival rates should be firmly avoided.
Internal borders for managing invasive marine species
Authors: Forrest Barrie M. , Gardner Jonathan P. A. , Taylor Michael D.
Published: August 2008

This study discusses theoretical and practical issues around the definition of internal borders for the management of marine pests, drawing on principles from freshwater and terrestrial pest management. Internal borders are defined as post‐border intervention points around which vector management and related activities, such as pest surveillance and incursion response, can be undertaken.

Internal borders can be identified in marine systems based on knowledge of natural barriers to the dispersal or establishment of pests, including planktonic life stages. We highlight opportunities to define internal borders at broad spatial scales according to oceanographic features or environmental conditions. At smaller spatial scales, habitat barriers based on relatively permanent features (e.g. substratum) could be defined for target organisms (or suites of similar organisms), especially those having both restricted habitat requirements and a limited planktonic duration.

Where internal borders are identified, risk‐based approaches can be used to determine vector management priorities, incorporating knowledge of the connectivity and strength of interactions between hubs of vector activity. However, greater biosecurity and biodiversity benefits may arise from approaches that combine risk‐based measures for target pests, with generic measures (e.g. vessel hull anti‐fouling) that are applied equally across all human‐mediated invasion pathways.

Case studies of high‐profile marine pests in New Zealand illustrate situations where the benefits of internal border management have been realized. The challenge now is to identify how practical it is to widely apply internal borders in a marine biosecurity context, and to identify new internal borders around which management can practically be undertaken.

Synthesis and applications. Effective control of pest species in marine environments is possible even when they become relatively widespread and established. The definition of internal borders provides an approach to marine biosecurity that assists in elucidating opportunities and priorities for management. Although marine systems are regarded as relatively open, the principles and approaches necessary for the successful management of marine pests are conceptually the same as those applied in freshwater systems and on land.
Characterization of apoptosis induced by grouper iridovirus in two newly established cell lines from barramundi, Lates calcarifer (Bloch)
Authors: Lai Y‐S , Chiou P P , Chen W‐J , Chen Y‐C , Chen C‐W , Chiu I‐S , Chen S‐D , Cheng Y‐H , Chang C‐Y
Published: August 2008

Two new cell lines have been established from the muscle and swim bladder tissues of barramundi, Lates calcarifer, and designated as BM (barramundi muscle) and BSB (barramundi swimbladder), respectively. The cells multiplied well at 28 °C in Leibovitz’s L‐15 medium supplemented with 10% foetal bovine serum, and have been continuously subcultured more than 100 times to date. Morphologically, BM cells were mostly fibroblastic, whereas BSB were mostly epithelial. Both cell lines were susceptible to grouper iridovirus (GIV) and displayed characteristics of apoptosis after viral infection. The induction of apoptosis was further assayed in GIV‐infected BM and BSB cells by various methods. The inhibition of cell growth by GIV was demonstrated by MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide] assay. Morphological observations revealed typical apoptotic features in the infected cells, including cell shrinkage and rounding, chromosome condensation and formation of apoptotic body‐like vesicles. Chromosome fragmentation was detected by DNA laddering and TUNEL assays. Finally, the appearance of phosphotidylserine on the outer leaflet of apoptotic cell membranes was confirmed by annexin V staining. This is the first report of apoptosis induced by GIV in fish cells.
Inactivated infectious haematopoietic necrosis virus (IHNV) vaccines
Authors: Anderson E , Clouthier S , Shewmaker W , Weighall A , LaPatra S
Published: August 2008

The inactivation dynamics of infectious haematopoietic necrosis virus (IHNV) by β‐propiolactone (BPL), binary ethylenimine (BEI), formaldehyde or heat and the antigenic and immunogenic properties of the inactivated vaccines were evaluated. Chemical treatment of IHNV with 2.7 mm BPL, 1.5 mm BEI or 50 mm formaldehyde abolished virus infectivity within 48 h whereas heat treatment at 50 or 100 °C rendered the virus innocuous within 30 min. The inactivated IHNV vaccines were recognized by rainbow trout, Oncorhynchus mykiss, IHNV‐specific antibodies and were differentially recognized by antigenic site I or antigenic site II IHNV glycoprotein‐specific neutralizing monoclonal antibodies. The BPL inactivated whole virus vaccine was highly efficacious in vaccinated rainbow trout challenged by waterborne exposure to IHNV 7, 28, 42 or 56 days (15 °C) after immunization. The formaldehyde inactivated whole virus vaccine was efficacious 7 or 11 days after vaccination of rainbow trout but performed inconsistently when tested at later time points. The other vaccines tested were not efficacious.
Expression of Insulin System in the Olfactory Epithelium: First Approaches to its Role and Regulation
Authors: Lacroix M.‐C. , Badonnel K. , Meunier N. , Tan F. , Poupon C. Schlegel‐Le , Durieux D. , Monnerie R. , Baly C. , Congar P. , Salesse R. , Caillol M.
Published: August 2008

Food odours are major determinants for food choice; their detection is influenced by nutritional status. Among different metabolic signals, insulin plays a major role in food intake regulation. The aim of the present study was to investigate a potential role of insulin in the olfactory mucosa (OM), using ex vivo tissues and in vitro primary cultures. We first established the expression of insulin receptor (IR) in rat olfactory mucosa. Transcripts of IR‐A and IR‐B isoforms, as well as IRS‐1 and IRS‐2, were detected in OM extracts. Using immunocytochemistry, IR protein was located in olfactory receptor neurones, sustentacular and basal cells and in endothelium of the lamina propria vessels. Moreover, the insulin binding capacity of OM was quite high compared to that of olfactory bulb or liver. Besides the main pancreatic insulin source, we demonstrated insulin synthesis at a low level in the OM. Interestingly 48 h of fasting, leading to a decreased plasmatic insulin, increased the number of IR in the OM. Local insulin concentration was also enhanced. These data suggest a control of OM insulin system by nutritional status. Finally, an application of insulin on OM, aiming to mimic postprandial insulin increase, reversibly decreased the amplitude of electro‐olfactogramme responses to odorants by approximately 30%. These data provide the first evidence that insulin modulates the most peripheral step of odour detection at the olfactory mucosa level.
Interleukin‐1β Release in the Supraoptic Nucleus Area During Osmotic Stimulation Requires Neural Function
Authors: Summy‐Long J. Y. , Hu S. , Long A. , Phillips T. M.
Published: August 2008

Interleukin (IL)‐1β is present throughout the magnocellular neuroendocrine system and co‐depletes with oxytocin and vasopressin from the neural lobe during salt‐loading. To examine whether IL‐1β is released from the dendrites/soma of magnocellular neurones during osmotic stimulation, microdialysis adjacent to the supraoptic nucleus (SON) in conscious rats was combined with immunocapillary electrophoresis and laser‐induced fluorescence detection to quantify cytokine in 5‐min dialysates collected before (0–180 min; basal), and after (180–240 min), hypertonic saline injected s.c. (1.5 m NaCl). Osmotic release of IL‐1β was compared after inhibiting local voltage‐gated channels for Na+ (tetrodotoxin) and Ca2+ (cadmium and nickel) or by reducing intracellular Ca2+ stores (thapsigargin). Immunohistochemistry combined with microdialysis was used to localise cytokine sources (IL‐1β+) and microglia (OX‐42+). Under conditions of microdialysis, the basal release of IL‐1β+ in the SON area was measurable and stable (pg/ml; mean ± SEM) from 0–60 min (2.2 ± 0.06), 60–120 min (2.32 ± 0.05) and 120–180 min (2.33 ± 0.06), likely originating locally from activated microglia (OX42+; IL‐1β+; ameboid, hypertrophied) and magnocellular neurones expressing IL‐1β. In response to osmotic stimulation, IL‐1β increased progressively in dialysates of the SON area by a mechanism dependent on intracellular Ca2+ stores sensitive to thapsigargin and, similar to dendritic secretion of oxytocin and vasopressin, required local voltage‐gated Na+ and Ca2+ channels for activation by osmoregulatory pathways from the forebrain. During osmotic stimulation, neurally dependent release of IL‐1β in the SON area likely upregulates osmosensitive cation currents on magnocellular neurones (observed in vitro by others), to facilitate dendritic release of neurohypophysial hormones.
Oestrogen Receptor α and β Immunoreactive Cells in the Suprachiasmatic Nucleus of Mice: Distribution, Sex Differences and Regulation by Gonadal Hormones
Authors: Vida B. , Hrabovszky E. , Kalamatianos T. , Coen C. W. , Liposits Z. , Kalló I.
Published: August 2008

Oestrogen regulates various aspects of circadian rhythm physiology. The presence of oestrogen receptors within the suprachiasmatic nucleus (SCN), the principal circadian oscillator, indicates that some actions of oestrogen on circadian functions may be exerted at that site. The present study analysed sex differences, topographic distribution, and neurochemical phenotype of neurones expressing the α and β subtypes of oestrogen receptors (ERα and ERβ) in the mouse SCN. We found that relatively few neurones in the SCN are immunoreactive (IR) for ERα (approximately 4.5% in females and 3% in males), but five‐ to six‐fold more SCN neurones express ERβ. ER‐IR neurones are primarily in the shell subdivision of the nucleus and show differences between the sexes, significantly greater numbers being found in females. Treatment of male or female gonadectomised mice with oestradiol benzoate for 24 h substantially reduced the number of ERβ‐IR neurones, but not ERα‐IR neurones. Double‐labelling immunocytochemical experiments to characterise the phenotype of the oestrogen‐receptive neurones showed the presence of the calcium‐binding proteins calretinin or calbindin D28K in approximately 12% and 10%, respectively, of ERα‐IR neurones. A higher proportion (approximately 38%) of ERβ‐IR neurones contains calbindin D28K; a few (approximately 2%) express calretinin or vasopressin. These double‐labelled cells appear primarily in the shell subdivision of the SCN. Neither vasoactive intestinal polypeptide‐ nor gastrin releasing peptide‐immunoreactivity was observed in ER‐IR neurones. These data indicate that the primary target cells for oestrogen are in the shell subdivision of the nucleus. The sexually differentiated expression and distribution of ERα and ERβ in various cell populations of the SCN suggest multiple modes of oestrogen signalling within this nucleus, which may modulate circadian functions.
The major and minor wall teichoic acids prevent the sidewall localization of vegetative dl‐endopeptidase LytF in Bacillus subtilis
Authors: Yamamoto Hiroki , Miyake Yukiko , Hisaoka Miharu , Kurosawa Shin‐ichirou , Sekiguchi Junichi
Published: August 2008

Cell separation in Bacillus subtilis depends on specific activities of dl‐endopeptidases CwlS, LytF and LytE. Immunofluorescence microscopy (IFM) indicated that the localization of LytF depended on its N‐terminal LysM domain. In addition, we revealed that the LysM domain efficiently binds to peptidoglycan (PG) prepared by chemically removing wall teichoic acids (WTAs) from the B. subtilis cell wall. Moreover, increasing amounts of the LysM domain bound to TagB‐ or TagO‐depleted cell walls. These results strongly suggested that the LysM domain specifically binds to PG, and that the binding may be prevented by WTAs. IFM with TagB‐, TagF‐ or TagO‐reduced cells indicated that LytF−6xFLAG was observed not only at cell separation site and poles but also as a helical pattern along the sidewall. Moreover, we found that LytF was localizable on the whole cell surface in TagB‐, TagF‐ or TagO‐depleted cells. These results strongly suggest that WTAs inhibit the sidewall localization of LytF. Furthermore, the helical LytF localization was observed on the lateral cell surface in MreB‐depleted cells, suggesting that cell wall modification by WTAs along the sidewall might be governed by an actin‐like cytoskeleton homologue, MreB.
Assembly of the translocase motor onto the preprotein‐conducting channel
Authors: Karamanou Spyridoula , Bariami Vassiliki , Papanikou Efrosyni , Kalodimos Charalampos G. , Economou Anastassios
Published: August 2008

Bacterial protein secretion is catalysed by the SecYEG protein‐conducting channel complexed with the SecA ATPase motor. To gain insight into the SecA–SecYEG interaction we used peptide arrays, thermodynamic quantification, mutagenesis and functional assays. Our data reveal that: (i) SecA binds with low affinity on several, peripheral, exposed SecYEG sites. This largely electrostatic association is modulated by temperature and nucleotides. (ii) Binding sites cluster in five major binding ‘regions’: three that are exclusively cytoplasmic and two that reach the periplasm. (iii) Both the N‐terminal and c‐terminal regions of SecA participate in binding interactions and share some sites. (iv) Several of these sites are essential for translocase catalysis. Our data provide residue‐level dissection of the SecYEG–SecA interaction. Two models of assembly of SecA on dimeric SecYEG are discussed.
Contact‐dependent growth inhibition requires the essential outer membrane protein BamA (YaeT) as the receptor and the inner membrane transport protein AcrB
Authors: Aoki Stephanie K. , Malinverni Juliana C. , Jacoby Kyle , Thomas Benjamin , Pamma Rupinderjit , Trinh Brooke N. , Remers Susan , Webb Julia , Braaten Bruce A. , Silhavy Thomas J. , Low David A.
Published: August 2008

Contact‐dependent growth inhibition (CDI) is a phenomenon by which bacterial cell growth is regulated by direct cell‐to‐cell contact via the CdiA/CdiB two‐partner secretion system. Characterization of mutants resistant to CDI allowed us to identify BamA (YaeT) as the outer membrane receptor for CDI and AcrB as a potential downstream target. Notably, both BamA and AcrB are part of distinct multi‐component machines. The Bam machine assembles outer membrane β‐barrel proteins into the outer membrane and the Acr machine exports small molecules into the extracellular milieu. We discovered that a mutation that reduces expression of BamA decreased binding of CDI+ inhibitor cells, measured by flow cytometry with fluorescently labelled bacteria. In addition, α‐BamA antibodies, which recognized extracellular epitopes of BamA based on immunofluorescence, specifically blocked inhibitor–target cells binding and CDI. A second class of CDI‐resistant mutants identified carried null mutations in the acrB gene. AcrB is an inner membrane component of a multidrug efflux pump that normally forms a cell envelope‐spanning complex with the membrane fusion protein AcrA and the outer membrane protein TolC. Strikingly, the requirement for the BamA and AcrB proteins in CDI is independent of their multi‐component machines, and thus their role in the CDI pathway may reflect novel, import‐related functions.
General stress response signalling: unwrapping transcription complexes by DNA relaxation via the sigma38 C‐terminal domain
Authors: Huo Yi‐Xin , Rosenthal Adam Z. , Gralla Jay D.
Published: August 2008

Escherichia coli responds to stress by a combination of specific and general transcription signalling pathways. The general pathways typically require the master stress regulator sigma38 (rpoS). Here we show that the signalling from multiple stresses that relax DNA is processed by a non‐conserved eight‐amino‐acid tail of the sigma 38 C‐terminal domain. By contrast, responses to two stresses that accumulate potassium glutamate do not rely on this short tail, but still require the overall C‐terminal domain. In vitro transcription and footprinting studies suggest that multiple stresses can target a poised RNA polymerase and activate it by unwrapping DNA from a nucleosome‐like state, allowing the RNA polymerase to escape into productive mode. This transition can be accomplished by either the DNA relaxation or potassium glutamate accumulation that characterizes many stresses.
A conserved anti‐repressor controls horizontal gene transfer by proteolysis
Authors: Bose Baundauna , Auchtung Jennifer M. , Lee Catherine A. , Grossman Alan D.
Published: August 2008

The mobile genetic element ICEBs1 is an integrative and conjugative element (a conjugative transposon) found in the Bacillus subtilis chromosome. The SOS response and the RapI‐PhrI sensory system activate ICEBs1 gene expression, excision and transfer by inactivating the ICEBs1 repressor protein ImmR. Although ImmR is similar to many characterized phage repressors, we found that, unlike these repressors, inactivation of ImmR requires an ICEBs1‐encoded anti‐repressor ImmA (YdcM). ImmA was needed for the degradation of ImmR in B. subtilis. Coexpression of ImmA and ImmR in Escherichia coli or co‐incubation of purified ImmA and ImmR resulted in site‐specific cleavage of ImmR. Homologues of immR and immA are found in many mobile genetic elements. We found that the ImmA homologue encoded by B. subtilis phage φ105 is required for inactivation of the φ105 repressor (an ImmR homologue). ImmA‐dependent proteolysis of ImmR repressors may be a conserved mechanism for regulating horizontal gene transfer.
Induction of Enzymes and Phenolic Compounds Related to the Natural Defence Response of Netted Melon Fruit by a Bio‐elicitor
Authors: Sánchez‐Estrada Alberto , Tiznado‐Hernández Martín Ernesto , Ojeda‐Contreras Angel Javier , Valenzuela‐Quintanar Ana Isabel , Troncoso‐Rojas Rosalba
Published: August 2008

Fungal disease in netted melon fruit is an important factor affecting their postharvest quality and therefore an important cause of large economic losses around the world. Among the alternatives to control fungal diseases, the induction of the natural defence response (NDR) in fruits is promising. The objective of this study was to induce the NDR in netted melon treated with a bio‐elicitor formulated from Fusarium oxysporum growth in a potato dextrose agar enriched with netted melon skin. Netted melon fruits (cv ‘Primo’) were not treated (C), untreated and inoculated with F. oxysporum (IN), treated with a bio‐elicitor (FES), or treated with the bio‐elicitor and inoculated (FES + IN). After treatments, fruits were stored for 8 days at 20°C with 90–92% relative humidity. Melon was sampled every 2 days at 20°C to evaluate the development of Fusarium rot symptoms as disease index percentage (DI), changes in phenolic compounds, changes in phenylalanine ammonia‐lyase (PAL) activity, chitinase activity (ChA) and β‐1,3‐glucanase activity (GA). It was found that DI in netted melon fruit was significantly reduced in the FES + IN as compared with the IN treatment. FES + IN and FES treatments showed the highest increase of phenolic acids. Higher levels of PAL activity were observed in the treatments IN, FES, and FES + IN with respect to C, after 4 days of storage. A large increase in ChA activity was observed in the treatments IN, FES and FES + IN after 6 days of storage. No differences in GA activity were found among FES, FES + IN and C treatments throughout storage. IN treatment showed the highest increase in GA activity after 4 days of storage. It is concluded that the bio‐elicitor activates the NDR as measured by the increase in phenolic acids synthesis, PAL and ChA enzymes activity, in a similar way as the infection by the living pathogen.
HOST RESPONSE TO COLORECTAL CANCER
Authors: Salama Paul , Platell Cameron
Published: August 2008

It has long been established that inflammation and immunity play critical roles in the pathogenesis, control and eventual metastasis of cancers. With the advent of more sophisticated animal models and immunohistochemical techniques a greater understanding of the immune system and its interactions has occurred. Individual immune cells are dynamic structures that have variable behaviour controlled by complex interactions in the tumour microenvironment. In the setting of colorectal cancer it was first observed that peritumoral inflammatory infiltrates were associated with improved prognosis. Immunohistochemistry has shown the individual cells types within these infiltrates. It now appears that an adaptive immune response, differentiated along the T‐helper 1 pathway controls tumour invasion and metastasis. Furthermore, the immune system exerts selection pressure leading to the evolution of tumour cell variants that can induce tolerance and disable adaptive immunity. These tumour cells then use the mechanisms of innate immunity to facilitate further growth, angiogenesis, invasion and eventual metastasis. These issues are investigated with particular relevance to colorectal cancer. Using the immune response to defeat CRC has been under intense investigation but has so far been unsuccessful. Nevertheless, researchers remain optimistic that immunotherapy will play an important role in the treatment of this common disease.
PANCREATICODUODENECTOMY FOR LOCALLY ADVANCED STOMACH CANCER: PRELIMINARY RESULTS
Authors: Chan Weng‐Hoong , Cheow Peng‐Chung , Chung Alexander Y. F. , Ong Hock‐Soo , Koong Heng‐Nung , Wong Wai‐Keong
Published: August 2008

Background:  Pancreaticoduodenectomy (PD) for locally advanced stomach cancer involving duodenum or/and pancreatic head was controversial and rarely carried out. It was mainly reported from the Japanese institutions.

Methods:  A review of prospective database from January 2003 to December 2006 of patients who had locally advanced stomach cancer involving duodenum or/and head of pancreas that precluded curative subtotal gastrectomy who underwent diagnostic laparoscopy or exploratory laparotomy to exclude peritoneal metastatic disease. Patients were advised to undergo neoadjuvant chemotherapy before PD.

Results:  Seven patients underwent PD during the above‐mentioned period. Only four patients had neoadjuvant chemotherapy before PD. The median operative time was 8 h (range 6–9 h). Five patients had combined tranverse colectomy done. There was no 30‐day operative mortality or re‐operation. Three patients developed controlled pancreatic leaks and fistulas that were successfully treated with conservative measures. The length of hospital stay was 10–53 days (median 15 days). Median survival was 13 months and 2‐year survival rate was 60%. Patients who received neoadjuvant chemotherapy seemed to have better survival rate (P = 0.039).

Conclusion:  Our initial experience has shown that with careful and stringent patients selection, PD for locally advanced stomach cancer can be carried out with acceptable morbidity and mortality. Early results for patients who received neoadjuvant chemotherapy showed trend towards prolonged survival. However, longer follow up and further patient recruitment are needed to confirm our initial optimistic findings.
DIAGNOSTIC VALUE OF ROUTINE AQUEOUS CONTRAST SWALLOW EXAMINATION AFTER OESOPHAGECTOMY FOR DETECTING LEAKAGE OF THE CERVICAL OESOPHAGOGASTRIC ANASTOMOSIS
Authors: Boone Judith , Rinkes Inne Borel , Van Leeuwen Maarten , Van Hillegersberg Richard
Published: August 2008

Background:  Water‐soluble contrast swallow examination is routinely carried out after oesophagectomy to detect leakage of the cervical oesophagogastric anastomosis. This study evaluated the diagnostic accuracy and clinical value.

Methods:  Patients with oesophageal carcinoma who underwent oesophagectomy with gastric conduit formation and a hand‐sewn cervical anastomosis between 1989 and 2003 were reviewed on outcome of routine aqueous contrast swallow examination (RACSE) and appearance of clinical anastomotic leakage.

Results:  An RACSE was carried out in 207 (82%) of 252 patients on postoperative day 8 (range 3–15). In 45 patients, no RACSE was executed, mainly because of a prolonged stay in intensive care unit. In 18 (9%) of 207 cases, the RACSE could not be interpreted by the radiologist. In 19 (53%) of 36 patients who developed a clinical leakage, the leak had already manifested clinically before the routine contrast examination was planned. Taken together, the false‐positive rate was 8%, the false‐negative rate 48%, sensitivity 52%, specificity 92%, positive predictive value 46% and negative predictive value 93%. No significant differences were found between the accuracy of RACSE in end‐to‐end or end‐to‐side cervical anastomoses.

Conclusion:  Given the very low sensitivity and low positive predictive value and given the fact that in 53% of patients with a clinical leak, the leakage had appeared clinically before the contrast swallow examination was routinely planned, we propose to abandon the routine contrast swallow examination after oesophagectomy to detect cervical anastomotic leakage. Alternatively, anastomotic integrity can be tested by drinking small amounts of water with simultaneous observation of the cervical wound.