bluuebunny/crossref_metadata_2025 huggingface dataset:
The Demosponge Amphimedon queenslandica: Reconstructing the Ancestral Metazoan Genome and Deciphering the Origin of Animal Multicellularity
Authors: Degnan Bernard M. , Adamska Maja , Craigie Alina , Degnan Sandie M. , Fahey Bryony , Gauthier Marie , Hooper John N.A. , Larroux Claire , Leys Sally P. , Lovas Erica , Richards Gemma S.
Published: December 2008

INTRODUCTION

Sponges are one of the earliest branching metazoans. In addition to undergoing complex development and differentiation, they can regenerate via stem cells and can discern self from nonself (“allorecognition”), making them a useful comparative model for a range of metazoan-specific processes. Molecular analyses of these processes have the potential to reveal ancient homologies shared among all living animals and critical genomic innovations that underpin metazoan multicellularity. Amphimedon queenslandica (Porifera, Demospongiae, Haplosclerida, Niphatidae) is the first poriferan representative to have its genome sequenced, assembled, and annotated. Amphimedon exemplifies many sessile and sedentary marine invertebrates (e.g., corals, ascidians, bryozoans): They disperse during a planktonic larval phase, settle in the vicinity of conspecifics, ward off potential competitors (including incompatible genotypes), and ensure that brooded eggs are fertilized by conspecific sperm. Using genomic and expressed sequence tag (EST) resources from Amphimedon, functional genomic approaches can be applied to a wide range of ecological and population genetic processes, including fertilization, dispersal, and colonization dynamics, host-symbiont interactions, and secondary metabolite production. Unlike most other sponges, Amphimedon produce hundreds of asynchronously developing embryos and larvae year-round in distinct, easily accessible brood chambers. Embryogenesis gives rise to larvae with at least a dozen cell types that are segregated into three layers and patterned along the body axis. In this article, we describe some of the methods currently available for studying A. queenslandica, focusing on the analysis of embryos, larvae, and post-larvae.
Dictyostelium discoideum: The Social Ameba
Authors: Gaudet Pascale , Fey Petra , Chisholm Rex
Published: December 2008

INTRODUCTION

Dictyostelium discoideum is a unicellular eukaryote often referred to as a “social ameba” because it can form a multicellular structure when nutrient conditions are limiting. D. discoideum and related organisms, known as the Dictyostelia, have been studied for almost 150 years. The cellular and molecular aspects of their multicellular lifestyle have been studied in detail, and general principles for cell-to-cell communication, intracellular signaling, and cytoskeletal organization during cell motility have been derived from this work and have been found to be conserved across all eukaryotes. The bacteriovore nature of the unicellular stage provides an excellent model in which to study phagocytosis and the mechanisms of bacterial virulence. D. discoideum has also been used successfully to explore the molecular basis of various human diseases, as well as the mechanisms of drug action and the pathways that lead to resistance to certain therapeutic agents. The availability of a complete genome sequence has further widened the scope of studies using D. discoideum. A large potential for secondary metabolism has become apparent, which opens the door to discovering new compounds with potential medical applications. Numerous putative orthologs of genes responsible for diseases in humans, but whose molecular functions are still uncharacterized, are present in the D. discoideum genome. Finally, the availability of community resources, including the genome database dictyBase and the Dicty Stock Center, makes D. discoideum an easily accessible and powerful model organism to study.
The Dogfish Scyliorhinus canicula: A Reference in Jawed Vertebrates
Authors: Coolen Marion , Menuet Arnaud , Chassoux Danièle , Compagnucci Claudia , Henry Sébastien , Lévèque Laurent , Da Silva Corinne , Gavory Frédérick , Samain Sylvie , Wincker Patrick , Thermes Claude , D’Aubenton-Carafa Yves , Rodriguez-Moldes Isabel , Naylor Gavin , Depew Michael , Sourdaine Pascal , Mazan Sylvie
Published: December 2008

INTRODUCTION

Due to their large size and long generation times, chondrichthyans have been largely ignored by geneticists. However, their key phylogenetic position makes them ideal subjects to study the molecular bases of the important morphological and physiological innovations that characterize jawed vertebrates. Such analyses are crucial to understanding the origin of the complex genetic mechanisms unraveled in osteichthyans. The small spotted dogfish Scyliorhinus canicula, a representative of the largest order of extant sharks, presents a number of advantages in this context. Due to its relatively small size among sharks, its abundance, and easy maintenance, the dogfish has been an important model in comparative anatomy and physiology for more than a century. Recently, revived interest has occurred with the development of large-scale transcriptomic and genomic resources, together with the establishment of facilities allowing massive egg and embryo production. These new tools open the way to molecular analyses of the elaborate physiological and sensory systems used by sharks. They also make it possible to take advantage of unique characteristics of these species, such as organ zonation, in analyses of cell proliferation and differentiation. Finally, they offer important perspectives to evolutionary developmental biology that will provide a better understanding of the origin and diversifications of jawed vertebrates. The dogfish whole-genome sequence, which may shortly become accessible, should establish this species as an essential shark reference, complementary to other chondrichthyan models. These analyses are likely to reveal an organism of an underestimated complexity, far from the primitive prototypical gnathostome anticipated in gradistic views.
Bone Marrow-Derived Macrophages (BMM): Isolation and Applications
Authors: Weischenfeldt Joachim , Porse Bo
Published: December 2008

INTRODUCTION

Bone marrow-derived macrophages (BMM) are primary macrophage cells, derived from bone marrow cells in vitro in the presence of growth factors. Macrophage colony-stimulating factor (M-CSF) is a lineage-specific growth factor that is responsible for the proliferation and differentiation of committed myeloid progenitors into cells of the macrophage/monocyte lineage. Mice lacking functional M-CSF are deficient in macrophages and osteoclasts and suffer from osteopetrosis. In this protocol, bone marrow cells are grown in culture dishes in the presence of M-CSF, which is secreted by L929 cells and is used in the form of L929-conditioned medium. Under these conditions, the bone marrow monocyte/macrophage progenitors will proliferate and differentiate into a homogenous population of mature BMMs. The efficiency of the differentiation is assessed using fluorescence-activated cell sorting (FACS) analysis of Mac-1 and 4/80 surface antigen expression. Once differentiated, the BMMs are suitable for numerous types of experimental manipulations, including morphological, gene expression, and physiological studies. For example, phagocytic cells such as macrophages have a unique ability to ingest microbes. We describe a test for the phagocytic efficiency of BMMs by exposing them to fluorescently labeled yeast zymosan bioparticles. Also, a method to deliver DNA or small interfering RNAs (siRNAs) into these hard-to-transfect cells is described. Finally, the proliferation of the BMMs is assayed using carboxyfluorescein succinimidyl ester (CFSE), a fluorescein derivative that partitions equally between daughter cells after cell division.
Isolation of Amphimedon Developmental Material
Authors: Leys Sally P. , Larroux Claire , Gauthier Marie , Adamska Maja , Fahey Bryony , Richards Gemma S. , Degnan Sandie M. , Degnan Bernard M.
Published: December 2008

INTRODUCTION

Fertilization occurs internally in Amphimedon and embryos are brooded in multiple chambers throughout the adult. Each chamber contains a mixture of developmental stages, from egg to late ring stages (i.e., prehatch late embryos). At the end of embryogenesis, swimming parenchymella larvae emerge from the adult. After several hours in the water column, the larvae settle and metamorphose into juvenile sponges. This protocol details how to obtain Amphimedon larvae and post-larvae/juveniles as well as embryos. Once isolated, these biological stages can be used for a variety of molecular and cellular analyses.
Selection of Dictyostelium Transformants
Authors: Gaudet Pascale , Fey Petra , Chisholm Rex
Published: December 2008

INTRODUCTION

Dictyostelium discoideum is a unicellular eukaryote often referred to as a social ameba because it can form a multicellular structure when nutrient conditions are limiting. General principles for cell-to-cell communication, intracellular signaling, and cytoskeletal organization during cell motility have been derived from cellular and molecular studies of Dictyostelium and have been found to be conserved across all eukaryotes. The availability of a complete genome database and stocks of wild-type and mutant strains make D. discoideum an accessible and powerful model organism. Dictyostelium is amenable to genetic manipulations that require the introduction of DNA into cells, such as gene knockout, overexpression, antisense RNA expression, RNA interference (RNAi)-mediated gene knockdown, and restriction-enzyme-mediated mutagenesis. Two commonly used methods for DNA-mediated transformation in Dictyostelium are calcium phosphate precipitation and electroporation. Transformants can then be selected in liquid media or on bacterial plates. The latter method reduces the chances of contamination because the cells are grown in buffered agar containing live or dead bacteria, rather than in a rich broth. This method also facilitates the isolation of clones from transformations because each transformant produces a single colony on the plate instead of the pools of transformants obtained in liquid culture. For gene ablation experiments, it is important to obtain a minimum of two independent clones with the same phenotype to exclude the possibility that the phenotype is due to a nonspecific mutation.
Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions. Protocol 1: Coexpression of Query Protein on NAPPA Slides
Authors: Link Andrew J. , LaBaer Joshua
Published: December 2008

INTRODUCTION

The Nucleic Acid Programmable Protein Array (NAPPA) approach for producing protein microarrays uses cell-free extracts to transcribe and translate cDNAs encoding target proteins directly onto glass slides. Following array preparation, interactions with a protein of interest (query protein) are detected either by probing an expressed NAPPA slide with the purified query protein or by coexpressing the query protein on the NAPPA slide at the same time that the target proteins are expressed. This protocol describes the coexpression method, which involves adding the gene for the query protein to the cell-free protein expression mix. The amount of query protein that is transcribed and translated from the corresponding plasmid DNA depends on the amount of plasmid DNA used and the size of the protein of interest, among other factors. If too little query protein is expressed, there may be no detectable binding signal. Excessive amounts of protein expression may generate nonspecific background signals. Because the optimum amount of query plasmid varies with each query protein, it is essential to assess empirically the optimal amount of query protein DNA to add to a coexpression experiment.
Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions. Protocol 2: Detection of Query Proteins on NAPPA Slides
Authors: Link Andrew J. , LaBaer Joshua
Published: December 2008

INTRODUCTION

The Nucleic Acid Programmable Protein Array (NAPPA) approach for producing protein microarrays uses cell-free extracts to transcribe and translate cDNAs encoding target proteins directly onto glass slides. Identification of protein interactions can be accomplished either by probing an expressed NAPPA slide with a purified protein of interest (the query protein) or by coexpressing the query protein on the NAPPA slide at the same time that the target proteins are expressed. This protocol describes detection of query protein following coexpression of the query protein on the NAPPA array.
Prognostic significance of L‐type amino acid transporter 1 (LAT1) and 4F2 heavy chain (CD98) expression in early stage squamous cell carcinoma of the lung
Authors: Kaira Kyoichi , Oriuchi Noboru , Imai Hisao , Shimizu Kimihiro , Yanagitani Noriko , Sunaga Noriaki , Hisada Takeshi , Ishizuka Tamotsu , Kanai Yoshikatsu , Endou Hitoshi , Nakajima Takashi , Mori Masatomo
Published: December 2008

The purpose of this study was to evaluate the prognostic value of L‐type amino acid transporter 1 (LAT 1) and 4F2 heavy chain (CD98) in patients with stage I squamous cell carcinoma of the lung. A total of 84 consecutive patients with completely resected pathologic stage I squamous cell carcinoma of the lung were retrospectively reviewed. All patients underwent resection of the tumor and the immunohistochemical analysis was done to determine the expression of LAT 1, CD98, Ki‐67 labeling index, vascular endothelial growth factor, and microvessel density. These pathological parameters were correlated with the prognosis of patients after complete resection of the tumor. A positive rate of LAT 1 expression (87%; 73/84) was significantly higher than that of CD98 expression (65%; 55/84) (P = 0.0018). Cooperative expression of LAT 1 and CD98 was recognized in 62% (52/84). LAT 1 expression was significantly correlated with CD98, Ki‐67 labeling index, vascular endothelial growth factor, and microvessel density. The 5‐year survival rates of the LAT 1‐positive and LAT1‐negative patients were 59% and 88%, respectively (P = 0.2186). Tumor cell proliferation and angiogenesis were not also (a) prognostic factor. However, the 5‐year survival rate of patients with both LAT 1 and CD98‐positivity (57%) was significantly worse than that of other patients (88%; P = 0.0136). Multivariate analysis confirmed that positive cooperative expression of LAT 1 and CD98 was an independent factor for predicting a poor prognosis. A cooperative expression of LAT 1 and CD98 is a significant pathological factor for predicting the poor prognosis in patients with resectable stage I squamous cell carcinoma of the lung. (Cancer Sci 2009; 100: 249–254)
Quantitative tests of liver function measure hepatic improvement after sustained virological response: results from the HALT‐C trial
Authors: EVERSON G. T. , SHIFFMAN M. L. , HOEFS J. C. , MORGAN T. R. , STERLING R. K. , WAGNER D. A. , DESANTO J. L. , CURTO T. M. , WRIGHT E. C. ,   THE HALT‐C TRIAL GROUP
Published: December 2008

Backgroud The impact of virologic response on hepatic function has not been previously defined.

Aim To determine the relationships of quantitative liver function tests (QLFTs) with virological responses to peginterferon (PEG) ± ribavirin (RBV) in patients with chronic hepatitis C and to use serial QLFTs to define the spectrum of hepatic improvement after sustained virological response (SVR).

Methods Participants (n = 232) were enrolled in the Hepatitis C Antiviral Long‐term Treatment against Cirrhosis (HALT‐C) Trial, had failed prior therapy, had bridging fibrosis or cirrhosis and were retreated with PEG/RBV. All 232 patients had baseline QLFTs; 24 patients with SVR and 68 nonresponders had serial QLFTs. Lidocaine, [24‐13C]cholate, galactose and99mTc‐sulfur colloid were administered intravenously; [2,2,4,2‐2H]cholate, [1‐13C]methionine, caffeine and antipyrine were administered orally. Clearances (Cl), breath13CO2, monoethylglycylxylidide (MEGX), perfused hepatic mass (PHM) and liver volume were measured.

Results Rates of SVR were 18–26% in patients with good function by QLFTs, but ≤6% in patients with poor function. Hepatic metabolism, measured by caffeinekelim(P = 0.02), antipyrinekelim(P = 0.05) and antipyrineCl(P = 0.02) and the portal circulation, measured by cholateCloral(P = 0.0002) and cholate shunt (P = 0.0003) and PHM (P = 0.03) improved after SVR.

Conclusion Hepatic dysfunction impairs the virological response to PEG/RBV. SVR improves hepatic metabolism, the portal circulation and PHM.
Psychopathology is associated with dyspeptic symptom severity in primary care patients with a new episode of dyspepsia
Authors: MUJAKOVIC S. , DE WIT N. J. , VAN MARREWIJK C. J. , FRANSEN G. A. J. , LAHEIJ R. J. F. , MURIS J. W. , SAMSOM M. , GROBBEE D. E. , JANSEN J. B. M. J. , KNOTTNERUS J. A. , NUMANS M. E.
Published: December 2008

Background  Personality and psychiatric disorders are reported to be more common in dyspeptic patients with severe complaints, but it remains unclear whether this association exists for patients with mild and moderate dyspepsia.

Aim  To study the association between dyspeptic symptom severity and psychopathology, major life events and coping ability in patients with a new episode of dyspepsia.

Methods  Dyspeptic symptom severity was measured using the validated eight symptom Veldhuyzen van Zanten questionnaire. Psychopathology was measured using the Symptom Check List‐90 (SCL 90). Major life events were measured with a modified version of the Social Readjustment Rating Scale (SRRS). Coping styles were measured by a short version of the Utrecht Coping Questionnaire, distinguishing six coping styles. Linear regression was used to assess the relationship between dyspepsia symptom severity and psychological factors.

Results  In all, 664 patients with a new episode of uninvestigated dyspepsia, aged >18 years were included. Dyspeptic symptom severity was positively correlated with the presence of depression (P < 0.01), somatization symptoms (P < 0.01), use of an active coping style (P < 0.01) and negatively correlated with age (P < 0.01).

Conclusions  Primary care patients consulting with dyspepsia have higher levels of depression and somatization especially at younger age. An active coping style is associated with dyspepsia symptom severity.
Co‐medication of statins and CYP3A4 inhibitors before and after introduction of new reimbursement policy
Authors: Devold Helene M. , Molden Espen , Skurtveit Svetlana , Furu Kari
Published: December 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• HMG‐CoA reductase inhibitors (statins) are frequently used drugs in the treatment of dyslipidaemia.

• Co‐medication with interacting drugs increases the risk of statin‐induced muscular side‐effects.

• Simvastatin exhibits particularly high interaction potential due to substantial metabolism via cytochrome P450 3A4 (CYP3A4).

WHAT THIS STUDY ADDS

• In June 2005, a new reimbursement policy was introduced by the Norwegian Medicines Agency stating that simvastatin should be prescribed as first‐line lipid‐lowering therapy.

• Following introduction of the new policy, the number of patients co‐medicated with simvastatin and CYP3A4 inhibitors almost doubled.

• A potential consequence is increased incidence of muscular side‐effects in the statin‐treated population.

AIMS

To assess the prevalence of co‐medication of statins and CYP3A4 inhibitors before and after introduction of a new Norwegian reimbursement policy, which states that all patients should be prescribed simvastatin as first‐line lipid‐lowering therapy.

METHODS

Data from patients receiving simvastatin, lovastatin, pravastatin, fluvastatin or atorvastatin in 2004 and 2006, including co‐medication of potent CYP3A4 inhibitors, were retrieved from the Norwegian Prescription Database covering the total population of Norway. Key measurements were prevalence of continuous statin use (two or more prescriptions on one statin) and proportions of different statin types among all patients and those co‐medicated with CYP3A4 inhibitors.

RESULTS

In 2004, 5.9% (n= 272 342) of the Norwegian population received two or more prescriptions on one statin compared with 7.0% (n= 324 267) in 2006. The relative number of simvastatin users increased from 39.7% (n= 112 122) in 2004 to 63.1% (n= 226 672) in 2006. A parallel increase was observed within the subpopulation co‐medicated with statins and CYP3A4 inhibitors, i.e. from 42.9% (n= 7706) in 2004 to 63.6% (n= 13 367) in 2006. For all other statins the number of overall users decreased to a similar extent to those co‐medicated with CYP3A4 inhibitors.

CONCLUSIONS

In both 2004 and 2006, the choice of statin type did not depend on whether the patient used a CYP3A4 inhibitor or not. Considering the pronounced interaction potential of simvastatin with CYP3A4 inhibitors, a negative influence of the new policy on overall statin safety seems likely.
Evaluation of cost of treatment of drug‐related events in a tertiary care public sector hospital in Northern India: a prospective study
Authors: Pattanaik Smita , Dhamija Punit , Malhotra Samir , Sharma Navneet , Pandhi Promila
Published: December 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Drug related events include ADRs, events due to patient or physician noncompliance, drug overdosage and drug interactions.

• Economic burden of management of drug related events are substantial and include both direct and indirect costs.

• Some data regarding cost of treatment of ADR exist from south and western India.

WHAT THIS STUDY ADDS

• An approximate cost of management of drug related events presenting to the emergency medical department in a tertiary care hospital over a period of 4 months.

• Compares the cost incurred in a public sector hospital to the projected cost of management of same events in a private sector hospital.

• Gives a rough estimate of economic burden on the health care system due to adverse drug events.

AIMS Drug related events (DREs) contribute significantly to hospital admissions. These are largely preventable events and require optimum use of the therapeutic agents. The study was conducted to analyze the cost of treatment of DREs.

PATIENTS & METHODS All visits to medical emergency department of a tertiary care public sector hospital in northern India were recorded in a prospective, non‐interventional manner over a period of 4 months. DREs were recognized and were followed up till their stay in the hospital. Data about the cost generating components of direct and indirect costs of treatment of DREs were collected. The projected cost of treatment of the same DREs in a private sector hospital was estimated and compared.

RESULTS Out of 1833 admissions, 92(5.01%) were due to DREs. Maximum cases were due to non compliance (66%) followed by ADR (28%) and drug overdose(6%). The common DREs leading to ED visits were cerebrovascular accident(19.44%), followed by accelerated hypertension(18.36%) and diabetic ketoacidosis(14.04%). Total cost of management of all the 92 DREs in our hospital was calculated to be INR17,37,339(€30,215). The direct cost was INR1,72,961(€3008) and the approximate indirect cost was INR15,64, 378(€27, 206). The projected cost of management of all the 92 DREs was estimated to be INR63,63,872(€1,01, 676) in a private sector hospital.

CONCLUSION The study shows that ADEs leading to emergency department visits and hospitalizations constitute a significant economic burden. Training of the patients and the prescribers may lessen the economic burden on the patient as well as the health care system.
The Stanford V regimen is effective in patients with good risk Hodgkin lymphoma but radiotherapy is a necessary component
Authors: Abuzetun Jamil Y. , Loberiza Fausto , Vose Julie , Bierman Philip , Greg Bociek R. , Enke Charles , Bast Martin , Weisenburger Dennis , Armitage James O. ,   for the Nebraska Lymphoma Study Group
Published: December 2008

One‐hundred and thirteen patients between the ages of 15 and 88 years with biopsy proven, untreated Hodgkin lymphoma were treated by physicians in the Nebraska Lymphoma Study Group using the Stanford V regimen (bleomycin, doxorubicin, etoposide, mechlorethamine, prednisone, vinblastine and vincristine) between January 1997 and January 2006. With a median follow‐up of all surviving patients of 63 months, the 5‐year overall survival (OS) was 84% and the 5‐year progression‐free survival (PFS) was 74%. Age >60 years, other than nodular sclerosing histological subtype, Ann Arbor Stage III/IV, and the presence of B‐symptoms significantly predicted treatment outcome. Patients with 0–2 of these factors had a 5‐year PFS and OS of 80% and 89% vs. 20% and 40% for patients with 3 or 4 factors. Patients who received radiotherapy had a superior 5‐year PFS (86% vs. 42%) and OS (96% vs. 53%). Patients with 0–2 adverse risk factors who received radiotherapy had an excellent treatment outcome with 5‐year PFS of 88% and a 5‐year OS of 97%. The Stanford V chemotherapy regimen has an excellent treatment outcome for good prognosis patients with Hodgkin lymphoma. The results are compromised when radiotherapy is not utilized.
Pregnancy‐induced thrombocytopenia and TTP, and the risk of fetal death, in Upshaw–Schulman syndrome: a series of 15 pregnancies in 9 genotyped patients
Authors: Fujimura Yoshihiro , Matsumoto Masanori , Kokame Koichi , Isonishi Ayami , Soejima Kenji , Akiyama Nobu , Tomiyama Junji , Natori Kazuhiko , Kuranishi Yasunobu , Imamura Yutaka , Inoue Nobumasa , Higasa Satoshi , Seike Masako , Kozuka Teruhiko , Hara Masamichi , Wada Hideo , Murata Mitsuru , Ikeda Yasuo , Miyata Toshiyuki , George James N.
Published: December 2008

Upshaw–Schulman syndrome (USS) is a congenital thrombotic thrombocytopenic purpura (TTP) due to mutations in the gene that encodes for ADAMTS13 (ADAMTS13), but its clinical signs may be mild or absent during childhood. We have identified 37 patients with USS (24 females, 13 males) belonging to 32 families. The nine women from six families who were diagnosed during their first pregnancy are the focus of this report. Six of the nine women had episodes of thrombocytopenia during childhood misdiagnosed as idiopathic thrombocytopenic purpura. Thrombocytopenia occurred during the second–third trimesters in each of their 15 pregnancies, with 16 babies (one twin pregnancy), often followed by TTP. Of 15 pregnancies, eight babies were stillborn or died soon after birth, and the remaining seven were all premature except one, who was born naturally following plasma infusions to the mother that had started at 8 weeks’ gestation. All nine USS women had severely deficient ADAMTS13 activity. ADAMTS13 analyses demonstrated that eight women were compound heterozygotes of Y304C/G525D (2 siblings), R125VfsX6/Q1302X (2 siblings), R193W/R349C (2 siblings), I178T/Q929X, and R193W/A606P; one woman was homozygous for R193W. Only the R193W mutation has been previously reported. These observations emphasize the importance of measuring ADAMTS13 activity in the evaluation of thrombocytopenia during childhood and pregnancy.
LONG‐TERM FOLLOW‐UP OF LOW‐DOSE EXTERNAL PITUITARY IRRADIATION FOR CUSHING'S DISEASE
Authors: LITTLEY M. D. , SHALET S. M. , BEARDWELL C. G. , AHMED S. R. , SUTTON M. L.
Published: December 2008

Twenty‐four patients (three male) with Cushing's disease, aged between 11 and 67 years, were treated with low‐dose external pituitary irradiation (20 Gy in eight fractions over 10–12 days) and followed for between 13 and 171 months (median 93 months). Eleven patients (46%) went into remission 4–36 months after irradiation, but five subsequently relapsed. Two of these received no further active treatment, one underwent successful pituitary surgery, one underwent a second course of low‐dose external irradiation (as yet unsuccessful) and one has been treated with metyrapone for a total of 75 months. One of the 13 patients who did not respond received a further course of low‐dose pituitary irradiation with prompt remission and two have received metyrapone for 41 months and 15 years without ill effect. One patient died from cerebrovascular disease. The remaining nine patients underwent bilateral adrenalectomy (one after unsuccessful pituitary surgery) with rapid resolution of hypercortisolism. Five of these patients have developed hyperpigmentation and elevated ACTH levels (range 505–1150 ng/1). A pituitary microadenoma has been demonstrated on CT scan in three and successfully removed by microadenomectomy. In the present series, the low incidence of radiation‐induced hypopituitarism and absence of other complications attributable to radiotherapy suggest that low‐dose pituitary irradiation may be a useful treatment option in selected patients. However, long‐term follow‐up has demonstrated a high relapse rate and failure to prevent Nelson's syndrome in adrenalectomized patients, indicating that it should not be used as primary treatment in preference to selective adenomectomy.
INHIBITION OF OESTRADIOL PRODUCTION BY EPIDERMAL GROWTH FACTOR IN HUMAN GRANULOSA CELLS OF NORMAL AND POLYCYSTIC OVARIES
Authors: MASON H. D. , MARGARA R. , WINSTON R. M. L. , BEARD R. W. , REED M. J. , FRANKS S.
Published: December 2008

Anovulation in women with polycystic ovary syndrome results from a disorder of FSH‐mediated follicular maturation which may involve paracrine modulation of FSH action by intra‐ovarian factors. Epidermal growth factor (EGF) is a potent inhibitor of FSH‐stimulated oestradiol production in the rat and has also been shown to inhibit aromatase activity in human granulosa cells obtained after superovulation. The purpose of this study was to investigate the action of EGF on granulosa cell function in human ovaries which had not been previously exposed to treatment with exogenous gonadotrophins and to compare the responses in tissue obtained from normal and from polycystic ovaries. Granulosa cells were obtained from antral follicles <10 mm in diameter after dissection of unstimulated normal or polycystic ovaries (PCO). Cells were pooled, washed, plated and incubated for 48h in the presence of 10‐7 M testosterone and various doses of human FSH. FSH dose responses were obtained with or without the addition of purified EGF (50 pg/ml). Testosterone in the absence of FSH resulted in a fourfold (range 2–7.5) increase in oestradiol accumulation in the medium after incubation of granulosa cells from both normal and polycystic ovaries. This increase was reversed by addition of EGF. FSH treatment stimulated a dose‐related increase in oestradiol regardless of the origin of the granulosa cells. The peak E2 response to FSH, obtained at a dose of 1–2.5 ng/ml was a 20‐fold increase above testosterone alone (range 4–55) in cells from PCO compared to sixfold (2–5.13) in cells from normal ovaries. The concurrent addition of EGF at 50 pg/ml caused an average 57% inhibition of the peak response to FSH. EGF also caused a dose‐dependent inhibition of oestradiol production stimulated by a fixed dose of FSH in cells from both normal and PCO. These data indicate that EGF inhibits FSH‐inducedoestradiol production by granulosa cells of both normal and polycystic ovaries and suggest that EGF, or its analogue, transforming growth factor α, which is known to be synthesized by theca cells, may have a role in human ovarian function.
TRANSSPHENOIDAL SURGERY FOR CUSHING'S DISEASE: DOES WHAT IS REMOVED DETERMINE THE ENDOCRINE OUTCOME?
Authors: BURKE C. W. , ADAMS C. B. T. , ESIRI M. M. , MORRIS C. , BEVAN J. S.
Published: December 2008

Fifty‐seven patients with pituitary‐dependent Cushing's syndrome and eight with Nelson's syndrome underwent transsphenoidal pituitary exploration, with removal of macroscopically abnormal tissue in 64 patients and detailed histology of this in 63. The cure rate by stringent criteria 1 month later was 48 (83%) of the 58 with assessable data, who were followed for 225 patient‐years. Two patients relapsed later, a rate of one per 112 patient‐years of follow‐up. In 27% of patients, the macroscopically abnormal tissue removed was histologically indistinguishable from normal pituitary gland but the cure rate was 82%, and a quarter of the patients in this group assessable for recovery of normal ACTH function gained it. Another 53% of biopsies showed corticotropin adenomas, and the cure rate in these was 89% though rather more (69%) recovered normal ACTH function. The remaining 20% of biopsies were consistent with corticotroph hyperplasia. The cure rate varied little whether the lesion was diffuse or localized, whether or not it was in the invasion zone/interlobar cleft, whether or not there was pituitary enlargement, or whether the surgery was radical or selective. Six patients, of whom three are cured, showed surgical or radiological evidence of invasion outside the pituitary fossa. The data are consistent with the idea that pituitary adenoma is merely the end stage of some other process in the corticotrophs, and cure often follows removal of a lesion other than adenoma. Of patients who were permanently cured, 47% regained normal ACTH function within 3 years of operation (none later), 53% remaining ACTH‐deficient at 3 years or more. Of all patients 48% acquired gonadotrophin deficiency, 28% have TSH deficiency and 25% permanent diabetes insipidus. All these pituitary function deficits were more common after radical surgery and in patients with normal histology. The literature contains so little objective data on these functions that we cannot say whethere the endocrine damage in our patients is exceptional or not.
THE EFFECT OF DANAZOL AND THE LHRH AGONIST ANALOGUE GOSERELIN (ZOLADEX) ON THE BIOLOGICAL ACTIVITY OF LUTEINIZING HORMONE IN WOMEN WITH ENDOMETRIOSIS
Authors: MAOURIS P. , DOWSETT M. , ROSE GILLIAN , EDMONDS D. K. , ROTHWELL CLAIRE , ROBERTSON W. R.
Published: December 2008

In an attempt to determine whether the suppression in oestradiol levels caused by danazol is due to an effect on the hypothalamic‐pituitary axis, we compared the endocrine effects of danazol with those of the LHRH (GnRH) agonist analogue goserelin. Serum levels of immunoreactive LH (I‐LH), FSH, 17β‐oestradiol (E2) and bioactive LH (B‐LH) (using a mouse Leydig cell bioassay), were measured in ten and 20 women with endometriosis treated with danazol and goserelin, respectively. I‐LH was measured both by radioimmunoassay (RIA) and immunoradiometric assay (IRMA). During 6 months of treatment with 600 mg of danazol daily, mean serum E2 decreased (P <0.05) to levels near the upper limit of the post‐menopausal range (to a mean (and 95% confidence interval of the mean) of 117 (65–169) pmol/1) whereas FSH, I‐LH (both by RIA and IRMA) and B‐LH levels were not significantly altered. During 6 months of treatment with monthly depot injections of 3–6 mg goserelin, mean serum E2 decreased (P<0.001) to well within the post‐menopausal range (to 23 (18–28) pmol/1). The mean FSH, I‐LH and B‐LH levels also decreased (P <0.05) during therapy with goserelin (from 3.9 (3–1–4.7) to 2.0 (1.6‐2.4) IU/1 for FSH, from 5.3 (4.5–6.1) to 1.9 (1.7–2.1) IU/1 for RIA‐LH, from 2.9 (2.5–3.3) to <0.5 (<0.5) IU/1 for IRMA‐LH and from 9.1 (7.1–11.1) to 2.9 (2.6–3.2) IU/1 for B‐LH). The bioactive to immunoreactive (B:I) RIA LH ratio was altered during the first month of treatment with both drugs, increasing with danazol from 1.6(1.3.1.9) to 2.0(1.7‐2.3) (P<0.05) and decreasing with goserelin from 1.7 (1.5‐1–9) to 1.4 (1.2–1.6) (P<0.05). The data indicate that danazol causes a relatively hypo‐oestrogenic state which cannot be attributed to a decrease in the immunoreactive gonadotrophin levels or to a decrease in the biological activity of LH and is associated with an increase in B: ILH ratio. In contrast, goserelin acts on the pituitary to produce suppression of gonadotrophin levels and effective down‐regulation of the pituitary.
Bone protective therapy in the young patient with fractures and chronic disease: what drug(s) should be given and for how long?
Authors: Cooper Mark
Published: December 2008

Fractures are fortunately rare in younger people even with severe chronic disease but when they occur they need to be carefully evaluated. When considering such a patient there needs to be an assessment of the risk of future fractures and their type, a search for potentially modifiable risk factors and an evaluation of the risks vs. benefits of bone protective therapy. These factors will have to be considered in the context of the underlying chronic disease and the way that this impacts on the patient. In those individuals where bone protective therapy is given a clear plan is needed relating to how long the treatment is used and what type of monitoring is warranted.
Seasonal variation in ornament expression, body size, energetic reserves, immune response, and survival in males of a territorial insect
Authors: CÓRDOBA‐AGUILAR A. , JIMÉNEZ‐CORTÉS J. G. , LANZ‐MENDOZA H.
Published: December 2008

Abstract 1. Seasonal variation in immune response has rarely been investigated in invertebrates and, therefore, we have studied this using territorial adult males of the damselfly, Hetaerina americana (Fabricius), in several generations, for a year and a half.

2. We investigated and related seasonal variation in red pigmented wing spot size (an ornamental trait) and body size to fat reserves using four immunity components – melanisation ability, phenoloxidase (PO) and nitric oxide (NO) activity, and haemolymph protein concentration – and survival ability after a bacterial immune challenge.

3. There was seasonal variation in spot expression, being more intense in the non‐winter months and less intense in winter months, and, to some extent, a similar pattern was found for NO. Although there was also variation in melanisation, PO activity, protein concentration and fat reserves, this was not consistently related to variation in spot size. Survival was lower in the winter than in non‐winter months.

4. Animals with larger spot and body size had greater values of melanisation, PO and NO activity, and protein concentration. The relation of spot and body size with fat reserves is still not clear.

5. Unlike other studies, ornament was not similarly correlated with all typical immune components (at best, mainly NO).

6. Similar to what occurs in vertebrates, survival is lower during the winter months. In the case of vertebrate studies, however, the same individual suffers these changes. In the American rubyspot, distinct cohorts demonstrated seasonal differences.
Modelling the current distribution of European diadromous fishes: an approach integrating regional anthropogenic pressures
Authors: LASSALLE GÉRALDINE , CROUZET PHILIPPE , ROCHARD ERIC
Published: December 2008

1. Twenty‐eight diadromous fish species occurred in Europe, North Africa and the Middle East in historical times. Their current distributions were assessed in terms of abundance classes (missing, rare, common and abundant) in 196 basins ranging from Morocco to northern Norway and from Greenland to Iran.

2. Their current distributions were modelled using abiotic, biotic, climatic and anthropogenic (regional anthropogenic pressures) variables. Anthropogenic variables were derived from characteristics of large dams (height, distance from the outlet, percentage of main stem river available downstream of dam) and human population density. These data were taken from the EEA Eldred 2.08 (European Lakes, Dams and Reservoirs Database) that deals comprehensively with large European dams and includes all obstacles of this type. To deal with ordinal response variables, we applied proportional odds models.

3. Twenty‐two species‐specific models were successfully built according to the reduction of deviance and the validation process, of which eight included one or more anthropogenic variables. No model could be established for six endemic or highly endangered species such as Acipenser sturio and Coregonus oxyrinchus.

4. Most response curves were easily interpretable since they were related to specific aspects of species’ ecology. Anthropogenic variables related to large dams impacted negatively on the distribution of diadromous fishes through the perturbation of river discharge patterns, the loss of river connectivity and the accessibility to essential habitats, particularly for Salmonid species that spawn in headwater streams. However, one species which can complete its life cycle using only the most downstream part of the basin, Liza ramada, was found to be favoured by the changes in hydrological regime. The bell‐shaped curves obtained from human population density for three diadromous species were connected on one side to a common settlement history of human and animal populations and on the other side to negative impacts of human activities arising at high population density.

5. Our approach can provide the basis for identifying special areas of conservation prior to planning restoration programmes at country or basin scales, as well as for more specialized studies focusing on one species only or at the local scale.
The role of modelling in achieving information systems success: UML to the rescue?
Authors: Larsen Tor J. , Niederman Fred , Limayem Moez , Chan Joyce
Published: December 2008

Much computer science literature addresses the mechanics of the Unified Modelling Language (UML) and requirements modelling, but little research has addressed the role of UML in the broader organizational and project development context. This study uses a socio‐technical approach to consider UML as a technology embedded in a social environment. In this study, project developers were interviewed in detail about their use of UML along with influences on their decisions to use this approach and the results of using it. Data were analyzed using causal mapping. Major findings included: (1) that definitions of success may differ by unit of analysis (e.g., developer, project, organization) and that the relationship among these definitions are complex; (2) a very large number of variables impacting project success were identified; (3) a number of important variables exist in complex (non‐linear) relationships with project success; and (4) the majority of interviewees linked the use of UML to project success.
Deletions in the desaturase2 gene in Israeli populations of Drosophila melanogaster: possible role in mate recognition
Authors: Zamorzaeva I. , Rashkovetsky E. , Nevo E. , Korol A.
Published: December 2008

Molecular analysis of the desaturase2 (desat2) gene responsible for female cuticular hydrocarbon synthesis, revealed that all tested Israeli populations of Drosophila melanogaster belong to the ‘Low’ type (low ratio of 5,9‐/7,11‐heptacosadiene) with a 16 bp deletion in the promoter region. Additional deletion leading to the appearance of the stop‐codon TGA was found in exon 1 in populations inhabiting opposite slopes at Nahal Oren canyon. Mating experiments between carriers of various desat2 alleles estimated the putative effect of desat2 status on courtship behaviour. The allele with an additional deletion in exon 1 seems to play some role in mating success, which is associated with shorter courtship latency and courtship duration. The appearance and maintenance of this mutant allele in populations inhabiting Nahal Oren canyon may reflect flies’ adaptation to peculiar microscale climatic conditions and may be associated with incipient sympatric differentiation.
Ribosome biogenesis is temperature‐dependent and delayed in Escherichia coli lacking the chaperones DnaK or DnaJ†
Authors: Al Refaii Abdalla , Alix Jean‐Hervé
Published: December 2008

In Escherichia coli strains carrying null mutations in either the dnaK or dnaJ genes, the late stages of 30S and 50S ribosomal subunit biogenesis are slowed down in a temperature‐dependent manner. At high temperature (44°C), 32S and 45S particles (precursors to 50S subunits) and 21S particles (precursors to 30S subunits) accumulate. The latter are shown by 3′5′ rapid amplification of cDNA ends analysis to contain unprocessed or partially processed 16S ribosomal RNA at the 5′ end, but the 3′ end was never processed. This implies that maturation of 16S ribosomal RNA starts at the 5′‐terminus, and that the 3′‐terminus is only trimmed at a later step. At normal temperatures (30°C−37°C), ribosome assembly in both mutants is not arrested but is significantly delayed, as shown by pulse‐chase analysis. Assembly defects are partially compensated by an overexpression of other heat‐shock proteins, which occurs in the absence of their negative regulator DnaK, or by a plasmid‐driven overexpression of GroES/GroEL, suggesting the involvement of a network of chaperones in ribosome biogenesis.
Adaptation of the bacterial membrane to changing environments using aminoacylated phospholipids
Authors: Roy Hervé , Dare Kiley , Ibba Michael
Published: December 2008

Fine‐tuning of the biophysical properties of biological membranes is essential for adaptation of cells to changing environments. For instance, to lower the negative charge of the lipid bilayer, certain bacteria add lysine to phosphatidylglycerol (PG) converting the net negative charge of PG (−1) to a net positive charge in Lys‐PG (+1). Reducing the net negative charge of the bacterial cell wall is a common strategy used by bacteria to resist cationic antimicrobial peptides (CAMPs) secreted by other microbes or produced by the innate immune system of a host organism. The article by Klein et al. in the current issue of Molecular Microbiology reports a new modification of the bacterial membrane, addition of alanine to PG, in Pseudomonas aeruginosa. In spite of the neutral charge of Ala‐PG, this modified lipid was found to be linked to several resistance phenotypes in P. aeruginosa. For instance, Ala‐PG increases resistance to two positively charged antibacterial agents, a β‐lactam and high concentrations of lactate. These findings shed light on the mechanisms by which bacteria fine‐tune the properties of their cell membranes by adding various amino acids on the polar head group of phospholipids.
Characterization of the individual glucose uptake systems of Lactococcus lactis: mannose‐PTS, cellobiose‐PTS and the novel GlcU permease
Authors: Castro Rute , Neves Ana R. , Fonseca Luís L. , Pool Wietske A. , Kok Jan , Kuipers Oscar P. , Santos Helena
Published: December 2008

According to previous reports, Lactococcus lactis imports glucose via two distinct phosphoenolpyruvate:phosphotransferase systems (mannose‐PTS and cellobiose‐PTS) and one or more unknown non‐PTS permease(s). GlcU was identified as the sole non‐PTS permease involved in the transport of glucose. Additionally, the biochemical properties of PTSMan, PTSCel and GlcU were characterized in double knockout mutants with glucose uptake restricted to a single system. Transport susceptibility to protonophores indicated that glucose uptake via GlcU is proton‐motive force dependent. Competition assays revealed a high specificity of GlcU for glucose. Furthermore, the permease has low affinity for glucose and displays strong preference for the β‐anomer as shown by the profiles of consumption of the two glucose anomers studied by 13C‐NMR. Similar kinetic properties were found for PTSCel, while PTSMan is a high‐affinity system recognizing equally well the two anomeric forms of glucose. Transcripts of the genes encoding the three transporters are present simultaneously in the parent strain NZ9000 as shown by reverse transcription‐PCR. Investigation of the distribution of GlcU homologues among bacteria showed that these proteins are restricted to the low‐GC Gram‐positive Firmicutes. This work completes the identification of the glucose transport systems in L. lactis MG1363.