bluuebunny/crossref_metadata_2025 huggingface dataset:
Jun NH
2
-terminal kinase (JNK) prevents nuclear β-catenin accumulation and regulates axis formation in
Xenopus
embryos
Authors: Liao Guanghong , Tao Qinghua , Kofron Matthew , Chen Juei-Suei , Schloemer Aryn , Davis Roger J. , Hsieh Jen-Chih , Wylie Chris , Heasman Janet , Kuan Chia-Yi
Published: October 2006

Jun NH
2
-terminal kinases (JNKs) regulate convergent extension movements in
Xenopus
embryos through the noncanonical Wnt/planar cell polarity pathway. In addition, there is a high level of maternal JNK activity spanning from oocyte maturation until the onset of gastrulation that has no defined functions. Here, we show that maternal JNK activation requires Dishevelled and JNK is enriched in the nucleus of
Xenopus
embryos. Although JNK activity is not required for the glycogen synthase kinase-3-mediated degradation of β-catenin, inhibition of the maternal JNK signaling by morpholino-antisense oligos causes hyperdorsalization of
Xenopus
embryos and ectopic expression of the Wnt/β-catenin target genes. These effects are associated with an increased level of nuclear and nonmembrane-bound β-catenin. Moreover, ventral injection of the constitutive-active
Jnk
mRNA blocks β-catenin-induced axis duplication, and dorsal injection of active
Jnk
mRNA into
Xenopus
embryos decreases the dorsal marker gene expression. In mammalian cells, activation of JNK signaling reduces Wnt3A-induced and β-catenin-mediated gene expression. Furthermore, activation of JNK signaling rapidly induces the nuclear export of β-catenin. Taken together, these results suggest that JNK antagonizes the canonical Wnt pathway by regulating the nucleocytoplasmic transport of β-catenin rather than its cytoplasmic stability. Thus, the high level of sustained maternal JNK activity in early
Xenopus
embryos may provide a timing mechanism for controlling the dorsal axis formation.
On structural transitions, thermodynamic equilibrium, and the phase diagram of DNA and RNA duplexes under torque and tension
Authors: Wereszczynski Jeff , Andricioaei Ioan
Published: October 2006

A precise understanding of the flexibility of double stranded nucleic acids and the nature of their deformed conformations induced by external forces is important for a wide range of biological processes including transcriptional regulation, supercoil and catenane removal, and site-specific recombination. We present, at atomic resolution, a simulation of the dynamics involved in the transitions from B-DNA and A-RNA to Pauling (P) forms and to denatured states driven by application of external torque and tension. We then calculate the free energy profile along a B- to P-transition coordinate and from it, compute a reversible pathway, i.e., an isotherm of tension and torque pairs required to maintain P-DNA in equilibrium. The reversible isotherm maps correctly onto a phase diagram derived from single molecule experiments, and yields values of elongation, twist, and twist-stretch coupling in agreement with measured values. We also show that configurational entropy compensates significantly for the large electrostatic energy increase due to closer-packed P backbones. A similar set of simulations applied to RNA are used to predict a novel structure, P-RNA, with its associated free energy, equilibrium tension, torque and structural parameters, and to assign the location, on the phase-diagram, of a putative force–torque-dependent RNA “triple point.”
Enzyme–microbe synergy during cellulose hydrolysis by
Clostridium thermocellum
Authors: Lu Yanpin , Zhang Yi-Heng Percival , Lynd Lee R.
Published: October 2006

Specific cellulose hydrolysis rates (g of cellulose/g of cellulase per h) were shown to be substantially higher (2.7- to 4.7-fold) for growing cultures of
Clostridium thermocellum
as compared with purified cellulase preparations from this organism in controlled experiments involving both batch and continuous cultures. This “enzyme–microbe synergy” requires the presence of metabolically active cellulolytic microbes, is not explained by removal of hydrolysis products from the bulk fermentation broth, and appears due to surface phenomena involving adherent cellulolytic microorganisms. Results support the desirability of biotechnological processes featuring microbial conversion of cellulosic biomass to ethanol (or other products) in the absence of added saccharolytic enzymes.
β-Arrestin2-mediated inotropic effects of the angiotensin II type 1A receptor in isolated cardiac myocytes
Authors: Rajagopal Keshava , Whalen Erin J. , Violin Jonathan D. , Stiber Jonathan A. , Rosenberg Paul B. , Premont Richard T. , Coffman Thomas M. , Rockman Howard A. , Lefkowitz Robert J.
Published: October 2006

The G protein-coupled receptor kinases (GRKs) and β-arrestins, families of molecules essential to the desensitization of G protein-dependent signaling via seven-transmembrane receptors (7TMRs), have been recently shown to also transduce G protein-independent signals from receptors. However, the physiologic consequences of this G protein-independent, GRK/β-arrestin-dependent signaling are largely unknown. Here, we establish that GRK/β-arrestin-mediated signal transduction via the angiotensin II (ANG) type 1A receptor (AT
1A
R) results in positive inotropic and lusitropic effects in isolated adult mouse cardiomyocytes. We used the “biased” AT
1A
R agonist [Sar
1
, Ile
4
, Ile
8
]-angiotensin II (SII), which is unable to stimulate G
α
q-mediated signaling, but which has previously been shown to promote β-arrestin interaction with the AT
1A
R. Cardiomyocytes from WT, but not AT
1A
R-deficient knockout (KO) mice, exhibited positive inotropic and lusitropic responses to both ANG and SII. Responses of WT cardiomyocytes to ANG were dramatically reduced by protein kinase C (PKC) inhibition, whereas those to SII were unaffected. In contrast, cardiomyocytes from β-arrestin2 KO and GRK6 KO mice failed to respond to SII, but displayed preserved responses to ANG. Cardiomyocytes from GRK2 heterozygous knockout mice (GRK2
+/−
) exhibited augmented responses to SII in comparison to ANG, whereas those from GRK5 KO mice did not differ from those from WT mice. These findings indicate the existence of independent G
α
q/PKC- and GRK6/β-arrestin2-dependent mechanisms by which stimulation of the AT
1A
R can modulate cardiomyocyte function, and which can be differentially activated by selective receptor ligands. Such ligands may have potential as a novel class of therapeutic agents.
Murine inner cell mass-derived lineages depend on Sall4 function
Authors: Elling Ulrich , Klasen Christian , Eisenberger Tobias , Anlag Katrin , Treier Mathias
Published: October 2006

Sall4 is a mammalian Spalt transcription factor expressed by cells of the early embryo and germ cells, an expression pattern similar to that of both Oct4 and Sox2, which play essential roles during early murine development. We show that the activity of Sall4 is cell-autonomously required for the development of the epiblast and primitive endoderm from the inner cell mass. Furthermore, no embryonic or extraembryonic endoderm stem cell lines could be established from
Sall4
-deficient blastocysts. In contrast, neither the development of the trophoblast lineage nor the ability to generate trophoblast cell lines from murine blastocysts was impaired in the absence of Sall4. These data establish Sall4 as an essential transcription factor required for the early development of inner cell mass-derived cell lineages.
Good genes sexual selection in nature
Authors: Byers John A. , Waits Lisette
Published: October 2006

Whether the mate sampling and choice performed by females in nature influences offspring performance is a controversial issue in theory and an open empirical question. Pronghorn (
Antilocapra americana
) females engage in an obvious and energetically expensive mate sampling process to identify vigorous males. Although individual females sample independently, their choices converge on a small proportion of males that sire most young. Offspring of attractive males were more likely to survive to weaning and to age classes as late as 5 years, resulting in a selection differential, calculated by expected differences in lifetime number of offspring weaned, of 0.32 against random mating. Enhanced survival to weaning appeared to be accomplished by faster growth rates. Females compensated for matings with a less attractive mate by elevating rates of milk delivery to their young. Because pronghorn males do not have costly ornaments, we conclude that female choice for good genes can exist in the absence of ornaments. Furthermore, female choice may be important and unrecognized as a force that can lower population genetic load.
MyoD-positive epiblast cells regulate skeletal muscle differentiation in the embryo
Authors: Gerhart Jacquelyn , Elder Justin , Neely Christine , Schure Jared , Kvist Tage , Knudsen Karen , George-Weinstein Mindy
Published: October 2006

MyoD mRNA is expressed in a subpopulation of cells within the embryonic epiblast. Most of these cells are incorporated into somites and synthesize Noggin. Ablation of MyoD-positive cells in the epiblast subsequently results in the herniation of organs through the ventral body wall, a decrease in the expression of Noggin, MyoD, Myf5, and myosin in the somites and limbs, and an increase in Pax-3–positive myogenic precursors. The addition of Noggin lateral to the somites compensates for the loss of MyoD-positive epiblast cells. Skeletal muscle stem cells that arise in the epiblast are utilized in the somites to promote muscle differentiation by serving as a source of Noggin.
A role for Dicer in immune regulation
Authors: Cobb Bradley S. , Hertweck Arnulf , Smith James , O'Connor Eric , Graf Daniel , Cook Terence , Smale Stephen T. , Sakaguchi Shimon , Livesey Frederick J. , Fisher Amanda G. , Merkenschlager Matthias
Published: October 2006

Micro RNAs (miRNAs) regulate gene expression at the posttranscriptional level. Here we show that regulatory T (T reg) cells have a miRNA profile distinct from conventional CD4 T cells. A partial T reg cell–like miRNA profile is conferred by the enforced expression of Foxp3 and, surprisingly, by the activation of conventional CD4 T cells. Depleting miRNAs by eliminating Dicer, the RNAse III enzyme that generates functional miRNAs, reduces T reg cell numbers and results in immune pathology. Dicer facilitates, in a cell-autonomous fashion, the development of T reg cells in the thymus and the efficient induction of Foxp3 by transforming growth factor β. These results suggest that T reg cell development involves Dicer-generated RNAs.
The price of the CD27–CD70 costimulatory axis: you can't have it all
Authors: Nolte Martijn A. , van Lier René A.W.
Published: October 2006

T cells require costimulatory signals for optimal proliferation, differentiation, and survival and thus to induce protective immune responses. Recent data, however, show that during chronic lymphocyte choriomeningitis virus (LCMV) infection, triggering of the costimulatory receptor CD27 by its ligand CD70 impedes neutralizing antibody production and leads to viral persistence. Thus, while being crucial for the induction of some adaptive effector pathways, costimulation may block the development of others. Pathogens may exploit this Achilles' heal to achieve persistence.
Sex in a cyclical parthenogen: mating behaviour of Chydorus sphaericus (Crustacea; Branchiopoda; Anomopoda)
Authors: DAMME KAY VAN , DUMONT HENRI J.
Published: October 2006

1. We describe the interactions during mating in Chydorus sphaericus, a cyclical parthenogenetic anomopod. Mating behaviour is more complex than previously assumed, with evidence for a diffusible chemical to which males react at the onset of mating, for reproductive isolation, and for postcopulatory mate guarding.

2. During mating, the male and female form a ‘mating cross’ that may be maintained for several hours, while copulation itself typically lasts less than a minute. Furthermore, males invariably attach to the right valve of females. Copulation involves intromission of the postabdomen between the valves, so that the gonopores approach the left ovarium.

3. This behaviour is reflected in the morphology of both sexes: males have a specialised anterior valve margin, postabdomen, first limb and rostrum, under selective pressure for successful mate guarding and copulation, while gamogenetic females have asymmetric ovaries, and a species‐specific setulation of the valves. Males of the structurally related Chydorus ovalis react to the presence of C. sphaericus, but fail to dock to females, suggesting a lock‐antilock element in the reproductive isolation of both species.

4. The morphological and ethological adaptations in C. sphaericus suggest that there is a strong selective pressure on mating behaviour in this cyclical parthenogen and specifically towards the formation of the ‘mating cross’.
Yeast application for desalting fibersol‐2
Authors: Bangoura Aboubacar Oumar , Jian Tang , He Qian
Published: October 2006

The following study made use of applying brewing yeast Saccharomyces cerevisiae for removing free glucose liberated during enzymatic hydrolysis of the pretreated cornstarch in the preparation of fibersol‐2. The fermentation condition was optimally realized at 20 °C, with 0.5% of yeast for 24 h without total soluble sugars dilution. The glucose removal rate was 98.27%, which is considered acceptable and economical, compared with other removal rates. The recovery of fibersol‐2 was 67.18% and the total dietary fibre content was 94%.
Phenolic composition of European cranberrybush (Viburnum opulus L.) berries and astringency removal of its commercial juice
Authors: Sedat Velioglu Y. , Ekici Lutfiye , Poyrazoglu Ender S.
Published: October 2006

Phenolic composition of the European cranberrybush (ECB) (Viburnum opulus L.) juice was determined using high‐performance liquid chromatography. The juice contained 2037 mg kg−1 chlorogenic acid, which was 54% of total phenolics, and several other phenolics such as (+)‐catechin, (−)‐epicatechin, cyanidin‐3‐glucoside, cyanidin‐3‐rutinoside and six different glucosides of quercetin. Because of its strong astringent taste, the juices were treated with various doses of two different types of activated carbons (Granucol Bi and Granucol Ge) in order to remove phenolic compounds. Results revealed that both types of activated carbons were equally effective on astringency removal (P < 0.01). A 20–30% reduction in total phenolics was achieved by application of 2.0–3.0 g L−1 activated carbon which also removed unpleasant taste and odour.
Low‐dose irradiation as a measure to improve microbiological quality of Turkish raw meat ball (cig kofte)
Authors: Vural Aydın , Aksu Harun , Erkan Mehmet E.
Published: October 2006

Raw meat ball is a traditional food, which is prepared by adding ground meat, spices and different supplements and consumed without any cooking process. The effect of low‐dose gamma irradiation application on microbiological quality of raw meat ball was examined. In this study, 1, 2, and 3 kGy irradiation doses have decreased or eliminated the microorganism counts in raw meat ball, parallel with the increased doses. Coliform bacteria counts were reduced under detectable value after application of 2 kGy irradiation doses; Staphylococcus aureus, sulphite‐reducing Clostridia, yeast and mould by the application of 3 kGy. It has been concluded that the low‐dose gamma irradiation applications have increased the hygienic quality of raw meat balls and possible public health risks can be prevented.
Short‐faced bats (Phyllostomidae: Stenodermatina): a Caribbean radiation of strict frugivores
Authors: Dávalos Liliana M.
Published: October 2006

Aim  To test the hypothesis that Caribbean Short‐faced bats descended from a single recent ancestor that originated in the continental Neotropics (Mexico, Central America and/or South America).

Location  The Neotropics, including the West Indies.

Methods  New mitochondrial cytochrome b and nuclear Rag2 sequences were combined with published molecular data to estimate phylogenetic relationships and sequence divergence among Short‐faced bats. The resulting phylogenies were compared with those compatible with the single‐origin hypothesis using two model‐based statistical tests. Confidence limits on sequence divergence were estimated using a parametric bootstrap.

Results  All molecular phylogenies revealed two independent Caribbean lineages and showed that continental Short‐faced bats share a recent common ancestor. Morphology‐based trees compatible with the single‐origin hypothesis were significantly worse at explaining the molecular data than any molecular phylogeny.

Main conclusions  The ancestor of all Short‐faced bats reached the Antilles in the Miocene, too recently to have used a proposed Oligocene land bridge, and well before the Pleistocene glaciations that are thought to have facilitated dispersal for many bats. After a long period of isolation, Short‐faced bats diversified quickly on the Caribbean islands. A single Short‐faced lineage then reached the continent and subsequently expanded its range and diversified into the four extant genera. Among bats, independent lineages of aerial insectivores and nectarivores have also recolonized the continent after evolving in the West Indies. The evidence for an insular origin of the short‐faced frugivorous radiation completes a dynamic model of Caribbean biogeography that encompasses an entire biological community.
The inappropriate use of HbA1c testing to monitor glycemia: is there evidence in laboratory data?
Authors: Akan Pinar , Cimrin Dilek , Ormen Murat , Kume Tuncay , Ozkaya Aygul , Ergor Gul , Abacioglu Hakan
Published: October 2006

Rationale, aims and objectives  Diabetes is a chronic illness and have relatively high prevalence. Glycemic control is fundamental to the management of diabetes. Hemoglobin A1c (HbA1c) is a commonly used laboratory test to monitor glycemia and to manage diabetes. This study aimed to assess the appropriateness of the frequency of HbA1c test order with respect to the commonly approved guidelines for monitoring glycaemia of patients.

Methods  To assess the rate of inappropriate test orders, laboratory records of HbA1c tests ordered between 2002 and 2004 were evaluated. Inappropriate orders were defined as any order for a given patient that takes place within a 29‐ or 89‐day‐period following the previous HbA1c order. The effects of various parameters, like ordering clinics, the first HbA1c level, or the on‐line availability of test results on test ordering were evaluated.

Results  Evaluation of test intervals showed that 10.3% of all orders and 33.8% of the inpatients’ orders were performed within 29 days, 35.5% of all orders and 55% of the inpatients’ orders were within 89 days.

Conclusions  Inappropriate laboratory utilization of HbA1c testing is very common especially in the inpatient clinics. We think that the application of the guidelines may decrease unnecessary health expenditure.
The Finnish national asthma programme: communication in asthma care – quality assessment of asthma referral letters
Authors: Tuomisto Leena E , Marina Erhola , Minna Kaila , Brander Pirkko E , Ritva Kauppinen , Hannu Puolijoki , Pertti Kekki
Published: October 2006

Aims and objectives  The Finnish National Asthma Programme, which was launched in year 1994, considered the management of asthma as a community problem. The role of the primary health care in the management of asthma was emphasized. Optimal asthma management includes good communication between health care professionals. Referral letters are an accepted tool for evaluation of the communication process. The aim of this study was to assess the quality of asthma‐related referral letters.

Methods  All non‐acute referral letters (n = 3176) to three pulmonary departments were screened in 2001 and all those related to asthma were included (n = 1289). The 14 previously derived asthma‐specific criteria were applied: occupation, smoking, known allergies, current medication, other diseases, onset of symptoms, wheezing, dyspnoea, specified dyspnoea, cough, specified cough, use of asthma medication, peak‐flow follow‐up or spirometry with bronchodilatation test as an attachment. The study group was prepared to accept the maximum of 30% of the referral letters to be of poor quality.

Results  Twenty‐one per cent of the referral letters were graded good, 34% satisfactory and 45% poor. Information on wheezing, smoking habits and current medication was mentioned in 44%, 42% and 41% of asthma letters respectively.

Conclusions  The Finnish National Asthma Programme calls for optimizing communication between doctors. The proportion of poor letters was 50% higher than the preset standard and clearly indicates a need for improvement. We found several issues, which need to be better communicated (smoking, lung function tests, wheezing, medication) when referring a patient with suspected asthma.
Critical role of ribavirin for the achievement of early virological response to HCV therapy in HCV/HIV‐coinfected patients
Authors: Ramos B. , Núñez M. , Rendón A. , Berdún M. A. , Losada E. , Santos I. , Echevarría S. , Ocampo A. , Miralles C. , Arazo P. , Barreiro P. , Romero M. , Labarga P. , Guardiola J. M. , Garcia‐Samaniego J. , Soriano V.
Published: October 2006

Summary.  The response to hepatitis C virus (HCV) therapy seems to be lower in HCV/HIV‐coinfected patients than in HCV‐monoinfected individuals. Given that most pivotal trials conducted in coinfected patients have used the combination of pegylated interferon (pegIFN) along with fixed low doses (800 mg/day) of ribavirin (RBV), it is unclear whether HIV itself and/or suboptimal RBV exposure could explain this poorer outcome. Two well‐defined end points of early virological response were evaluated in Peginterferon Ribavirina España Coinfección (PRESCO), a multicentre trial in which the combination of pegIFN plus RBV (1000 mg if body weight <75 kg and 1200 mg if >75 kg) was prescribed to coinfected patients. For comparisons, we used unpublished data from early kinetics in two other large trials, one performed in HIV‐negative patients [Pegasys International Study Group (PISG)] in which RBV 1000–1200 mg/day was used and another [AIDS Pegasys Ribavirin Coinfection Trial (APRICOT)] in which HIV‐positive patients received fixed low RBV doses (800 mg/day). A total of 348 HCV/HIV‐coinfected patients from the PRESCO trial were analysed as well as all patients treated with pegIFN plus RBV, who completed 12 weeks of therapy in the comparative studies (435 in PISG and 268 in APRICOT). Negative serum HCV‐RNA at week 4 (which has the highest positive predictive value of sustained virological response, SVR) was attained in 33.3%, 31.2% and 13% of treated patients with HCV genotype 1, respectively, in PRESCO, PISG and APRICOT. For HCV genotypes 2/3, responses were 83.7%, 84.2% and 37%, respectively. A decline lower than 2 log10 at week 12 (which has the highest negative predictive value of SVR) was seen in 25.5%, 19.5% and 37% of HCV genotype‐1‐infected patients, and in 2.1%, 2.9% and 12% of genotypes‐2/3‐infected patients, respectively. Prescription of high RBV doses enhances the early virological response to HCV therapy in HCV/HIV‐coinfected patients, with results approaching those seen in HCV‐monoinfected patients.
Transcriptional slippage controls production of type III secretion apparatus components in Shigella flexneri
Authors: Penno Christophe , Hachani Abderrahman , Biskri Latefa , Sansonetti Philippe , Allaoui Abdelmounaïm , Parsot Claude
Published: October 2006

During transcription, series of approximately 9 As or Ts can direct RNA polymerase to incorporate into the mRNA nucleotides not encoded by the DNA, changing the reading frame downstream from the slippage site. We detected series of 9 or 10 As in spa13, spa33 and mxiA encoding type III secretion apparatus components. Analysis of cDNAs indicated that transcriptional slippage occurs in spa13, mxiA and spa33. Changes in the reading frame were confirmed by using plasmids carrying slippage sites in the 5′ part of lacZ. Slippage is required for production of Spa13 from two overlapping reading frames and should lead to production of truncated MxiA and Spa33 proteins. Complementation of spa13 and mxiA mutants with plasmids carrying altered sites indicated that slippage in spa13 is required for assembly of the secretion apparatus and that slippage sites in spa13 and mxiA have not been selected to encode Lys residues or to produce two proteins endowed with different activities. The presence of slippage sites decreases production of Spa13 by 70%, of MxiA and Spa33 by 15% and of Spa32 (encoded downstream from spa13) by 50%. These results suggest that transcriptional slippage controls protein production by reducing the proportion of mRNA translated into functional proteins.
Neutrophilic eccrine hidradenitis in non‐small cell lung cancer
Authors: Lee Seung Gyu , In Seung Gyun , Shin Jeong Hyun , Choi Gwang Seong , Kim Yoo Chan
Published: October 2006

A 75‐year‐old man was treated for non‐small cell lung cancer (NSCLC) (stage IIIb) with three‐cycle chemotherapy (cisplatin and paclitaxel) for 3 weeks. No skin lesions were observed during the first and second chemotherapy cycles. After the third chemotherapy cycle, he developed slightly pruritic skin eruptions on the anterior chest. He was treated for 1 month with topical steroid without improvement, and was referred to our dermatologic department.

Skin examination revealed slightly elevated, dusky, erythematous, edematous patches on the anterior chest in a bilateral distribution (Fig. 1).

Dusky, erythematous, edematous patches on the anterior chest

image

Histologic examination of a biopsy specimen taken from the cutaneous lesions showed a neutrophilic infiltrate surrounding the secretory eccrine glands. The epithelial cells of the eccrine ducts and coils were not affected. There was a mild superficial perivascular infiltrate of mainly lymphocytes. These changes were consistent with a diagnosis of neutrophilic eccrine hidradenitis (Fig. 2).

An inflammatory infiltrate involving eccrine coils in the dermis without vasculitis (hematoxylin and eosin, ×40). Inset: neutrophils surrounding eccrine coils (hematoxylin and eosin, ×400)

image

He was treated with dapsone, 50 mg/day, and methylprednisolone, 8 mg/day, for 2 weeks, and his skin lesions improved. Two months later the erythematous eruptions recurred on the chest, back, and extremities without the use of any chemotherapeutic agent, and were treated effectively with the same drugs.
Epidemiology of pemphigus in Macedonia: A 15‐year retrospective study (1990–2004)
Authors: V’lckova‐Laskoska Marija T. , Laskoski Dimitri S. , Kamberova Suzana , Caca‐Biljanovska Nina , Volckova Nadja
Published: October 2006

Background  Pemphigus is an autoimmune blistering skin disease mediated by auto‐antibodies directed against desmoglein proteins. There are only a few epidemiological studies on pemphigus. Our objective was to determine the epidemiological features of pemphigus in Macedonia, and to compare the results with those reported elsewhere.

Methods  Diagnosis in all cases was confirmed by histopathology and direct immunofluorescence. Binomial distribution testing and Fisher's exact‐test at the 0.01 level of significance were used to determine if particular demographic groups were over‐/ under‐represented among the pemphigus patients.

Results  One hundred and thirty‐three new pemphigus cases were diagnosed in Macedonia from 1990–2004. The average annual incidence was 0.44/100,000 inhabitants (SD = 0.17). The incidence doubled to 0.89 in 2001 during the local armed unrest. The disease did not affect either gender to a greater extent. The average annual incidence was 0.51 for ethnic Macedonians. Roma (Gypsies) had a statistically significantly higher incidence of pemphigus at 2.4 cases/100,000 individuals. Ethnic Albanians had statistically significantly lower incidence of 0.1 cases/100,000 individuals. The most common variant was pemphigus vulgaris (77.4%).

Conclusions  The annual incidence for pemphigus in Macedonia is 0.44 cases/100,000 inhabitants. Most common form was pemphigus vulgaris. An epidemiological peak occurred in 2001 during the local armed conflict. Macedonian Roma had a sixfold higher incidence of pemphigus compared with the overall population; ethnic Albanians had a fourfold lower incidence.
Characterizing the specific coaggregation between Actinobacillus actinomycetemcomitans serotype c strains and Porphyromonas gingivalis ATCC 33277
Authors: Suzuki N. , Nakano Y. , Kiyoura Y.
Published: October 2006

A visual coaggregation study showed specific interspecies coaggregation between an Actinobacillus actinomycetemcomitans serotype c strain and Porphyromonas gingivalis strains ATCC 33277 and 381. We mutagenized A. actinomycetemcomitans SUNYaB 67 (serotype c) with transposon IS903φkan and isolated three transposon insertion mutants that had a reduced ability to aggregate with P. gingivalis ATCC 33277. The three transposon insertions in the mutant strains mapped to the genes at ORF12, ORF13 and ORF16 of the gene cluster responsible for producing serotype c‐specific polysaccharide antigen (SPA). Western blot analysis with serotype c‐specific antibody showed that these strains did not produce the high‐molecular‐mass smear of SPA. Furthermore, two SPA‐deficient mutants and an SPA‐producing mutant were constructed. The two SPA‐deficient mutants were deficient for ORF12 and ORF14, which are necessary for the synthesis of serotype c‐SPA, and the SPA‐producing mutant was deficient for ORF17, which is not related to SPA synthesis. The ORF12‐ and ORF14‐deficient mutants showed reduced ability to aggregate with P. gingivalis ATCC 33277, while the ORF17‐deficient mutant aggregated with ATCC 33277 to the same extent as wild‐type SUNYaB 67. Our findings suggest that serotype c‐SPA of A. actinomycetemcomitans mediates coaggregation with P. gingivalis ATCC 33277.
Retinoic acid‐inducible gene‐I is induced in gingival fibroblasts by lipopolysaccharide or poly IC: possible roles in interleukin‐1β, ‐6 and ‐8 expression
Authors: Kubota K. , Sakaki H. , Imaizumi T. , Nakagawa H. , Kusumi A. , Kobayashi W. , Satoh K. , Kimura H.
Published: October 2006

Retinoic acid‐inducible gene‐I (RIG‐I) is a member of the DExH family of proteins, and little is known of its biological function in the oral region. We previously reported that interleukin 1β (IL‐1β) induced RIG‐I expression in gingival fibroblasts. In this study, we studied the mechanism of RIG‐I expression induced by lipopolysaccharide (LPS) or double‐stranded RNA (dsRNA) in gingival fibroblasts. We also addressed the role of RIG‐I in the expression of IL‐1β, IL‐6 and IL‐8 in gingival fibroblasts stimulated with LPS or dsRNA. We stimulated cultured human gingival fibroblasts with LPS or dsRNA, and examined the expression of RIG‐I mRNA and protein. The effect of cycloheximide, a protein synthesis inhibitor, on RIG‐I induction by these stimuli was examined. The expression of IL‐1β, IL‐6 and IL‐8 in gingival fibroblasts transfected with RIG‐I cDNA stimulated with LPS or dsRNA was examined. LPS or dsRNA induced the expression of mRNA and protein for RIG‐I in concentration‐ and time‐dependent manners. We also examined the localization of RIG‐I, and found that it was expressed in cytoplasm. Cycloheximide did not suppress the LPS or dsRNA‐induced RIG‐I expression. Introduction of RIG‐I cDNA into gingival fibroblasts resulted in enhanced expression of IL‐1β, IL‐6 and IL‐8; moreover, overexpression of RIG‐I stimulated with LPS or dsRNA synergistically increased expression of IL‐1β, IL‐6 and IL‐8. RIG‐I may have important roles in the innate immune response in the regulation of IL‐1β, IL‐6 and IL‐8 expression in gingival fibroblasts in response to LPS and dsRNA.
Asthma and allergy medication use and costs among pediatric primary care patients on asthma controller therapy
Authors: Sazonov‐Kocevar Vasilisa , Laforest Laurent , Travier Noemie , Yin Donald D. , Ganse Eric Van
Published: October 2006

As observational studies in children initiating GINA‐Step 3 therapies are scarce, we evaluated outcomes and costs in a primary care cohort. Two‐yr retrospective cohort study included French children (age: 6–14) continuously followed in BKL‐Thalès database who received ≥2 consecutive prescriptions for GINA‐Step 3 therapy (=addition of montelukast or other controllers (‘other’), such as increasing inhaled‐corticosteroid dose (hICS), adding long‐acting β agonist (LABA), or ICS + LABA). After matching on gender and propensity score, medication use [rescue (short‐acting β agonists), acute (antibiotics (AB), oral corticosteroids (OCS)), allergy (antihistamines, nasal steroids) and other respiratory] was estimated via mean number of prescriptions and mean cost (per child/per month), and cost trends. During 12‐month follow‐up, children adding montelukast (n = 71) vs. ‘other’ (n = 213) had similar asthma rescue/acute and allergy medication use. Subgroup with asthma and allergic rhinitis (A + AR) adding montelukast used less OCS and AB (p = 0.014). Two‐yr cost trends suggest stable asthma/allergy medication use in montelukast group (€0.83) compared with increase in ‘other’ (€5.39), which was driven by nasal steroid use [€0.32 (‘other’) vs. €−0.04 (montelukast), p = 0.0013]. In subgroup with A + AR decline in asthma/allergy medication use in montelukast group (€−0.47) vs. increase in ‘other’ (€11.05), p = 0.015, was driven by differences in AB and OCS (p = 0.04) and nasal steroid use (p = 0.001). Concomitant asthma/allergy medication use was similar in children adding montelukast or ‘other’ controllers (hICS, LABA, ICS + LABA), while children with allergic rhinitis on montelukast used less AB. Concomitant medication costs after addition of montelukast remained stable, while ‘other’ group experienced increase, especially in children with concomitant allergic rhinitis.
Towards endothelial cell‐specific transgene expression in pigs: characterization of the pig ICAM‐2 promoter
Authors: Godwin James W. , Fisicaro Nella , D'Apice Anthony J. F. , Cowan Peter J.
Published: October 2006

Abstract:  Background:  Targeting protective gene expression to porcine endothelium has obvious advantages in xenotransplantation, but no endothelial cell‐specific promoters have yet been used successfully in transgenic pigs. We have previously reported that a human intercellular adhesion molecule‐2 (ICAM‐2) gene promoter fragment functioned efficiently in transgenic mice but not pigs, suggesting that it lacked important transcriptional signals. In this study, we cloned and characterized regulatory elements of the pig ICAM‐2 gene.

Methods:  Various segments of the pig ICAM‐2 gene upstream region and first intron were cloned into a luciferase reporter vector and assayed for promoter activity in vitro. Putative regulatory elements were analysed by site‐directed mutagenesis.

Results:  A 0.90‐kb pig ICAM‐2 promoter fragment had strong activity in pig endothelial cells but not in non‐endothelial cells. Deletion analysis revealed that the majority of promoter activity was specified by a 0.48‐kb sub‐fragment with significant homology to the human ICAM‐2 promoter. Conserved positive‐acting elements included binding sites for GATA and Ets transcription factors, and a palindromic octamer (P8) that has been implicated in the endothelial specificity of several genes. Significant enhancer activity was identified within the first intron of the pig ICAM‐2 gene. Mutational analysis was used to show that a second P8 site in the first intron was essential for enhancer activity.

Conclusions:  The pig and human ICAM‐2 promoters exhibit many similarities, but the pig ICAM‐2 gene, unlike its human and mouse homologs, contains P8 sites in both the promoter and first intron. The enhancer activity associated with the intronic P8 site suggests that it may be the key to achieving strong endothelial cell‐specific transgene expression in pigs.
NK cells promote peritoneal xenograft rejection through an IFN‐γ‐dependent mechanism
Authors: Lin Ming‐Lee , Zhan Yifan , Nutt Stephen L. , Brady Jason , Wojtasiak Magdalena , Brooks Andrew G. , Lew Andrew M.
Published: October 2006

Abstract:  Background:  Natural killer (NK) cells have emerged as major players in anti‐viral and anti‐tumour immune responses. Like cytotoxic T lymphocytes (CTL), they express perforin and are potent secretors of γ‐interferon (IFN‐γ). However, there is conflicting evidence about their role in mediating rejection of xenogeneic tissue.

Methods:  A pig‐to‐mouse peritoneal cell model of xenotransplantation was used to investigate the effect of NK deficiency on xenograft recovery and the possible mechanisms behind this NK‐mediated graft rejection. γc−/−RAG−/− mice were used as a model of NK deficiency. Additionally, NK cells were depleted in RAG−/− mice using anti‐asialo GM1. The contributions of IFN‐γ, perforin and NKT cells were studied using knock‐out mice that were depleted in vivo of T cells. Mice were injected with 107 pig cells intraperitoneally and peritoneal fluid was assessed 5 days later for xenograft recovery and phenotypic analysis. The requirement for NK cells for xenograft rejection was also assessed using luciferase‐transfected porcine cells in a renal subcapsular model of transplantation.

Results:  Pig cell recovery was enhanced in both γc−/−RAG−/− and NK‐depleted RAG−/− mice when compared with RAG−/− control mice. IFN‐γ−/− mice depleted of T cells also demonstrated superior graft survival compared with their B6 counterparts. However, there were minimal graft survival differences between Pfp−/− and B6 control mice. Similarly, a deficiency in NKT cells did not improve pig xenograft recovery from the peritoneum of these mice.

Conclusions:  Therefore, we conclude that NK cells, but not NKT cells, are important mediators of xenograft rejection in the peritoneal cavity, and that their role may be unmasked in the absence of T cells. The mechanism for this xenorejection appears to involve IFN‐γ but is perforin independent.
Research ethics and the development of medical biotechnology
Authors: Persson Anders
Published: October 2006

Abstract:  Commercial funding is of most importance for the development of new biomedical technologies such as xenotransplantation. The dependence of such funding may, however, carry certain risks from an ethical point of view. In this article some of these risks are exemplified and it is argued that the adherence to basic research ethical norms are of vital importance for the field of xenotransplantation to develop properly.
A new liquid, intravenous immunoglobulin product (IGIV 10%) highly purified by a state‐of‐the‐art process
Authors: Teschner W. , Butterweck H. A. , Auer W. , Muchitsch E.‐M. , Weber A. , Liu S.‐L. , Wah P.‐S. , Schwarz H.‐P.
Published: October 2006

Background and Objectives  The ultimate goal was to generate an industrial‐scale process suitable to produce a high‐yield, safe and stable immunoglobulin G (IgG) preparation for intravenous administration, which is ready to use for customer convenience. This new liquid 10% IgG preparation (IGIV 10%) was compared to Gammagard SD, a licenced lyophilized immunoglobulin in biochemical and preclinical testing.

Materials and Methods  The new process, which includes three dedicated virus clearance steps, is a streamlined combination of the currently applied and well‐established manufacturing procedures. The biochemical characterization is done by standard methods focusing on purity, integrity and functionality of the preparation. Efficacy is demonstrated in vivo by mouse protection testing and in vitro by opsonization and protein A affinity chromatography. Pharmacokinetics in rats is evaluated after a single intravenous dose. The anaphylactoid potential is determined in rats and in guinea pigs, while thrombogenicity is assessed in a rabbit model. The influence of the products on vital functions is tested on dogs, while acute toxicity studies are carried out on mice and rats.

Results  The biochemical characterization data demonstrate the high purity of monomeric IgG in the product. The mouse protection test showed that the protective activity against systemic bacterial infections of IGIV 10% is at least as good as the reference Gammagard SD. This result is supported by the broad spectrum of antibodies in high titres against bacteria and viruses and the high functional integrity of the IgG molecule (≥ 90% functionally intact IgG) in IGIV 10%. The opsonic activity of all IGIV 10% lots is similar to the one of the reference Gammagard SD. In safety and thrombogenicity studies, no adverse effects of IGIV 10% were observed. Pharmacokinetic studies showed no statistically significant differences between the two products. In the acute toxicity animal studies, IGIV 10% compared favourably to the reference Gammagard SD.

Conclusions  The new manufacturing process enables the production of a highly purified IgG preparation for intravenous administration. The product has an IgG subclass distribution similar to plasma and contains a broad spectrum of functionally intact antibodies. Preclinical studies demonstrate that the liquid IGIV 10% combines excellent qualities of efficacy, safety and tolerability.
An Argentine Equine Herpesvirus Strain with Special Restriction Patterns Protect Mice Challenged with a Pathogenic Strain
Authors: Galosi C. M. , Barbeito C. G. , Martin Ocampos G. P. , Martinez J. P. , Ayala M. A. , Corva S. G. , Fuentealba N. A. , Gimeno E. J.
Published: October 2006

Equine herpesvirus 1 (EHV‐1) was first isolated in Argentina in 1979. This strain SPv has especial restriction patterns, but a previous study demonstrated that SPv did not modify its growth in cell culture. In addition, it showed low virulence in the mouse respiratory model consistently with results found in female BALB/C at different state of gestation. This study evaluates in a mouse respiratory model, if primary infection with SPv strain protects animals from subsequent challenge with a pathogenic strain. Body weight loss was not observed in mice intranasally inoculated with SPv strain and challenged with HH1 Japanese strain. The SPv primary infection does not completely prevent clinical presentation by HH1 infection but the SPv inoculated animals recovered more quickly, with less intense and less persistent histological lesions. The challenge infection caused a rapid and prolonged increase in anti‐EHV‐1 antibodies in the mice previously infected with SPv, along with a more rapid reduction of viral titres in lungs. In this work it was demonstrated that this EHV‐1 strain constitute a good immunogen. These results show that this SPv strain could be considered to produce an EHV‐1 vaccine.
Molecular Characterization of MG Isolates Using RAPD and PFGE Isolated from Chickens in Brazil
Authors: Mettifogo E. , Buzinhani M. , Buim M. R. , Piantino Ferreira A. J. , Kleven S. H. , Timenetsky J.
Published: October 2006

In the present study, 27 primers were screened under different cycles by random amplified polymorphic DNA (RAPD) method. Mathematical models were used for analysis of the genetic relationships among strains, including vaccinal, reference strains and nine field isolates previously characterized as Mycoplasma gallisepticum (MG)F by RAPD and pulsed field gel electrophoresis (PFGE). The PFGE was considered as laborious, expensive and time‐consuming than RAPD method. These methods improved the typeability for epidemiological studies of MG with regard to differentiation from vaccinal and field strains.
Dynamics of Infestation of Cattle and Pasture by Gastrointestinal Nematodes in an Atlantic Temperate Environment
Authors: Nogareda C. , Mezo M. , Uriarte J. , Lloveras J. , Cordero del Campillo M.
Published: October 2006

The objective of the present study was to determine the dynamics of infestation of cattle and pasture by gastrointestinal nematodes in a mild humid environment in northwestern Spain. For this, infestation of pasture by free‐living stages (L3), dynamics of faecal egg output, blood pepsinogen levels and worm burden in slaughtered animals were quantified. The results showed a high degree of annual variability, which was dependent on weather conditions. The seasons were clearly defined in the study area, with mild humid winters and relatively dry summers registered throughout the years of the study. Infestation of pasture by larvae varied from year to year, peaking during August in the first year, between August and December in the second year, and during October in the third year. The annual variation was mainly due to weather conditions, particularly the amount of rain in summer. The patterns of faecal egg output were similar in the first and third grazing seasons, with maximum levels observed in May/June; however, in the second year, the peak was reached in October. Blood pepsinogen levels increased from pasture turnout (March/April) until the end of the grazing season (November/December), reaching maximum values from August/September onwards. The nematode parasite species identified at necropsy wereOstertagia osteragi, O. lyrata, Cooperia oncophora, C. macmasteri, C. punctataandTrichuris ovis, withO. ostertagi and C. oncophorapredominating. In faecal cultures, the following genera were also identified:Haemonchus, Trichostrongylus, Nematodirus, Bunostomum, OesophagostomumandStrongyloides. There was a significant correlation (r = 0.97,P < 0.01) between worm burden (Ostertagiaspp.) and pasture infestation (OstertagiaL3) 3 weeks prior to slaughter of the calves, and also between blood pepsinogen levels and pasture infestation byOstertagiaL3 (r = 0.33,P < 0.02). Correlations between worm burden and faecal egg output and between blood pepsinogen level and faecal egg output were not significant. The results obtained in the present study confirm that there is annual variability in the time‐course of nematodosis in cattle, and demonstrate the importance of weather, particularly summer rainfall, in an Atlantic temperate environment.
Development of aDrosophilaseizure model forin vivohigh‐throughput drug screening
Authors: Stilwell Geoff E. , Saraswati Sudipta , Littleton J. Troy , Chouinard Scott W.
Published: October 2006

An important application of model organisms in neurological research has been to identify and characterise therapeutic approaches for epilepsy, a recurrent seizure disorder that affects > 1% of the human population. Proconvulsant‐treated rodent models have been widely used for antiepileptic drug discovery and development, but are not suitable for high‐throughput screening. To generate a genetically tractable model that would be suitable for large‐scale, high‐throughput screening for antiepileptic drug candidates, we characterized aDrosophilachemical treatment model using the GABAAreceptor antagonist picrotoxin. This proconvulsant, delivered toDrosophilalarvae via simple feeding methods suitable for automated screening, generated robust generalised seizures with lethality occurring at doses between 0.3 and 0.5 mg/mL. Electrophysiological analysis of CNS motor neuron output in picrotoxin‐treated larvae revealed generalised seizures within minutes of drug exposure. At subthreshold doses for seizure induction, picrotoxin produced an increased frequency of motor neuron action potential bursting, indicating that CNS GABAergic transmission regulates patterned activity. Mutants in theDrosophila RdlGABAAreceptor are resistant to picrotoxin, confirming that seizure induction occurs via a conserved GABAAreceptor pathway. To validate the usefulness of this model forin vivodrug screening, we identified several classes of neuroactive antiepileptic compounds in a pilot screen, including phenytoin and nifedipine, which can rescue the seizures and lethal neurotoxicity induced by picrotoxin. The well‐defined actions of picrotoxin inDrosophilaand the ease with which compounds can be assayed for antiseizure activity makes this genetically tractable model attractive for high‐throughputin vivoscreens to identify novel anticonvulsants and seizure susceptibility loci.