bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of JTP‐70902, a p15INK4b‐inductive compound, as a novel MEK1/2 inhibitor
Authors: Yamaguchi Takayuki , Yoshida Takayuki , Kurachi Reina , Kakegawa Junya , Hori Yoshikazu , Nanayama Toyomichi , Hayakawa Kazuhide , Abe Hiroyuki , Takagi Koichi , Matsuzaki Youichirou , Koyama Makoto , Yogosawa Shingo , Sowa Yoshihiro , Yamori Takao , Tajima Nobuyuki , Sakai Toshiyuki
Published: September 2007

The INK4 family members p16INK4a and p15INK4b negatively regulate cell cycle progression by inhibition of cyclin‐dependent kinase (CDK) 4/6. Loss of p16INK4a functional activity is frequently observed in tumor cells, and is thought to be one of the primary causes of carcinogenesis. In contrast, despite the biochemical similarity to p16INK4a, the frequency of defects in p15INK4b was found to be lower than in p16INK4a, suggesting that p15INK4b‐inductive agents may be useful for tumor suppression. Here we report the discovery of a novel pyrido‐pyrimidine derivative, JTP‐70902, which exhibits p15INK4b‐inducing activity in p16INK4a‐inactivated human colon cancer HT‐29 cells. JTP‐70902 also induced another CDK‐inhibitor, p27KIP1, and downregulated the expression of c‐Myc and cyclin D1, resulting in G1 cell cycle arrest. MEK1/2 was identified by compound‐immobilized affinity chromatography as the molecular target of JTP‐70902, and this was further confirmed by the inhibitory activity of JTP‐70902 against MEK1/2 in kinase assays. JTP‐70902 suppressed the growth of most colorectal and some other cancer cell lines in vitro, and showed antitumor activity in an HT‐29 xenograft model. However, JTP‐70902 did not inhibit the growth of COLO320 DM cells; in these, constitutive extracellular signal‐regulated kinase phosphorylation was not detected, and neither p15INK4b nor p27KIP1 induction was observed. Moreover, p15INK4b‐deficient mouse embryonic fibroblasts were found to be more resistant to the growth‐inhibitory effect of JTP‐70902 than wild‐type mouse embryonic fibroblasts. These findings suggest that JTP‐70902 restores CDK inhibitor‐mediated cell cycle control by inhibiting MEK1/2 and exerts a potent antitumor effect. (Cancer Sci 2007; 98: 1809–1916)
A randomized trial of effectiveness and cost‐effectiveness of observed versus unobserved administration of buprenorphine–naloxone for heroin dependence
Authors: Bell James , Shanahan Marian , Mutch Carolyn , Rea Felicity , Ryan Anni , Batey Robert , Dunlop Adrian , Winstock Adam
Published: September 2007

Aims To compare the effectiveness and cost‐effectiveness of unobserved versus observed dosing of patients seeking treatment of heroin dependence.

Design Randomized controlled trial and cost‐effectiveness analysis.

Setting Specialist out‐patient drug treatment centres in Australia.

Participants Heroin users seeking maintenance treatment.

Intervention Participants were allocated randomly to observed or unobserved dosing for 3 months. All subjects received buprenorphine–naloxone and weekly clinical reviews.

Measurements Primary end‐points were retention in treatment and heroin use at 3 months. Costs of treatment were measured (in Australian dollars, AU$) and cost‐effectiveness compared. Secondary outcomes included quality of life, psychological symptoms and use of non‐opioid drugs.

Findings A total of 119 subjects were randomized and analysed. At 3 months, 33/58 (57%) randomized to unobserved treatment, and 37/61 (61%) observed were retained (log‐rank χ2 = 0.04, df = 1,P = 0.84). On an intention‐to‐treat analysis, reductions in days of heroin use in the preceding month, from baseline to 3 months, did not differ significantly; 18.5 days (95% CI: 21.8–15.3) and 22.0 days (95% CI: 24.3–19.7), respectively (Mann–WhitneyU = 807.5,P = 0.13). The mean cost for the unobserved group was AU$1663 (95% CI 1308–2017) per treatment episode, significantly less than the mean cost for the observed group at AU$2138 (95% CI 1713–2562).

Conclusions Retention and heroin use was not significantly different between observed and unobserved dosing groups. Attendance for observed dosing was not associated with worse retention. Treatment with close clinical monitoring, but no observation of dosing, was significantly cheaper and therefore significantly more cost‐effective.
Intravenous and intranasal heroin‐dependent treatment‐seekers: characteristics and treatment outcome
Authors: Highfield David A. , Schwartz Robert P. , Jaffe Jerome H. , O'Grady Kevin E.
Published: September 2007

Aims  This study compared the characteristics of intravenous (i.v.) and intranasal (i.n.) heroin users seeking methadone treatment, and their response to treatment.

Participants  A total of 319 heroin‐dependent adults.

Design  Participants were assigned randomly to receive interim methadone treatment or to a waiting list control on a 3 : 2 basis. Analyses were conducted by dividing participants into two groups based on their route of heroin ingestion: i.v. or i.n.

Setting  A methadone clinic in Baltimore City, Maryland.

Intervention  Interim methadone treatment consisted of providing an adequate and stable dose of methadone, but no psychosocial services, to heroin‐dependent adults for up to 120 days while they awaited an opening for comprehensive methadone treatment.

Measures  Addiction Severity Index, Texas Christian University AIDS Risk Assessment, a questionnaire on treatment entry and a urine drug test were collected at baseline and at entry into a comprehensive treatment program, or at 120 days after baseline assessment, whichever came first.

Findings  At baseline, over 60% of participants were i.n. users and had been for an average of over 12 years; i.v. users, compared to i.n. users, were more likely to have ever used cocaine, to have used cocaine in the past 30 days, to have more medical complications and to report more income generated from criminal behavior. Both i.v. and i.n. users reduced their self‐reported days of heroin use, cocaine use and days of criminal activity in response to interim methadone treatment.

Conclusions  Despite differences in baseline characteristics, i.n. and i.v. heroin‐dependent individuals did not differ in their response to interim methadone treatment.
Population structure of montane bamboo and causes of its decline in Echuya Central Forest Reserve, South West Uganda
Authors: Bitariho Robert , McNeilage Alastair
Published: September 2007

Montane bamboo is of immense importance to the people living adjacent to Echuya Forest Reserve. It is used for building poles, bean‐staking and basket weaving. The bamboo in Uganda occurs mostly in protected areas. Over the past 50 years, the bamboo forest cover has been declining. This study aimed at determining bamboo density and distribution and the possible causes of its decline within Echuya. We used exploratory inventories, bamboo assessment plots and village interviews to determine bamboo population structure and possible causes of its decline in Echuya. Bamboo forest decline in Echuya may be blamed on several factors that interplay together. Poor harvest methods especially during the clear cutting of bamboo for bean‐stakes and stakes for house wefts seems to be the remote cause of the bamboo forest decline. Other causes for the bamboo forest decline are damages caused by insect borers and climber loads on the bamboo stems. The remote causes have been exacerbated by the invasion of secondary forest tree species such as Macaranga kilimandscharica that have slowly taken over areas previously occupied by the bamboo forest. Seventy per cent of the local people interviewed agree that over‐harvesting of bamboo is the major reason for its decline.
A survey of emergency airway management in the United Kingdom
Authors: RATNAYAKE B. , LANGFORD R. M.
Published: September 2007

A postal questionnaire aiming to ascertain the availability of cricothyrotomy and transtracheal ventilation equipment, and experience of its use, was sent to all tutors of the Royal College of Anaesthetists in the United Kingdom and Ireland. The response rate was 74.9%. Almost half of the respondents had experience of cricothyrotomy. The “Portex Minitrach” (58.6%) was the most commonly used preformed device, followed by the ‘William‐Cook’ cannula (8.5%). The 14‐G intravenous cannula (21.1%) was the most frequently improvised cricothyrotomy device. Seventy six percent of all theatres had equipment available for such emergency airway management. Sixty five percent of Portex‐Minitrach insertions resulted in complications, more than half of which were serious (total failure to cannulate (17.1%), multiple attempts (20.7%), pneumothorax (8.5%) and severe bleeding (7.3%)). Major complication rates of the ‘William‐Cook’ device and 14‐G intravenous cannula were 27.3% and 22.2% respectively. Seventy five percent of patients reported had eventual successful airway management with full recovery, while 9.6% had partial recovery and 15.4% died. Formal training of emergency airway management was practised in 14.2% of anaesthetic departments, while 73.6% had informal training. Twelve percent of the departments did not specifically teach their trainees the skill of emergency airway management.
Lack of efficacy of intra‐articular opioids for analgesia after day‐case arthroscopy
Authors: WRENCH I. J. , TAYLOR P. , HOBBS G. J.
Published: September 2007

A randomised double‐blind placebo‐controlled trial was undertaken to assess the analgesic efficacy of intra‐articular opioids following arthroscopy of the knee. At the completion of surgery, patients received an intra‐articular injection of: morphine 1 mg, buprenorphine 30 μg or 0.9% saline. There were no differences in pain scores among groups for the first 24 h postoperatively. We have found no clinical evidence for a peripherally‐mediated opioid analgesic effect.
The airways and anaesthesia II
Authors: BURWELL D. R. , JONES G.
Published: September 2007

We pointed out in the first of these two articles [1] that the commonest cause of an anaesthetic disaster in young healthy patients is a loss of airway patency then a failure to intubate occurring unexpectedly in the absence of head or neck pathology. Upper airway obstruction is a very common complication of general anaesthesia and all anaesthetists must be trained in the management of this problem. Less obvious are the changes that can occur in the lower airways which can impair gas exchange by increasing ventilalion‐perfusion mismatch. This article is concerned with these pathophysiological changes that occur during general anaesthesia.
Detection of quantitative trait loci for body weights and conformation traits in Beijing ducks
Authors: Huang Y. , Haley C. S. , Hu S. , Hao J. , Wu C. , Li N.
Published: September 2007

Quantitative trait loci (QTL) for body weights and conformation traits were detected in Beijing ducks. Traits included body weights (BW) at hatching and at 1–7 weeks of age; lengths of the body (BL), keel bone (KBL), shank (SL) and neck (NL) at 7 weeks of age; width of breast (BTW) at 7 weeks; and girths of shank (SG) and breast (BG) at 7 weeks. Using a half‐sib analysis with a multiple‐QTL model, linkage between the phenotypic traits and 95 microsatellite markers was studied. Six genome‐wide suggestive QTL for three body weights and two conformation traits were identified in CAU1, CAU2, CAU6 and CAU12. Chromosome‐wide significant QTL influencing one body weight trait and one conformation trait were located in CAU4 and CAU10 respectively. Twelve chromosome‐wide suggestive QTL for six body weight traits and four conformation traits were found in seven linkage groups (CAU1, CAU2, CAU3, CAU4, CAU6, CAU10 and CAU12). In addition, the QTL in CAU6 at 21 and 73 cM jointly affected SG and explained 10.6% of the phenotypic variation. This study provides the first evidence for QTL involved in body weights and conformation traits in ducks, and will stimulate further investigations into the genetic architecture of these traits in this species.
Correlation of serum IGF‐I and IGFBP‐1 and ‐3 to cardiovascular risk indicators and early carotid atherosclerosis in healthy middle‐aged men
Authors: Boquist S. , Ruotolo G. , Skoglund‐Andersson C. , Tang R. , Björkegren J. , Bond M. G. , De Faire U. , Brismar K. , Hamsten A.
Published: September 2007

Objectives  IGF‐I, IGFBP‐1 and IGFBP‐3 are putative mediators in cardiovascular disease. The present study examined (i) the correlations of circulating IGF‐I, IGFBP‐1 and IGFBP‐3 to established cardiovascular risk factors and signs of early atherosclerosis as reflected by ultrasound measurement of common carotid intima–media thickness (IMT), and (ii) whether serum concentrations of these analytes are modulated during alimentary lipaemia.

Design  Cross‐sectional clinical study.

Patients  A biobank and clinical database based on 96 healthy Caucasian men, aged 50 years, with an apolipoprotein (apo) E3/E3 genotype, who had originally undergone investigations of postprandial lipoprotein metabolism was used for the study.

Measurements  Total IGF‐I, IGFBP‐1 and IGFBP‐3 were determined in serum by radioimmunoassay (RIA). Free IGF‐I was measured by a commercial two‐site immunoradiometric assay (IRMA).

Results  In multivariate analyses, fasting serum free IGF‐I correlated inversely with IMT and accounted for 5% of the variation in multiple R2. When fasting serum IGFBP‐1 was entered in the models instead of IGF‐I, IGFBP‐1 correlated positively with IMT and accounted for 6% of the variation in IMT. IGFBP‐3 and total IGF‐I were unrelated to IMT. There were no associations between free IGF‐I and cardiovascular risk factors, whereas IGFBP‐1 behaved like a component of the insulin resistance syndrome. Serum free IGF‐I increased and IGFBP‐1 decreased postprandially.

Conclusion  The data indicate that serum free IGF‐I and IGFBP‐1 are implicated in early atherosclerosis.
Modified‐release hydrocortisone for circadian therapy: a proof‐of‐principle study in dexamethasone‐suppressed normal volunteers
Authors: Newell‐Price J. , Whiteman M. , Rostami‐Hodjegan A. , Darzy K. , Shalet S. , Tucker G. T. , Ross R. J. M.
Published: September 2007

Background  All existing long‐term glucocorticoid replacement therapy is suboptimal as the normal nocturnal rise and waking morning peak of serum cortisol is not reproduced.

Aim  To test whether it is possible to reproduce the normal overnight rise and morning peak in serum cortisol using an oral delayed and sustained release preparation of hydrocortisone (Cortisolds).

Subjects and methods  Six healthy normal male volunteers attended on two occasions, in a single‐dose, open‐label, nonrandomized study. Endogenous cortisol secretion was suppressed by administration of dexamethasone. Cortisolds (formulation A or B) was administered at 2200 h on day 1. Blood samples for measurement of cortisol were taken from 2200 h every 30 min until 0700 h, then hourly until 2200 h on day 2. Fifteen body mass index (BMI)‐matched control subjects had serum cortisol levels measured at 20‐min intervals for 24 h. Serum cortisol profiles and pharmacokinetics after Cortisolds were compared with those in controls.

Results  Formulations A and B were associated with delayed drug release (by 2 h and 4 h, respectively), with median peak cortisol concentrations at 4·5 h (0245 h) and 10 h (0800 h), respectively, thereby reproducing the normal early morning rise in serum cortisol. Total cortisol exposure was not different from controls.

Conclusions  For the first time we have shown that it is possible to mimic the normal circadian rhythm of circulating cortisol with an oral modified‐release formulation of hydrocortisone, providing the basis for development of physiological circadian replacement therapy in patients with adrenal insufficiency.
Effects of serum TSH and FT4 levels and the TSHR‐Asp727Glu polymorphism on bone: the Rotterdam Study
Authors: Van Der Deure Wendy M. , Uitterlinden André G. , Hofman Albert , Rivadeneira Fernando , Pols Huibert A. P. , Peeters Robin P. , Visser Theo J.
Published: September 2007

Background  TSH and thyroid hormone may have independent effects on bone. In this study we investigated the association of TSH and free T4 (FT4) with different bone parameters in human subjects. TSH and FT4 are known to be associated with body mass index (BMI) and a higher BMI gives a higher bone mineral density (BMD). Thus, we aimed to determine whether the effects of TSH and FT4 on bone are mediated by BMI. As TSH exerts its biological effect through the TSH receptor (TSHR), the TSHR gene might be a candidate gene affecting bone mass. The TSHR‐Asp727Glu polymorphism is associated with lower TSH levels. We therefore examined the association of this polymorphism with bone parameters.

Method  Genotypes were determined by Taqman assay in 4934 elderly Caucasian men and women of the Rotterdam Study, of whom BMD and bone geometry data were available. Serum thyroid parameters were available in a random set of 1327 subjects,.

Results  Femoral neck BMD as well as narrow neck BMD and cortical thickness increased with serum TSH. However, FT4 was more strongly and negatively associated with bone parameters. Regression models showed BMI‐dependent and ‐independent effects of both TSH and FT4 on bone. Carriers of the TSHR‐Glu727 allele had a 2·3% higher femoral neck BMD.

Conclusion  In line with the effect of TSH on bone in mice, serum TSH shows a positive trend with BMD in human subjects, a finding that is strengthened by the association between the TSHR‐Asp727Glu polymorphism and femoral neck BMD. However, serum FT4 has a much greater influence on bone than TSH.
The degree of external genitalia virilization in girls with 21‐hydroxylase deficiency appears to be influenced by the CAG repeats in the androgen receptor gene
Authors: Rocha Rosana O. , Billerbeck Ana Elisa C. , Pinto Emília M. , Melo Karla F. S. , Lin Chin J. , Longui Carlos A. , Mendonca Berenice B. , Bachega Tânia A. S. S.
Published: September 2007

Background  Women with 21‐hydroxylase deficiency present much variability in external genitalia virilization, even among those with similar impairments of 21‐hydroxylase (21OH) activity.

Objective  To evaluate if the number of CAG (nCAG) repeats of the androgen receptor gene influences the degree of external genitalia virilization in women with CYP21A2 mutations, grouped according to impairment of 21OH activity.

Patients  The nCAG was determined in 106 congenital adrenal hyperplasia (CAH) patients and in 302 controls. The patients were divided, according to their CYP21A2 genotypes, into Groups A and B, which confer total and severe impairment of 21OH activity, respectively.

Methods  The inactivation pattern of the X‐chromosome was studied through genomic DNA digestion with Hpa II. The CAG repeat region was amplified by polymerase chain reaction (PCR) and analysed by GeneScan.

Results  The nCAG and the frequency of severe skewed X‐inactivation did not differ between normal women and patients. The nCAG median in genotype A was 20·7 (IQR 2·3) for Prader I + II, 22·5 (3·6) for Prader III and 21 (2·9) for Prader IV + V (P < 0·05 for Prader III and Prader IV + V). The nCAG median in genotype B was 21·3 (1·1) for Prader I + II, 20·5 (2·9) for Prader III and 22 (2·8) for Prader IV + V (P > 0·05). A significant difference was found regarding the nCAG median in patients presenting Prader III from genotypes A and B.

Conclusions  We observed great variability in the degree of external genitalia virilization in both CYP21A2 genotypes, and we showed that the CAG repeats of the androgen receptor gene influences this phenotypic variability.
Cognitive impairment is prevalent in pseudohypoparathyroidism type Ia, but not in pseudopseudohypoparathyroidism: possible cerebral imprinting of Gsα
Authors: Mouallem M. , Shaharabany M. , Weintrob N. , Shalitin S. , Nagelberg N. , Shapira H. , Zadik Z. , Farfel Z.
Published: September 2007

Objective  Pseudohypoparathyroidism type Ia (PHP‐Ia) is a hereditary disorder characterized by resistance to multiple hormones that work via cAMP such as PTH and TSH, accompanied by typical skeletal features including short stature and brachydactyly, termed Albright hereditary osteodystrophy (AHO). In affected kindreds, some members may have AHO but not hormone resistance; they are termed as pseudopseudohypoparathyroidism (PPHP). The molecular basis for the disorder is heterozygous inactivating mutation of the Gsα gene. In affected families, subjects with both PHP‐Ia and PPHP have the same Gsα mutations. The skeletal features common to PPHP and PHP‐Ia are presumably caused by tissue‐specific Gsα haploinsufficiency. Other features that distinguish between PPHP and PHP‐Ia, such as the multihormone resistance, are presumably caused by tissue‐specific paternal imprinting of Gsα. This suggests that major differences in phenotype between PHP‐Ia and PPHP point to specific tissues with Gsα imprinting. One such major difference may be cognitive function in PHP‐Ia and PPHP.

Design  Description of a large family with PHP‐Ia and PPHP.

Patients  Eleven affected subjects with PHP‐Ia or PPHP in one family.

Measurements  Cognitive impairment (CI) was defined by a history of developmental delay, learning disability and the Wechsler intelligence scale.

Results  CI occurred only in the five PHP‐Ia but not in the six PPHP subjects. Hypothyroidism which occurred in all PHP‐Ia subjects was apparently not the cause of CI as it was mild, and was treated promptly. Analysis of additional Israeli cases, and the published cases from the literature, all with documented Gsα mutations, revealed that CI is prevalent in PHP‐Ia [60 of 77 subjects (79%)] but not in PPHP [3 of 30 subjects (10%)] (P < 1 × 10−6).

Conclusion  We suggest that Gsα is imprinted in the brain.
An assessment of catches and harvest of recreational shore angling in the north of Portugal
Authors: RANGEL M. O. , ERZINI K.
Published: September 2007

Abstract  Recreational shore angling in the Atlantic Ocean between Moledo and Aveiro (Portugal) was examined using roving creel surveys (March/September 2001). Cooperation was high (90% of 2310 anglers approached). At least 39 species of fish were caught at a rate of approximately 0.5 fish angler h−1. An estimated 7319 kg of Dicentrarchus labrax (L.), with 45.6% below the minimum legal size, and 2040 kg of sea breams (genus Diplodus), correspond to 5.75% and 1.19% of the commercial landings in the same geographical area, respectively. The results shed light on a number of issues relevant to integrated coastal management, including temporal and spatial distribution of fishing effort, species caught, sizes of fish, catch rates, and factors influencing catches and angler satisfaction. Whilst the results suggest that the catches of sea breams and sea bass by day‐time recreational shore angling in northern Portugal are small compared with commercial fishing, other recreational activities, such as boat fishing and spear‐fishing, must be assessed.
PAX6 and SOX2‐dependent regulation of the Sox2 enhancer N‐3 involved in embryonic visual system development
Authors: Inoue Masashi , Kamachi Yusuke , Matsunami Hideyuki , Imada Katsumi , Uchikawa Masanori , Kondoh Hisato
Published: September 2007

Sox2 is universally expressed in the neural and placodal primordia in early stage embryos, and this expression depends on various phylogenetically conserved enhancers having different regional and temporal specificities. The enhancer N‐3 was identified as a regulator of the Sox2 gene active in the diencephalon, optic vesicle, and after the contact of the vesicle with the ectoderm, in the lens placodal surface area, suggesting its involvement in embryonic visual system development. A 36‐bp minimal essential core sequence was defined in the 568‐bp‐long enhancer N‐3, which in a tetrameric form emulates the original enhancer activity. The core sequence comprises a SOX‐binding sequence and a non‐canonical PAX6 (Paired domain) binding sequence, and is activated by the synergistic action of SOX2 and PAX6 in transfected cells. The SOX and PAX6 binding sequences of the N‐3 core are arranged with the same orientation and spacing as the DC5 sequence of the δ‐crystallin enhancer previously demonstrated to be cooperatively bound by SOX2 and PAX6. The N‐3 core sequence was also bound by these factors in a cooperative fashion, but with a higher threshold of these factors’ levels than DC5, and the enhancer effect of the tetrameric sequence activated by exogenous SOX2 and PAX6 was less pronounced than that of DC5. The observations suggest that gene activation mechanisms that depend on the cooperative interaction of SOX2 and PAX6 but with different thresholds of the factor levels are crucial for the regulation of visual system development.
Depth migration anisotropy analysis in the time domain
Authors: Kumar Chandan , Sen Mrinal K. , Ferguson Robert J.
Published: September 2007

In areas of complex geology such as the Canadian Foothills, the effects of anisotropy are apparent in seismic data and estimation of anisotropic parameters for use in seismic imaging is not a trivial task. Here we explore the applicability of common‐focus point (CFP)‐based velocity analysis to estimate anisotropic parameters for the variably tilted shale thrust sheet in the Canadian Foothills model. To avoid the inherent velocity‐depth ambiguity, we assume that the elastic properties of thrust‐sheet with respect to transverse isotropy symmetry axis are homogeneous, the reflector below the thrust‐sheet is flat, and that the anisotropy is weak. In our CFP approach to velocity analysis, for a poorly imaged reflection point, a traveltime residual is obtained as the time difference between the focusing operator for an assumed subsurface velocity model and the corresponding CFP response obtained from the reflection data. We assume that this residual is due to unknown values for anisotropy, and we perform an iterative linear inversion to obtain new model parameters that minimize the residuals. Migration of the data using parameters obtained from our inversion results in a correctly positioned and better focused reflector below the thrust sheet. For traveltime computation we use a brute force mapping scheme that takes into account weakly tilted transverse isotropy media. For inversion, the problem is set up as a generalized Newton's equation where traveltime error (differential time shift) is linearly dependent on the parameter updates. The iterative updates of parameters are obtained by a least‐squares solution of Newton's equations. The significance of this work lies in its applicability to areas where transverse isotropy layers are heterogeneous laterally, and where transverse isotropy layers are overlain by complex structures that preclude a moveout curve fitting.
Pathogenesis of haemophilic synovitis: clinical aspects
Authors: HOOTS W. K. , RODRIGUEZ N. , BOGGIO L. , VALENTINO L. A.
Published: September 2007

Summary.  Arthropathy remains a major cause of morbidity in patients with haemophilia. Frequent bleeding into the joints leads to joint damage with resultant contractures, joint deformities and arthritis. This in turn leads to muscle atrophy, limited physical activity, osteoporosis and disability. Even though several studies of prophylactic factor replacement for persons with severe haemophilia demonstrate improved joint function, this therapy is still not readily available to most people with haemophilia around the world and a universal treatment protocol has not been used. In this article, we discuss key issues in the treatment of severe haemophilia: the optimal timing of initiation and termination of therapy, dosing options and goals of therapy. The options for countries where prophylaxis is not readily available are also discussed. Most studies are small and not randomized making consensus treatment recommendations difficult to formulate. Randomized, clinical trials are needed to provide the answers regarding the optimal treatment of patients with severe haemophilia.
Diagnosis of haemophilic synovitis
Authors: GODDARD N. J. , MANN H.
Published: September 2007

Summary.  The pathophysiology of haemophilic synovitis is complex and not as well understood on a molecular level as we have thought. The repeated bleeding episodes and synovial hypertrophy occurring within the joint. Iron and other red‐cell debris are phagocytized, and hydrolytic enzymes are produced leading to the destruction of articular cartilage is a simplistic model. However, how iron stimulates this destructive cascade remains speculative and poorly understood. Diagnostic options include radiographic examination using horizontal beam technique, which may reveal a fat blood fluid level, and computed tomography scanning or magnetic resonance imaging.
Prevention of haemophilic synovitis: prophylaxis
Authors: LEE C. A.
Published: September 2007

Summary.  It has been proven that early prophylactic therapy can prevent bleeding and arthropathy. Numerous retrospective non‐randomized cohort studies have demonstrated that prophylaxis, if started early in life, is associated with a considerable reduction of the mean number of joint bleeds and the rate of joint deterioration. It is quite extraordinary that despite the considerable evidence base it has been considered necessary by investigators to pursue the ideal of the controlled randomized trial and expose children to the risk of cerebral bleed. This questionable ethical approach is driven by the reluctance of the ‘willingness to pay’ but it is important that patients are not subjected to unnecessary investigation at either the behest of the Cochrane Database or those who control the financing of haemophilia care.
Female red squirrels fit Williams’ hypothesis of increasing reproductive effort with increasing age
Authors: DESCAMPS SÉBASTIEN , BOUTIN STAN , BERTEAUX DOMINIQUE , GAILLARD JEAN‐MICHEL
Published: September 2007

Williams predicted that reproductive effort should increase as individuals age and their reproductive value declines. This simple prediction has proven difficult to test because conventional measures of energy expenditure on reproduction may not be a true reflection of reproductive effort.

We investigated age‐specific variation in female reproductive effort in a stable population of North American red squirrels where energy expenditure on reproduction is likely to reflect actual reproductive effort. We used seven measures of reproductive effort spanning conception to offspring weaning.

We found that females completed growth by age 3 and that reproductive value decreased after this age likely because of reproductive and survival senescence. We therefore, predicted that reproductive effort would increase from age 3 onwards.

The probability of breeding, litter mass at weaning, and likelihood of territory bequeathal were all lower for 1‐ and 2‐year‐old females than for females older than 3 years, the age at which growth is completed. That growing females are faced with additional energetic requirements might account for their lower allocation to reproduction as compared with older females.

The probability of attempting a second reproduction within the same breeding season and the propensity to bequeath the territory to juveniles increased from 3 years of age onwards, indicating an increase in reproductive effort with age. We think this increase in reproductive effort is an adaptive response of females to declining reproductive values when ageing, thereby supporting Williams’ prediction.
Portion size selection of fruits and vegetables by 9‐ to 10‐year‐old children in Liverpool
Authors: Mullarkey D. , Johnson B. , Hackett A.
Published: September 2007

Background  It is important to promote the intake of fruit and vegetables but it is not clear how children interpret the term ‘portion’. The aim of this study was to describe portion sizes of various fruits and vegetables selected by primary school children.

Methods  A cross‐sectional study collected data on children’s self‐selected portions of fruits and vegetables. Forty‐three boys and 52 girls aged 9–10 years attending after school food clubs in Liverpool took part. Children placed a ‘portion’ of each fruit and vegetable into an empty bowl which was weighed using digital scales accurate to 1 g.

Results  Mean portion sizes ranged from 126 g for satsumas to 61 g for peas but there were no significant differences between boys and girls. Portions of fruits were larger than portions of vegetables and although there were statistically significant correlations between the portion sizes selected by individual children they were not particularly strong.

Conclusions  These children had all taken part in activities promoting ‘5‐a‐day’ but their perception of ‘a portion’ varied enormously. It is recommended that children receive targeted activities to help them improve their understanding of what constitutes a ‘portion’.
Cryo‐fluorescence microscopy facilitates correlations between light and cryo‐electron microscopy and reduces the rate of photobleaching
Authors: SCHWARTZ CINDI L. , SARBASH VASILY I. , ATAULLAKHANOV FAZOIL I. , MCINTOSH J. RICHARD , NICASTRO DANIELA
Published: September 2007

Fluorescence light microscopy (LM) has many advantages for the study of cell organization. Specimen preparation is easy and relatively inexpensive, and the use of appropriate tags gives scientists the ability to visualize specific proteins of interest. LM is, however, limited in resolution, so when one is interested in ultrastructure, one must turn to electron microscopy (EM), even though this method presents problems of its own. The biggest difficulty with cellular EM is its limited utility in localizing macromolecules of interest while retaining good structural preservation. We have built a cryo‐light microscope stage that allows us to generate LM images of vitreous samples prepared for cryo‐EM. Correlative LM and EM allows one to find areas of particular interest by using fluorescent proteins or vital dyes as markers within vitrified samples. Once located, the sample can be placed in the EM for further study at higher resolution. An additional benefit of the cryo‐LM stage is that photobleaching is slower at cryogenic temperatures (−140°C) than at room temperature.
A handheld support system to facilitate stereological measurements and mapping of branching structures
Authors: GARDI J.E. , WULFSOHN D. , NYENGAARD J.R.
Published: September 2007

BranchSampler is a system for computer‐assisted manual stereology written for handheld devices running Windows CE. The system has been designed specifically to streamline data collection and optimize sampling of tree‐like branching structures, with particular aims of reducing user errors, saving time, and saving data in formats suited for further analysis in other software, for example, a spreadsheet. The system can be applied in a wide range of applications, from biomedical science to agriculture and horticulture. It can be applied for sampling nested generations of lung bronchioles and renal arterioles or for collection and optimizing sampling of crops for precision agriculture. Although the system has been designed specifically for sampling branching structures, it is sufficiently flexible to be used for other applications involving nested stereological designs. We describe the system specifications, software and Graphical User Interface development, functionality and application of the handheld system using four examples: (a) sampling monkey lung bronchioles for estimation of diameter and wall thickness (b) sampling rat kidney for estimating number of arteries and arterioles in a specific generation (c) mapping fruit (apple) tree yield in an orchard and (d) estimating the total leaf surface area of chrysanthemum plants in a greenhouse.
Antimicrobial susceptibility and mechanism of resistance to fluoroquinolones in Staphylococcus intermedius and Staphylococcus schleiferi
Authors: INTORRE L. , VANNI M. , DI BELLO D. , PRETTI C. , MEUCCI V. , TOGNETTI R. , SOLDANI G. , CARDINI G. , JOUSSON O.
Published: September 2007

The objective of the present study was to determine the antimicrobial susceptibility of 136 canine isolates of Staphylococcus intermedius and 10 canine isolates of S. schleiferi subspecies coagulans to 16 fluoroquinolones (FQs), and to investigate the mechanisms of resistance in the nonsusceptible isolates. Of the 136 of S. intermedius tested 98.5% were susceptible to all 16 FQs whereas only 40% of the 10 isolates of S. schleiferi subspecies coagulans were susceptible. Two isolates of S. intermedius and six isolates of S. schleiferi, were found to be resistant to 13 out of 16 FQs, while they retained their susceptibility to fourth generation FQs such as gatifloxacin, moxifloxacin and trovafloxacin. Sequencing of the quinolone‐resistance determining regions of gyrA and grlA genes showed that in S. intermedius, dichotomous resistance to FQs was associated with the occurrence of one alteration in GyrA‐84 and one in GrlA‐80, while in S. schleiferi the same pattern of resistance was observed in isolates showing these changes only in gyrA. This study is the first to screen FQs of the second, third and fourth generation for antimicrobial resistance in clinical isolates of S. intermedius and S. schleiferi of canine origin, and to describe mutations in gyrA and grlA associated with FQ resistance in these bacterial species.
Landscape genetics of the self‐compatible forest herbGeum urbanum: effects of habitat age, fragmentation and local environment
Authors: VANDEPITTE K. , JACQUEMYN H. , ROLDÁN‐RUIZ I. , HONNAY O.
Published: September 2007

To investigate the role of habitat fragmentation, fragment age and local environment in shaping the genetics of plant populations, we examined the genetic structure of the self‐compatible forest herbGeum urbanumusing microsatellite markers. A historical land‐use reconstruction assigned the studied populations to two age classes: populations in primary forest fragments, and populations in secondary fragments. Local environmental conditions were quantified on the basis of the herb‐layer community composition. A stepwise general linear model revealed that levels of within‐population genetic diversity were best explained by population size, landscape connectivity and the interaction between both. Connectivity was positively correlated with the genetic diversity of small populations, but did not significantly affect the diversity of large populations. Contrary to what we expected, secondary‐forest populations showed lower divergence relative to populations located in primary patches. Small populations were genetically more diverged compared to large populations. Mantel tests showed no significant isolation by distance and no significant correlation between habitat similarity and genetic differentiation. We conclude that gene flow has probably prevented founder events from being reflected in the present genetic structure ofG. urbanum. Gene flow towards low‐connectivity populations, however, seemed to be insufficient to counteract the effects of drift in small populations.
Mycoplasma genitalium: an efficient strategy to generate genetic variation from a minimal genome
Authors: Ma Liang , Jensen Jørgen S. , Myers Leann , Burnett Judy , Welch Mary , Jia Qiuyao , Martin David H.
Published: September 2007

Mycoplasma genitalium, a human pathogen associated with sexually transmitted diseases, is unique in that it has smallest genome of any known free‐living organism. The goal of this study was to investigate if and how M. genitalium uses a minimal genome to generate genetic variations. We analysed the sequence variability of the third gene (MG192 or mgpC) of the M. genitalium MgPa adhesion operon, demonstrated that the MG192 gene is highly variable among and within M. genitalium strains in vitro and in vivo, and identified MG192 sequence shifts in the course of in vitro passage of the G37 type strain and in sequential specimens from an M. genitalium‐infected patient. In order to establish the origin of the MG192 variants, we examined nine genomic loci containing partial copies of the MgPa operon, known as MgPar sequences. Our analysis suggests that the MG192 sequence variation is achieved by recombination between the MG192 expression site and MgPar sequences via gene cross‐over and, possibly, also by gene conversion. It appears plausible that M. genitalium has the ability to generate unlimited variants from its minimized genome, which presumably allows the organism to adapt to diverse environments and/or to evade host defences by antigenic variation.
Expression of the Fis protein is sustained in late‐exponential‐ and stationary‐phase cultures of Salmonella enterica serovar Typhimurium grown in the absence of aeration
Authors: Cróinín Tadhg Ó , Dorman Charles J.
Published: September 2007

The classic expression pattern of the Fis global regulatory protein during batch culture consists of a high peak in the early logarithmic phase of growth, followed by a sharp decrease through mid‐exponential growth phase until Fis is almost undetectable at the end of the exponential phase. We discovered that this pattern is contingent on the growth regime. In Salmonella enterica serovar Typhimurium cultures grown in non‐aerated SPI1‐inducing conditions, Fis can be detected readily in stationary phase. On the other hand, cultures grown with standard aeration showed the classic Fis expression pattern. Sustained Fis expression in non‐aerated cultures was also detected in some Escherichia coli strains, but not in others. This novel pattern of Fis expression was independent of sequence differences in the fis promoter regions of Salmonella and E. coli. Instead, a clear negative correlation between the expression of the Fis protein and of the stress‐and‐stationary‐phase sigma factor RpoS was observed in a variety of strains. An rpoS mutant displayed elevated levels of Fis and had a higher frequency of epithelial cell invasion under these growth conditions. We discuss a model whereby Fis and RpoS levels vary in response to environmental signals allowing the expression and repression of SPI1 invasion genes.
Borrelia burgdorferi adhesins identified using in vivo phage display
Authors: Antonara Styliani , Chafel Rebecca M. , LaFrance Michelle , Coburn Jenifer
Published: September 2007

Borrelia burgdorferi, the agent of Lyme disease, disseminates from the site of deposition by Ixodes ticks to cause systemic infection. Dissemination occurs through the circulation and through tissue matrices, but the B. burgdorferi molecules that mediate interactions with the endothelium in vivo have not yet been identified. In vivo selection of filamentous phage expressing B. burgdorferi protein fragments on the phage surface identified several new candidate adhesins, and verified the activity of one adhesin that had been previously characterized in vitro. P66, a B. burgdorferi ligand for β3‐chain integrins, OspC, a protein that is essential for the establishment of infection in mammals, and Vls, a protein that undergoes antigenic variation in the mammal, were all selected for binding to the murine endothelium in vivo. Additional B. burgdorferi proteins for which no functions have been identified, including all four members of the OspF family and BmpD, were identified as candidate adhesins. The use of in vivo phage display is one approach to the identification of adhesins in pathogenic bacteria that are not easily grown in the laboratory, or for which genetic manipulations are not straightforward.
Ultrastructural evidence for communication between intramuscular vagal mechanoreceptors and interstitial cells of Cajal in the rat fundus
Authors: Powley T. L. , Wang X.‐y. , Fox E. A. , Phillips R. J. , Liu L. W. C. , Huizinga J. D.
Published: September 2007

Abstract  To assess whether afferent vagal intramuscular arrays (IMAs), putative gastrointestinal mechanoreceptors, form contacts with interstitial cells of Cajal of the intramuscular type (ICC‐IM) and to describe any such contacts, electron microscopic analyses were performed on the external muscle layers of the fundus containing dextran‐labelled diaminobenzidin (DAB)‐stained IMAs. Special staining and embedding techniques were developed to preserve ultrastructural features. Within the muscle layers, IMA varicosities were observed in nerve bundles traversing major septa without contact with ICC‐IM, contacting unlabelled neurites and glial cells. IMA varicosities were encountered in minor septa in contact with ICC‐IM which were not necessarily in close contact with muscle cells. In addition, IMA varicosities were observed within muscle bundles in close contact with ICC‐IM which were in gap junction contact with muscle cells. IMAs formed varicosities containing predominantly small agranular vesicles, occasionally large granular vesicles and prejunctional thickenings in apposition to ICC‐IM processes, indicating communication between ICC and IMA via synapse‐like contacts. Taken together, these different morphological features are consistent with a hypothesized mechanoreceptor role for IMA‐ICC complexes. Intraganglionic laminar ending varicosities contacted neuronal somata and dendrites in the myenteric plexus of the fundus, but no contacts with ICC associated with Auerbach’s plexus were encountered.
The hallucinogenic herbSalvia divinorumand its active ingredient salvinorin A reduce inflammation‐induced hypermotility in mice
Authors: Capasso R. , Borrelli F. , Zjawiony J. , Kutrzeba L. , Aviello G. , Sarnelli G. , Capasso F. , Izzo A. A.
Published: September 2007

Abstract The hallucinogenic plantSalvia divinorumhas been used for medical treatments of gastrointestinal disorders. Here, we evaluated the effect of a standardized extract from the leaves ofSalvia divinorum(SDE) and of its active ingredient salvinorin A on motilityin vivo, both in physiological states and during croton oil‐induced intestinal inflammation. SDE (1–100 mg kg−1) significantly inhibited motility only in inflamed, but not in control, mice. In control mice, salvinorin A (0.01–10 mg kg−1) significantly inhibited motility only at the highest doses tested (3 and 10 mg kg−1) and this effect was not counteracted by naloxone or by the κ‐opioid receptor (KOR) antagonist nor‐binaltorphimine. Inflammation significantly increased the potency of salvinorin A (but not of the KOR agonist U‐50488) in reducing motility. The inhibitory effects of both salvinorin A and U‐50488 in inflamed mice were counteracted by naloxone or by nor‐binaltorphimine. We conclude that salvinorin A may reduce motility through activation of different targets. In physiological states, salvinorin A, at high doses, inhibited motility through a non‐KOR mediated mechanism. Gut inflammation increased the potency of salvinorin A; this effect was mediated by KOR, but it was not shared by U‐50488, thus suggesting that salvinorin A may have target(s) other than KOR in the inflamed gut.
Pemphigus vulgaris and pemphigus foliaceus: similar prognosis?
Authors: Zaraa I. , Mokni M. , Hsairi M. , Boubaker S. , Sellami M. , Zitouni M. , Makni S. , Dhahri A. Ben Osman
Published: September 2007

Background  Pemphigus is a rare, life‐threatening, acquired autoimmune bullous dermatosis. The prognosis of pemphigus foliaceus (PF) is usually regarded as more favourable than that of pemphigus vulgaris (PV). Our study aims to compare the clinical course of PV and PF in 37 patients.

Patients and Methods  We conducted a retrospective study over a period of eight years (1994–2001). The patients were referred during this period and were followed until December 2003. PF and PV were included based on clinical, histological and immunopathological criteria.

Results  In our study there was no significant difference between PF group and PV group concerning; age, sex, duration of the disease, presence of disseminated lesions, treatment, healing time, remission, relapse, complications, death and follows up duration. The survival graph showed no difference between the two groups for the first two relapses. There was a tendency to significance concerning an additional treatment and relapses frequency in the PF group.

Conclusions  Few studies in the literature were interested in the evolution of the two forms of pemphigus. They showed that the two populations share the same clinical course; nevertheless they revealed the frequency of partial remission, failed treatment, relapses, necessity of high dose of corticosteroids, and difficulties of discontinuing treatment in PF. Our study, suggests that PF and PV may share the same clinical course.