bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of regulated passive membrane conductance in action potential–firing fast- and slow-twitch muscle
Authors: Pedersen Thomas Holm , Macdonald William Alexander , de Paoli Frank Vincenzo , Gurung Iman Singh , Nielsen Ole Bækgaard
Published: September 2009

In several pathological and experimental conditions, the passive membrane conductance of muscle fibers (Gm) and their excitability are inversely related. Despite this capacity of Gm to determine muscle excitability, its regulation in active muscle fibers is largely unexplored. In this issue, our previous study (Pedersen et al. 2009. J. Gen. Physiol. doi:10.1085/jgp.200910291) established a technique with which biphasic regulation of Gm in action potential (AP)-firing fast-twitch fibers of rat extensor digitorum longus muscles was identified and characterized with temporal resolution of seconds. This showed that AP firing initially reduced Gm via ClC-1 channel inhibition but after ∼1,800 APs, Gm rose substantially, causing AP excitation failure. This late increase of Gm reflected activation of ClC-1 and KATP channels. The present study has explored regulation of Gm in AP-firing slow-twitch fibers of soleus muscle and compared it to Gm dynamics in fast-twitch fibers. It further explored aspects of the cellular signaling that conveyed regulation of Gm in AP-firing fibers. Thus, in both fiber types, AP firing first triggered protein kinase C (PKC)-dependent ClC-1 channel inhibition that reduced Gm by ∼50%. Experiments with dantrolene showed that AP-triggered SR Ca2+ release activated this PKC-mediated ClC-1 channel inhibition that was associated with reduced rheobase current and improved function of depolarized muscles, indicating that the reduced Gm enhanced muscle fiber excitability. In fast-twitch fibers, the late rise in Gm was accelerated by glucose-free conditions, whereas it was postponed when intermittent resting periods were introduced during AP firing. Remarkably, elevation of Gm was never encountered in AP-firing slow-twitch fibers, even after 15,000 APs. These observations implicate metabolic depression in the elevation of Gm in AP-firing fast-twitch fibers. It is concluded that regulation of Gm is a general phenomenon in AP-firing muscle, and that differences in Gm regulation may contribute to the different phenotypes of fast- and slow-twitch muscle.
Keeping Vulnerable Children Safe from Pertussis Preventing Nosocomial Pertussis Transmission in the Neonatal Intensive Care Unit
Authors: Greer Amy L. , Fisman David N.
Published: September 2009

Objective.

To examine the impact of different acellular pertussis booster vaccination strategies on the probability of a nosocomial pertussis outbreak occurring and the distribution of outbreak sizes observed for each intervention strategy.

Setting.

Neonatal intensive care unit.

Methods.

We developed a stochastic, agent-based simulation model to examine the impact of booster vaccination strategies for pertussis on health care-related transmission.

Results.

Our results demonstrate that healthcare worker booster vaccination decreases the probability of secondary transmission from 49% (base case, no boosting) to 2% (if 95% of healthcare workers are boosted) and decreases final outbreak size. Boosting family caregivers did not have a clinically significant impact on nosocomial disease transmission.

Conclusion.

The provision of booster vaccine to healthcare workers in the neonatal intensive care unit substantially reduces the risk of hospital-centered pertussis outbreaks in a manner that enhances the health of hospitalized children. A formal health economic analysis of this finding is currently under way. Policies to protect patient safety in pediatric facilities should include compliance with the United States Advisory Committee on Immunization Practices, which recommends provision of pertussis booster vaccination to healthcare workers.
Differential expression of phosphorylated extracellular signal‐regulated kinase 1/2, phosphorylated p38 mitogen‐activated protein kinase and nuclear factor‐κB p105/p50 in chronic inflammatory skin diseases
Authors: WANG Shuang , UCHI Hiroshi , HAYASHIDA Sayaka , URABE Kazunori , MOROI Yoichi , FURUE Masutaka
Published: September 2009

The keratinocytes actively participate in the cutaneous immune responses. Dysregulation and abnormal expression of inflammatory mediators or their receptors in keratinocytes are relevant to the pathogenesis of chronic inflammatory skin diseases. The mechanism of long‐lasting inflammatory processes is related with the activation of nuclear factor (NF)‐κB and mitogen‐activated protein kinase (MAPK), which play a crucial role in the immune responses. There are potential interaction points between these two pathways. The aim of this study is to investigate the differences in expression levels and distributions of phosphorylated extracellular signal‐regulated kinase (ERK)1/2, phosphorylated p38 MAPK and NF‐κB p105/p50 in chronic inflammatory skin diseases. An immunohistochemical staining technique was employed to measure the expression of these molecules in 25 cases of lichen planus, 22 cases of psoriasis, 26 cases of chronic eczema, seven cases of prurigo and seven cases of normal skin. We observed that the expression of phosphorylated ERK1/2, phosphorylated p38 MAPK and NF‐κB p105/p50 was significantly more augmented in the lesional epidermis of all the inflammatory skin diseases than those in normal skin (P < 0.05), and the number of positive keratinocytes was significantly more in lichen planus than that in other inflammatory diseases (P < 0.001). Moreover, the positive keratinocytes of these three molecules were more widely distributed in the entire layer of the epidermis in lichen planus than those in other diseases. We concluded that ERK1/2, p38 MAPK and NF‐κB p105/p50 might play important roles in the pathophysiology of chronic inflammatory skin diseases.
Case of pigmented neurofibroma with hypertrichosis with no association to neurofibromatosis
Authors: NA Chan Ho , SONG In Guk , CHUNG Byoung Soo , SHIN Bong Seok
Published: September 2009

Pigmented neurofibroma (PNF) is a rare cutaneous tumor that has been observed in patients with or without neurofibromatosis (NF). This tumor is histologically characterized by the coexistence of scattered melanin‐laden cells and benign spindle cells with neural differentiations. Hypertrichosis is the excessive growth of hair on non‐androgen‐dependent areas of the body. It has been reported that hypertrichosis may sometimes overlie a neurofibroma. We highlight a case of PNF with hypertrichosis on a 17‐year‐old woman with no associated NF. We also discuss the possible underlying pathogenic mechanism of a localized hypertrichosis in PNF patients.
Guidelines for management of atopic dermatitis
Authors: SAEKI Hidehisa , FURUE Masutaka , FURUKAWA Fukumi , HIDE Michihiro , OHTSUKI Mamitaro , KATAYAMA Ichiro , SASAKI Rikako , SUTO Hajime , TAKEHARA Kazuhiko ,   COMMITTEE for GUIDELINES for the MANAGEMENT of ATOPIC DERMATITIS of JAPANESE DERMATOLOGICAL ASSOCIATION
Published: September 2009

Atopic dermatitis (AD) is a chronic relapsing eczematous skin disease characterized by pruritus and inflammation and accompanied by cutaneous physiological dysfunction (dry and barrier‐disrupted skin). Most of the patients have atopic diathesis. A standard guideline for the management (diagnosis, severity classification and therapy) of AD has been established. In our guideline, the necessity of dermatological training is emphasized in order to assure diagnostic skill and to enable evaluation of the severity of AD. The definitive diagnosis of AD requires the presence of all three features: (i) pruritus; (ii) typical morphology and distribution; and (iii) chronic and chronically relapsing course. For the severity classification of AD, three elements of eruption (erythema/acute papules, exudation/crusts and chronic papules/nodules/lichenification) are evaluated in the most severely affected part of each of the five body regions (head/neck, anterior trunk, posterior trunk, upper limbs and lower limbs). The areas of eruption on the five body regions are also evaluated, and both scores are totaled (maximum 60 points). The present standard therapies for AD consist of the use of topical corticosteroids and tacrolimus ointment as the main treatment for the inflammation, topical application of emollients to treat the cutaneous physiological dysfunction, systemic antihistamines and anti‐allergic drugs as adjunctive treatments for pruritus, avoidance of apparent exacerbating factors, psychological counseling and advice about daily life. Tacrolimus ointment (0.1%) and its low‐density ointment (0.03%) are available for adult patients and 2–15‐year‐old patients, respectively. The importance of the correct selection of topical corticosteroids according to the severity of the eruption is also emphasized. Furthermore, deliberate use of oral cyclosporine for severe recalcitrant adult AD is referred.
Micropropagation by tissue culture triggers differential expression of infectious endogenous Banana streak virus sequences (eBSV) present in the B genome of natural and synthetic interspecific banana plantains
Authors: CÔTE FRANÇOIS X. , GALZI SERGE , FOLLIOT MICHEL , LAMAGNÈRE YANNICK , TEYCHENEY PIERRE‐YVES , ISKRA‐CARUANA MARIE‐LINE
Published: September 2009

The genome of Musa balbisiana spp. contains several infectious endogenous sequences of Banana streak virus (eBSV). We have shown previously that in vitro micropropagation triggers the activation of infectious eBSOLV (endogenous sequences of Banana streak Obino l'Ewai virus) in the synthetic tetraploid interspecific hybrid FHIA21 (AAAB). In this work, we show that another synthetic tetraploid (AAAB) hybrid and two natural triploid (AAB) plantains are equally prone to the activation of infectious eBSOLV during tissue culture. These results are a strong indication that such activation is a general phenomenon in interspecific Musa cultivars, whether synthetic or natural. We also report the first in‐depth study of the correlation between the duration of tissue culture and the level of activation of infectious eBSOLV, and show that specific and common activation patterns exist in these banana plants. We hypothesize that these patterns result from the concomitant activation of infectious eBSOLV and a decrease in the virus titre in neoformed plantlets, resulting from cell multiplication outcompeting virus replication. We provide experimental data supporting this hypothesis. No activation of infectious eBSGFV (endogenous sequences of Banana streak Goldfinger virus) by tissue culture was observed in the two natural AAB plantain cultivars studied here, whereas such activation occurred in the AAAB synthetic hybrid studied. We demonstrate that this differential activation does not result from differences in the structure of eBSGFV, as all banana genomes harbour eaBSGFV‐7.
Exploitation of Encephalartos transvenosus outside and inside Mphaphuli Cycads Nature Reserve, Limpopo Province, South Africa
Authors: Ravele A. M. , Makhado R. A.
Published: September 2009

This study has investigated the use and threats of Encephalartos transvenosus outside and inside Mphaphuli Cycads Nature Reserve, Limpopo Province, South Africa. Data were collected from 2005 to 2006 through social and ecological surveys. Semi‐structured interviews were conducted with a total of 40 people per household, sampled from four selected villages around the reserve. E. transvenosus is used for medicinal purposes, decorations, cultural activities and as food. Youth and adult persons harvest E. transvenosus for income generation, while children and aged people mainly use it for subsistence purposes. The survival of E. transvenosus in the wild is uncertain due to various threats, such as illegal collection, habitat destruction, fire and grazing. Some conservation measures are also discussed.
Effects of allergen and trigger factor avoidance advice in primary care on asthma control: a randomized‐controlled trial
Authors: Bobb C. , Ritz T. , Rowlands G. , Griffiths C.
Published: September 2009

Background
Allergy contributes significantly to asthma exacerbation, yet avoidance of triggers, in particular allergens, is rarely addressed in detail in regular asthma review in primary care.

Objective
To determine whether structured, individually tailored allergen and trigger avoidance advice, given as part of a primary care asthma review, improves lung function and asthma control.

Methods
In a randomized‐controlled trial 214 adults with asthma in six general practices were either offered usual care during a primary care asthma review or usual care with additional allergen and trigger identification (by skin prick testing and structured allergy assessment) and avoidance advice according to a standardized protocol by trained practice nurses. Main outcome measures were lung function, asthma control, asthma self‐efficacy.

Results
Both intervention groups were equivalent in demographic and asthma‐related variables at baseline. At 3–6‐month follow‐up, patients receiving the allergen and trigger avoidance review showed significant improvements in lung function (assessed by blinded research nurses) compared with those receiving usual care. Significantly more patients in the intervention group than in the control group showed improvements in forced expiratory volume in 1 s 15%. No significant differences were found in self‐report measures of asthma control. Asthma‐specific self‐efficacy improved in both groups but did not differ between groups.

Conclusions
Allergen and trigger identification and avoidance advice, given as part of a structured asthma review delivered in primary care by nurses results in clinically important improvements in lung function but not self‐report of asthma control.

Trial Registration
ISRCTN45684820.

Cite this as: C. Bobb, T. Ritz, G. Rowlands and C. Griffiths, Clinical & Experimental Allergy, 2010 (40) 143–152.
Modelling intercrop management impact on runoff and erosion in a continuous maize cropping system: Part II. Model Pareto multi‐objective calibration and long‐term scenario analysis using disaggregated rainfall
Authors: Laloy E. , Bielders C. L.
Published: September 2009

In order to evaluate the impact of intercrop management on runoff and erosion in a continuous maize cropping system, the plot scale, continuous and process‐based erosion model (CREHDYS) developed previously must be calibrated accounting for its two main outputs: runoff and sediment loss. To do that, a global Pareto multi‐objective calibration was applied to these two potentially conflicting objectives, considering daily runoff and periodical erosion rates, for two sites with different slopes and soil textures. This revealed a trade‐off between both objectives. The large resulting Pareto uncertainty regarding parameters did not translate into a large predictive uncertainty of daily runoff but resulted into a large uncertainty on erosion prediction. Globally, model results were satisfactory with regard to daily runoff prediction (Nash‐Sutcliffe index varying within the Pareto solution set from 0.65 to 0.91 for calibration and 0.64 to 0.77 for validation period) and relatively satisfactory for periodical erosion. However, the small number of available data points (three) for model validation in terms of periodical erosion prediction was not sufficient to ensure a proper validation. The calibrated model was in turn used to perform a scenario analysis of the long‐term hydrological and erosive impact of inter‐cropping period management in a continuous maize cropping system, using disaggregated rainfall. The long‐term simulations mainly revealed that, with regard to the erosion prevention during the inter‐cropping period, planting a winter cover crop is a better option than reduced tillage with a cultivator (0–12 cm), even if the cover is destroyed early (1 January). As compared with the situation of a bare heavily crusted soil with two semi‐permanent wheel tracks, reduced tillage led to an erosion reduction from 90 to 97%, an early cover destruction (1 January) to an erosion decrease from 92 to 98% and a cover destroyed on 1 March or later to an average soil loss reduction from 96 to 99%.
Beyond satisfaction, what service users expect of inpatient mental health care: a literature review
Authors: HOPKINS J. E. , LOEB S. J. , FICK D. M.
Published: September 2009

Accessible summary

Inpatient care is a necessary part of the care continuum for some mental health service users. This inpatient care is increasingly burdened by fewer beds, shorter lengths of stay and fewer resources.

Service users expect to have a safe environment in which to receive mental health treatment. They emphasize the expectation that this treatment will involve developing relationships with nurses and other staff.

They expect their inpatient care to provide one‐on‐one counselling, self‐help groups, educational sessions, informal opportunities for communication with knowledgeable and empathetic staff, and discharge planning.

If nurses are to meet these expectations, attention will need to be given to: providing nursing students with opportunities to practice individual counseling; creating a welcoming environment for new nurses to inspire them to select mental health nursing as their specialty and to encourage them to remain in the field; limiting the number of patients assigned to each nurse; and refocusing nurses away from office work and administrative duties and towards more contact with patients.

To provide efficient and effective inpatient mental health services, it is imperative to not only ascertain if service users are satisfied with the care received from nurses, but also the degree to which initial expectations are being met. Ten reports of primary research on service users' experiences, perceptions and expectations of inpatient mental health care were examined to understand what service users' expect of inpatient mental health care and the implications for nursing practice. The World Health Organization's description of responsiveness to service users' non‐medical expectations of care was used as a framework for retrieving literature and organizing the research outcomes. Responsiveness includes seven categories of healthcare performance ranging from respect for the dignity of the person, to adequacy of amenities, and choice of provider. Service users expect to form interpersonal relationships with nurses; however, non‐clinical responsibilities serve as barriers which consume considerable available nursing time that otherwise could be spent developing therapeutic relationships. In addition, inpatient programming ideas are identified for the provision of better services. Hospitals' expectations of mental health nurses will need to be reconsidered if these nurses are to provide the time and resources necessary to meet current service users' expectations.
Chapter 2: Source pigs
Authors: Schuurman Henk‐Jan
Published: September 2009

Abstract:  An islet xenotransplantation product includes live cells from a non‐human source, in this study, a pig source. A live product cannot be subjected to conventional disinfection, and therefore the source pig must be depleted of infectious agents that can transmit to the recipient and cause disease. Among other requirements, regulatory guidances specify that donor animals fulfill the designated pathogen‐free (DPF) status. Donor pigs fulfilling DPF status are generally bred and maintained in biosecure facilities, where they are shielded from the outside by filtered air, disinfected water, and irradiated food that is certified free of any mammalian protein. All materials are autoclaved upon entry, and personnel enter by shower‐in/shower‐out, and are wearing special clothes. The operation of such facilities is in compliance with Current Good Manufacturing Practices. To ensure DPF status, pigs are brought into such facilities via Cesarean section, and the source pigs are kept as a closed herd. The DPF status cannot be realized for endogenous viruses, such as porcine endogenous retrovirus. Therefore, regulatory authorities require patient monitoring after xenotransplantation. Considering the infectious pathogen status and necessary regulatory compliance, it is recommended that organ procurement be conducted at the animal facility and that cell manufacturing facilities be located nearby. To enable assessment of as‐yet unknown pathogens long after xenotransplantation, regulatory guidances mandate archiving donor materials for at least 50 yr. As this is essentially a public health issue, governmental institutions are urged to be responsible for the archive.

Table of Contents

Introduction

Designated pathogen‐free status

Biosecure barrier facility

Organ retrieval and islet manufacturing

Alternatives
Chapter 3: Pig islet product manufacturing and release testing
Authors: Korbutt Gregory S.
Published: September 2009

Abstract:  This chapter provides recommendations on pig islet product manufacturing and release testing to scientific and corporate programs interested in future clinical studies using xenogeneic porcine pancreatic islet cell products for the treatment of type 1 diabetes.To facilitate control of manufacturing as well as reproducibility and consistency of product lots, the manufacturing process, and the manufacturing facility must be in compliance with current Good Manufacturing Practices regulations. Data must be provided to demonstrate that islet products can be consistently prepared that would meet basic lot release requirements. To facilitate product safety: (i) materials used in the manufacturing process, including the pig pancreas, must be free of adventitious agents; (ii) islets must be manufactured using aseptic processing; and (iii) final product must undergo tests for sterility, mycoplasma (if cultured) and endotoxin. Safety specifications for pig islet product release include a negative Gram stain and an endotoxin content of <5.0 EU/kg recipient body weight. Product post‐release assessments must include sterility cultures on the final product. Because results for sterility are available only retrospectively, a plan of action must be in place for patient notification and treatment in case the sterility culture results are positive for contamination. Product characterization information must address important aspects of lot release testing such as identity/purity (cell composition), quantity [islet equivalents (IE), cell number] and potency (insulin secretory capacity, oxygen consumption rate corrected for DNA or transplant bioassay in immunoincompetent diabetic mice). This information is also critical to demonstrate manufacturing control and product consistency across multiple islet preparations (lots). Providing islet products containing an islet mass sufficient to restore euglycemia in trial participants (≥10 000 IE/kg) requires pooling of islets from multiple donor pancreata (two to four from adult donors and seven to 10 from neonatal donors). Demonstration of product consistency across products from individual pancreata would warrant release testing to be performed on a sample of the pooled product. As product development and clinical trials advance, the increasingly more detailed specifications of potency assays on adult porcine islet products are expected to be predictive of post‐transplant glycemic control. The immaturity of fetal and neonatal porcine islet tissue precludes the use of in vitro insulin secretion as a potency test as part of lot release testing; another measure of potency appropriate to fetal and neonatal cells will need to be developed for product release testing and evaluation of aliquots of these products in mouse transplant bioassays should be performed to provide meaningful post‐release information.

Table of Contents

Introduction

Animal source

Pancreas release criteria

Pig islet product manufacturing

Pig islet product release testing

Conclusion
Chapter 4: Pre‐clinical efficacy and complication data required to justify a clinical trial
Authors: Cooper David K. C. , Casu Anna
Published: September 2009

Abstract:  Pre‐clinical efficacy and complication data required to justify a porcine islet transplantation clinical trial have been considered. To be relevant to clinical islet transplantation, pre‐clinical data should be obtained in a pig‐to‐diabetic non‐human primate (NHP) model. In view of the difficulties of maintaining immunosuppressed NHPs for long periods of time, flexibility must be allowed with regard to the following recommendations. A pre‐clinical trial would be deemed a success if five of eight consecutive experiments in NHPs with proven diabetes meet the following guidelines.

1. The fasting and non‐fasting blood glucose levels have been maintained in the NHP at approximately <150 mg/dl (>8.3 mmol/l) and <200 mg/dl (11.1 mmol/l), respectively, throughout a 6‐month follow‐up period (preferably with observations in one or two NHPs extending to 12 months).

2. After the transplantation of adult or fetal/neonatal pig islets, no or greatly reduced exogenous insulin has been required after the development of a state of normoglycemia (that may take approximately the first 4 weeks for adult and 12 weeks for fetal/neonatal islets, respectively), except under exceptional, temporary circumstances.

3. Sequential intravenous glucose tolerance tests [at 1, 3, 6 (and 12) months post‐transplantation] or arginine stimulation tests have indicated a significant response to glucose in the form of porcine C‐peptide in the absence of a significant response of NHP C‐peptide.

4. Histologic study of the native pancreas (if present) after necropsy indicates no or minimal insulin‐positive β cells, and histologic examination of the liver or other site of islet transplantation indicates multiple viable insulin‐positive cells.

5. Numerous or serious life‐threatening complications have not been associated with the transplantation procedure, immunosuppressive regimen, tolerance‐inducing regimen, or encapsulation of the islets.

Pre‐clinical trials in which the recipient NHPs do not require continuing immunosuppressive therapy (e.g., when encapsulated islets have been transplanted) may possibly be considered acceptable as a basis for a clinical trial with slightly less stringent requirements with regard to points (1) and (2), but would be expected to provide evidence that points (3), (4), and (5) had been achieved.

Table of Contents

Introduction

Induction of diabetes in the non‐human primate potential recipient

Porcine islet transplantation

Immunotherapeutic protocol

Determination of success of islet transplantation

Conclusions
Structural and Histochemical Studies on the Teleostean Bulbus Arteriosus
Authors: Leknes I. L.
Published: September 2009

The structure and histochemical properties of the bulbus arteriosus in two species from an evolutionary old teleost family, Characidae, and in three modern teleosts, family Cichlidae, are described. The bulbar wall was composed of an outer layer, a middle layer and a strongly folded inner layer covered by a thick, granule‐rich endothelial cell layer towards the lumen. One of the cichlid species (Thorichthys meeki) was injected intraperitoneally with horse ferritin; the endothelial cell layer of the heart atrium and ventricle displayed high ability to endocytose ferritin particles from the blood stream, but the corresponding layer in the bulbus arteriosus displayed no such uptake. This finding suggests that the bulbar endothelial cell layer plays no scavenger or immunological blood cleansing roles in this species. The bulbar endothelial cell granules were strongly coloured by periodic acid–Schiff (PAS) in the present cichlids, but weakly coloured by PAS in the present characids. These cell layers were uncoloured by alkaline carmine in ethanol in both cichlids and characids. The negative carmine test combined with a positive PAS test for the bulbar endothelial cell layer in the present cichlids indicates that these cells contain only small amounts of polysaccharides. The weak PAS‐colouring for the bulbar endothelial cell layer in characids indicates a very low content of sugars in these cells. These findings together with the fact that this cell layer in the present cichlids and characids was nearly uncoloured when treated with orcein, Heidenhain’s Azan or Schmorl’s solutions for elastic materials suggest that the bulbar endothelial granules do not play any role in the blood cleansing or in the rebuilding or maintenance of the ground substance or elastic material in the bulbar wall. Probably, the granules in the bulbar endothelial cell layer in the present species contain mainly proteins, connected to some PAS‐positive polysaccharides to enhance their solubility.
Initiation of Steroidogenesis Precedes Expression of Cholesterologenic Enzymes in the Fetal Mouse Testes
Authors: Büdefeld T. , Jezek D. , Rozman D. , Majdic G.
Published: September 2009

Sexual differentiation is a carefully regulated process that ultimately results in a development of the male or female phenotype. Proper development of the male phenotype is dependent upon the action of testosterone and anti‐mullerian hormone. Leydig cells start to produce testosterone around day 12.5 in the fetal mouse testis, and continue to produce high levels of this hormone throughout gestation. In the present study, we examined whether expression of lanosterol 14α‐demethylase (cyp51) and cytochrome P450 NADPH reductase, both involved in the cholesterol production, occurs simultaneously with proteins required for the production of steroid hormones. Immunocytochemical staining with the antibodies against cyp51, cytochrome P450 NADPH reductase, steroidogenic acute regulatory protein (StAR) and 3beta‐hydroxysteroid dehydrogenase I (3β‐HSD I) was used to determine the ontogeny of expression of these four proteins. As expected, 3β‐HSD I and StAR proteins were detected on day 12.5 p.c., while expression of cyp51 and NADPH cytochrome P450 reductase appeared 1 day later, on day 13.5. Thereafter, the expression of all four proteins remained strong throughout gestation. Results of this study suggest that initial steps of steroid hormone production in murine Leydig cells are mostly dependent on exogenously derived cholesterol, while from day 13.5 onwards, mouse Leydig cells are able to synthesize cholesterol and are therefore not dependent on exogenous cholesterol resources.
Anatomicohistological Characteristics of the Female Genital Organs of the White‐lipped Peccary (Tayassu pecari) in the Peruvian Amazon
Authors: Mayor P. , Bodmer R. E. , Schettini L. , Mariño G. O. , López‐Béjar M.
Published: September 2009

This study examined the anatomical and histological characteristics of the genital organs of the female white‐lipped peccary in the wild in different reproductive stages, collected by rural hunters in the North‐eastern Peruvian Amazon. Mean ovulation rate was 2.12 ± 0.83 follicles and litter size was 1.78 ± 0.41 embryos or fetuses per pregnant female, resulting in a low rate of reproductive wastage, averaging 0.33 ± 0.66 (16.04%) oocytes or embryos per pregnancy. The ovulation rate and the anatomical performance of the uterus could limit the prolificacy of this species. Females in follicular phase showed follicular waves suggesting the synchronous growth of a cohort of follicles. Different uterine and vaginal epithelium features changed in accordance with the reproductive state of the female. Pregnant females and females in the luteal phase presented a significant proliferation of endometrial uterine glands, characterized by hyperplasia and branching of endometrial glands, and increase in the proportion of cervical epithelial cells with periodic acid‐schiff (PAS)‐positive granules compared with that in females in the follicular phase. Females in the follicular phase showed a more developed vaginal epithelium (in thickness and in layer composition) than females in the luteal phase and pregnant females.
7‐difluoromethyl‐5,4′‐dimethoxygenistein, a novel agent protecting against vascular endothelial injury caused by oxidative stress
Authors: Wang Li , Zheng Xing , Xiang Hong‐Lin , Fu Xiao‐Hua , Cao Jian‐Guo
Published: September 2009

1. Genistein is known to protect the vascular endothelium. However, genistein exhibits poor bioavailability, which limits its use in the treatment of cardiovascular diseases. 7‐Difluoromethyl‐5,4′‐dimethoxygenistein (dFMGEN), prepared by the difluoromethylation and alkylation of genistein, is a new active chemical entity. The protective effects of dFMGEN against vascular endothelial injury caused by oxidative stress were investigated in the present study.

2. Human umbilical vein endothelial cells were treated with either genistein (10 μmol/L) or various concentrations of dFMGEN (0.1, 0.3, 1, 3 and 10 μmol/L) for 30 min before exposure to 1 mmol/L H2O2 for 24 h. The generation of reactive oxygen species (ROS) was assessed by fluorescence flow cytometry, the release of lactate dehydrogenase (LDH) was examined by biochemical assay, cell viability was measured by the 3‐(4,5‐dimethyl‐2 thiazoyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide assay, cell apoptosis was detected by flow cytometry and the expression of caspase 3 was examined by western blot analysis.

3. Pretreatment with 0.1, 0.3, 1, 3 and 10 μmol/L dFMGEN decreased the generation of ROS and the release of LDH in H2O2‐exposed vascular endothelial cells, enhanced cell viability in a concentration‐dependent manner over the concentration range 0.1–10 μmol/L, suppressed H2O2‐induced apoptosis of vascular endothelial cells and downregulated the expression of caspase 3. The protective effect of 10 μmol/L dFMGEN against oxidative stress‐induced endothelial injury was stronger than that of 10 μmol/L genistein.

4. The results of the present study suggest that dFMGEN can protect against vascular endothelial injury caused by oxidative stress.
Association of exon 9 but not intron 8 VDR polymorphisms with occult HBV infection in south‐eastern Iranian patients
Authors: Arababadi Mohammad Kazemi , Pourfathollah Ali Akbar , Jafarzadeh Abdollah , Hassanshahi Gholamhossein , Rezvani Mohammad Ebrahim
Published: September 2009

Background and Aim:  Occult hepatitis B infection (OBI) is characterized as a form of hepatitis in which, despite the absence of detectable hepatitis B surface antigen (HBsAg), hepatitis B virus DNA (HBV–DNA) is present in a patient's peripheral blood. Investigators believe that divergent genetics and immunological parameters vary between resistant individuals and patients with OBI. Vitamin D3 and its known receptor appear to be involved in antiviral immune responses. Therefore, because OBI is a form of viral infection, the aim of this study was to investigate the association between polymorphisms in intron 8 and exon 9 of the vitamin D receptor (VDR) with OBI.

Methods:  In this experimental study, the plasma samples of 3700 blood donors were collected and tested for HBsAg and anti‐HBs using ELISA. The HBsAg–/anti‐HBc+ samples were selected and screened for HBV–DNA using polymerase chain reaction (PCR). HBV–DNA‐positive samples assigned as OBI cases and PCR–restricted fragment length polymorphism.

Results:  The results of the current study demonstrated that 352 (9.5%) of 3700 blood samples were HbsAg‐/anti‐HBc+. HBV–DNA was detected in 57/352 (16.1%) of HBsAg–/anti‐HBc+ samples. Our results showed a significant difference in the T/T allele of exon 9 of VDR, but any differences were also observed in the other examined alleles.

Conclusion:  The polymorphisms in the T/T allele of exon 9 of VDR is possibly associated with OBI, thus it can be concluded that VDR and its functional polymorphisms are likely to be related to sensitivity and resistance of the immune system to HBV in OBI patients.
Usefulness of non‐invasive markers for predicting liver cirrhosis in patients with chronic hepatitis B
Authors: Lee Kwang Gyun , Seo Yeon Seok , An Hyonggin , Um Soon Ho , Jung Eun Suk , Keum Bora , Yim Hyung Joon , Jeen Yoon Tae , Chun Hoon Jai , Kim Chang Duck , Ryu Ho Sang
Published: September 2009

Background and Aim:  Recently, various non‐invasive blood markers and indices have been studied to overcome the limitations of liver biopsy, such as its invasiveness and sampling errors. However, the majority of these studies have focused on patients with chronic hepatitis C. Accordingly, this study was performed to evaluate the significances of various non‐invasive serum markers in terms of predicting the presence of liver cirrhosis in chronic hepatitis B.

Methods:  We included 125 chronic hepatitis B patients who had undergone liver biopsy. Fibrosis stage was assessed using the METAVIR scoring system (F0–F4), which defines liver cirrhosis as F4. In addition, we measured various blood markers at times of liver biopsy.

Results:  Thirty four of the 125 patients (27.2%) were rated as F4 by liver biopsy. Age, platelet, white blood cells, aspartate aminotransferase (AST), alanine aminotransferase, haptoglobin, apolipoprotein‐A1 (Apo‐A1), collagen‐IV, hyaluronic acid, α2‐macroglobulin, matrix metalloproteinase‐2, and YKL‐40 were significantly different between patients with chronic hepatitis and those with liver cirrhosis. However, multivariate analysis showed that only platelet, AST, haptoglobin, and Apo‐A1 independently predicted the presence of liver cirrhosis. Having identified these four factors, we devised a system, which we refer to as platelet count, AST, haptoglobin, and Apo‐A1 (PAHA). The area under the receiver‐operating characteristics (AUROC) of PAHA indices for the presence of liver cirrhosis was 0.924 (95% confidence interval, 0.877–0.971), which was significantly greater than the AUROC of other indices of fibrosis.

Conclusion:  The devised PAHA system was found to be useful for predicting the presence of liver cirrhosis in patients with chronic hepatitis B.
Chronic pancreatitis in Chinese children: Etiology, clinical presentation and imaging diagnosis
Authors: Wang Wei , Liao Zhuan , Li Zhao‐Shen , Shi Xin‐Gang , Wang Luo‐Wei , Liu Feng , Wu Ren‐Pei , Zheng Jian‐Ming
Published: September 2009

Background and Aims:  There is a paucity of literature regarding the clinical profile of chronic pancreatitis (CP) in children. The aims of this retrospective study were to determine the etiology and clinical presentation, and to present our experience in diagnosing CP in children in China.

Methods:  Clinical data of children who were treated for CP at Changhai Hospital from January 1997 to August 2006 were reviewed.

Results:  A total of 427 CP patients presented to our center. There were 42 (9.8%) children with CP, including 21 males and 21 females, with a mean age of 11.7 years at the first onset. The main etiological factor was idiopathic (73.8%). Of the patients, 78.5% had episodes of mild to moderate abdominal pain and 54.8% had multiple (≥ 4) episodes. The mean duration of symptoms prior to the diagnosis was 41.6 months and a definite diagnosis was not made until 2 years later in 57.1% of these patients. The positive rates of ultrasound (US), computed tomography (CT) and magnetic resonance cholangiopancreatography (MRCP) (or magnetic resonance imaging [MRI]) for detecting ductal changes and intraductal stones or pancreatic calcification were 51.4% and 45.4%, 71.4% and 87.5%, 80.0% and 61.5%, respectively.

Conclusion:  The main etiological factor of Chinese children with CP is idiopathic. The main symptom in these patients is multiple episodes of mild to moderate abdominal pain, which often lead to a delay in the definite diagnosis. CT and MRCP (or MRI) should be used as the first investigation in the evaluation of these cases.
MicroRNA‐224 is upregulated in HepG2 cells and involved in cellular migration and invasion
Authors: Li Qiong , Wang Ge , Shan Jin‐Lu , Yang Zhi‐Xiang , Wang Hong‐Zhong , Feng Jin , Zhen Ji‐Jun , Chen Chuan , Zhang Zhi‐Min , Xu Wen , Luo Xi‐Zhong , Wang Dong
Published: September 2009

Background and Aim:  MicroRNAs are a class of small non‐coding RNAs that negatively regulate the expression of their target genes. The aim of the present study was to explore the effects of microRNA on biological behaviors of HepG2 cells and further analyze its characteristics.

Methods:  We detected different expression profiles of miRNAs in HepG2 and L02 cell lines by microRNA microarray. Northern blot, quantitative real‐time polymerase chain reaction, methylthiazolyl tetrazolium, fluorescence‐activated cell sorting, scratch wound, transwell migration and Matrigel invasion assays and western blot were carried out to determine whether or not microRNA‐224 (miR‐224) can influence the biological behaviors of HepG2 cells.

Results:  MiR‐224 was significantly upregulated in HepG2 cells. Cell proliferation, migration and invasion, but not cell cycles, were altered after changing the expression of miR‐224. Taking invasion and migration as a breakthrough, a close relationship between the expression of miR‐224 and its proteins such as PAK4 and MMP9, which were involved in the invasion of tumor, was found.

Conclusions:  Overexpression of miR‐224 was involved in the malignant phenotype of HepG2 cells, and it may be an important factor in regulating the migration and invasion of HepG2 cells.
Short‐term celecoxib to regress long‐term persistent gastric intestinal metaplasia after Helicobacter pylori eradication
Authors: Hung Kuei‐Hsiang , Yang Hsiao‐Bai , Cheng Hsiu‐Chi , Wu Jiunn‐Jong , Sheu Bor‐Shyang
Published: September 2009

Background and Aim:  The intestinal metaplasia (IM) has overexpressions of cyclooxygenase‐2 (COX‐2) and β‐catenin. This pilot study assessed whether celecoxib, a selective COX‐2 inhibitor, could regress IM that persisted long term after Helicobacter pylori eradication.

Methods:  Thirty‐three patients with H. pylori eradication were enrolled in the present study due to the persistence of IM after a 3‐year follow up. These patients received celecoxib 200 mg/day for 8 weeks, and were serially checked for levels of blood urea nitrogen and creatinine once per 2 weeks. After 8‐week celecoxib treatment, IM regression was assessed by panendoscopy. The gastric specimens, taken before and after celecoxib, were immunochemically stained for COX‐2 and β‐catenin.

Results:  The intention‐to‐treat and per‐protocol analyses to the rates of IM regression by 8‐week celecoxib treatment were 24.2% (8/33) and 28.6% (8/28), respectively. All enrolled patients had no renal impairment. Even in the patients without total IM regression, mean IM scores in the antrum decreased after 8‐week celecoxib treatment (P = 0.007). The patients with complete regression of IM after 8‐week celecoxib treatment had a significantly lower COX‐2 expression, but not β‐catenin expression, at enrollment than those patients without IM regression (P = 0.031).

Conclusion:  Short‐term celecoxib treatment can be safe and promising to regress long‐term persistent IM after H. pylori eradication.
Interstitial cells of Cajal are present in human extrahepatic bile ducts
Authors: Ahmadi Omid , De L Nicholson Martha , Gould Maree L , Mitchell Allan , Stringer Mark D
Published: September 2009

Background and Aims:  Interstitial cells of Cajal (ICC) are distributed with smooth muscle throughout the gastrointestinal tract and are involved in regulating motility. ICC were recently discovered in the wall of the human gallbladder. This study sought to determine whether ICC are present in human bile ducts.

Methods:  Biliary tract samples were obtained from several sources: surgical specimens (n = 16, 11 women, mean age 61 years); archival post‐mortem specimen (n = 1, 86 years, man); and cadavers (n = 2, 68 and 80 years, men). Paraffin‐embedded sections (3 µm) from the gallbladder (fundus, body and neck) and both extrahepatic and intrahepatic bile ducts were investigated. A double immunofluorescence protocol using polyclonal and monoclonal c‐kit antibodies and mast cell tryptase was used to distinguish c‐kit‐positive cells with typical ICC morphology from c‐kit‐positive mast cells. Small bowel samples were used as positive controls. ICC in the gallbladder were confirmed by ultrastructural study.

Results:  c‐kit‐positive cells with characteristic ICC morphology were identified in the subepithelial and muscular layers of the gallbladder and extrahepatic bile ducts. They were most prominent within the muscle layer of the extrahepatic bile ducts where they were organized into loosely arranged laminae running parallel to circular smooth muscle fibers. ICC were not found in intrahepatic bile ducts.

Conclusion:  This study demonstrates for the first time that ICC are present in human extrahepatic bile ducts where they are more densely aggregated than in the gallbladder. This cellular network is likely to be involved in biliary tract motility and its related disorders.
Compound Astragalus and Salvia miltiorrhiza extract inhibits cell invasion by modulating transforming growth factor‐β/Smad in HepG2 cell
Authors: Liu Xin , Yang Yan , Zhang Xiaoxiang , Xu Shengxin , He Shufang , Huang Weijuan , Roberts Michael S
Published: September 2009

Background and Aims:  Compound Astragalus and Salvia miltiorrhiza extract (CASE) is made up of astragalosides, astragalus polysaccharide and salvianolic acids extracted from Astragalus membranaceus Bunge (Leguminosae) and Salvia miltiorhiza Bunge (Lamiaceae) with a standard ratio. Previous reports showed that CASE inhibited hepatic fibrosis by mediating transforming growth factor (TGF)‐β/Smad signaling. This study further investigated the effect of CASE on hepatoma HepG2 cells stimulated by TGF‐β1 and its potential action mechanisms by TGF‐β/Smad signaling.

Methods:  Cell proliferation was studied by MTT assay and cell invasion was evaluated by measuring cell migration through Matrigel. Protein expression in hepatoma HepG2 cells stimulated by TGF‐β1 was analyzed by western blotting and plasminogen activator inhibitor type 1 (PAI‐1) transcriptional activity in HepG2 cells was evaluated.

Results:  CASE (40 µg/mL) markedly suppressed cell invasion triggered by TGF‐β1. Smad3 phosphorylation at the linker region (pSmad3L) and Samd2 phosphorylation at the C‐terminal region (pSmad2C) were significantly reduced by CASE. Mild elevated Smad3 phosphorylation at C‐terminal (pSmade3C) region was enhanced by CASE at 20 µg/mL. In addition, treatment of CASE decreased the level of Smad2/3/4 complex at 80 µg/mL, but upregulated the expression of Smad7 in a dose‐dependent manner. CASE also showed inhibitory effect on PAI‐1 transcriptional activity.

Conclusion:  All these results suggest that CASE exerts anti‐HepG2 cell invasion effect by modulating TGF‐β/Smad signaling.
Oral health status in epileptic children
Authors: Gurbuz Taskin , Tan Huseyin
Published: September 2009

Background:  The aim of the present study was to evaluate the oral hygiene status and dental treatment requirements in children with epilepsy.

Methods:  The treatment group consisted of 211 children with epilepsy (120 boys and 91 girls, 4–15 years old, mean age 7.85 ± 2.98 years). The control group consisted of healthy children, matched by age and gender. Clinical features of the patients were obtained from hospital records. Clinical examinations were conducted, under standard light, using a plane buccal mirror, a dental probe and air drying to evaluate caries experience and to record the periodontal health of each child. Statistical analysis was performed using χ2 test, Fisher exact test and anova.

Results:  The number of decayed and missing teeth, the degree of abrasion and periodontal indexes were significantly worse in patients with epilepsy, compared to the control group (P < 0.001). Gingival enlargement was documented in 42% of patients on valproate monotherapy compared to only in 16% of patients on phenobarbital. Dental caries and halitosis were the most common oral disorders. Generalized tonic–clonic seizures often cause minor oral injuries and traumatized anterior teeth.

Conclusions:  Epileptic children are at an increased risk of developing caries and gingivitis compared with healthy subjects.