Isolation of peptides from porcine intestinal tissue that induce extracellular acidification in CHO cells: identification of the active peptide as IGF-I and characterization of a fragment of calponin H1 processed at a dibasic site.
Chinese hamster ovary (CHO) cells are widely used as hosts for receptor expression and pharmacological studies. However, several endogenous receptor populations are present on these cells. Intestinal tissue extracts were found to induce strong extracellular acidification responses (ECAR) in CHO cells, yet several pure hormonal peptides, such as VIP, secretin, CCK, GIP, and galanin were ineffective. It is not known, which are the active compounds in the extracts that can stimulate the extracellular acidification in CHO cells. These active substances may be ligands for yet unknown receptors that are present natively in this cell type. We therefore decided to identify the active compound(s) by isolation from intestinal extract and structural characterization. Using chromatographic separations in combination with microphysiometry we have purified and characterized one such bioactive ligand. Structural analysis indicated that the isolated peptide was identical to insulin-like growth factor I (IGF-I). In the intestine, IGF-I is present in low amounts and has previously been detected only with radioimmunoassays. The results indicate that CHO cells express functional receptors for IGF-I. Among the peptides extracted from the intestine IGF-I is probably the strongest stimulator of ECAR in CHO cells. Moreover, IGF-I acts synergistically with other factors present in the crude tissue extract. Additionally, a fragment of calponin H1 (residues 1-43), previously not described at the protein level, was identified in the IGF-I containing fractions. The fragment was characterized by mass spectrometry and found to be N-terminally modified by acetylation suggesting that the whole protein bears the same posttranslational modification.


Delayed endometrial maturation induced by daily administration of the antiprogestin RU 486: a potential new contraceptive strategy.
Our objective was to determine if a progesterone antagonist might interdict the development of a secretory endometrium. Eleven normally cycling women not at risk for pregnancy received RU 486 (1 mg/day orally) or placebo throughout one menstrual cycle in a randomized, double-blind, crossover fashion. Estradiol, progesterone, and placental protein 14 were measured every 3 days; luteinizing hormone was measured until the midcycle surge was detected. An endometrial biopsy was performed on luteal phase day 7 to 9 and interpreted with Noyes' criteria. Differences between treatment groups were analyzed by the Student t test. RU 486 delayed ovulation, retarded endometrial maturation, and reduced peak levels of placental protein 14 without affecting gonadal steroid production. The abnormalities in endometrial morphology and function are similar to those seen in infertile women with luteal phase defects. We hypothesize that this regimen of antiprogestin administration may prevent implantation and offer a novel strategy for fertility control.


A Method to Detect Presence of Chest Compressions during Resuscitation using Transthoracic Impedance.
Interruptions in chest compressions during treatment of out-of-hospital cardiac arrest are associated with lower likelihood of successful resuscitation. Real-time automated detection of chest compressions may improve CPR administration during resuscitation, and could facilitate application of next-generation ECG algorithms that employ different parameters depending on compression state. In contrast to accelerometer sensors, transthoracic impedance (TTI) is commonly acquired by defibrillators. We sought to develop and evaluate the performance of a TTI-based algorithm to automatically detect chest compressions. Five-second TTI segments were collected from patients with out-of-hospital cardiac arrest treated by one of four defibrillator models. Segments with and without chest compressions were collected prior to each of the first four defibrillation shocks (when available) from each case. Patients were divided randomly into 40% training and 60% validation groups. From the training segments, we identified spectral and time-domain features of the TTI associated with compressions. We used logistic regression to predict compression state from these features. Performance was measured by sensitivity and specificity in the validation set. The relationship between performance and TTI segment length was also evaluated. The algorithm was trained using 1859 segments from 460 training patients. Validation sensitivity and specificity were >98% using 2727 segments from 691 validation patients. Validation performance was significantly reduced using segments shorter than 3.2 s. A novel method can reliably detect the presence of chest compressions using TTI. These results suggest potential to provide real-time feedback in order to improve CPR performance or facilitate next-generation ECG rhythm algorithms during resuscitation.


A succinylcholine-like action of an Ipomoea carnea Jacq subsp. fistulosa (Mart. ex choisy) extract.
An aqueous alcoholic extract of Ipomoea carnea subsp. fistulosa was tested in vitro on isolated preparations of the frog rectus abdominis muscle and the rabbit perfused heart as well as in vivo in the chicken. The extract and succinylcholine (suxamethonium) contracted the frog rectus abdominis muscle and this action was blocked by tubocurarine, 1.5 micrograms/ml. Both extract and succinylcholine produced bradycardia of the rabbit heart which was antagonized by atropine, 20 micrograms/ml. Like succinylcholine, the extract produced spastic paralysis in chickens. It is suggested that the extract possesses a depolarizing action similar to that of succinylcholine.


Essential oil composition and anti Acanthamoeba studies of Teucrium ramosissimum.
The aim of the present study was to evaluate the chemical composition of the essential oil obtained from the aerial parts of T. ramosissimum by hydrodistillation and to investigate their anti-Acanthamoeba activity. Identification and quantification were realized by gas chromatography-mass spectrometry (GC-MS) and gas chromatography with flame ionization detection by (GC-FID). Sixty-eight compounds representing 97.78% of the essential oil were identified, of which δ-cadinene (18.63%), δ-cadinol (18.70%), β-eudesmol (12.13%), γ-gurjunene (4.34%) and 8-cedrene (3.99%) were the main compounds. This essential oil contained a complex mixture consisting mainly on sesquiterpenes (80.62%) and monoterpene fractions (14.34%). The findings of the anti-Acanthamoeba assay indicate that T. ramosissimum essential oil have a good activity with an IC50 = 25.73 ± 0.75 μg/mL.


Residence background and fertility in Greater Bombay.
Abstract The residence background of wives who migrate to metropolitan areas plays an important role in determining their fertility. From the data collected during 1966, relating to 7,872 currently married women of Greater Bombay, an attempt was made to establish differentials in marital fertility by residence background of the wives. This was categorized into three groups - non-migrants, urban migrants and rural migrants. It was observed that rural migrant wives exhibited significantly higher fertility compared with the other two groups, and this was explained by their lower educational attainment. Between the non-migrant and the urban migrant wives the latter consistently showed lower fertility for all age groups up to 40, while there was a reversal in the age group 40 and above, where non-migrants exhibited lower fertility. The urban migrant wives showed a somewhat higher level of education, most likely on account of selectivity, compared to the non-migrants. However, presence of a sizeable number of Parsee wives, characterized by a distinct urban culture and considerably lower fertility, was largely responsible for the low fertility of the non-migrant wives in the age group 40 and above. The variable that has emerged as the most influential in creating fertility differentials is education of the wife, which is shown to be negatively associated with the level of fertility. Wife's education explains to a large extent the observed fertility differentials by residence background.


[Ultrasound imaging of the urethral sphincter].
The integrity and functional capacity of the urethral sphincter is one of the important prerequisites of urinary incontinence in women. Urodynamic investigations revealed repeatedly that the maximum closure pressure in the median portion of the urethra corresponds to the maximal thickness of the external urethral sphinctor (rhabdosphincter urethrae). This striated muscle is adapted to maintain a relatively steady tonus which assists the closure mechanism of the urethra [4]. In the submitted study the authors focused attention on the ultrasonic visualization of the internal urethral sphincter in order to assess the relationship between the size of this sphincter and the stress type of incontinence (genuine stress incontinence-GSI). The investigation comprised thirty women with confirmed GSI and a control group of thirty asymptomatic volunteers. During perineal ultrasonic examination of women in a supine position by means of an ACUSON 128 XP 10 apparatus using a convex probe with a frequency of 5 MHz the authors recorded statistically significant differences in the areas and maximal thickness of the urethral sphincter in women with stress incontinence and symptom-free women. From the results ensues that the size of this muscle is much smaller in women suffering from GSI.


[Surveillance and prevention of infections after open heart surgery].
From October 1987 to December 1989, bacteriological surveillance was done after open heart surgery in 100 patients with cardiovascular diseases (54 with congenital, 44 with rheumatic, and 2 with other types of heart disease). The time of bypass was 58 to 234 minutes (mean = 98.7 minutes). The positive rate of culture was 5% for blood, 40% for sputum, and 15% for pericardial drainage. Many types of bacteria were separated. Most of them were conditional pathogenic bacteria. Postoperative infection was found in 9 patients, (bacteremia in 1 and pulmonary infection in 8.) Tracheostomy was done in 5 cases. In discussion, the authors think that the low defensive ability of the patients with serious heart disease is usually further damaged by bacterial invasion after open heart surgery and that bacterial contamination should be reduced in order to prevent postoperative infection.


Characterization of an amphioxus paired box gene, AmphiPax2/5/8: developmental expression patterns in optic support cells, nephridium, thyroid-like structures and pharyngeal gill slits, but not in the midbrain-hindbrain boundary region.
On the basis of developmental gene expression, the vertebrate central nervous system comprises: a forebrain plus anterior midbrain, a midbrain-hindbrain boundary region (MHB) having organizer properties, and a rhombospinal domain. The vertebrate MHB is characterized by position, by organizer properties and by being the early site of action of Wnt1 and engrailed genes, and of genes of the Pax2/5/8 subfamily. Wada and others (Wada, H., Saiga, H., Satoh, N. and Holland, P. W. H. (1998) Development 125, 1113-1122) suggested that ascidian tunicates have a vertebrate-like MHB on the basis of ascidian Pax258 expression there. In another invertebrate chordate, amphioxus, comparable gene expression evidence for a vertebrate-like MHB is lacking. We, therefore, isolated and characterized AmphiPax2/5/8, the sole member of this subfamily in amphioxus. AmphiPax2/5/8 is initially expressed well back in the rhombospinal domain and not where a MHB would be expected. In contrast, most of the other expression domains of AmphiPax2/5/8 correspond to expression domains of vertebrate Pax2, Pax5 and Pax8 in structures that are probably homologous - support cells of the eye, nephridium, thyroid-like structures and pharyngeal gill slits; although AmphiPax2/5/8 is not transcribed in any structures that could be interpreted as homologues of vertebrate otic placodes or otic vesicles. In sum, the developmental expression of AmphiPax2/5/8 indicates that the amphioxus central nervous system lacks a MHB resembling the vertebrate isthmic region. Additional gene expression data for the developing ascidian and amphioxus nervous systems would help determine whether a MHB is a basal chordate character secondarily lost in amphioxus. The alternative is that the MHB is a vertebrate innovation.


Cytochalasin D induces changes in cell shape and promotes in vitro chondrogenesis: a morphological study.
One of the initial events required for the expression of cartilage-specific macromolecules in monolayer cultures is the reversion to the initial round shape of chondrocytes. Thus, considerable research efforts have focused on developing reliable procedures to maintain a round morphology of cultured chondrocytes. Our study focuses on evaluating the response of dedifferentiated fetal rat chondrocytes to cytochalasin D, an actin-disrupting agent, with special emphasis on the morphological events. Immediately after exposure to the drug, cells round up but flatten again after removing the agent. However, immunocytochemical procedures revealed a disorganization of microfilaments and intermediate filaments. Phase-contrast and scanning electron microscopic observations revealed that on day 6 of culture, cells located at the top of the cell layer adopted a spherical morphology. Prominent differences were noted in control cultures where cells had to aggregate prior to overt chondrogenesis. Transmission electron microscopy confirmed the round morphology of the cells situated at the top layer but also revealed the presence of cell contacts between the cells. In addition, cells located at the central part of the cell layer displayed a typical morphology of mature chondrocytes, separated by an extensive extracellular matrix. These morphological changes occurred parallel to the expression of type II collagen and chondroitin sulfate, both hallmarks of the chondrocyte phenotype strong in experimental cultures, relatively weak in control cultures, and only restricted on areas of polygonal cellular aggregates. Furthermore, [35S]-sulfate incorporation into sulfated glycosaminoglycans increased rapidly with the period of culture to a maximum after 7 days and was then two-fold in treated cultures. Taken together, these findings indicated that cytochalasin D stimulates chondrogenesis in response to modification of cytoskeleton architecture and the subsequent rounding up of the cells.


Human growth hormone treatment of children with growth failure and normal growth hormone levels by immunoassay: lack of correlation with somatomedin generation.
Children with growth failure, normal growth hormone responses to stimulation tests, and low somatomedin levels are being recognized with increasing frequency. Sixteen patients, aged 2 to 17 years, with these findings were studied. When treated with human growth hormone, the mean plasma somatomedin-C levels of these patients increased from 0.19 +/- 0.05 (SD) U/mL to 2.29 +/- 1.11 U/mL (P less than .001) and mean growth velocity increased from 3.6 +/- 1.6 (SD) cm/yr to 7.4 +/- 2.9 cm/yr at 8 months of treatment (P less than .001). The effect of human growth hormone therapy on the whole group was statistically significant, but the effect on individuals was highly variable. There was no correlation between magnitude of the increase in somatomedin-C and growth response (r = .26, NS). Thus, the long-term growth-promoting effect of human growth hormone therapy in this group of patients could not be predicted from the magnitude of the somatomedin-C responses.


[Identification of a very short YCAII allele in the northern Polish population].
The YCAII system is one of the Y-chromosomal STR markers with dinucleotide repetitive units and due to the duplication on the chromosome, its amplification provides two polymorphic products. Due to the high polymorphism, YCAII was included in the Y-STR Haplotype Reference Database (http://www.ystr.org) and significantly increased discrimination power of the collected haplotypes. During studies on the Y-STR polymorphism in the Polish population, a very short YCAII allele with relatively low stutter was identified. Sequencing revealed the presence of 11 dinucleotide repetitive units characteristic of this system. Among 3998 extended haplotypes collected in the database, this allele was found only in the population of south-western Germany. Similarities between the haplotypes possessing the YCAII*11 allele suggest their common origin.


Complex actions of serotonergic agonists on cold-stimulated TSH secretion in male rats.
The effects of serotonergic activation on cold-stimulated thyrotropin (TSH) and prolactin secretion were studied in male rats. Peripheral injections of both 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), a 5-HT1A receptor agonist, and 1-(3-chlorophenyl)piperazine (m-CPP), a 5-HT1 agonist, decreased TSH levels. The action of 8-OH-DPAT was antagonized by (+/-)-pindolol, which is known to have 5-HT1 antagonist activity, but not by metergoline or ketanserin. The action of m-CPP was antagonized by ketanserin but not by metergoline. TSH levels were not affected by a 5-HT3 receptor agonist, 2-methyl-5-HT, or by a 5-HT3 antagonist, MDL 72222. Infusion of 8-OH-DPAT into the anterior third ventricle increased TSH levels; 5-HT tended to increase TSH levels, but the effect was not significant. Inversely, infusion of 5-HT, 8-OH-DPAT or m-CPP into the posterior third ventricle decreased TSH levels. The action of 5-HT was counteracted by metergoline, ketanserin and (+/-)-pindolol. Unexpectedly, m-CPP infusion into the anterior third ventricle also inhibited TSH secretion. The prolactin-elevating effects of 5-HT, 8-OH-DPAT and m-CPP were neither consistent nor site-specific. In conclusion, stimulation of both 5-HT1 and 5-HT2 receptors may inhibit TSH secretion, but the exact mechanism underlying the site-dependent action of 5-HT and 8-OH-DPAT on TSH secretion remains to be identified.


From hospital to the community: the influence of deinstitutionalization on discharged long-stay psychiatric patients.
The aim of the present study is to clarify the influence of deinstitutionalization on discharged long-stay patients based on a review of recently published literature. English-language studies were searched from PubMed and Embase for the years 2000-2012, with the terms 'long-stay' or 'chronic mentally ill' and 'deinstitutionalis(z)ation' or 'community' or 'discharged' in the title and/or abstract. The present study defined long-stay as hospitalization >6 months. A total of 14 articles were identified. The majority of the participants suffered from schizophrenia. The most frequently assessed outcomes were social functioning, psychiatric symptoms and quality of life (QOL)/participant attitudes towards the environment. For social functioning, most of the studies found favorable changes. As regards psychiatric symptoms, stability or at least some improvements were detected. For QOL/participant attitudes towards the environment, positive changes, at least at some time-points, were noted. Significant deterioration was rare in all of the three outcomes. The majority of the participants showed enhancements in social functioning despite the fact that functional impairment is common in persons with schizophrenia, which indicates that even long-stay patients could achieve better functioning by deinstitutionalization. Although the stability in symptoms might be due to continuous schizophrenia course, moving to the community may also lead to improvement. The favorable changes in QOL/participants' attitudes towards the environment are in line with other studies.


Shift work at a modern offshore drilling rig.
The oil and gas exploration and production offshore units are classified as hazardous installations. Work in these facilities is complex, confined and associated with a wide range of risks. The continuous operation is secured by various shift work patterns. The objective of this study was to evaluate how offshore drilling workers perceived shift work at high seas and its impacts on their life and working conditions. The main features of the studied offshore shift work schedules are: long time on board (14 to 28 days), extended shifts (12 hours or more per day), slow rotation (7 to 14 days in the same shift), long sequence of days on the night shift (7 to 14 days in a row) and the extra-long extended journey (18 hours) on shift change and landing days. Interviews revealed a wide range of stressors caused by the offshore shift work, as well as difficulties to conciliate work with family life. It was observed that changes of the family model, leading to role conflicts and social isolation, work in a hazardous environment, perceiving poor sleep when working at night shifts and the imbalance between the expected and actual rewards are the major stressors for the offshore drilling workers.


Clinical effectiveness and soft tissue compatibility of a temporary restorative material.
The purpose of this six-week study was to evaluate in vivo the effectiveness of Systemp.inlay as a temporary restorative material. Subjects with the proper indication for a class II restoration were entered into the study. Twenty-three subjects and twenty-nine temporary restorations were evaluated at time of placement, three and six weeks. Two faculty members were calibrated to evaluate the material. Following the six-week trial, the temporary restorative material was removed and the test teeth were restored with a definitive restorative material. The temporary restorative material provided mechanical and anticariogenic protection of tooth structure while preventing dental sensitivity and patient discomfort. It promoted soft tissue health as evidenced by the enhanced plaque and gingival indices found associated with the temporarily restored teeth. Its ease of placement and removal makes it an attractive alternative for the dental practitioner. The change in color of the material over the six-week duration of the study may limit its use to posterior teeth and to lingual/palatal surfaces of anterior teeth. Ease of placement and removal and retention of the temporary restorative material were extremely good. Marginal and surface integrity of the material was maintained and no or minimal sensitivity was reported throughout the study. The material did not promote the accumulation of plaque nor result in gingival inflammation. The color change noted was not of clinical importance. Therefore, this temporary restorative material was demonstrated to be clinically effective in Class II cavity preparations.


Neurological deficit secondary to a pre-sacral abscess with epidural extension up to L3: A case report and literature review.
Isolated epidural abscesses are uncommon lesions. Surgical treatment may be difficult due to the extension of these lesions. We present a case of a pelvic abscess spreading along the path of the sciatic nerve to the gluteus muscles and the lumbar canal, causing neurological compression; requiring surgical treatment with three simultaneous approaches.


Kolya. Antiphonal development between adult and child.
The film Kolya (1996) directed by Jan Sverak, illustrates the antiphonal calling and answering development between man and boy who are both thrown and drawn together. Amid the larger context of the "Velvet Revolution" in Czechoslovakia, each supports the quieter revolution in the development of the other. The intraplay and interplay deepen with consideration of the father-son collaboration between director (son) and playwright-actor (father) which gave rise to this remarkable picture.


In-Line Film Coating Thickness Estimation of Minitablets in a Fluid-Bed Coating Equipment.
Film coating thickness of minitablets was estimated in-line during coating in a fluid-bed equipment by means of visual imaging. An existing, commercially available image acquisition system was used for image acquisition, while dedicated image analysis and data analysis methods were developed for this purpose. The methods were first tested against simulated minitablet's images and after that examined on a laboratory-scale fluid-bed Wurster coating process. An observation window cleaning mechanism was developed for this purpose. Six batches of minitablets were coated in total, using two different dispersions, where for the second dispersion coating endpoint was determined based on the in-line measurement. Coating thickness estimates were calculated from the increasing size distributions of the minitablet's major and minor lengths, assessed from the acquired images. Information on both the minitablet's average band and average cap coating thicknesses was obtained. The in-line coating thickness estimates were compared to the coating thickness weight gain calculations and the optical microscope measurements as a reference method. Average band coating thickness estimate was found the most accurate in comparison to microscope measurements, with root mean square error of 1.30 μm. The window cleaning mechanism was crucial for the accuracy of the in-line measurements as was evident from the corresponding decrease of the root mean square error (9.52 μm, band coating thickness). The presented visual imaging approach exhibits accuracy of at least 2 μm and is not susceptible to coating formulation or color variations. It presents a promising alternative to other existing techniques for the in-line coating thickness estimation.


A review of one-to-one dietetic obesity management in adults.
This review outlines the evidence and proposed professional consensus on one-to-one dietetic weight management for UK adults. New opportunities and challenges are arising for dietitians as the status afforded to obesity management increases. It is timely therefore to reflect on dietetic practice and consider the associated evidence. Despite a long standing history of working in obesity, little is known about the specific nature of UK dietetic practice, making it difficult to determine whether services are evolving and meeting recommended guidelines. It is essential that an evaluation of dietetic practice is undertaken and, importantly, that findings are published and disseminated to the profession. Assessment is the foundation of good weight management. However, given the complexity of obesity, the practice of completing both an assessment and instigating treatment during a brief consultation may not be feasible. The practitioner's interpersonal skills are central to treatment outcomes and merit regular review and comprehensive training. The ability to express empathy is a key skill, which is particularly important given the widespread weight bias in healthcare. Evidence supports multicomponent interventions combining diet, physical activity and behaviour modification. Following stabilisation of eating, as well as improvements to diet quality, the evidence supports the 600-kcal deficit, meal replacements and, in carefully selected individuals, very low energy diets. A weight maintenance component to management is linked with reduced weight regain, although the optimal frequency, duration and content of sessions are unclear. For many services, this higher intensity provision will likely require a multidisciplinary, multi-agency approach.


Long-term treatment with N-acetylcysteine, but not caloric restriction, protects mesenchymal stem cells of aged rats against tumor necrosis factor-induced death.
The survival of mesenchymal stem cells (MSCs) to tumor necrosis factor alpha (TNFalpha) stimulation was evaluated after a long-term antioxidant treatment, or caloric restriction, in aged rats. MSCs were isolated from bone marrow of 30-month-old rats which orally received N-acetylcysteine in the last 18 months. The necrotic cell death-induced in vitro by TNFalpha, determined by trypan blue exclusion, was markedly attenuated in MSCs obtained from treated vs. control aged rats (percent mean+/-SEM: 10.9+/-2.17 vs. 17.8+/-0.53; p<0.05). Also, the proliferation rate of MSCs from control, but not N-acetylcysteine-treated, aged rats evaluated up to 2 weeks was significantly higher than that of MSCs from younger (4-month-old) rats. No significant effect was observed relative to the parameters investigated when the aged rats were previously subjected to a hypocaloric diet for 18 months. In conclusion, a prolonged supplementation with N-acetylcysteine in rats can increase resistance to necrotic death of MSCs and may also counteract an excessive rate of MSC proliferation.


Low prevalence of disability among patients with inflammatory bowel diseases a decade after diagnosis.
Disability is defined by chronic limitations that preclude the ability to engage in usual daily activities. Studies of disability in patients with inflammatory bowel disease (IBD) have focused on work and employment, with few descriptions of more general disability among multiple domains. We examined disability and the factors associated with it a decade after diagnosis in a population-based cohort of IBD patients. We interviewed 125 patients with Crohn's disease (CD) and 119 with ulcerative colitis (UC) from the population-based Manitoba IBD Cohort study a median of 12.3 years after diagnosis. Disability was assessed by using 2 validated measures. Disease activity was assessed semiannually, and long-term activity was defined as symptoms of active IBD at more than 65% of semiannual assessments. Mean levels of disability were significantly higher among patients with CD than those with UC (P < .01). On the basis of the Work and Social Adjustment Scale, rates of disability were 19% among patients with CD vs 11% among those with UC (P < .05). Results from the World Health Organization Disability Assessment Schedule v.2 and the Work and Social Adjustment Scale correlated (r = 0.58 for patients with CD and 0.60 for those with UC; P < .01). Disability was associated with reduced quality of life. Long-term active disease and a lifetime history of major depression were associated with disability, whereas history of IBD-related surgeries or hospitalizations was not. A minority of patients with IBD have significant disability after a decade of disease, although a higher proportion of patients with CD are disabled than those with UC. Long-term active disease and psychological factors are important predictors of disability. Depression should be treated as aggressively as the IBD itself.


The national free delivery policy in Nepal: early evidence of its effects on health facilities.
Nepal faces the challenge of high levels of poverty, difficult access to health facilities and poor, though improving, health indicators. In response, in the past 5 years it has been experimenting with a range of approaches to removing user fees. Access to health care is now enshrined as a constitutional right for all. This article examines the latest policy, which was introduced in January 2009: free delivery care across the country. The study objective was to understand the effects of the policy on health facilities. Study methods included structured forms to retrieve financial and activity data from national, district and facility records (comparing 10 months before implementation with 10 months after). These were supplemented by semi-structured interviews with key informants at different levels of the health system. Findings include that utilization of services (at the facilities visited) continues to rise, with caesareans proportionate to the general growth in deliveries. Funds for the free delivery policy ('Aama') are found to be adequate to cover the main costs of services, with some surplus which can be invested in staff and in improving services. The system for reimbursing facilities is operating without undue delay and there is satisfaction with the flexibility of use of resources which it allows and the additional incentives for staff. The main concerns relate to wider systemic issues-in particular, understaffing in some key posts and areas, and dwindling general revenues for the facilities, especially through loss of wider user fee revenues. This may explain the ongoing charges for patients, which both facilities and patients report. It will be challenging to build on the gains of the past few years and sustain them, at the same time as merging the separate free care funding streams.


High mobility group proteins cHMG1a, cHMG1b, and cHMGI are distinctly distributed in chromosomes and differentially expressed during ecdysone dependent cell differentiation.
The mammalian high mobility group proteins HMGI/Y and HMG1/2 are thought to play an architectural role in assembly of nucleoprotein structures. Counterparts to these proteins have recently been found in the cells of the Dipteran insect Chironomus. In this report we investigate the distribution of three abundant HMG proteins in interphase giant chromosomes of the midge, Chironomus. By means of the indirect immunofluorescence technique the cHMG1b and cHMGI proteins were localized in chromosomal puffs, suggesting their involvement in the organization of transcriptionally active chromatin. In contrast, the highly abundant protein cHMG1a was rather uniformly distributed in the chromosomes. The cHMGI protein, but not cHMG1a or cHMG1b, was detected in nucleoli, which may indicate a role in the transcription of ribosomal genes. The regions of the interphase chromosomes containing AT-rich DNA did not contain higher levels of the cHMGI and cHMG1b proteins. A correlation between the specific location of these proteins in chromatin and their synthesis and turnover rates was observed.


The fingerprints of dioxin-like bromocarbazoles and chlorocarbazoles in selected forest soils in Germany.
The occurrence of bromocarbazoles and chlorocarbazoles was studied in 86 forest soil samples from different regions in Germany. Carbazole, 3-chlorocarbazole, 3-bromocarbazole and 3,6-dibromocarbazole were qualitatively detected in the humic layer of 59 soil samples with bromocarbazoles reported here for the first time in soil. Furthermore, the halogenated carbazoles, PCDD/Fs and PCBs were detected in the humic and mineral soil horizons (0-5 cm and 5-10 cm) of a subset of 11 soil samples subjected to quantitative analysis. Concentrations ranged from 0.6 to 267.6 ng/g (carbazole); 0.2-7.2 ng/g (3-bromocarbazole); 0.0-9.1 ng/g (3-chlorocarbazole); 0.2-19.8 ng/g (3,6-dibromocarbazole); 0.4-67.6 ng/g (3,6-dichlorocarbazole); 0.0-0.7 ng/g (PCDDs); 0.0-0.3 ng/g (PCDFs) and 0.0-33.7 ng/g (PCBs). Concentrations decreased with depth and correlated positively to total organic carbon (TOC). When it was based on TOC%, an increase in concentration with depth was observed in most soil samples. With respect to dioxin-like toxicity, 3-bromocarbazole, 3-chlorocarbazole, 3,6-dibromocarbazole and 3,6-dichlorocarbazoles caused induction of CYP1A1-dependent EROD activity in HII4E rat hepatoma cell line. Their relative effect potency after 72 h exposure ranged from 0.00005 to 0.00013 and was directly related to the degree of halogenation with 2,3,7,8-tetrachlorodibenzo-p-dioxin as reference. Furthermore, their contribution to overall soil dioxin-like toxicity was not significant in comparison to PCDD/Fs and PCBs though the sum toxic equivalency was limited to three halogenated carbazole congeners. Bromocarbazoles and chlorocarbazoles are emerging dioxin-like toxic environmental contaminants with potential for wide distribution occurring simultaneously with PCDD/Fs and PCBs.


[Cutaneous adverse reactions to chemotherapy and their management].
Chemotherapy is an essential component in the multidisciplinary management of many cancers. Skin reactions to chemotherapeutics are rather frequent, and their morbidity may be significant. It is wise to identify them with precision in order to chose the most adequate preventive and therapeutic measures. The dermatopathology assessment is helpful for the diagnostic.


Glucocorticoid-induced central diabetes insipidus in a case of malignant lymphoma.
A 37-year-old man was diagnosed as malignant lymphoma infiltrating in the central nervous system with hypopituitrism and secondary glucocorticoid deficiency. In this case, plasma arginine vasopressin (AVP) increased, but glucocorticoid administration decreased plasma AVP and increased urine volume with a reduction in urinary osmolality, indicative of the presence of glucocorticoid-induced diabetes insipidus. At the terminal stage, plasma AVP did not increase in response to the withdrawal of glucocorticoid and urine volume remained decreased, suggesting the presence of masked diabetes insipidus. Autopsy showed an infiltration of lymphoid cells around the cerebral ventricles and necrosis in the hypothalamo-hypophyseal system. These findings suggested that glucocorticoid might centrally play an important role in the regulation of AVP release, and its deficiency potentiated AVP release.


Decontamination of minimally invasive surgical endoscopes and accessories.
(1) Infections following invasive endoscopy are rare and are usually of endogenous origin. Nevertheless, infections do occur due to inadequate cleaning and disinfection and the use of contaminated rinse water and processing equipment. (2) Rigid and flexible operative endoscopes and accessories should be thoroughly cleaned and preferably sterilized using properly validated processes. (3) Heat tolerant operative endoscopes and accessories should be sterilized using a vacuum assisted steam sterilizer. Use autoclavable instrument trays or containers to protect equipment during transit and processing. Small bench top sterilizers without vacuum assisted air removal are unsuitable for packaged and lumened devices. (4) Heat sensitive rigid and flexible endoscopes and accessories should preferably be sterilized using ethylene oxide, low temperature steam and formaldehyde (rigid only) or gas plasma (if appropriate). (5) If there are insufficient instruments or time to sterilize invasive endoscopes, or if no suitable method is available locally, they may be disinfected by immersion in 2% glutaraldehyde or a suitable alternative. An immersion time of at least 10 min should be adopted for glutaraldehyde. This is sufficient to inactivate most vegetative bacteria and viruses including HIV and hepatitis B virus (HBV). Longer contact times of 20 min or more may be necessary if a mycobacterial infection is known or suspected. At least 3 h immersion in glutaraldehyde is required to kill spores. (6) Glutaraldehyde is irritant and sensitizing to the skin, eyes and respiratory tract. Measures must be taken to ensure glutaraldehyde is used in a safe manner, i.e., total containment and/or extraction of harmful vapour and the provision of suitable personal protective equipment, i.e., gloves, apron and eye protection if splashing could occur. Health surveillance of staff is recommended and should include a pre-employment enquiry regarding asthma, skin and mucosal sensitivity problems and lung function testing by spirometry. (7) Possible alternative disinfectants to glutaraldehyde include peracetic acid (0.2-0.35%), chlorine dioxide (700-1100 ppm) and superoxidized water. These are very effective, killing vegetative bacteria, including mycobacteria, and viruses in 5 min and bacterial spores in 10 min. An endorsement of compatibility with endoscopes, accessories and processing equipment is required from both the solution/device manufacturer and the endoscope manufacturer. Other important considerations are stability, cost and safety from the user and environmental standpoints. (8) Cleaning and disinfection or sterilization should be undertaken by trained staff in a dedicated area, e.g., SSD or TSSU. A suitable training programme is described. (9) If endoscopes are processed by immersion in disinfectants, harmful residues must be removed by thorough rinsing. Sterile or bacteria free water is essential for rinsing all invasive endoscopes and accessories to prevent recontamination. (10) If an automated washer disinfector is used it must be effective, non-damaging, reliable, easy to use and its performance regularly monitored. (11) If used, washer disinfectors and other processing equipment should be disinfected on a regular basis, i.e., between patients or at the start of each session. This will prevent biofilm formation and recontamination of instruments during rinsing. Disinfection should include the water treatment system, if present. (12) To comply with the Medical Devices Directive, manufacturers are obliged to provide full details on how to decontaminate the reusable devices they supply. This should include details of compatibility with heat, pressure, moisture, processing chemicals and ultrasonics. (13) The Infection Control Team should always be involved in the formulation and implementation of decontamination policies. Wherever possible, the national good practice guidelines produced by the Medical Devices Agency and/or professional societies shoul


Structure, function, and evolution of the beta-thymosin/WH2 (WASP-Homology2) actin-binding module.
beta-thymosins are acknowledged G-actin sequesterers. However, in the recent years, the conserved beta-thymosins/WH2 actin-binding module, has been identified in a large number of proteins that all interact with actin and play diverse functions in cell motility. The functional evolution of the WH2 domain has been approached by a combination of structural and biochemical methods, using thymosin beta4 (Tbeta4) and Ciboulot, a 3 beta-thymosin repeat protein from Drosophila as models. Ciboulot binds actin like Tbeta4 but promotes actin assembly like profilin. The first repeat of Ciboulot (D1) has the profilin function of the whole protein. The crystal structure of Ciboulot-actin shows that the major interaction with G-actin lies in the N-terminal amphipathic helix of D1. By point mutagenesis the sequestering activity of Tbeta4 can be changed into a profilin activity. ((1)H, (15)N)-NMR studies show that the functional switch from inhibition to promotion of actin assembly is linked to a change in the dynamics of interaction of the central and C-terminal regions of the WH2 domain with subdomains 1 and 2 of G-actin. Further systematic mutagenesis studies have been performed by engineering a series of chimeras of Ciboulot and Tbeta4. Proteins displaying either profilin function or enhanced sequestering activity compared to Tbeta4 have been characterized. The results provide insight into the structural basis for the regulation of the multiple functions of the WH2 domain.


Correlation of aqueous and lipid solubilities with flux for prodrugs of 5-fluorouracil, theophylline, and 6-mercaptopurine: A Potts-Guy approach.
The Potts and Guy equation that has been used to predict permeability coefficients for molecules being delivered from aqueous vehicles has been transformed to accommodate lipid vehicles that are less polar than skin, and polar vehicles that are less polar than water. Solubilities in pH 4.0 aqueous buffer (SAQ), solubilities in isopropyl myristate (SIPM), and molecular weights (MW) of prodrugs of 5-fluorouracil (5-FU), theophylline (Th), and 6-mercaptopurine (6-MP) have been regressed against their fluxes from suspensions in IPM (JM). Seven series (n = 39) of alkylcarbonyloxymethyl (ACOM), alkyloxycarbonyl (AOC), alkylcarbonyl (AC), and alkylaminocarbonyl (AAC) prodrugs were used to determine the best fit to the transformed Potts and Guy equation (eq 6): log JM = x + y log SIPM + (1 - y)log SAQ - z MW. The estimated values for x, y, and z were -0. 193, +0.525, and +0.00364, respectively, with r 2 = 0.945 for n = 39. Inclusion of a miscellaneous series comprised of the parent drugs and a branched alkyl chain prodrug gave an equally good fit only if 6-MP was excluded from the analysis. The best performer (largest JM) in each series was usually correctly identified. The values for x, y, and z were consistent with values obtained by Potts and Guy, but the inclusion of the (l - y)log SAQ term in eq 6 and the value for y, shows that water solubility is almost as important as lipid solubility in predicting flux. There were no significant changes in predicted log JM or xi for each series if their log JM or xi were calculated using y and z coefficients obtained for solutions to eq 6 from which the data for the series had been excluded. This suggests that the data from all the series is homogeneous. Data from Kasting, Smith, and Cooper for SIPM, SPG, and MW of unrelated molecules were regressed against their fluxes from propylene glycol (PG) using eq 7: log JM = x + y log SIPM + (1 - y) log SPG - z MW. The estimated values for x, y, and z were -1.673, +0.599, and +0.00595, respectively, with r 2 = 0.852 for n = 28. These values for x, y, and z are also consistent with those previously reported by Potts and Guy, and, together with the results for fluxes from IPM, show the general utility of the transformed Potts and Guy equation in predicting flux from vehicles other than water and in showing the importance of solubility in a polar solvent as well as a nonpolar solvent in predicting flux.


Orientation and training: preparing agency administrators and staff to replicate an HIV prevention intervention.
Effective orientation and training are fundamental to the successful implementation of any intervention because they communicate the critical first impressions of the intervention and the skills needed to conduct it. When research-based HIV prevention interventions are translated into practice, issues arise that require adaptation and expansion of the basic functions of orientation and training. This article identifies some of these issues by drawing on the experience of researchers in the Replicating Effective Programs (REP) project. The purpose, structure, and instructional approach of the orientation and training for administrators, staff, and volunteers are described in depth for one project, with comparisons and additional examples from others. Based on these descriptions, critical issues for orientation and training for replication are presented. These include extending orientation and training to a broad audience within the adopting agency, allocating sufficient time to ensure understanding of the intervention, and planning for staff turnover.


The relationship between basal metabolic rate (BMR) and concentrations of plasma thyroid hormones in fasting cockerels.
A correlation between the Basal Metabolic Rate (BMR) and the level of rT3, and occasionally between BMR and T3 or T4 was found in 12 month fasting cockerels. The birds were fasted for 48 hrs and BMR was measured eight times (before fasting, at 6, 12, 24, 30, 36, and 48 hrs of fasting, and 4 hours after fasting). Blood samples for plasma collection were taken immediately after measuring the BMR. During starvation a decrease in BMR was observed. After refeeding BMR returned to the starting level. The decrease in BMR was accompanied by an increase in rT3 and T4 plasma levels. Between BMR and levels of T4 and rT3 negative coefficients of correlation were observed (r = -0.20 and r = -0.42, respectively). Contrary to this, the T3 level declined and was correlated with BMR (r = 0.62). After refeeding, the T3 level rapidly increased against the control value. Moreover, a high coefficient of correlation (r = -0.39) was found between the level of T3 and rT3. The data show that the reduction in plasma T3 level and increase in the rT3 one during starvation may be due to inhibition of deiodination of T4 to T3, since rT3 is a competitive inhibitor of this reaction. The presented results support the suggestion that in birds T3 is the metabolically active thyroid hormone, and rT3 antagonizes this effect.


Prevention of cardiovascular diseases. Coronary artery disease, congestive heart failure, and stroke.
Cardiovascular disease leads to significant morbidity and mortality in the older population. Results of risk reduction can be dramatic in terms of patient survival and quality of life. This article reviews evidence for cardiovascular risk factors and disease prevention in older adults. Interventions which reduce morbidity and mortality from coronary artery disease, heart failure, and cerebrovascular disease in the elderly population are examined. Attention is given to the role of cardiovascular disease in older women and in minorities, subsets not well-represented in many studies.


Desiderata for product labeling of medical expert systems.
The proliferation and increasing complexity of medical expert systems raise ethical and legal concerns about the ability of practitioners to protect their patients from defective or misused software. Appropriate product labeling of expert systems can help clinical users to understand software indications and limitations. Mechanisms of action and knowledge representation schema should be explained in layperson's terminology. User qualifications and resources available for acquiring the skills necessary to understand and critique the system output should be listed. The processes used for building and maintaining the system's knowledge base are key determinants of the product's quality, and should be carefully documented. To meet these desiderata, a printed label is insufficient. The authors suggest a new, more active, model of product labeling for medical expert systems that involves embedding 'knowledge of the knowledge base', creating user-specific data, and sharing global information using the Internet.


Effect of YM992, a novel antidepressant with selective serotonin re-uptake inhibitory and 5-HT 2A receptor antagonistic activity, on a marble-burying behavior test as an obsessive-compulsive disorder model.
YM992 ((S)-2-[[(7-fluoroindan-4-yl)oxy]methyl]morpholine) monohydrochloride is a novel antidepressant with selective serotonin (5-hydroxytryptamine, 5-HT) re-uptake inhibition and 5-HT(2A) receptor antagonistic activity. The effects of YM992 and two selective 5-HT re-uptake inhibitors (SSRIs) were studied in a marble-burying behavior test as a model of an obsessive-compulsive disorder (OCD) in mice at doses of 5, 10 and 15 mg/kg, i.p. YM992 and fluoxetine significantly inhibited marble-burying behavior at a dose of 15 mg/kg (i.p.) without affecting spontaneous locomotor activities. Citalopram also significantly inhibited the behavior at doses of 5, 10 and 15 mg/kg (i.p.) without affecting spontaneous locomotor activities. These results suggest that YM992, as well as SSRIs, may exhibit anti-OCD activity in addition to an antidepressive effect in clinical use.


A global census of nitrogenase diversity.
The global diversity of nitrogen-fixing microorganisms was assessed through construction and analysis of an aligned database of 16,989 nifH sequences. We conclude that the diversity of diazotrophs is still poorly described and that many organisms remain to be discovered. Our analyses indicate that diversity is not distributed evenly across phylogenetic groups or across environments and that some of the most diverse assemblages and environments remain the most poorly characterized. The majority of OTUs were rare, falling in the long tail of the frequency distribution. The most dominant OTUs fell into either the Cyanobacteria or the α, β, and γ Proteobacteria, and five of these dominant OTUs do not have any representatives cultivated in isolation. Soils contained the greatest diversity of nifH sequences of all of the environments surveyed. Cluster III, which is dominated by nifH sequences from obligate anaerobes, was found to contain the greatest diversity of all nifH lineages and is also the group for which diversity is the least sampled. Our findings provide context for ongoing efforts to explore diazotroph diversity, indicating specific groups and environments that remain poorly characterized.


The Relationship between Spiritual Intelligence and Resiliency of Rehabilitation Staff.
This research aims to determine the relationship between spiritual intelligence and resiliency of rehabilitation staff. The reliable and validated spiritual intelligence and CD-RISC resiliency and demographic scales were used to collect data. There is a significant relationship between spiritual intelligence and the resiliency of rehabilitation staff (r = 0.38). Results showed that by increasing spiritual intelligence, resilience could increase.


Thoracoscopic repair of congenital diaphragmatic hernia in children.
Minimal invasive surgery (MIS) has been first proposed in case of delayed congenital diaphragmatic hernia (CDH). Since then, about 32 cases of thoracoscopic CDH approach in newborns have been published. Conditions of thoracoscopy are reviewed and enlightened with our preliminary series. The advantages of thoracoscopy versus a laparoscopic approach are detailed. Since 1999, all children presenting with CDH after the immediate neonatal period were offered a MIS approach. We started treating stable newborns suffering a CDH by thoracoscopic procedures in 2003. In cases of late presentations, 10 thoracoscopies and 1 laparoscopy were performed. Among them, 4 patients suffered from an incarcerated hernia. One conversion to a thoracotomy, 1 video-assisted thoracic surgery (VATS), and 2 conversions to laparoscopies were required for the reduction of hernia contents. In those last cases, the hernia defects were sutured, coming back to the thoracoscopic approach. Six newborns with neonatal diagnosis of CDH were primarily treated by thoracoscopy. Four procedures were completed without difficulty. The diaphragm was approximated with interrupted 2/0 nonabsorbable sutures. On the lateral part of the defect, in which there is a lack of diaphragm against the ribs, plegetted rib-anchoring stitches were used. A wide defect requiring a patch needed for conversion to a thoracotomy; in this case, we encountered a very rare pericardial defect and had difficulties in reducing the liver. In another case, we went to a VATS with a less than 2-cm opening to insert a Gore Tex patch, which was required. Thoracoscopy for delayed CDH repair seems to be easy and feasible with good results. A combined procedure with both thoracoscopy and laparoscopy has proven its usefulness in case of incarcerated hernia. Thoracoscopic CDH repair in newborns is not feasible in every case due to the patient's conditions. Criteria for eligibility need more cases to be evaluated. The advantages and disadvantages of thoracoscopy versus laparoscopy are reviewed. Nevertheless, the thoracoscopic approach seems easier.


The transcription factor HOXC9 regulates endothelial cell quiescence and vascular morphogenesis in zebrafish via inhibition of interleukin 8.
The transcription factor HOXC9 belongs to the homeobox gene family acting as developmental morphogen in several species. HOXC9 is EXPRESSED in different vascular beds in vivo. Yet vascular functions of HOXC9 have not been studied. This study was aimed at characterizing HOXC9 functions in human vascular endothelial cells and in zebrafish vascular development. HOXC9 was abundantly expressed in resting human umbilical vein endothelial cells and was downregulated by hypoxia. Overexpression of HOXC9 inhibited endothelial cell proliferation, migration, and tube formation in vitro. Expression profiling and chromatin immunoprecipitation experiments in human umbilical vein endothelial cells identified interleukin 8 as the major HOXC9 target and demonstrated the direct binding of HOXC9 to the interleukin 8 promotor. HOXC9 overexpression led to reduced endothelial interleukin 8 production, whereas HOXC9 silencing increased interleukin 8. The antimigratory and antiangiogenic effect of HOXC9 overexpression could be rescued by external interleukin 8 administration. Corresponding to the cellular experiments, endothelial-specific overexpression of HOXC9 and morpholino-based interleukin 8 loss-of-function experiments inhibited zebrafish vascular development. The data identify HOXC9 as an endothelial cell active transcriptional repressor promoting the resting, antiangiogenic endothelial cell phenotype in an interleukin 8-dependent manner.


Surgery versus radical endotherapies for early cancer and high grade dysplasia in Barrett's oesophagus.
Barrett's oesophagus is one of the most common premalignant lesions in the world. Currently the mainstay of therapy is surgical management of advanced cancer but this has improved the five-year survival very little in the last 30 years. As a consequence, improved survival relies on early detection through endoscopic surveillance programmes. Success of this strategy relies on the fact that late stage premalignant lesions or very early cancers can be cured by intervention. Currently there is considerable controversy over which method is best: i.e. conventional open surgery or endotherapy (techniques involving endoscopy). We used data from randomised controlled trials to examine the effectiveness of endotherapies compared with surgery, in people with Barrett's Oesophagus; those with early neoplasias (defined as high grade dysplasia (HGD), and those with early cancer (defined as carcinoma in-situ, superficially invasive, early cancer or superficial cancer T-1m (T1-a) and T-1sm (T1-b)). We used the Cochrane highly sensitive search strategy to identify randomized trials in MEDLINE, EMBASE, CENTRAL, ISI Web of Science, EBMR, Controlled Trials mRCT and ISRCTN and LILACS, in July and August 2008. Types of studies: randomised controlled trials comparing endotherapies with surgery in the treatment of high grade dysplasia (HGD), or early cancer. All cellular types of cancer were included (i.e. adenocarcinomas, squamous cell carcinomas and more unusual types) but will be discussed separately. patients of any age and either gender with a histologically confirmed diagnosis of early neoplasia (HGD and early cancer) in Barrett's or squamous lined oesophagus.Types of interventions; endotherapies (the intervention) compared with surgery (the control), all with curative intent. Reports of studies which meet the inclusion criteria for this review would have been analysed using the methods detailed in Appendix 9. We did not identify any studies which met the inclusion criteria. This Cochrane review has indicated that there are no randomised control trials to compare management options in this vital area, therefore trials should be undertaken as a matter of urgency. The problems with such randomised methods are standardising surgery and endotherapies in all sites; standardising histopathology in all centres; assessing which patients are fit or unfit for surgery; and making sure there are relevant outcomes for the study i.e. no progression of high grade dysplasia or long term survival i.e. over five years.


Treatment of recurrent thymic tumors.
Surgery is the cornerstone of therapy for recurrent thymic tumors. The pattern of recurrence is, however, less defined. Between 1966 and 1988, we operated on 83 patients with thymoma, 11 of whom underwent surgery for recurrence (group I). In 1989, we initiated a prospective multimodality protocol and have enrolled 128 patients with 9 (7%) recurrences since (group II). In group I, 1 patient was originally at stage I, 2 were at stage II, 5 at stage III, and 3 at stage IV. The patients underwent 1 (#10) or 2 (#1) reoperations and 5 showed histological progression of malignancy. One patient died postoperatively, 6 died of disease, and 3 are alive and disease free 18 to 22 years after the first operation. In group 2, no patient was originally at stage I, 1 was at stage II, 4 were at stage III, and 4 at stage IV. Reoperation (5 patients) was followed by chemotherapy and 2 showed histological progression of disease. One patient died after 2 years, and 4 patients are alive after 6 to 11 years. All recurrent tumors were thymomas with cortical differentiation. Early onset of recurrence was a negative prognostic factor. Thymomas can recur also at early stages. A multimodality approach is indicated also for early stage lesions based on histology.


Carbon-Dot and Quantum-Dot-Coated Dual-Emission Core-Satellite Silica Nanoparticles for Ratiometric Intracellular Cu(2+) Imaging.
Copper (Cu(2+)) is physiologically essential, but excessive Cu(2+) may cause potential risk to plants and animals due to the bioaccumulative properties. Hence, sensitive recognition is crucial to avoid overintake of Cu(2+), and visual recognition is more favored for practical application. In this work, a dual-emission ratiometric fluorescent nanoprobe was developed possessing the required intensity ratio, which can facilitate the sensitive identification of Cu(2+) by the naked eye. The probe hybridizes two fluorescence nanodots (quantum dots (QDs) and carbon dots (CDs)). Although both of them can be viable fluorescence probes for metal ion detection, rarely research has coupled this two different kinds of fluorescence material in one nanosensor to fabricate a selectively ratiometric fluorescence probe for intracellular imaging. The red emitting CdTe/CdS QDs were capped around the silica microsphere to serve as the response signal label, and the blue-emitting CDs, which is insensitive to the analyte, were covalently attached to the QDs surface to act as the reference signal. This core-satellite hybrid sphere not only improves the stability and brightness of QDs significantly but also decreases the cytotoxicity toward HeLa cells tremendously. Moreover, the Cu(2+) could quench the QDs emission effectively but have no ability for reduction of the CDs emission. Accordingly, a simple, efficient, and precise method for tracing Cu(2+) was proposed. The increase of Cu(2+) concentration in the series of 0-3 × 10(-6) M was in accordance with linearly decrease of the F650/F425 ratio. As for practical application, this nanosensor was utilized to the ratiometric fluorescence imaging of copper ions in HeLa cells.


MiR-543 Promotes Migration, Invasion and Epithelial-Mesenchymal Transition of Esophageal Cancer Cells by Targeting Phospholipase A2 Group IVA.
The aim of this study was to investigate the roles of miR-543 and phospholipase A2 group IVA (PLA2G4A) in cell mobility and the invasiveness cascade in esophageal squamous cell carcinoma (ESCC) and to validate the interactive relationship between miR-543 and PLA2G4A. Microarray analysis showed the different expression levels of PLA2G4A in two ESCC cell lines (KYSE30 and KYSE180). The expression levels of miR-543 and PLA2G4A in ESCC tissues were confirmed by qRT-PCR and Western blotting. The targeted relationship between miR-543 and PLA2G4A was studied and verified by a luciferase activity assay. Then, the invasion and metastasis ability of ESCC cell lines transfected with miR-543 mimics, miR-543 inhibitor, or PLA2G4A and miR-543 mimics were analyzed separately by Transwell migration and invasion assays. In addition, the roles of miR-543 and PLA2G4A in the expression of E-cadherin and vimentin were also investigated. PLA2G4A up-regulated the level of E-cadherin and down-regulated the level of vimentin, which curbed ESCC cell mobility and invasion. In ESCC cells, the expression of miR-543 was significantly higher, whereas the expression of PLA2G4A was markedly lower. MiR-543 facilitated ESCC cell mobility and invasion by repressing PLA2G4A. MiR-543 enhanced the cell mobility and the invasiveness cascade in ESCC cells via the down-regulation of PLA2G4A expression.


Interleukin-22 and rheumatoid arthritis: emerging role in pathogenesis and therapy.
Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by synovial inflammation and resultant progressive joint damage. It has become increasingly evident that cytokines play an important role in the pathogenesis of RA. Interleukin-22 (IL-22) is a member of the IL-10 cytokine family. Recent findings suggest that not only the expression of IL-22 is abnormal both in RA patients and in arthritis mice but also the aberrant IL-22 performs significantly in disease onset of RA. In this paper, we focus on the critical role of IL-22 in RA. Hopefully, the information obtained may lead to a better understanding of the pathogenesis and development of novel therapeutic strategies for this systemic autoimmune disease.


Increasing bile duct injury and decreasing utilization of intraoperative cholangiogram and common bile duct exploration over 14 years: an analysis of outcomes in New York State.
During laparoscopic cholecystectomy (LC), common bile duct (CBD) visualization either directly or with cholangiography (IOC) is less routine. Cholangiography can be used to identify and possibly prevent bile duct injury (BDI), which is a dreaded complication of cholecystectomy. The purpose of our study was to evaluate the trend of IOC/CBD exploration and BDI during LC for benign disease. A state-wide database (SPARCS) was used to identify all LC for benign biliary non-obstructive and obstructive disease between 2000 and 2014 in the state of New York. ICD-9 and CPT codes were used to identify IOC/CBD exploration and BDI. Multivariable logistic regression models were used in examining the linear trend in the risk of complication, 30-day readmission, 30-day ED visits, and BDI among all cholangiogram patients after controlling for possible confounding factors. During 2000-2014, 391,945 patients underwent laparoscopic cholecystectomy. The trend of IOC/CBD exploration performed significantly decreased for LC overall (12.37-10.44%, relative risk = 0.98, p <.0001) and particularly, in the outpatient setting (10.77-7.52%, relative risk = 0.96, p value <.0001). Among patients with IOC, overall complication rate, 30-day readmission rate, and 30-day ED visit rates increased. When looking at overall complication rate, there was an increase by about 4% per year (relative risk = 1.04, p value <.0001). After controlling for confounding factors, the complication risk and 30-day ED visit risk increased through years, while the 30-day readmission risk did not have significant change. Risk of BDI also increased significantly (p = 0.03). In an era of laparoscopy, the rate of IOC/CBD exploration during LC has significantly decreased, while BDI significantly increased.


Analytical requirements, perspectives and limits of immunological methods for drugs in hair.
The analytical requirements for analysis of drugs in hair are sensitivity in the range of picograms per milligram of hair, specificity for lipophilic drugs and absence of matrix effects with hair digests. These requirements are met by immunoassays which are also inexpensive, rapid and easy to use. However, in applying immunoassays to hair testing, certain limitations of the assay and of interpretation of assay results should be kept in perspective. These limitations are illustrated in this review with examples of the analysis of opiates in hair from patients and opiate addicts. The first requirement for immunological analysis of hair digests is that the digest must not denature the antibody proteins of the immunoassay reagents. For this reason enzymatic digests are better for immunological assay than chemical digests. Strongly acidic or alkaline digests must be brought to a neutral pH before immunoassay. Immunoassays used for analysis of hair should be calibrated with spiked hair digest standards to correct for possible matrix effects. The second requirement is that the immunoassay have the sensitivity and specificity to detect the drug in hair. Drugs of abuse are found in hair in the range of 10 pg-10 ng/mg hair. Radioimmunoassays are capable of detection and quantitation in this concentration range. Although the mechanism of drug incorporation into hair is not known, it is now apparent that primarily the parent drug and lipophilic metabolites are found in hair. For example, the ratio of cocaine/benzoylecgonine averages 10 (range 2-50) in published reports of analysis of hair from cocaine users. Therefore, immunoassays which are highly sensitive for the parent drug are required and results of immunoassays should be expressed as equivalents. When spiking standards for calibration of hair digest immunoassays, parent drug known to be present in hair should be used, e.g. cocaine not benzoylecgonine. With immunoassays which are specific for the lipophilic metabolite found in hair such as 6-MAM, differential radioimmunoassay can be used to discriminate between medical and illicit sources for the opiate drugs found in hair. Because of the low concentrations of drugs encountered in hair, immunoassays for hair have been used at cutoff concentrations at their limits of detection. The limit of detection (LOD) has been determined by calculating the mean and standard deviation (S.D.) for the assay response for a number of negative hair samples. The cutoff was then set at a distance of 2, 3, or 5 S.D.s from the mean response.(ABSTRACT TRUNCATED AT 400 WORDS)


Mechanosensor polycystin-1 potentiates differentiation of human osteoblastic cells by upregulating Runx2 expression via induction of JAK2/STAT3 signaling axis.
Polycystin-1 (PC1) has been proposed as a chief mechanosensing molecule implicated in skeletogenesis and bone remodeling. Mechanotransduction via PC1 involves proteolytic cleavage of its cytoplasmic tail (CT) and interaction with intracellular pathways and transcription factors to regulate cell function. Here we demonstrate the interaction of PC1-CT with JAK2/STAT3 signaling axis in mechanically stimulated human osteoblastic cells, leading to transcriptional induction of Runx2 gene, a master regulator of osteoblastic differentiation. Primary osteoblast-like PC1-expressing cells subjected to mechanical-stretching exhibited a PC1-dependent increase of the phosphorylated(p)/active form of JAK2. Specific interaction of PC1-CT with pJAK2 was observed after stretching while pre-treatment of cells with PC1 (anti-IgPKD1) and JAK2 inhibitors abolished JAK2 activation. Consistently, mechanostimulation triggered PC1-mediated phosphorylation and nuclear translocation of STAT3. The nuclear phosphorylated(p)/DNA-binding competent pSTAT3 levels were augmented after stretching followed by elevated DNA-binding activity. Pre-treatment with a STAT3 inhibitor either alone or in combination with anti-IgPKD1 abrogated this effect. Moreover, PC1-mediated mechanostimulation induced elevation of Runx2 mRNA levels. ChIP assays revealed direct regulation of Runx2 promoter activity by STAT3/Runx2 after mechanical-stretching that was PC1-dependent. Our findings show that mechanical load upregulates expression of Runx2 gene via potentiation of PC1-JAK2/STAT3 signaling axis, culminating to possibly control osteoblastic differentiation and ultimately bone formation.


Community and participation for general practice: perceptions of general practitioners and community nurses.
A central theme of health policy has concerned the public's participation in primary health care services, both as individual consumers and collectively as communities. In the U.K. primary care increasingly centres on general practice. This paper reports an exploratory study undertaken with practice teams in inner city Sheffield about community participation. The study design was influenced by a grounded theory approach to data collection and analysis. The main data collected was from interviews of 23 general practitioners, 20 practice nurses, 11 health visitors and 7 district nurses. Concepts of community and participation were explored by interview with these primary care workers. Typologies of community and participation for general practice are presented in the findings, along with associated strategic positions and political tensions. The discussion highlights a number of tensions and issues concerning community participation when primary care is organized around general practice.


New observations on the epidemiology and pathogenesis of sporotrichosis.
Five uncommon cases of skin sporotrichosis are presented, including a small family outbreak with interesting epidemiological findings and two patients treated with corticosteroids; we attributed the severity of the lesions to the steroids. Mice were used to confirm our hypothesis on the pathogenesis. The pathological findings in case of sporotrichosis in a dog are presented. A group of 359 pulmonary patients was tested with two different antigens of Sporotrichum schenkii.


The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.
Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.


Calcium and bone metabolism disorders during pregnancy and lactation.
Pregnancy and lactation cause a substantial increase in demand for calcium that is met by different maternal adaptations within each period. Intestinal calcium absorption more than doubles during pregnancy, whereas the maternal skeleton resorbs to provide most of the calcium content of breast milk during lactation. These maternal adaptations also affect the presentation, diagnosis, and management of disorders of calcium and bone metabolism. Although some women may experience fragility fractures as a consequence of pregnancy or lactation, for most women, parity and lactation do not affect the long-term risks of low bone density, osteoporosis, or fracture.


Distinct phenotype in maternal uniparental disomy of chromosome 14.
We report on the occurrence of maternal uniparental disomy for chromosome 14 (mUPD14) in a 4-year-old girl with a de novo Robertsonian translocation, 45,XX,t (13q,14q). The child has arrested hydrocephalus, short stature, minor anomalies, small hands with hyperextensible joints, and mild to moderate developmental delay. Comparison of her phenotype with those of three previously described individuals show some common distinct traits which suggest a mUPD14 syndrome.


Anaesthetic properties of pregnanolone emulsion. A comparison with alphaxolone/alphadolone, propofol, thiopentone and midazolam in a rat model.
The anaesthetic properties of pregnanolone emulsion, a new steroid preparation for intravenous administration, were evaluated in rats and compared to those of Althesin, propofol, thiopentone and midazolam. The therapeutic index was found to be 50% higher than that of Althesin and 6-8 times higher than those of propofol, thiopentone and midazolam. Induction with pregnanolone emulsion was reliable and fast, with only minor excitatory side effects, although induction time was longer compared to Althesin, propofol and thiopentone. Movement of unstimulated rats during sleep was similar to that seen with Althesin but movement on painful stimulation was less than with propofol and thiopentone. Recovery was rapid and smooth, and the proportion of sleeping time to recovery time was higher than found for the other four anaesthetics. No cumulative effects were seen on repeated administration and there were no venous sequelae. The results indicate that pregnanolone emulsion may provide a short-acting, less cumulative and less toxic alternative to existing intravenous anaesthetic agents.


A Leader-Led Supervision Group as a Model for Practicing Genetic Counselors.
With the stage set by the overview of supervision models in the previous paper, thispaper now presents the development and evolution of a currently ongoing leader-ledsupervision group for experienced genetic counselors. I discuss the procedures forgetting started; the creation and maintenance of the contract; typical issues and themesconsidered; the format for case presentation; and the overall growth of the group and itsmembers.


Improved Function and Maturation of Dendritic Cells Stimulated by Recombinant pp65 Protein: An in-vitro Design.
Stimulated dendritic cells (DCs) have been shown to be effective in the induction of specific immune cells. Also, the CMV pp65 plays an important role in CMV life cycle and immune recognition. To assess the effect of CMV pp65 on the maturity and function of dendritic cells. Splenic DCs were treated with non-cytotoxic concentrations of the pp65 and analyzed for MHC II, CD86, and CD40 expression by flow cytometry. Then, ROR-γ, GATA3, T-bet, and FOXP3 gene expression levels were evaluated in T cells co-cultured with DCs using Real time-PCR. Finally, the effects of pp65 on allogenic T-cell responses in mixed lymphocyte culture (MLR), and the release of cytokines were investigated by ELISA and flow cytometry. The phagocytosis rate was significantly lower in the pp65-treated DCs than the non-stimulated DCs. There were significant differences in the raised level of CD40, CD86, and CCR7 in DCs as maturation markers. Furthermore, ROR-γ, and T-bet overexpression in T cells of the pp65-treated group compared with the non-stimulated group was observed. Significant differences were observed in the levels of IL-2, IL-6, IL-17, IL-22, TNF-α, and IFN-γ in pp65-stimulated groups compared with the non-stimulated DCs. The pp65-treated DCs can induce differentiation and functional activity of the cellular immune system, including Th17, and Th1, but not other major T-cell subsets such as Tregs, and Th2 population.


Use of YouTube as a Resource for Surgical Education-Clarity or Confusion.
The use of video in surgical education and training is an innovative means to teach and expose learners to surgical procedures. The accessibility of YouTube provides a favourable platform for educational content; however, the current lack of peer review may mean that content is lacking or inaccurate. We reviewed articles that examined the quality of videos describing surgical procedures, and especially those found on the sharing platform "YouTube". In particular, studies investigating videos for their benefit to surgical trainees were included. A literature review was performed to determine the educational quality of surgical videos found on YouTube. Articles were included, which reviewed the quality of videos describing surgical procedures, their accuracy, and their utility for surgical trainees to complement traditional surgical education. The studies reviewed suggested that there is a lack of comprehensive material currently found on YouTube across multiple surgical disciplines. However, the popularity of YouTube among surgical trainees suggests that it is in the interest of surgical colleges and surgical educators to participate in sharing videos on this platform. YouTube videos are inevitably used as supplementary tools in surgical education. To harness the popularity and accessibility of YouTube for educational purposes, a peer-reviewed YouTube video library may be necessary.


Pathological study of the sural nerve in Fabry's disease.
A man with Fabry's disease, who died at 52, suffered from lancinating limb pains between the age of 20 and 27, and from severe arthralgia between 20 and 40. The sural nerve showed a severe loss of fibers (1,614 myelinated fibers/mm2), chiefly affecting the small myelinated fibers. Inclusions of both homogeneous and lamellated appearance were present in the perineurial cells, fibroblasts and vessel walls. Teased fibers displayed moderate remyelination, regeneration and the presence of globular focal myelin thickening. The transient time course of painful manifestations in this case is stressed in view of a possible pathogenesis of pain in Fabry's disease. It appears that small-fiber neuropathy is not directly related to the occurrence of pain symptoms, since the prevalent loss of small myelinated fibers was seen in the absence of actual pain. The small sensory neurons in the dorsal root ganglia, that are especially affected by lipid accumulation, probably give rise to abnormal discharges producing pain via the prevalence of the small-fiber versus the large-fiber input to the dorsal horn of the spinal cord (gate control theory). Thus pain fits may spontaneously abate, as in our case, if degeneration and loss of the small sensory neurons overstep a critical level.


Predictors of success with the use of donor sperm.
This study was undertaken to assess the effect of multiple factors that influence the success rate and time to conception among couples undergoing donor sperm insemination. A retrospective analysis of 960 cycles of frozen donor sperm insemination was performed at the University of Florida. Cycle pregnancy rates and cumulative probability of pregnancy were compared using several variables. The pregnancy rate was 12.1% per treatment cycle, and the cumulative probability of pregnancy exceeded 80% for the entire cohort. Seventy percent of pregnancies resulted in a liveborn infant. Age had a profound impact on the cycle pregnancy rate. The cycle pregnancy rates for women younger than 30 years, between the ages of 30 and 35 years, between the ages of 35 and 40 years, and older than 40 years were 15.8%, 14.6%, 8.2%, and 0%, respectively. There was a trend toward higher cycle pregnancy rates in women with prior pregnancies versus women without prior pregnancies of 14.4% and 12.3%, respectively. Parity had no effect on the cycle pregnancy rate or the cumulative probability of pregnancy. There was a trend toward higher cumulative probability of pregnancy in women whose partners were azoospermic versus oligospermic. There was no difference in pregnancy rates obtained with the Percoll wash gradient versus the Isolate gradient. At >20 million total motile sperm per insemination, there was no threshold above which the pregnancy rate was improved. The most significant influence on pregnancy rates in the donor sperm insemination program at the University of Florida was maternal age. Nulligravidity and a diagnosis of mild oligospermia in the man may have a negative impact on pregnancy rates.


A Ga2O3 underlayer as an isomorphic template for ultrathin hematite films toward efficient photoelectrochemical water splitting.
Hematite photoanodes for photoelectrochemical (PEC) water splitting are often fabricated as extremely-thin films to minimize charge recombination because of the short diffusion lengths of photoexcited carriers. However, poor crystallinity caused by structural interaction with a substrate negates the potential of ultrathin hematite photoanodes. This study demonstrates that ultrathin Ga2O3 underlayers, which were deposited on conducting substrates prior to hematite layers by atomic layer deposition, served as an isomorphic (corundum-type) structural template for ultrathin hematite and improved the photocurrent onset of PEC water splitting by 0.2 V. The benefit from Ga2O3 underlayers was most pronounced when the thickness of the underlayer was approximately 2 nm. Thinner underlayers did not work effectively as a template presumably because of insufficient crystallinity of the underlayer, while thicker ones diminished the PEC performance of hematite because the underlayer prevented electron injection from hematite to a conductive substrate due to the large conduction band offset. The enhancement of PEC performance by a Ga2O3 underlayer was more significant for thinner hematite layers owing to greater margins for improving the crystallinity of ultrathin hematite. It was confirmed that a Ga2O3 underlayer was applicable to a rough conducting substrate loaded with Sb-doped SnO2 nanoparticles, improving the photocurrent by a factor of 1.4. Accordingly, a Ga2O3 underlayer could push forward the development of host-guest-type nanocomposites consisting of highly-rough substrates and extremely-thin hematite absorbers.


Cellular pathology changes in rat skin following intradermal injection of nerve growth factor: neutrophil-dependent and -independent events.
Nerve growth factor (NGF) regulates the survival and development of specific populations of neurones and is involved in wound healing. A further area of study relating to the role of neurotrophins in the mature animal has concerned the possibility that NGF may be a pivotal mediator of inflammation and pain. It has previously been shown that injection of intradermal NGF can result in a neutrophil-dependent hyperalgesia in the rat. The purpose of the present study was to examine the pathological consequence of NGF injected intradermally into mature rat skin and to examine further the role of neutrophils. Standard histopathology techniques (H & E) were employed to determine inflammatory cell counts. Circulating neutrophils were depleted using an anti-rat neutrophil antiserum and results were compared to treatment with vehicle controls. Saline-pretreated rats exhibited normal circulating neutrophil numbers and the dorsal skin showed a significant increase of neutrophil and macrophages at 3 and 5 h and lymphocytes at 5 h after NGF treatment. By comparison, skin sites from neutrophil-depleted rats did not demonstrate a significant increase in neutrophil and macrophage accumulation after NGF administration. All NGF-treated sites, independent of pretreatment, demonstrated abnormal muscle fibre morphology and proliferation of the muscle sarcolemmal nuclei after NGF injection, indicative of tissue injury. In addition, oedema and some fibroplasia were also noted. Furthermore, fibrin production was increased at 3 and 5 h after NGF administration. It is suggested that NGF has a damaging effect on rat muscle which is independent of accumulating neutrophil and other inflammatory cells. In conclusion, the findings indicate a link between NGF-induced neutrophil and macrophage accumulation, as the increase in dermal macrophages was not observed in neutrophil-depleted rats. The results also suggest that NGF can have a profound effect on rat muscle and that this effect may be related to muscle regeneration.


Effect of graft-versus-host disease on anti-tumor immunity.
BCL1, a spontaneous B cell leukemia of BALB/c origin, is rejected by C.B-20 (Ighb, H-40b) but not BALB/c (Igha, H-40a) mice. Adoptive transfer of C.B-20 anti-BCL1 effector cells specific for the minor histocompatibility Ag H-40a protects irradiated C.B-20 but not BALB/c recipients. Because C.B-20 donor cells could potentially generate graft-vs-host disease (GVHD) in BALB/c recipients, we investigated the possibility that GVHD prevents the anti-tumor effect. GVHD was induced in (C.B-20 X B10.D2)F1 [H-2d, H-40b X H-2d,H-40b] recipients after injection of B10.D2-primed C.B-20 donor cells. GVHD was indicated by the histologic appearance of tissue sections from C.B-20----F1 livers, target organs of GVHD, which showed a marked mononuclear cell infiltrate around the portal tracts and central veins. In addition, splenic lymphocytes from these mice had altered CD4/CD8 ratios and were unable to respond to the polyclonal activators Con A and LPS. The mitogen unresponsiveness was at least partially due to the presence of a suppressor cell, because proliferation of normal spleen cells to Con A and LPS was suppressed upon addition of C.B-20----F1 spleen cells. Further immune dysfunction was evident by the inability of T cells from mice with GVHD to generate a CTL response to H-2 alloantigens. Addition of C.B-20----F1 spleen cells to F1 responder cells at the induction of culture did not prevent generation of CTL, indicating that a suppressor cell was not responsible for the lack of CTL activity. In this setting of GVHD, F1 recipients were able to reject BCL1 upon adoptive transfer of C.B-20 anti-BCL1 effector cells. These data indicate that GVHD-induced immune dysfunction does not inhibit the activity of antileukemia T cells.


Spirulina maxima prevents fatty liver formation in CD-1 male and female mice with experimental diabetes.
The dietary administration of 5% Spirulina maxima (SM) during four weeks to diabetic mice, starting one week after a single dose of alloxan, 250 mg/Kg body weight, prevented fatty liver production in male and female animals. The main action of SM was on triacylglycerol levels in serum and liver. There was also a moderate hypoglycemia in male mice. The thiobarbituric acid reactive substances also decreased in serum and liver after SM administration. There was also a decrease in the percentage of HDL in diabetic mice that was reverted by the SM administration. The sum of LDL + VLDL percentages was also partially normalized in diabetic animals by the SM administration. An additional observation was the lower incidence of adherences between the liver and the intestine loops in the diabetic mice treated with SM compared with diabetic mice without SM. Male and female mice showed differences to diabetes susceptibility and response to SM, the female being more resistant to diabetes induction by alloxan and more responsive to the beneficial effects of SM. It is worth future work of SM on humans looking for better quality of life and longer survival of diabetic patients.


Heparin neutralization by an extract of the human placenta: measurements and the concept of placental barrier to heparin.
A soluble extract from human placenta was found to neutralize heparin in a factor Xa-dependent heparin assay system. In comparison with serum the antiheparin activity of placenta tissue is 10 times higher (extract from 1 g of placenta neutralizes 8.94 +/- 3.31 IU of heparin; 1 ml of serum approximately 1 IU). The substance neutralizing heparin has not been identified, but it has been found that it is nondialyzable, is not adsorbed on heparin-Sepharose, and does not act progressively. It was suggested that the substance(s) responsible for the antiheparin effect in vitro might contribute to the failure of heparin to penetrate the placenta in vivo (placental barrier to heparin).


Inhibition of phosphodiesterase 9A reduces cytokine-stimulated in vitro adhesion of neutrophils from sickle cell anemia individuals.
Leukocyte adhesion to vessel walls may initiate vaso-occlusion in sickle cell anemia (SCA); however, the extent to which inflammation participates in this mechanism is not understood. This in vitro study investigated whether inflammatory molecules, commonly augmented in SCA, can affect neutrophil adhesive properties and whether cyclic guanosine monophosphate (cGMP)-elevating agents can inhibit such adhesion. Effects of Interleukin 8 (IL-8), tumor necrosis factor-α (TNF-α), granulocyte macrophage-colony stimulating factor (GM-CSF) cytokines, BAY 73-6691 [phosphodiesterase (PDE)-9A-inhibitor], and BAY 41-2271 (guanylate-cylase stimulator) on the adhesive properties of neutrophils from healthy control (CON) and steady-state SCA individuals were determined using static-adhesion assays. SCA neutrophils demonstrated increased adhesive properties, compared to CON neutrophils; IL-8, TNF-α and GM-CSF increased CON neutrophil adhesion and further increased SCA neutrophil adhesion to fibronectin (FN). The PDE9A inhibitor, BAY-73-6691, significantly reduced basal CON neutrophil and SCA neutrophil adhesion; this was accompanied by decreased SCA neutrophil surface expressions of the L-selectin and CD11b adhesion molecules. BAY-73-6691 also significantly reduced cytokine-stimulated CON neutrophil and SCA neutrophil adhesion to FN; however, this was not accompanied by alterations in adhesion-molecule presentation. The chronic inflammatory nature of SCA may contribute to leukocyte adhesive functions in SCA. Furthermore, elevation of leukocyte cGMP may be an interesting approach for inhibition of leukocyte adhesion to the vessel wall, even in the presence of inflammatory stimuli.


Impact of Early (<24 h) Surgical Decompression on Neurological Recovery in Thoracic Spinal Cord Injury: A Meta-Analysis.
The impact of surgical timing in neurological recovery in thoracic and thoracolumbar traumatic spinal cord injury (tSCI) is still a subject of discussion. While in cervical tSCI one may expect a beneficial effect of early intervention within 24 h, especially in complete cases, this has not yet been demonstrated clearly for thoracic tSCI. This study addresses neurological improvement after early and late surgery for thoracic and thoracolumbar tSCI. A systematic search retrieved 14 publications of observational studies reporting outcome measurements after surgery in 1075 patients with thoracic and thoracolumbar tSCI from PubMed and Embase databases. Surgery was considered early within 24 h and late thereafter. An improvement of at least one and two grades on the American Spinal Injury Association Impairment Scale (ASIA) was evaluated. The Meta-Analyses and Systematic Reviews of Observational Studies guidelines were followed. Improvement rates were summarized using individual patient data in a Bayesian random effects model and compared for those with early and late surgery. In the qualitative analysis, six of seven studies, which investigated the effect of surgical timing, observed a significant effect of early surgery on at least one ASIA grade improvement. Quantitative analysis in 948 patients with thoracic and thoracolumbar tSCI data, however, did not reveal a significant increase in odds of ≥1 ASIA grade recovery in early surgery (66.8% [95% confidence interval (CI): 45.0-87.8%] compared with late surgery (48.9% [95% CI: 25.1-70.7%; odds ratio (OR) 2.2 (95% CI: 0.6-14.0]). This study did not observe a significant beneficial effect of surgical decompression within 24 h in patients with thoracic and thoracolumbar tSCI.


From Genetic Predisposition to Molecular Mechanisms of Autoimmune Primary Adrenal Insufficiency.
Autoimmune Addison's disease (AAD) is a complex disease that results from the interaction of a predisposing genetic background with still unknown environmental factors. Pathogenic variants in the autoimmune regulator (AIRE) gene are responsible for autoimmune polyendocrine syndrome type 1, of which AAD is a major disease component. Among the genetic factors for isolated AAD and autoimmune polyendocrine syndrome type 2, a key role is played by HLA class II genes: HLA-DRB1*0301-DQA1*0501-DQB1*0201 and DRB1*04-DQA1*0301-DQB1*0302 are positively, and DRB1*0403 is negatively, associated with genetic risk for AAD. The MHC class I chain-related gene A (MICA) allele 5.1 is strongly and positively associated with AAD. Other gene polymorphisms contribute to the genetic risk for AAD, including CIITA (MHC class II transactivator), the master regulator of MHC class II expression, cytotoxic T-lymphocyte antigen-4 (CTLA-4), PTPN22, STAT4, PD-L1, NALP1, FCRL3, GPR174, GATA3, NFATC1, CYP27B1 and the vitamin D receptor.


Evaluation of concentration of voriconazole in aqueous humor after topical and oral administration in horses.
To determine penetration of topically and orally administered voriconazole into ocular tissues and evaluate concentrations of the drug in blood and signs of toxicosis after topical application in horses. 11 healthy adult horses. Each eye in 6 horses was treated with a single concentration (0.5%, 1.0%, or 3.0%) of a topically administered voriconazole solution every 4 hours for 7 doses. Anterior chamber paracentesis was performed and plasma samples were collected after application of the final dose. Voriconazole concentrations in aqueous humor (AH) and plasma were measured via high-performance liquid chromatography. Five horses received a single orally administered dose of voriconazole (4 mg/kg); anterior chamber paracentesis was performed, and voriconazole concentrations in AH were measured. Mean +/- SD voriconazole concentrations in AH after topical administration of 0.5%, 1.0%, and 3.0% solutions (n = 4 eyes for each concentration) were 1.43 +/- 0.37 microg/mL, 2.35 +/- 0.78 microg/mL, and 2.40 +/- 0.29 microg/mL, respectively. The 1.0% and 3.0% solutions resulted in significantly higher AH concentrations than the 0.5% solution, and only the 3.0% solution induced signs of ocular toxicosis. Voriconazole was detected in the plasma for 1 hour after the final topically administered dose of all solutions. Mean +/- SD voriconazole concentration in AH after a single orally administered dose was 0.86 +/- 0.22 microg/mL. Results indicated that voriconazole effectively penetrated the cornea in clinically normal eyes and reached detectable concentrations in the AH after topical administration. The drug also penetrated noninflamed equine eyes after oral administration. Low plasma concentrations of voriconazole were detected after topical administration.


ATP-binding Cassette Exporters: Structure and Mechanism with a Focus on P-glycoprotein and MRP1.
Proteins that belong to the ATP-binding cassette superfamily include transporters that mediate the efflux of substrates from cells. Among these exporters, P-glycoprotein and MRP1 are involved in cancer multidrug resistance, protection from endo and xenobiotics, determination of drug pharmacokinetics, and the pathophysiology of a variety of disorders. To review the information available on ATP-binding cassette exporters, with a focus on Pglycoprotein, MRP1 and related proteins. We describe tissue localization and function of these transporters in health and disease, and discuss the mechanisms of substrate transport. We also correlate recent structural information with the function of the exporters, and discuss details of their molecular mechanism with a focus on the nucleotide-binding domains. Evaluation of selected publications on the structure and function of ATP-binding cassette proteins. Conformational changes on the nucleotide-binding domains side of the exporters switch the accessibility of the substrate-binding pocket between the inside and outside, which is coupled to substrate efflux. However, there is no agreement on the magnitude and nature of the changes at the nucleotide- binding domains side that drive the alternate-accessibility. Comparison of the structures of Pglycoprotein and MRP1 helps explain differences in substrate selectivity and the bases for polyspecificity. P-glycoprotein substrates are hydrophobic and/or weak bases, and polyspecificity is explained by a flexible hydrophobic multi-binding site that has a few acidic patches. MRP1 substrates are mostly organic acids, and its polyspecificity is due to a single bipartite binding site that is flexible and displays positive charge.