Methadone dose increase and abstinence reinforcement for treatment of continued heroin use during methadone maintenance.
Although methadone maintenance is an effective therapy for heroin dependence, some patients continue to use heroin and may benefit from therapeutic modifications. This study evaluated a behavioral intervention, a pharmacological intervention, and a combination of both interventions. Throughout the study all patients received daily methadone hydrochloride maintenance (initially 50 mg/d orally) and weekly counseling. Following baseline treatment patients who continued to use heroin were randomly assigned to 1 of 4 interventions: (1) contingent vouchers for opiate-negative urine specimens (n = 29 patients); (2) methadone hydrochloride dose increase to 70 mg/d (n = 31 patients); (3) combined contingent vouchers and methadone dose increase (n = 32 patients); and (4) neither intervention (comparison standard; n = 28 patients). Methadone dose increases were double blind. Vouchers had monetary value and were exchangeable for goods and services. Groups not receiving contingent vouchers received matching vouchers independent of urine test results. Primary outcome measure was opiate-negative urine specimens (thrice weekly urinalysis). Contingent vouchers and a methadone dose increase each significantly increased the percentage of opiate-negative urine specimens during intervention. Contingent vouchers, with or without a methadone dose increase, increased the duration of sustained abstinence as assessed by urine screenings. Methadone dose increase, with or without contingent vouchers, reduced self-reported frequency of use and self-reported craving. In patients enrolled in a methadone-maintainence program who continued to use heroin, abstinence reinforcement and a methadone dose increase were each effective in reducing use. When combined, they did not dramatically enhance each other's effects on any 1 outcome measure, but they did seem to have complementary benefits.


A retrospective study of the management of vulvodynia.
Vulvodynia is characterized by chronic vulvar pain caused by sexual intercourse and often results in female sexual dysfunction. Because the causes of vulvodynia are not clear, many patients do not receive optimal treatment. Recently, gabapentin and botulinum toxin A have both been shown to be effective treatments for vulvodynia. In this study, we retrospectively analyzed the clinical outcomes of botulinum toxin A and gabapentin treatment for chronic pain in women with this condition. Seventy-three women with vulvar pain were administered either gabapentin (n=62) or botulinum toxin A (n=11) injections. Effectiveness was measured by use of a visual analogue scale (VAS). We analyzed the treatment method, treatment duration, success of treatment, and side effects or adverse reactions. Pain levels in both groups significantly decreased after treatment. In the gabapentin group, the VAS score decreased from 8.6 before treatment to 3.2 after treatment (p<0.001). The VAS score in the botulinum toxin A group was reduced from 8.1 to 2.5 (p<0.001). Side effects for both therapies were few and subsided with treatment with general antibiotics and nonsteroidal antiinflammatory drugs. Gabapentin and botulinum toxin A are safe and effective treatments for vulvodynia. This condition can cause sexual dysfunction and affect quality of life. However, with proper management, satisfactory outcomes for women with vulvodynia can be achieved.


Clinical presentation and follow-up of 30 patients with congenital nephrogenic diabetes insipidus.
Congenital nephrogenic diabetes insipidus is characterized by insensitivity of the distal nephron to arginine vasopressin. Clinical knowledge of this disease is based largely on case reports. For this study, data were collected on clinical presentation and during long-term follow-up of 30 male patients with congenital nephrogenic diabetes insipidus. The majority of patients (87%) were diagnosed within the first 2.5 yr of life. Main symptoms at clinical presentation were vomiting and anorexia, failure to thrive, fever, and constipation. Three older patients were diagnosed as a result of events not directly related to the disease. Except for a possibly milder phenotype in patients with a G185C mutation, no clear relationship between clinical and genetic data could be found. Most patients were on hydrochlorothiazide-amiloride treatment without significant side effects. Two patients suffered from severe hydronephrosis with a small rupture of the urinary tract after a minor trauma, and two patients experienced episodes of acute urine retention. Height SD scores for age remained below the 50th percentile in the majority of patients, whereas weight for height SD scores showed a catch-up after several years of underweight.


Enforced expression of miR-125b affects myelopoiesis by targeting multiple signaling pathways.
MicroRNAs (miRNAs) are small, noncoding RNAs that regulate gene expression by sequence-specific targeting of multiple mRNAs. Although lineage-, maturation-, and disease-specific miRNA expression has been described, miRNA-dependent phenotypes and miRNA-regulated signaling in hematopoietic cells are largely unknown. Combining functional genomics, biochemical analysis, and unbiased and hypothesis-driven miRNA target prediction, we show that lentivirally over-expressed miR-125b blocks G-CSF-induced granulocytic differentiation and enables G-CSF-dependent proliferation of murine 32D cells. In primary lineage-negative cells, miR-125b over-expression enhances colony-formation in vitro and promotes myelopoiesis in mouse bone marrow chimeras. We identified Stat3 and confirmed Bak1 as miR-125b target genes with approximately 30% and 50% reduction in protein expression, respectively. However, gene-specific RNAi reveals that this reduction, alone and in combination, is not sufficient to block G-CSF-dependent differentiation. STAT3 protein expression, DNA-binding, and transcriptional activity but not induction of tyrosine-phosphorylation and nuclear translocation are reduced upon enforced miR-125b expression, indicating miR-125b-mediated reduction of one or more STAT3 cofactors. Indeed, we identified c-Jun and Jund as potential miR-125b targets and demonstrated reduced protein expression in 32D/miR-125b cells. Interestingly, gene-specific silencing of JUND but not c-JUN partially mimics the miR-125b over-expression phenotype. These data demonstrate coordinated regulation of several signaling pathways by miR-125b linked to distinct phenotypes in myeloid cells.


Molecular mechanisms of cytokine-induced neuroprotection: NFkappaB and neuroplasticity.
Since the first attempts to understand the mechanisms of learning, memory, development, and other instances of neuroplasticity, gene expression has been an attractive explanation for the persistence of such processes. It has been hypothesized that changes in the levels of expression of a gene, or a coordinated set of genes, would be necessary for dramatic structural changes like the growth of new neurites. And more subtle biochemical changes at existing synapses might also result from an alteration in the array of gene products being manufactured in the relevant cells. However, a great deal of what is classified as neuroplasticity is dependent on primary changes in electrophysiological activity or other conditions at synapses. Therefore, a seminal question to those interested in the molecular underpinnings of neuroplasticity is that of signal transduction: How do changes in synaptic activity get communicated to the nucleus? To many who learn about the regulation of the transcription factor NFkappaB with this question in mind, its utility seems clear. Furthermore, NFkappaB is an important signaling factor for cytokines that appear to participate in several pathological conditions (e.g., Parkinson's disease, multiple sclerosis, and depression), so understanding its mechanisms of action and its relationship to other elements of cytokine signaling may be fundamental to determining the role of the inflammatory system in psychiatric and neurodegenerative conditions. For these reasons, NFkappaB has garnered considerable attention in various aspects of neuroplasticity, from long-term potentiation to the most dramatic forms of plasticity: cell birth and death. In a few cases, elegant experimental design has resulted in convincing evidence for the involvement of NFkappaB in specific phenomena. However, the complexity of this transcription factor-including confusion over what exactly is meant by "NFkappaB"-has led to some misleading conclusions, as well. This chapter highlights some of the potential red herrings to be encountered in the study of NFkappaB and will summarize the data and interpretations in which some degree of confidence can be placed. The final answers will depend on the application of models and tools only now in development.


Cognitive-behavioural therapy for severe and recurrent bipolar disorders: randomised controlled trial.
Efficacy trials suggest that structured psychological therapies may significantly reduce recurrence rates of major mood episodes in individuals with bipolar disorders. To compare the effectiveness of treatment as usual with an additional 22 sessions of cognitive-behavioural therapy (CBT). We undertook a multicentre, pragmatic, randomised controlled treatment trial (n=253). Patients were assessed every 8 weeks for 18 months. More than half of the patients had a recurrence by 18 months, with no significant differences between groups (hazard ratio=1.05; 95% CI 0.74-1.50). Post hoc analysis demonstrated a significant interaction (P=0.04) such that adjunctive CBT was significantly more effective than treatment as usual in those with fewer than 12 previous episodes, but less effective in those with more episodes. People with bipolar disorder and comparatively fewer previous mood episodes may benefit from CBT. However, such cases form the minority of those receiving mental healthcare.


Lessons Still Being Learned: Acute Pulmonary Tumor Embolus During Renal Cell Carcinoma Resection.
Renal cell carcinoma (RCC) is the most common primary renal neoplasm and is associated with the intraluminal growth into the venous system with possible extension into the inferior vena cava or even right heart. Intraoperative pulmonary embolism is a complication of resection of RCC, which may be mitigated by the use of the cardiopulmonary bypass with or without deep hypothermic circulatory arrest. We present a case of unexpected pulmonary embolism diagnosed during RCC surgery. The case highlights the central importance of intraoperative transesophageal echocardiography use and the need for proper preoperative planning for the use of cardiopulmonary bypass.


Mortality and stroke after amaurosis fugax.
One hundred and ten patients who had presented with amaurosis fugax and were treated medically were studied retrospectively. Follow up from the time of presentation was six to nineteen years with a median of eight years. The mortality and frequency of strokes in this group was compared with populations matched for age and sex. Life expectancy in patients with amaurosis fugax was reduced. Ischaemic heart disease was the most frequent cause of death and occurred at a greater rate than in the general population (p less than 0.01). The indicence of stroke was higher than in the Oxfordshire Community Stroke Project 1981-1983 (p less than 0.01). Comparing our results to those published for cerebral transient ischaemic attacks, patients with amaurosis fugax have a similar mortality rate but probably have a lower incidence of stroke. Patients with amaurosis fugax who have an occluded or narrowed proximal internal carotid artery have a greater risk of subsequent stroke than those with a normal carotid artery, or an arteriogram (p less than 0.01).


Cancer staging and survival rates: an analysis.
The interrelation between tumor staging and survival is an inverse one; patients with more widespread disease tend to have a shorter survival. Using this as a basis, a relationship was given between the logarithm of the percent survival at five years and the tumor stage at diagnosis. The description yielded an excellent fit for six reported neoplasms. For four other malignancies, the curves revealed a shape which resembled that of a multitarget model. The simpler description of logarithm of survival versus stage could be viewed as the limiting value of a multitarget model when the target number equals 1.


Mesogen polarity effects on biaxial nematics. Centrally located dipoles.
We investigate the phase organisation of thermotropic biaxial Gay-Berne (GB) mesogens yielding a biaxial nematic (Nb) phase upon endowing them with a central point dipole. We study the effects of changing the strength and orientation of the dipole on the phase behaviour, and in particular we examine, using molecular dynamics (MD) simulations, the possibility of improving the stability of the Nb phase. After mapping the boundaries of the Nb phase, we find that the strength of the embedded dipole is the parameter with the strongest influence on the mesogenic properties, while its orientation plays a minor role. For these central dipole systems, we find that the Nb phase organisation is stable only for mesogens with relatively weak dipole moments, while it disappears if electrostatic interactions become comparable in magnitude with dispersion interactions.


Amphotericin B-loaded natural latex dressing for treating Candida albicans wound infections using Galleria mellonella model.
Amphotericin B (AmB) is the gold standard for antifungal drugs. However, AmB systemic administration is restricted because of its side effects. Here, we report AmB loaded in natural rubber latex (NRL), a sustained delivery system with low toxicity, which stimulates angiogenesis, cell adhesion and accelerates wound healing. Physicochemical characterizations showed that AmB did not bind chemically to the polymeric matrix. Electronic and topographical images showed small crystalline aggregates from AmB crystals on the polymer surface. About 56.6% of AmB was released by the NRL in 120 h. However, 33.6% of this antifungal was delivered in the first 24 h due to the presence of AmB on the polymer surface. The biomaterial's excellent hemo- and cytocompatibility with erythrocytes and human dermal fibroblasts (HDF) confirmed its safety for dermal wound application. Antifungal assay against Candida albicans showed that AmB-NRL presented a dose-dependent behavior with an inhibition halo of 30.0 ± 1.0 mm. Galleria mellonella was employed as an in vivo model for C. albicans infection. Survival rates of 60% were observed following the injection of AmB (0.5 mg.mL-1) in G. mellonella larvae infected by C. albicans. Likewise, AmB-NRL (0.5 mg.mL-1) presented survival rates of 40%, inferring antifungal activity against fungus. Thus, NRL adequately acts as an AmB-sustained release matrix, which is an exciting approach, since this antifungal is toxic at high concentrations. Our findings suggest that AmB-NRL is an efficient, safe, and reasonably priced ($0.15) dressing for the treatment of cutaneous fungal infections.


Potential Activity, Size, and Structure of Sulfate-Reducing Microbial Communities in an Exposed, Grazed and a Sheltered, Non-Grazed Mangrove Stand at the Red Sea Coast.
After oxygen, sulfate is the most important oxidant for the oxidation of organic matter in mangrove forest soils. As sulfate reducers are poor competitors for common electron donors, their relative success depends mostly on the surplus of carbon that is left by aerobic organisms due to oxygen depletion. We therefore hypothesized that sulfate-cycling in mangrove soils is influenced by the size of net primary production, and hence negatively affected by mangrove degradation and exploitation, as well as by carbon-exporting waves. To test this, we compared quantitative and qualitative traits of sulfate-reducing communities in two Saudi-Arabian mangrove stands near Jeddah, where co-occurring differences in camel-grazing pressure and tidal exposure led to a markedly different stand height and hence primary production. Potential sulfate reduction rates measured in anoxic flow-through reactors in the absence and presence of additional carbon sources were significantly higher in the samples from the non-grazed site. Near the surface (0-2 cm depth), numbers of dsrB gene copies and culturable cells also tended to be higher in the non-grazed sites, while these differences were not detected in the sub-surface (4-6 cm depth). It was concluded that sulfate-reducing microbes at the surface were indeed repressed at the low-productive site as could be expected from our hypothesis. At both sites, sulfate reduction rates as well as numbers of the dsrB gene copies and viable cells increased with depth suggesting repression of sulfate reduction near the surface in both irrespective of production level. Additionally, sequence analysis of DNA bands obtained from DGGE gels based on the dsrB gene, showed a clear difference in dominance of sulfate-reducing genera belonging to the Deltaproteobacteria and the Firmicutes between sampling sites and depths.


Exploring multimorbidity profiles in middle-aged inpatients: a network-based comparative study of China and the United Kingdom.
Multimorbidity is better prevented in younger ages than in older ages. This study aims to identify the differences in comorbidity patterns in middle-aged inpatients from China and the United Kingdom (UK). We utilized 184,133 and 180,497 baseline hospitalization records in middle-aged populations (40-59 years) from Shaanxi, China, and UK Biobank. Logistic regression was used to calculate odds ratios and P values for 43,110 unique comorbidity patterns in Chinese inpatients and 21,026 unique comorbidity patterns in UK inpatients. We included the statistically significant (P values adjusted by Bonferroni correction) and common comorbidity patterns (the pattern with prevalence > 1/10,000 in each dataset) and employed network analysis to construct multimorbidity networks and compare feature differences in multimorbidity networks for Chinese and UK inpatients, respectively. We defined hub diseases as diseases having the top 10 highest number of unique comorbidity patterns in the multimorbidity network. We reported that 57.12% of Chinese inpatients had multimorbidity, substantially higher than 30.39% of UK inpatients. The complete multimorbidity network for Chinese inpatients consisted of 1367 comorbidities of 341 diseases and was 2.93 × more complex than that of 467 comorbidities of 215 diseases in the UK. In males, the complexity of the multimorbidity network in China was 2.69 × more than their UK counterparts, while the ratio was 2.63 × in females. Comorbidities associated with hub diseases represented 68.26% of comorbidity frequencies in the complete multimorbidity network in Chinese inpatients and 55.61% in UK inpatients. Essential hypertension, dyslipidemia, type 2 diabetes mellitus, and gastritis and duodenitis were the hub diseases in both populations. The Chinese inpatients consistently demonstrated a higher frequency of comorbidities related to circulatory and endocrine/nutritional/metabolic diseases. In the UK, aside from these comorbidities, comorbidities related to digestive and genitourinary diseases were also prevalent, particularly the latter among female inpatients. Chinese inpatients exhibit higher multimorbidity prevalence and more complex networks compared to their UK counterparts. Multimorbidity with circulatory and endocrine/nutritional/metabolic diseases among both Chinese and UK inpatients necessitates tailored surveillance, prevention, and intervention approaches. Targeted interventions for digestive and genitourinary diseases are warranted for the UK.


The effect of food supplements on completely edentulous women rehabilitated with complete dentures: A randomized controlled trial.
Neglected oral health is a major issue, especially in women of developing countries, leading to early loss of teeth which may further lead to malnutrition, degradation of overall health, and increased chances of osteoporosis. Thus, the aim of this study was to assess the effect of food supplement on masticatory performance, nutritional status, electromyography (EMG) (masseter and temporalis), and bone mineral density (BMD) among women rehabilitated with complete denture. Hospital based randomized controlled trial. A randomized controlled trial with 106 women of 45-65 years rehabilitated with complete denture (56 received food supplement and 50 did not receive food supplement) and 52 healthy control was conducted. The outcomes were assessed at baseline and 3 and 6 months of follow up (after complete denture fabrication). Outcomes were measured via masticatory performance, nutritional status (hemoglobin, serum calcium, albumin, and Vitamin D level), EMG of masseter and temporalis muscles, and BMD. Friedman's analysis of variance test was used as a nonparametric test, and the Statistical Package for the Social Sciences version 21.0 at a significance level of 0.05 was used for statistical analysis. A statistically significant change was observed during follow up for the group with food supplement for BMD, EMG, and masticatory performance. When biochemical parameters were assessed during follow up, no statistically significant change was observed for both groups (with and without food supplement), except for serum calcium level in group which received food supplement. It was found that the magnitude of effect was remarkably meager in food supplement group which could be perhaps due to less time given for follow up period. Longer duration of trials would yield better results.


Neonatal exposure to a glyphosate-based herbicide alters the histofunctional differentiation of the ovaries and uterus in lambs.
The aim of the present study was to compare the effect of oral and subcutaneous exposure to a glyphosate-based herbicide (GBH) on the female reproductive system, specifically in the ovaries and uterus of prepubertal lambs. To this end, ewe lambs were exposed to a s.c. (n: 5) or an oral (n: 5) environmentally relevant dose of GBH (2 mg/kg/day) or to vehicle (controls, n: 12), from postnatal day (PND) 1 to PND14. Serum glyphosate and aminomethylphosphonic acid (AMPA) concentrations were measured on PND15 and PND45. The ovaries and uterus were obtained and weighed on PND45. Ovarian follicular dynamics and uterine morphological features were determined by picrosirius-hematoxylin staining. The proliferation marker Ki67 was evaluated by immunohistochemistry in ovarian and uterine samples. Glyphosate but not AMPA was detected in serum of exposed lambs on PND15, whereas neither glyphosate nor AMPA were detected on PND45. Controls were negative for glyphosate and AMPA on PND15 and PND45. GBH exposure did not affect ovarian or uterine weight. However, on PND45, the ovary of GBH-exposed lambs showed altered follicular dynamics, increased proliferation of granulosa and theca cells, and decreased mRNA expression of FSHR and GDF9, whereas their uterus showed decreased cell proliferation but no alterations in the histomorphology or gene expression. In conclusion, GBH exposure altered the ovarian follicular dynamics and gene expression, and the proliferative activity of the ovaries and uterus of lambs. It is noteworthy that all the adverse effects found in the ovaries and uterus of both GBH-exposed groups were similar, independently of the administration route.


Breast-conserving radiotherapy with simultaneous integrated boost; field-in-field three-dimensional conformal radiotherapy versus inverse intensity-modulated radiotherapy - A dosimetric comparison: Do we need intensity-modulated radiotherapy?
To examine the feasibility of improving breast-conserving radiotherapy with simultaneous integrated boost (SIB) and analyzing the efficiency of forward versus inverse intensity-modulated radiotherapy (IMRT) techniques in providing the same. Three-dimensional conformal radiotherapy (3DCRT) field-in-field (FIF) plans with simultaneous and sequential boost and IMRT SIB plans were generated for the datasets of 20 patients who had undergone breast-conserving surgery. The 3 plans were compared dosimetrically for efficiency in terms of planning target volume (PTV) coverage (PTV 95%), homogeneity and conformity, dose delivered to ipsilateral/contralateral lungs (I/L: V10, V20, C/L: Vmean, V5), heart and contralateral breast (Vmean, V30 for heart and Vmean, V1, V5 for C/L breast). The FIF 3DCRT plan with SIB (PLAN B) was more homogeneous than the classical technique with sequential boost (PLAN A). There were less hot spots in terms of Dmax (63.7 ± 1.3) versus Dmax (68.9 ± 1), P < 0.001 and boost V107%, B (0.3 ± 0.7) versus A (3.5 ± 5.99), P = 0.001. The IMRT SIB (PLAN C) did not provide any significant dosimetric advantage over the 3DCRT SIB technique. IMRT SIB plan C was associated with increased dose to contralateral lung in-terms of V5 (10.35 +/- 18.23) vs. (1.13 +/- 4.24), P = 0.04 and Vmean (2.12 ± 2.18) versus Vmean (0.595 ± 0.89), P = 0.008. There was 3-fold greater exposure in terms of Monitor Unit (MU) (1024.9 ± 298.32 versus 281.05 ± 20.23, P < 0.001) and treatment delivery time. FIF 3DCRT SIB provides a dosimetrically acceptable and technically feasible alternative to the classical 3DCRT plan with sequential boost for breast-conserving radiotherapy. It reduces treatment time by 2 weeks. IMRT SIB does not appear to have any dosimetric advantage; it is associated with significantly higher doses to contralateral lung and heart and radiation exposure in terms of MU.


Podoplanin expression in peritumoral keratinocytes predicts aggressive behavior in extramammary Paget's disease.
Recent studies have demonstrated podoplanin expression in several tumors, which has been associated with lymph node metastasis and poor prognosis. Podoplanin expression in peritumoral cells such as cancer-associated fibroblasts also correlates with tumor progression in several cancers. However, podoplanin expression and its association with extramammary Paget's disease (EMPD) remain unclear. In this study, we examined whether the presence of podoplanin expression in tumor cells or peritumoral basal keratinocytes correlated with aggressive behavior in patients with EMPD and investigated the mechanisms of podoplanin-mediated tumor invasion in this disorder. Skin samples of 37 patients with EMPD were investigated by immunohistochemical analysis. The functions of podoplanin in keratinocytes were examined in vitro by RT-PCR and with invadopodia gelatin-degradation assays using HaCaT cells. Podoplanin was not identified in tumor cells in all cases. Podoplanin expression in peritumoral basal keratinocytes was observed in 25 patients (67.6%). In in situ EMPD, 50% of cases (9 in 18) exhibited podoplanin-positive keratinocytes, whereas 84.2% (16 in 19) demonstrated positive staining in invasive EMPD (P<0.05). Podoplanin expression in peritumoral keratinocytes was also associated with tumor thickness (P<0.005). By immunohistochemical analysis, podoplanin-positive peritumoral keratinocytes were found to be negative for E-cadherin, one of the major adhesion molecules of keratinocytes, which might contribute to tumor invasion into the dermis through a crack in the basal cell layer induced by down-regulation of cell adhesion therein. We further found that podoplanin-positive keratinocytes exhibited invadopodia, which are thought to function in the migration of cancer cells through tissue barriers, indicating that podoplanin-positive peritumoral basal keratinocytes might assist tumor invasion by degrading the extracellular matrix. The presence of podoplanin expression in peritumoral keratinocytes correlates with aggressive behavior in EMPD and might therefore serve as a useful prognostic marker for patients with EMPD.


Activated charcoal effectively removes inhaled anesthetics from modern anesthesia machines.
If a malignant hyperthermia-susceptible patient is to receive an anesthetic, an anesthesia machine that has been used previously to deliver volatile anesthetics should be flushed with a high fresh gas flow. Conflicting results from previous studies recommend flush times that vary from 10 to 104 minutes. In a previously proposed alternative decontamination technique, other investigators placed an activated charcoal filter in the inspired limb of the breathing circuit. We placed activated charcoal filters on both the inspired and expired limbs of several contaminated anesthesia machines and measured the time needed to flush the machine so that the delivered concentrations of isoflurane, sevoflurane, and desflurane would be <5 parts per million (ppm). We next simulated the case for which malignant hyperthermia is diagnosed 90 minutes after induction of anesthesia and measured how well activated charcoal filters limit further exposure. Activated charcoal filters decrease the concentration of volatile anesthetic delivered by a contaminated machine to an acceptable level in <2 minutes. The concentrations remained well below 5 ppm for at least 60 minutes. When malignant hyperthermia is diagnosed after induction of anesthesia, we found that with charcoal filters in place, the current anesthesia machine may be used for at least 67 minutes before the inspired concentration exceeds 5 ppm. Activated charcoal filters provide an alternative approach to the 10 to 104 minutes of flushing that are normally required to prepare a machine that has been used previously to deliver a volatile anesthetic.


Physical compatibility of cisatracurium with selected drugs during simulated Y-site administration.
The physical compatibility of cisatracurium with selected drugs during simulated Y-site administration was studied. Study drugs were selected based on the lack of physical compatibility data with cisatracurium and their use in intensive care units. Test admixtures were prepared by mixing 2.5-mL samples of varying concentrations of calcium gluconate, diltiazem, esomeprazole, regular insulin, nicardipine, pantoprazole, and vasopressin with either 2.5 mL of normal saline 0.9% (control) or 2.5 mL of cisatracurium (experimental) to simulate a 1:1 Y-site ratio. Drug infusions were prepared at the maximum concentrations used clinically. Physical compatibility of the admixtures was determined by visual and turbidimetric assessments performed in triplicate immediately after mixing and at 15, 30, and 60 minutes. Visual incompatibility was defined as a change in color, the formation of haze or precipitate, the presence of particles, or the formation of gas in the experimental groups compared with the controls. Disturbances invisible to the naked eye were determined by assessing changes in turbidity of experimental admixtures compared with the controls. None of the admixtures exhibited visual changes when mixed with cisatracurium. Six of the seven admixtures exhibited turbidimetric compatibility with cisatracurium. Pantoprazole admixtures demonstrated a significant difference in turbidimetric assessment between the control and experimental groups when mixed with cisatracurium (p < 0.001). Calcium gluconate, diltiazem hydrochloride, esomeprazole, regular insulin, nicardipine hydrochloride, and vasopressin demonstrated physical compatibility with cisatracurium over 60 minutes during simulated Y-site administration. Cisatracurium and pantoprazole should not be coadministered due to a significant difference in turbidity between control and experimental samples.


Institutional geographies in dying: nurses' actions and observations on dying spaces inside and outside intensive care units.
This paper articulates the geographies associated with intensive care nursing work with dying patients and their families. Six focus groups were conducted with 27 registered critical care nurses who practice in hospitals in a mid-western city in the United States. The analysis is structured by three emerging themes (i) the importance of a 'good' and 'sacred' place, (ii) the body as mapped by medical specialties, and (iii) problems with procedurally driven suspension of 'do not resuscitate' orders beyond intensive care units (ICUs). Recommendations describe the need for institutional recognition of the moral importance of strong relationships between nurses, clients, and their families, and nurses' wide-ranging roles in bridging the various spatial domains of intensive care.


Repair of bone defects using synthetic mimetics of collagenous extracellular matrices.
We have engineered synthetic poly(ethylene glycol) (PEG)-based hydrogels as cell-ingrowth matrices for in situ bone regeneration. These networks contain a combination of pendant oligopeptide ligands for cell adhesion (RGDSP) and substrates for matrix metalloproteinase (MMP) as linkers between PEG chains. Primary human fibroblasts were shown to migrate within these matrices by integrin- and MMP-dependent mechanisms. Gels used to deliver recombinant human bone morphogenetic protein-2 (rhBMP-2) to the site of critical- sized defects in rat crania were completely infiltrated by cells and were remodeled into bony tissue within five weeks. Bone regeneration was dependent on the proteolytic sensitivity of the matrices and their architecture. The cell-mediated proteolytic invasiveness of the gels and entrapment of rhBMP-2 resulted in efficient and highly localized bone regeneration.


Mitochondrial Dysfunction and Its Relationship with mTOR Signaling and Oxidative Damage in Autism Spectrum Disorders.
Mitochondria are organelles that play a central role in processes related to cellular viability, such as energy production, cell growth, cell death via apoptosis, and metabolism of reactive oxygen species (ROS). We can observe behavioral abnormalities relevant to autism spectrum disorders (ASDs) and their recovery mediated by the mTOR inhibitor rapamycin in mouse models. In Tsc2(+/-) mice, the transcription of multiple genes involved in mTOR signaling is enhanced, suggesting a crucial role of dysregulated mTOR signaling in the ASD model. This review proposes that the mTOR inhibitor may be useful for the pharmacological treatment of ASD. This review offers novel insights into mitochondrial dysfunction and the related impaired glutathione synthesis and lower detoxification capacity. Firstly, children with ASD and concomitant mitochondrial dysfunction have been reported to manifest clinical symptoms similar to those of mitochondrial disorders, and it therefore shows that the clinical manifestations of ASD with a concomitant diagnosis of mitochondrial dysfunction are likely due to these mitochondrial disorders. Secondly, the adenosine triphosphate (ATP) production/oxygen consumption pathway may be a potential candidate for preventing mitochondrial dysfunction due to oxidative stress, and disruption of ATP synthesis alone may be related to impaired glutathione synthesis. Finally, a decrease in total antioxidant capacity may account for ASD children who show core social and behavioral impairments without neurological and somatic symptoms.


Fabrication of targeted and pH responsive lysozyme-hyaluronan nanoparticles for 5-fluorouracil and curcumin co-delivery in colorectal cancer therapy.
Green nanotechnology is considered a promising method to construct functional materials with significant anticancer activity, while overcoming the shortcomings of traditional synthesis process complexity and high organic solvents consumption. Thus, in this study, we report for the first time the rational design and green synthesis of functionalized 5-fluorouracil and curcumin co-loaded lysozyme-hyaluronan composite colloidal nanoparticles (5-Fu/Cur@LHNPs) for better targeted colorectal cancer therapy with minimized side effects. The functionalized 5-Fu/Cur@LHNPs exhibit stabilized particle size (126.1 nm) with excellent homogeneity (PDI = 0.1), favorable colloidal stabilities, and excellent re-dispersibility. In vitro cell experiments illustrate that the cellular uptake of 5-Fu/Cur@LHNPs was significantly improved and further promoted a higher apoptosis ratio of HCT-116 cells. Compared with the control group, the 5-Fu/Cur@LHNPs formulation group achieved effective inhibition (60.1 %) of colorectal tumor growth. The alcohol-free self-assembly method to construct 5-Fu/Cur@LHNPs is simple and safe for a translational chemotherapy drug, also to promote more robust delivery systems for treating colorectal cancer.


Knowledge, attitudes, beliefs and behaviour intentions for three bowel management practices in intensive care: effects of a targeted protocol implementation for nursing and medical staff.
Bowel management protocols have the potential to minimize complications for critically ill patients. Targeted implementation can increase the uptake of protocols by clinicians into practice. The theory of planned behaviour offers a framework in which to investigate clinicians' intention to perform the behaviour of interest. This study aimed to evaluate the effect of implementing a bowel management protocol on intensive care nursing and medical staffs' knowledge, attitude, subjective norms, perceived behavioural control, behaviour intentions, role perceptions and past behaviours in relation to three bowel management practices. A descriptive before and after survey using a self-administered questionnaire sent to nursing and medical staff working within three intensive care units before and after implementation of our bowel management protocol (pre: May - June 2008; post: Feb - May 2009). Participants had significantly higher knowledge scores post-implementation of our protocol (pre mean score 17.6; post mean score 19.3; p = 0.004). Post-implementation there was a significant increase in: self-reported past behaviour (pre mean score 5.38; post mean score 7.11; p = 0.002) and subjective norms scores (pre mean score 3.62; post mean score 4.18; p = 0.016) for bowel assessment; and behaviour intention (pre mean score 5.22; post mean score 5.65; p = 0.048) for administration of enema. This evaluation, informed by the theory of planned behaviour, has provided useful insights into factors that influence clinician intentions to perform evidence-based bowel management practices in intensive care. Addressing factors such as knowledge, attitudes and beliefs can assist in targeting implementation strategies to positively affect clinician behaviour change. Despite an increase in clinicians' knowledge scores, our implementation strategy did not, however, significantly change clinician behaviour intentions for all three bowel management practices. Further research is required to explore the influence of opinion leaders and organizational culture on clinicians' behaviour intentions related to bowel management for intensive care patients.


Diode-pumped Yb:GSO femtosecond laser.
Compact femtosecond laser operation of Yb:Gd(2)SiO(5) (Yb:GSO) crystal was demonstrated under high-brightness diode-end-pumping. A semiconductor saturable absorption mirror was used to start passive mode-locking. Stable mode-locking could be realized near the emission bands around 1031, 1048, and 1088 nm, respectively. The mode-locked Yb:GSO laser could be tuned from one stable mode-locking band to another with adjustable pulse durations in the range 1~100 ps by slightly aligning laser cavity to allow laser oscillations at different central wavelengths. A pair of SF10 prisms was inserted into the laser cavity to compensate for the group velocity dispersion. The mode-locked pulses centered at 1031 nm were compressed to 343 fs under a typical operation situation with a maximum output power of 396 mW.


Female sexual function evaluation of the tension-free vaginal tape (TVT) and transobturator suburethral tape (TOT) incontinence surgery: results of a prospective study.
Significantly more women reporting stress urinary incontinence (SUI) or low urinary tract symptoms complained of sexual dysfunction than a general healthy female population. The use of a tension-free vaginal tape (TVT), placed transvaginally under the mid-urethra, ensures to reacquire continence but could affect sexual function. This study aimed to determine if a suburethral tape for the correction of SUI could interfere with sexual activity. Of 108 patients, 37 underwent positioning of a tension free vaginal tape through a retropubic approach (TVT), and 71 through a transobturator approach from outside to inside (TOT). All patients enrolled had a complete urodynamic study and were invited to answer the Female Sexual Function Index questionnaire and our personal questionnaire, before surgery, after 1 month, and then at 3-month intervals. Of the 108 patients, 67% of the women experienced incontinence during intercourse, 96% during penetration, and 4% on orgasm. The cure rate for SUI was 97.1%. Sixty-two women (87%) with TOT placement and 31 (84%) with TVT were satisfied with the operation as regards sexual function, 68 (96%) and 29 (78%), respectively, with TOT and TVT had an improvement of urinary symptoms with resolution of urinary leakage during intercourse, after a 12.3-month follow-up. Just one patient treated with a TOT approach and two with the TVT complained of low grade of leakage. Of the 101 sexually active women, 90.1% reported a significant improvement in their sexual life, 9.9% referred a poor sexual activity not due to surgical intervention. In the surgical treatment of SUI, it is important to think about the patients' future sexual life and inform them that the great part of women can expect to improve their quality of life.


Drug susceptibility testing of Malassezia furfur strains to antifungal agents.
The minimal inhibitory concentration of the three most commonly used antifungal agents--ketoconazole, fluconazole and intraconazole against 28 strains of Malassezia furfur, was studied. The strains were recovered from patients with pityriasis versicolor, pityriasis simplex capitis and dermatitis seborrhoides and were cultured on Dixon's agar. Diagnostic sensitivity test agar was used to determine the minimal inhibitory concentrations of the antifungal agents. The established minimal inhibitory concentration of ketoconazole against Malassezia furfur was 0.02 mg/l, the minimal inhibitory concentration of intraconazole--0.05 mg/l and that of fluconazole--0.09 mg/l. The results of the study are consistent with the data reported by other European authors in their studies. On the basis of the accumulated data it is concluded that the tested antifungal agents are effective in the treatment of infections caused by Malassezia furfur but ketoconazole is superior to intraconazole and fluconazole in the treatment of these infections. This is the first study on the drug susceptibility of Malassezia furfur in Bulgaria.


GLUT-4 protein and citrate synthase activity in distally or proximally denervated rat soleus muscle.
The potential role of neurotrophic factors in the decline of glucose transporter (GLUT-4) protein levels and citrate synthase (CS) activity was studied by comparing distally with proximally denervated juvenile rat soleus muscle. Severing of the tibial nerve produced distal (long stump) or proximal (short stump) denervation. GLUT-4 levels and CS activities were measured at 24-h intervals for up to 96 h after denervation. No differences were observed in GLUT-4 or CS activity between soleus muscles left with short or long nerve stumps at any time point. However, within just 24 h, denervation decreased (P < 0.05). GLUT-4 and CS (67.4 +/- 3.3 and 63.4 +/- 1.7% of innervated control values, respectively). Both parameters continued to decline up to 96 h (44.4 +/- 3.1 and 48.7 +/- 4.0%, respectively). There was a significant correlation between the GLUT-4 protein level and CS activity over this 96-h period of denervation (r = 0.653, P < 0.001). A similar response in the 24-h denervated soleus of adult rats was observed. In contrast, 24-h denervation of red gastrocnemius (type IIa fibers) left with a long nerve stump resulted in a prevention of the decline of GLUT-4 and CS seen in red gastrocnemius left with a short nerve stump in both juvenile and adult animals. These results suggest that unlike type IIa muscles, the decline in GLUT-4 level and CS activity in type I soleus muscle after denervation results from a lack of coordinated electrical activity but likely does not involve a neurotrophic agent. These results also support the hypothesis that there is coregulation of decreased expression of GLUT-4 protein and CS activity in this model of reduced neuromuscular activity.


Late-onset porphyrias: what are they?
Porphyrias are inherited disorders of heme biosynthesis. ALA dehydratase porphyria (ADP) and congenital erythropoietic porphyria (CEP) are autosomal recessive porphyrias, and are typically expressed at birth or in childhood. However, a few cases of late-onset recessive porphyrias have been reported. Recently we encountered a late-onset ADP patient who developed symptoms of acute porphyria when he was 63 years old. This was accompanied by polycythemia vera. It was concluded that he developed the porphyria because an abnormal ALAD allele was clonally expanded by polycythemia vera. Upon reviewing the literature, a few cases of late-onset CEP were found to be also associated with hematologic abnormalities suggestive of myelodysplastic syndrome (MDS), another clonal disorder. These findings suggest that these late-onset porphyrias may be heterozygous for their gene defects, but clinical expression may be elicited if there is a loss of heterozygosity, either by a clonal expansion of the porphyric allele or by a loss of function mutation in the other allele.


Bacteremia caused by a recently described novel Desulfovibrio species.
An obligately anaerobic, fastidious, slowly growing, spiral, gram-negative bacterium was isolated from the blood of a 75-year-old man with acute onset of pyrexia. The patient responded rapidly to appropriate antibiotic therapy. Extensive investigation failed to detect a focus for the infection. Phenotypically, the organism was consistent with Desulfovibrio species. Microscopic investigation revealed an organism with a vibrioid or spirillioid morphology with rapidly progressive motility by means of a single polar flagellum. Biochemically, the organism produced large amounts of H2S and contained desulfovirdin. The 16S rRNA gene sequence of the organism was found to be most similar to those of members of the genus Desulfovibrio, with identical sequence homology to the newly proposed species described by Tee et al. (W. Tee, M. Dyall-Smith, W. Woods, and D. Eisen, J. Clin. Microbiol. 34:1760-1764, 1996). This is a second unrelated isolation of this novel species from two widely different locations in Australia. The two isolates show some phenotypic differences, indicating that they are different strains of the same species.


Protecting America's secrets while maintaining academic freedom.
The terrorist attacks of September 11, 2001, and the subsequent anthrax mail attacks, have had a profound impact on Americans' personal and professional lives and have sparked an active debate regarding the delicate balance between the need for national security and the pursuit of academic freedom. Although academic freedom can be defined in many ways, there are four primary tenets of freedom in an academic environment: freedom to research, freedom to publish, freedom to teach, and freedom to speak. Each of these tenets has come under attack in the wake of September 11, 2001. In this report the author further defines academic freedom and reflects upon recent events that have had a real or perceived impact on this freedom, including (1) attempts to categorize and restrict some research as "sensitive," (2) implementation of export control laws and select agent regulations, (3) limitations on the publication of research findings, (4) prohibition of certain foreign nationals from collaborating with U.S. researchers and receiving education and training in U.S. colleges and universities, and (5) restraint of faculty free speech. The author offers some suggestions as to how academia might achieve a proper balance between protecting our national security while promoting and maintaining academic freedom.


Sex differences in clinical response to olanzapine compared with haloperidol.
There is current disagreement over whether men and women respond differently to typical or atypical antipsychotic medications. This study reanalyzed a large international clinical trial of olanzapine (Olz) compared with haloperidol (Hal) to test for sex differences in treatment response, controlling for illness chronicity and menopausal status. We hypothesized that women would show a greater response to either medication than men, particularly among first admission, premenopausal women. DSM-III-R schizophrenia inpatients (700 women and 1295 men) were randomly assigned to a 6-week trial of Olz vs. Hal. Longitudinal random effect models were used to test for interactions of sex with medication, chronicity and menopausal status on treatment response. Findings showed that women on olanzapine had a greater drop in overall symptomatology by week 4 than any other group, and their level of symptomatology remained lower throughout the 6-week trial. The sex differences in treatment response in olanzapine compared with haloperidol were, in part, dependent on chronicity and, in women, menopausal status. That is, first episode women on haloperidol exhibited an increase in symptomatology over the 6-week trial compared to their male counterparts, while multiply hospitalized women had a better treatment response on haloperidol than their male counterparts. Women on olanzapine had a significantly better treatment response than men, regardless of chronicity. Finally, premenopausal women had a significantly better treatment response than postmenopausal women, regardless of treatment and chronicity.


Natural and probe fluorescence of lymphocytes of patients with pulmonary tuberculosis exposed to radiation as a result of the Chernobyl nuclear accident.
Parameters of natural and probe fluorescence of peripheral blood lymphocytes have been studied in 25 normal persons and 72 patients suffering from extensive forms of pulmonary tuberculosis who live in various radioecological conditions. It was found that in cell suspension the intensity of fluorescence of protein and the membrane-bound merocyanine 540 dye was 2.0-3.2 and 3.9-7.1 times higher, respectively, for the lymphocytes of the examined patients. For lymphocytes of the patients who live in radionuclide-contaminated areas fluorescence of reduced pyridine nucleotides and some fluorescent probes (ANS, DPGT, the ratio of eximer to monomer fluorescence of pyrene) and the merocyanine-photosensitized cell death were also found to increase. These can indicate some changes of cell membranes and reduction of the oxidation resistance of lymphocytes. There was not any substantial dependence on the changes in the parameters determined in the patients calculated radiation doses was revealed. It was found that biophysical tests, in particular, photosensitized cell death, are informative for estimation of severity of the tuberculous process and prognosis of its outcome in patients who live in adverse radioecological conditions.


[Excessive left retroperitoneal abscess of an unknown aetiology perforating into the peritoneal cavity: a case report].
Retroperitoneal abscesses present a relatively uncommon complication of diseases of various abdominal organs, although most commonly they are related to acute appendicitis of retrocoecal location. The paper presents the case of a healthy patient in whom an excessive left retroperitoneal abscess developed, perforated into the peritoneal cavity, and almost perforated through the skin in XIIth intercostal space. The patient had no abdominal symptoms or signs, but complained from slight pain in the left lumbar area. The diagnosis was established based on abdominal computed tomography scanning and an operative treatment by laparotomy. The evacuation and drainage of the abscess was effective and the patient recovered. Bacteriological examination of the pus from the abdominal cavity revealed single colonies of anaerobic Fusobacterium species. The cause of the occurrence of the abscess remained unknown.


Health care costs associated with child maltreatment: impact on medicaid.
To estimate the increased Medicaid expenditures associated with child maltreatment. Data on child maltreatment were collected from the National Survey of Child and Adolescent Well-Being, a nationally representative sample of cases investigated or assessed by local Child Protective Services agencies between October 1999 and December 2000. Medicaid claims data for 2000 to 2003 were obtained from the Medicaid Analytic Extract (MAX). Children from the National Survey of Child and Adolescent Well-Being who had Medicaid were matched to the MAX data by Social Security number or birthdate, gender, and zip code. Propensity score matching was used to select a comparison group from the MAX data. Two-part regression models were used to estimate the impact of child maltreatment on expenditures. Data with individual identifiers were obtained under confidentiality agreements with the collecting agencies. Children who were identified as maltreated or as being at risk of maltreatment incurred, on average, Medicaid expenditures that were >$2600 higher per year compared with children not so identified. This finding accounted for ~9% of all Medicaid expenditures for children. Child maltreatment imposes a substantial financial burden on the Medicaid system. These expenses could be partially offset by increased investment in child maltreatment prevention.


Ortho Isomeric Mn(III) N-Alkyl- and Alkoxyalkylpyridylporphyrins-Enhancers of Hyaluronan Degradation Induced by Ascorbate and Cupric Ions.
High levels of hyaluronic acid (HA) in tumors correlate with poor outcomes with several types of cancers due to HA-driven support of adhesion, migration and proliferation of cells. In this study we explored how to enhance the degradation of HA into low-molecular fragments, which cannot prevent the immune system to fight tumor proliferation and metastases. The physiological solution of HA was exposed to oxidative degradation by ascorbate and cupric ions in the presence of either one of three ortho isomeric Mn(III) substituted N-alkyl- and alkoxyalkylpyridylporphyrins or para isomeric Mn(III) N-methylpyridyl analog, commonly known as mimics of superoxide dismutase. The changes in hyaluronan degradation kinetics by four Mn(III) porphyrins were monitored by measuring the alteration in the dynamic viscosity of the HA solution. The ortho compounds MnTE-2-PyP5+ (BMX-010, AEOL10113), MnTnBuOE-2-PyP5+ (BMX-001) and MnTnHex-2-PyP5+ are able to redox cycle with ascorbate whereby producing H2O2 which is subsequently coupled with Cu(I) to produce the •OH radical essential for HA degradation. Conversely, with the para analog, MnTM-4-PyP5+, no catalysis of HA degradation was demonstrated, due to its inertness towards redox cycling with ascorbate. The impact of different Mn(III)-porphyrins on the HA decay was further clarified by electron paramagnetic resonance spectrometry. The ability to catalyze the degradation of HA in a biological milieu, in the presence of cupric ions and ascorbate under the conditions of high tumor oxidative stress provides further insight into the anticancer potential of redox-active ortho isomeric Mn(III) porphyrins.


In vivo NMR imaging of tissue sodium in the intact cat before and after acute cerebral stroke.
The first in vivo nuclear magnetic resonance (NMR) images of tissue sodium in the intact animal are presented. The distribution of sodium in the normal cat's head is described. An experimental stroke was surgically induced. Sodium NMR imaging showed a pronounced focal increase in cerebral sodium concentration 9 hr after ligation of the middle cerebral artery. The method appears to be very sensitive for early detection of infarction. The measured increase in the regional sodium NMR signal probably reflects both a true increase in concentration of brain sodium and an increased NMR visibility of the sodium nucleus in the region of the infarction.


Mechanisms on boron-induced alleviation of aluminum-toxicity in Citrus grandis seedlings at a transcriptional level revealed by cDNA-AFLP analysis.
The physiological and biochemical mechanisms on boron (B)-induced alleviation of aluminum (B)-toxicity in plants have been examined in some details, but our understanding of the molecular mechanisms underlying these processes is very limited. In this study, we first used the cDNA-AFLP to investigate the gene expression patterns in Citrus grandis roots responsive to B and Al interactions, and isolated 100 differentially expressed genes. Results showed that genes related to detoxification of reactive oxygen species (ROS) and aldehydes (i.e., glutathione S-transferase zeta class-like isoform X1, thioredoxin M-type 4, and 2-alkenal reductase (NADP+-dependent)-like), metabolism (i.e., carboxylesterases and lecithin-cholesterol acyltransferase-like 4-like, nicotianamine aminotransferase A-like isoform X3, thiosulfate sulfurtransferase 18-like isoform X1, and FNR, root isozyme 2), cell transport (i.e., non-specific lipid-transfer protein-like protein At2g13820-like and major facilitator superfamily protein), Ca signal and hormone (i.e., calcium-binding protein CML19-like and IAA-amino acid hydrolase ILR1-like 4-like), gene regulation (i.e., Gag-pol polyprotein) and cell wall modification (i.e., glycosyl hydrolase family 10 protein) might play a role in B-induced alleviation of Al-toxicity. Our results are useful not only for our understanding of molecular processes associated with B-induced alleviation of Al-toxicity, but also for obtaining key molecular genes to enhance Al-tolerance of plants in the future.


Exacerbation of Epilepsy by Astrocyte Alkalization and Gap Junction Uncoupling.
Seizures invite seizures. At the initial stage of epilepsy, seizures intensify with each episode; however, the mechanisms underlying this exacerbation remain to be solved. Astrocytes have a strong control over neuronal excitability and the mode of information processing. This control is accomplished by adjusting the levels of various ions in the extracellular space. The network of astrocytes connected via gap junctions allows a wider or more confined distribution of these ions depending on the open probability of the gap junctions. K+ clearance relies on the K+ uptake by astrocytes and the subsequent diffusion of K+ through the astrocyte network. When astrocytes become uncoupled, K+ clearance becomes hindered. Accumulation of extracellular K+ leads to hyperexcitability of neurons. Here, using acute hippocampal slices from mice, we uncovered that brief periods of epileptiform activity result in gap junction uncoupling. In slices that experienced short-term epileptiform activity, extracellular K+ transients in response to glutamate became prolonged. Na+ imaging with a fluorescent indicator indicated that intercellular diffusion of small cations in the astrocytic syncytium via gap junctions became rapidly restricted after epileptiform activity. Using a transgenic mouse with astrocyte-specific expression of a pH sensor (Lck-E2GFP), we confirmed that astrocytes react to epileptiform activity with intracellular alkalization. Application of Na+/HCO3- cotransporter blocker led to the suppression of intracellular alkalization of astrocytes and to the prevention of astrocyte uncoupling and hyperactivity intensification both in vitro and in vivo Therefore, the inhibition of astrocyte alkalization could become a promising therapeutic strategy for countering epilepsy development.SIGNIFICANCE STATEMENT We aimed to understand the mechanisms underlying the plastic change of forebrain circuits associated with the intensification of epilepsy. Here, we demonstrate that first-time exposure to only brief periods of epileptiform activity results in acute disturbance of the intercellular astrocyte network formed by gap junctions in hippocampal tissue slices from mice. Moreover, rapid clearance of K+ from the extracellular space was impaired. Epileptiform activity activated inward Na+/HCO3- cotransport in astrocytes by cell depolarization, resulting in their alkalization. Our data suggest that alkaline pH shifts in astrocytes lead to gap junction uncoupling, hampering K+ clearance, and thereby to exacerbation of epilepsy. Pharmacological intervention could become a promising new strategy to dampen neuronal hyperexcitability and epileptogenesis.


Motivation to change, coping, and self-esteem in adolescent anorexia nervosa: a validation study of the Anorexia Nervosa Stages of Change Questionnaire (ANSOCQ).
Understanding motivation to change is a key issue in both the assessment and the treatment of eating disorders. Therefore, sound instruments assessing this construct are of great help to clinicians. Accordingly, the present study analysed the psychometric properties of the Anorexia Nervosa Stages of Change Questionnaire (ANSOCQ), including its relation to coping style and self-esteem. N = 92 adolescents referred to an eating disorders outpatient clinic meeting criteria for anorexia nervosa gave written informed consent to participate in this study and completed the ANSOCQ, the Eating Disorder Inventory, the Eating Attitudes Test, the Body Image Questionnaire, two questionnaires measuring Self-Related Cognitions and the Coping Across Situations Questionnaire. After a treatment period of nine months, clinical anorexia nervosa diagnosis and the body mass index were re-assessed. In addition to exploratory factor analysis, correlational analysis was used to test for the convergent validity of the ANSOCQ and logistic regression analysis was used to test its predictive validity. The ANSOCQ had good psychometric properties. Factor analysis yielded two meaningful factors labelled as 'weight gain and control' and 'attitudes and feelings'. Internal consistencies of the two factors amounted to Cronbach's alpha = .87 and .76, respectively. Significant correlations with other scales measuring eating disorder psychopathology were indicative of meaningful construct validity. Higher motivation to change was related to higher self-esteem and a more active coping style. Higher (positive) ANSOCQ total scores predicted remission of anorexia nervosa after nine months of treatment. A higher score on 'attitudes and feelings' was a protective factor against drop-out from intervention. The ANSOCQ is a clinically useful instrument for measuring motivation to change in adolescents with AN. Two factorial dimensions explain most of the variation. Self-esteem and coping style are relevant additional constructs for the understanding of the motivation to change in anorexia nervosa. NCT02828956. Retrospectively registered July 2016.


Chronic Remote Ischemic Conditioning Is Cerebroprotective and Induces Vascular Remodeling in a VCID Model.
Vascular contributions to cognitive impairment and dementia (VCID) make up 50% of the cases of dementia. The purpose of this study was to determine the effect of chronic remote ischemic conditioning (C-RIC) on improving long-term (6 months) outcomes and cerebral blood flow (CBF) and collateral formation in a mouse model of VCID. Adult C57BL/6J male mice (10 weeks) were randomly assigned to four different groups: (1) sham-bilateral carotid artery stenosis (BCAS), (2) BCAS + sham RIC, (3) BCAS+C-RIC for 1 month (1MO), and (4) BCAS+C-RIC-4 months (4MO). CBF, cognitive impairment, and functional outcomes were performed up for 6 months after BCAS surgery. The expression of CD31, α-SMA, and myelin basic protein (MBP) was assessed by immunohistochemistry (IHC). Additional set of mice were randomized to sham, BCAS, and BCAS+C-RIC. The cerebrovascular angioarchitecture was studied with micro-CT. RIC therapy for either 1 or 4 months significantly improved CBF, new collateral formation, functional and cognitive outcomes, and prevented white matter damage. There was no difference between C-RIC for 1 or 4 months; IHC studies at 6 months showed an increase in brain CD31 and α-SMA expression indicating increased angiogenesis and MBP indicating preservation of white matter in animals receiving RIC. One month of daily RIC is as effective as 4 months of daily RIC in improving CBF, angiogenesis, and long-term functional outcomes (6 months) in a VCID model. This suggests that 1 month of RIC is sufficient to reduce cognitive impairment and induce beneficial cerebrovascular remodeling.


GHRH agonist MR-409 protects β-cells from streptozotocin-induced diabetes.
Patients with type 1 diabetes (T1D) suffer from insufficient functional β-cell mass, which results from infiltration of inflammatory cells and cytokine-mediated β-cell death. Previous studies demonstrated the beneficial effects of agonists of growth hormone-releasing hormone receptor (GHRH-R), such as MR-409 on preconditioning of islets in a transplantation model. However, the therapeutic potential and protective mechanisms of GHRH-R agonists on models of T1D diabetes have not been explored. Using in vitro and in vivo models of T1D, we assessed the protective propertie of the GHRH agonist, MR409 on β-cells. The treatment of insulinoma cell lines and rodent and human islets with MR-409 induces Akt signaling by induction of insulin receptor substrate 2 (IRS2), a master regulator of survival and growth in β-cells, in a PKA-dependent manner. The increase in cAMP/PKA/CREB/IRS2 axis by MR409 was associated with decrease in β-cell death and improved insulin secretory function in mouse and human islets exposed to proinflammatory cytokines. The assessment of the effects of GHRH agonist MR-409 in a model of T1D induced by low-dose streptozotocin showed that mice treated with MR-409 exhibited better glucose homeostasis, higher insulin levels, and preservation of β-cell mass. Increased IRS2 expression in β-cells in the group treated with MR-409 corroborated the in vitro data and provided evidence for the underlying mechanism responsible for beneficial effects of MR-409 in vivo. Collectively, our data show that MR-409 is a novel therapeutic agent for the prevention and treatment of β-cells death in T1D.


Effects of grade of oocyte-cumulus complex and the interactions between grades on the production of blastocysts in the cow, ewe and lamb.
There is an association between the size of the cumulus investment and the in vitro developmental ability of the oocyte-cumulus complex (OCC) that provides a basis for the selection of OCCs. However, the value of selection is confounded by humoral interactions between OCCs that influence the development of OCCs of other grade(s). This study examined the effect of size of the cumulus investment (OCC grade) and the interactions between grades on the developmental ability of oocytes collected from the cow, ewe and lamb. OCCs were classified into A, B and C grades on visual assessment of the number of cumulus cell layers or left unselected (Unselected). In the cow, there were 669 +/- 228 to 4763 +/- 228 cells per OCC whereas comparable figures in the ewe and lamb were 593 +/- 252 to 3716 +/- 252 and 366 +/- 228 to 3263 +/- 228 respectively (A > Unselected > B > C; Experiment 1). In Experiment 2, OCCs were made to mature within grade and the efficiency of blastocyst production and blastocyst quality was compared with that obtained in the Unselected group. Grade was associated with significant (p < 0.05) differences in cleavage rate, blastocyst production rate and the mean number of nuclei per embryo (generally A > B > C across animal types). However, the performance of A grade OCCs in the cow and lamb did not differ significantly from that obtained in the Unselected group whereas in the ewe, A grade OCCs were significantly (p < 0.05) better. Furthermore, the performance of the Unselected group was significantly (p < 0.05) better than that of the combined grades (A + B + C) in the cow but there were no differences in either the ewe or lamb. It is concluded that (i) interactions between OCCs of different grade influence the developmental ability of OCCs in the cow and, to a lesser extent, the lamb, (ii) selection of OCCs in the cow and lamb would lead to the exclusion of many OCCs that have the ability to develop into blastocysts and (iii) selection in the ewe would improve the efficiency of blastocyst production although its value is limited by the low percentage of A grade OCCs.


A let-7 MicroRNA-sensitive vesicular stomatitis virus demonstrates tumor-specific replication.
Creation of potent oncolytic viruses (OVs) suitable for the clinic may require new strategies in virus design. Replication-competent viruses facilitate a variety of approaches to achieving tumor specificity. Altered expression of microRNAs is a common hallmark of cancer that we demonstrate can be used to alter expression of a potent wild-type viral gene to achieve tumor-specific replication of an engineered vesicular stomatitis virus (VSV). Incorporation of let-7 microRNA complementary sequences within VSV eliminates undesirable replication and associated toxicity in normal cells but permits growth in cancer cells in vitro and in vivo. This is proof of concept that viruses designed to exploit the differential microRNA expression in cancer cells is a viable approach, potentially useful in optimizing oncolytic viral gene expression for maximal antitumor activity and safety.


A comparison of support needs between rural and urban family caregivers providing palliative care.
The present study examined the support needs for urban and rural family caregivers of a palliative family member using a cross-sectional telephone survey in northeastern Ontario, Canada (n = 140; 70 urban, 70 rural). Support needs identified as most important by both the groups were informational. Rural caregivers reported greater unmet needs in tangible support (P = .01). No differences were observed between the groups for emotional or informational support needs (P = .25 and P = .35, respectively). Rural and urban caregivers perceived care for care recipients as accessible (mean accessibility score 1.9, standard deviation [SD] = 0.09 and 1.7, SD = 0.7, respectively, P = .20); the majority indicated that when needed, services were easily and quickly obtained. Although there are similarities in the formal care experiences, rural caregivers experience greater unmet needs in receiving support for instrumental activities.


Characterization of biomechanical properties of cells through dielectrophoresis-based cell stretching and actin cytoskeleton modeling.
Cytoskeleton is a highly dynamic network that helps to maintain the rigidity of a cell, and the mechanical properties of a cell are closely related to many cellular functions. This paper presents a new method to probe and characterize cell mechanical properties through dielectrophoresis (DEP)-based cell stretching manipulation and actin cytoskeleton modeling. Leukemia NB4 cells were used as cell line, and changes in their biological properties were examined after chemotherapy treatment with doxorubicin (DOX). DEP-integrated microfluidic chip was utilized as a low-cost and efficient tool to study the deformability of cells. DEP forces used in cell stretching were first evaluated through computer simulation, and the results were compared with modeling equations and with the results of optical stretching (OT) experiments. Structural parameters were then extracted by fitting the experimental data into the actin cytoskeleton model, and the underlying mechanical properties of the cells were subsequently characterized. The DEP forces generated under different voltage inputs were calculated and the results from different approaches demonstrate good approximations to the force estimation. Both DEP and OT stretching experiments confirmed that DOX-treated NB4 cells were stiffer than the untreated cells. The structural parameters extracted from the model and the confocal images indicated significant change in actin network after DOX treatment. The proposed DEP method combined with actin cytoskeleton modeling is a simple engineering tool to characterize the mechanical properties of cells.


Phylogenetic characterization of Escherichia coli O157 : H7 based on IS629 distribution and Shiga toxin genotype.
Shiga toxin (stx)-producing Escherichia coli O157 : H7 is a prominent food-borne pathogen. Symptoms in human infections range from asymptomatic to haemorrhagic colitis and haemolytic uraemic syndrome, and there is a need for methods that yield information that can be used to better predict clinical and epidemiological outcomes. IS629 is an insertion sequence notable for its prevalence and variable distribution in the chromosome of E. coli O157 : H7, which has been exploited for subtyping and strain characterization. In particular, IS629 distribution is closely aligned with the major phylogenetic lineages that are known to be distinctive in their genome structure and virulence potential. In the present study, a comprehensive subtyping method in which IS629-typing was combined with stx genotyping was developed using a conventional PCR approach. This method consisted of a set of 32 markers based on the unique distribution of IS629 in the three major phylogenetic lineages of E. coli O157 : H7 and six additional markers to determine the stx genotype, a key virulence signature associated with each lineage. The analysis of IS629 loci variation with the 32 markers allowed us to determine the IS629 distribution profile (IDP), phylogenetic lineage and genetic relatedness of the 31 E. coli O157 : H7 strains examined. An association between IDP typing and stx genotype was observed. The use of both IDP and the stx genotype for strain characterization provided confirmative and complementary data in support of lineage placement of closely related strains. In addition, IS629/stx profiles were in agreement with strain segregation based on LSPA-6 (lineage-specific polymorphism assay) and PFGE subtyping, demonstrating its potential as a subtyping and strain tracking method.


Microbial community structure and sulfur biogeochemistry in mildly-acidic sulfidic geothermal springs in Yellowstone National Park.
Geothermal and hydrothermal waters often contain high concentrations of dissolved sulfide, which reacts with oxygen (abiotically or biotically) to yield elemental sulfur and other sulfur species that may support microbial metabolism. The primary goal of this study was to elucidate predominant biogeochemical processes important in sulfur biogeochemistry by identifying predominant sulfur species and describing microbial community structure within high-temperature, hypoxic, sulfur sediments ranging in pH from 4.2 to 6.1. Detailed analysis of aqueous species and solid phases present in hypoxic sulfur sediments revealed unique habitats containing high concentrations of dissolved sulfide, thiosulfate, and arsenite, as well as rhombohedral and spherical elemental sulfur and/or sulfide phases such as orpiment, stibnite, and pyrite, as well as alunite and quartz. Results from 16S rRNA gene sequencing show that these sediments are dominated by Crenarchaeota of the orders Desulfurococcales and Thermoproteales. Numerous cultivated representatives of these lineages, as well as the Thermoproteales strain (WP30) isolated in this study, require complex sources of carbon and respire elemental sulfur. We describe a new archaeal isolate (strain WP30) belonging to the order Thermoproteales (phylum Crenarchaeota, 98% identity to Pyrobaculum/Thermoproteus spp. 16S rRNA genes), which was obtained from sulfur sediments using in situ geochemical composition to design cultivation medium. This isolate produces sulfide during growth, which further promotes the formation of sulfide phases including orpiment, stibnite, or pyrite, depending on solution conditions. Geochemical, molecular, and physiological data were integrated to suggest primary factors controlling microbial community structure and function in high-temperature sulfur sediments.


Experiences and challenges faced by community mental health workers when providing care to people with mental health conditions: A qualitative descriptive study.
The global prevalence of mental health conditions and the associated wide treatment gaps have led to increased demand for quality mental healthcare services. In Singapore, despite a shift towards a joint provision of mental healthcare in hospitals and the community, experiences of mental healthcare teams in such hospital-community partnerships have remained unclear. Through a qualitative descriptive approach, this study explored community mental health workers' experiences and perceptions of working with partners from tertiary hospitals and other community organisations. Eighteen participants were individually interviewed with a semi-structured question guide through video conferencing. This study was guided by the 32-item checklist from the Consolidated Criteria for Reporting Qualitative Research. Data were subjected to Braun and Clark's six-step thematic analysis. Three themes emerged: (i) the need for supportive partners, due to institutional and legal challenges faced in the participants' liaison with hospital partners and other community organisations; (ii) the need for a supportive organisation, whose presence and importance were revealed by the interviews; and (iii) the necessity of advanced training, identified by the participants as a contributor to their professional growth. The findings highlighted the importance of inter-sectoral communication and positive effects of a robust organisational support system. Accordingly, hospitals and community mental healthcare organisations should consider strengthening networking and boosting case discussions through regular inter-sectoral meetings that would prioritise the clients' continuity of care. Furthermore, opportunities for advanced trainings should be created for community-based mental healthcare workers.


Somatostatin receptor subtype 1 might be a predictor of better response to therapy in medullary thyroid carcinoma.
Medullary thyroid carcinoma (MTC) is a malignant neoplasm of parafollicular cells. Because it is a neuroendocrine tumor, it has known somatostatin receptors (SSTRs). The actual frequencies of the SSTR subtypes and their potential influences (by binding with endogenous somatostatin) on MTC cell proliferation have not been fully elucidated to date. The present study evaluated the occurrence of SSTR subtypes 1, 2, 3 and 5 as well as the possible role that each subtype plays in the clinical evolution of patients with MTC. This retrospective, longitudinal study analyzed thyroid surgical material from 42 patients with MTC. Immunohistochemical staining was performed with monoclonal antibodies against subtypes 1, 2, 3 and 5 of SSTR. The histological material was classified as negative, focal positive or diffuse positive, in relation to each of the SSTR subtypes. The initial response to treatment, clinical course and patient mortality rate were assessed and related to the presence of SSTR subtypes. The most prevalent SSTR subtype was SSTR 3, which was found in 81% of the patients, when considering any pattern of positivity. However, subtype 2 had the lowest number of positive patients, with 28.6% demonstrating any positive pattern. Subtypes 1 and 5 had an intermediate prevalence of positivity, with subtype 1 present in 45.2% of the patients and subtype 5 positive in 54.8% of the patients, when considering any pattern of positivity. The presence of STR 1, in the form of diffuse positivity, independently predicted a better response to the initial therapy, with a hazard ratio (HR) of 4.80 (p = 0.03). This is the first study to show the correlation of the presence of SSTR1, detected by monoclonal immunohistochemical techniques, and better response to initial treatment and possibly better long-term clinical response in patients with MTC. In addition, these patients had low positivity rates for SSTR2, which might explain the low sensitivity of diagnostic and limited therapeutic response to octrotide based radioisotopes.


The negative impact of cadmium on nitrogen transformation processes in a paddy soil is greater under non-flooding than flooding conditions.
Nitrification and denitrification are two important processes in the nitrogen (N) cycle. Under heavy-metal pollution with water management of paddy soils, these two processes are not well understood. This study aimed to examine the effect of cadmium (Cd) on N transformation under flooding and non-flooding conditions. A paddy soil was incubated under two water regimes (flooding and non-flooding) and four Cd levels (0, 2, 5 and 10 mg kg-1). The availability of Cd was higher in the non-flooding than flooding conditions. Cadmium contamination significantly (p ≤ 0.05) decreased the copy number of archaeal and bacterial amoA genes, bacterial nirS, nirK and nosZ genes under both conditions with the decrease being greater under non-flooding. High level of Cd (10 mg kg-1) was more toxic in non-flooding than flooding conditions to the nitrifiers and denitrifiers, which in turn decreased N transformation through microbially-mediated processes. Its contamination decreased N2O emission initially under both water regimes but the effect was greater under the non-flooding condition. However, the non-significant stimulatory effect of Cd on N2O emission was observed during the late phase. The microbial community structure was changed with time and water regimes. Irrespective of water regime, the dominated fungal phyla were Ascomycota and Basidiomycota while the dominated bacteria phyla were Actinobacteria, Proteobacteria, Firmicutes and Acidobacteria. In summary, water regimes and Cd bioavailability changed soil N transformations via microbial mediated processes.


Spironolactone versus eplerenone for the treatment of idiopathic hyperaldosteronism.
The aim of this prospective, randomised, open-label, blinded-end point study was to compare the efficacy and safety of eplerenone versus spironolactone in patients with bilateral idiopathic hyperaldosteronism (IHA). After a 2-week washout period, 34 patients with IHA were assigned to receive either spironolactone 25 mg b.i.d. (n = 17) or eplerenone 25 mg b.i.d. (n = 17) for 24 weeks. If the patients' blood pressure (BP) was not < 140/90 mmHg, the doses were gradually increased up to 400 mg for spironolactone and 200 mg for eplerenone. If the patients' BP remained uncontrolled, a daily dose of hydrochlorothiazide 12.5 mg was added at week 16. The primary outcome was the percentage of patients with BP < 140/90 mmHg at 16 weeks (i.e., with aldosterone antagonist monotherapy). The patients' BP was normalised in 13 out of 17 (76.5%) and 14 out of 17 (82.4%) patients in the spironolactone and eplerenone groups, respectively (p = 1.00). Systolic BP decreased more rapidly with eplerenone. Serum potassium levels were normalised (> 3.5 mmol/l) in all patients at 4 weeks. Mild hyperkalaemia was observed in two patients receiving 400 mg of spironolactone and in three patients receiving 150 mg of eplerenone. Two patients presented with bilateral painful gynaecomastia at the end of week 16 while receiving 400 mg of spironolactone. Switching spironolactone to 150 mg of eplerenone daily resulted in resolution of gynaecomastia and also maintained BP control. At the end of the study, 19 patients were on eplerenone and 15 were on spironolactone. However, this did not affect the primary end point, because the switch from spironolactone to eplerenone (in two patients) occurred at the end of week 16. It was concluded that eplerenone was as effective as spironolactone in reducing BP in patients with IHA. The risk of mild hyperkalaemia was similar with both drugs.


[The duration of psychosomatic rehabilitation: provisions, influencing factors, recommendations].
The decision of the German legislator to limit rehabilitation programme participation under the health-insurance scheme to a duration of three weeks had been oriented less by rehabilitation-scientific findings rather than by economic considerations. This is the background for discussing the question to what extent medical experience and empirical data are providing new information as to the durations of rehabilitation necessary and feasible in the field of psychosomatic rehabilitation and as to measures capable of supporting its effectiveness without extending the duration of therapy.


Neutron Scattering Measurements of Carbon Dioxide Adsorption in Pores within the Marcellus Shale: Implications for Sequestration.
Shale is an increasingly viable source of natural gas and a potential candidate for geologic CO2 sequestration. Understanding the gas adsorption behavior on shale is necessary for the design of optimal gas recovery and sequestration projects. In the present study neutron diffraction and small-angle neutron scattering measurements of adsorbed CO2 in Marcellus Shale samples were conducted on the Near and InterMediate Range Order Diffractometer (NIMROD) at the ISIS Pulsed Neutron and Muon Source, STFC Rutherford Appleton Laboratory along an adsorption isotherm of 22 °C and pressures of 25 and 40 bar. Additional measurements were conducted at approximately 22 and 60 °C at the same pressures on the General-Purpose Small-Angle Neutron Scattering (GP-SANS) instrument at Oak Ridge National Laboratory. The structures investigated (pores) for CO2 adsorption range in size from Å level to ∼50 nm. The results indicate that, using the conditions investigated densification or condensation effects occurred in all accessible pores. The data suggest that at 22 °C the CO2 has liquid-like properties when confined in pores of around 1 nm radius at pressures as low as 25 bar. Many of the 2.5 nm pores, 70% of 2 nm pores, most of the <1 nm pores, and all pores <0.25 nm, are inaccessible or closed to CO2, suggesting that despite the vast numbers of micropores in shale, the micropores will be unavailable for storage for geologic CO2 sequestration.


ADHESIVENESS OF SPIDER SILK.
Moths, by virtue of the loose scales that cover their wings and bodies, are admirably adapted to elude capture by orb-weaving spiders. Rather than sticking to the web, they may simply lose some of their scales to the viscid threads, and then fly on. Other insects, covered with detachable hairs or waxy powder, are similarly protected against entrapment. Quantitative data are presented on the adhesiveness of spider thread to insect cuticles oq various kinds.


Work of a nurse in a health centre treatment room.
In six months there were 1,704 attendances at the treatment room of a small health centre. The attendance rate for the population registered with the health centre doctors was 448 per 1,000 patients per year. Females between 15 and 44 years and males under 15 had the highest attendance rates. There were 256 casual attenders, 58 (23%) of whom were referred to a doctor or hospital for further advice or treatment.It is suggested that in a health centre treatment room about six hours of nursing time a week for every 1,000 patients is required, and that a case can be made out for some of the routine work of casualty departments being done in health centres.


[An observational study of 'assertive outreach' with remission as outcome measure].
Systems of intensive care such as the Function Assertive Community Treatment (Function-act) are advocated because they are reported to have achieved very promising results. However, comparative studies aimed at measuring the efficacy of such systems are mainly of short duration and the outcome measure is nearly always the amount of care provided. To investigate, via an observational study, changes that occurred in the number of patients going into remission and in the amount of care provided before and after the introduction of f-act. f-act was introduced in Maastricht and the surrounding area (population 220,000) at the beginning of 2002. Cumulative routine measurements of the effect of care in the region had been performed since 1998. These measurements were used to compare the proportion of patients in remission in two non-overlapping patients cohorts, each covering a 4-year period, namely 1998-2001 and 2002-2005. results The proportion of patients who made the transition to remission increased from 19% in the period before the introduction of f-act to 31% in the period after its introduction (or = 2.21; 95% ci: 1.03-4.78), but after controlling for confounders the difference was no longer statistically significant. In routine clinical practice f-act can reduce the intensity of psychopathological symptoms, possibly because the care provided under f-act is less fragmented. However, more research is needed before the claimed beneficial effects of f-act can be proved conclusively.


Role of endothelin-1 in age-dependent cerebrovascular hypotensive responses after brain injury.
This study was designed to compare the effect of fluid percussion brain injury (FPI) on the hypotensive cerebrovascular response in newborn and juvenile pigs as a function of time postinsult and to determine the role of endothelin-1 (ET-1) in any age-dependent differences in hypotensive cerebrovascular regulation after injury. Ten minutes of hypotension (10-15 ml blood/kg) decreased mean arterial blood pressure uniformly in both groups ( approximately 45%). In the newborn, hypotensive pial artery dilation (PAD) was blunted within 1 h, remained diminished for at least 72 h, but was resolved within 168 h postinjury (66 +/- 4, 69 +/- 4, 71 +/- 4, and 64 +/- 4% inhibition at 1, 4, 8, and 72 h post-FPI). During normotension, regional cerebral blood flow (rCBF) was decreased by FPI, and hypotension further reduced the already decremented rCBF for at least 72 h. Cerebrospinal fluid (CSF) ET-1 was increased from 26 +/- 4 to 206 +/- 25 pg/ml within 72 h post-FPI, whereas an ET-1 antagonist partially restored impaired hypotensive PAD and altered hypotensive rCBF. In contrast, hypotensive PAD and altered CBF were only inhibited for 4 h post-FPI in the juvenile (56 +/- 3 and 34 +/- 4% inhibition at 1 and 4 h post-FPI). CSF ET-1 was only increased from 27 +/- 4 to 67 +/- 9 pg/ml at 4 h, whereas the concentration returned to preinjury value by 8 h post-FPI. ET-1 antagonism similarly partially restored impaired hypotensive PAD and altered hypotensive rCBF. These data show that FPI disturbs cerebral autoregulation during hypotension both to a greater magnitude and for a longer duration in the newborn than in the juvenile. These data suggest that the greater FPI-induced ET-1 release in the newborn could contribute to age-dependent differences in impaired hypotensive cerebral autoregulation after FPI.


Embryonal carcinoma cells transfected with ZP3 genes differentially glycosylate similar polypeptides and secrete active mouse sperm receptor.
Mouse and hamster sperm receptors, called mZP3 (approximately 83,000 Mr) and hZP3 (approximately 56,000 Mr), respectively, are glycoproteins located in the ovulated egg zona pellucida. Certain of the glycoprotein O-linked oligosaccharides are essential for sperm receptor activity. Here, we transfected mouse embryonal carcinoma (EC) cells with mZP3 and hZP3 genes placed under control of a constitutive promoter. Transfected cells synthesized and secreted large amounts of the glycoproteins, called EC-mZP3 and EC-hZP3. Although the primary structures of mZP3 and hZP3 polypeptides (44,000 Mr) are very similar to one another, EC-mZP3 (approximately 83,000 Mr) and EC-hZP3 (approximately 49,000 Mr) were glycosylated to very different extents, such that they resembled their egg counterparts. Like egg mZP3, EC-mZP3 inhibited binding of sperm to ovulated eggs and induced sperm to acrosome-react in vitro. In addition, large numbers of sperm bound to aggregates of mZP3-transfected EC cells in vitro. On the other hand, unlike egg hZP3, EC-hZP3 did not exhibit either sperm receptor or acrosome reaction-inducing activity, and sperm failed to bind to aggregates of hZP3-transfected EC cells. Thus, transfected EC cells not only express sperm receptor genes, but also discriminate between very similar polypeptides with respect to glycosylation and, in the case of mZP3, add specific oligosaccharides essential for biological activity. In addition, the results demonstrate that EC cells can serve as a source for large amounts of functional mouse sperm receptor.


Modulation of Neutrophil Extracellular Trap and Reactive Oxygen Species Release by Periodontal Bacteria.
Oral bacteria are the main trigger for the development of periodontitis, and some species are known to modulate neutrophil function. This study aimed to explore the release of neutrophil extracellular traps (NETs), associated antimicrobial proteins, and reactive oxygen species (ROS) in response to periodontal bacteria, as well as the underlying pathways. Isolated peripheral blood neutrophils were stimulated with 19 periodontal bacteria. NET and ROS release, as well as the expression of NET-bound antimicrobial proteins, elastase, myeloperoxidase, and cathepsin G, in response to these species was measured using fluorescence-based assays. NET and ROS release was monitored after the addition of NADP (NADPH) oxidase pathway modulators and inhibitors of Toll-like receptors (TLRs). Moreover, bacterial entrapment by NETs was visualized microscopically, and bacterial killing was assessed by bacterial culture. Certain microorganisms, e.g., Veillonella parvula and Streptococcus gordonii, stimulated higher levels of ROS and NET release than others. NETs were found to entrap, but not kill, all periodontal bacteria tested. NADPH oxidase pathway modulators decreased ROS production but not NET production in response to the bacteria. Interestingly, TLR inhibitors did not impact ROS and NET release. These data suggest that the variability in the neutrophil response toward different bacteria may contribute to the pathogenesis of periodontal diseases by mechanisms such as bacterial avoidance of host responses and activation of neutrophils. Moreover, our results indicate that bacterium-stimulated NET release may arise in part via NADPH oxidase-independent mechanisms. The role of TLR signaling in bacterium-induced ROS and NET release needs to be further elucidated.


A Novel Inorganic Ni-La2O3 Composite with Superfast and Versatile Water Purification Behavior.
Water-purifying materials are of vital importance for providing sanitary water, especially in epidemic and disaster areas and for wilderness survival. Here, we report a novel inorganic composite with a fast, reusable, and pleiotropic water purification ability. The composite of heterogeneous Ni-La2O3 nanofibers was developed using an easy, low-cost, and large-scale production process, i.e., an electrospinning technique. Because of the unique heterostructure formed by introducing nickel nanoparticles into the La2O3 host, the nanocomposite fibers can rapidly remove various detrimental micropollutants with absorption rate constants hundreds of times greater than those of inorganic materials and activated carbons. In addition, the nanocomposite can be recycled by a soft washing procedure at ambient temperature and demonstrates a much more stable performance than the state-of-the-art activated carbon materials. Furthermore, the nanofibers have an excellent germicidal effect against the high-risk human pathogen Escherichia coli. Our work introduces a promising inorganic absorbent suitable for purifying life-sustaining water supplies in emergency situations and alleviating worldwide water shortages.


Evaluation of a cement incus replacement prosthesis in a temporal bone model.
In reconstruction of the ossicular chain for a damaged incus, it is important that the incus replacement prosthesis (IRP) length is ideal in order to provide optimal tension between the tympanic membrane or malleus and stapes head to achieve the best post-operative hearing result. Even though the length of commercially available IRPs can be adjusted, it still may be difficult to achieve clinically. We describe experiments in a human temporal bone model using dental cement as an IRP after removal of the incus. This cement IRP (CIRP) hardens in situ and becomes the length of the gap to be spanned so that tension should be ideal. Two different CIRPs were studied; one was a conventional rod-type CIRP connecting either the umbo or mid-malleus handle to the stapes head. The second was a Y-shaped CIRP (Y-CIRP), connecting two sites on the malleus to the stapes head. The wide Y-CIRP connected the malleus head and umbo to the stapes head, while the narrow Y-CIRP connected the malleus neck and mid-handle to the stapes head. The acoustic performance of these experimental CIRPs was studied using a laser Doppler vibrometer system in 12 fresh human temporal bones. The CIRP demonstrated better acoustic performance than conventional IRPs studied previously in the same model. While all the CIRPs showed similar function below 2.0 kHz, the narrow Y-CIRP appeared best above 3.0 kHz. A prosthesis of this type may have an acoustic advantage over conventional IRPs.


Surface segregation and stability of core-shell alloy catalysts for oxygen reduction in acid medium.
Density functional theory is used for the evaluation of surface segregation, trends for dissolution of Pt surface atoms in acid medium, and oxygen reduction reaction activity of core-shell materials, containing a monolayer of platinum over a monometallic or bimetallic core. Two groups of cores are investigated: Pt/X with X = Ir, Au; Pd, Rh, Ag; Co, Ni, Cu; and Pt/Pd(3)X, with X = Co, Fe, Cr, V, Ti, Ir, Re. It is found that all the 4d and 5d pure cores may serve as stable cores, and their beneficial effect on the Pt monolayer may be further tuned by alloying the core to another element, here chosen from 3d or 5d groups. The Pd(3)X cores enhance the stability of the surface Pt atoms both in vacuum and under adsorbed oxygen; however the high oxygen philicity of some of the X elements induces their surface segregation that may cause surface poisoning with oxygenated species and their dissolution in acid medium.