Abdominal adipose tissue cytokine gene expression: relationship to obesity and metabolic risk factors.
Adipose tissue is a major source of inflammatory and thrombotic cytokines. This study investigated the relationship of abdominal subcutaneous adipose tissue cytokine gene expression to body composition, fat distribution, and metabolic risk during obesity. We determined body composition, abdominal fat distribution, plasma lipids, and abdominal subcutaneous fat gene expression of leptin, TNF-alpha, IL-6, PAI-1, and adiponectin in 20 obese, middle-aged women (BMI, 32.7 +/- 0.8 kg/m2; age, 57 +/- 1 yr). A subset of these women without diabetes (n = 15) also underwent an OGTT. In all women, visceral fat volume was negatively related to leptin (r = -0.46, P < 0.05) and tended to be negatively related to adiponectin (r = -0.38, P = 0.09) gene expression. Among the nondiabetic women, fasting insulin (r = 0.69, P < 0.01), 2-h insulin (r = 0.56, P < 0.05), and HOMA index (r = 0.59, P < 0.05) correlated positively with TNF-alpha gene expression; fasting insulin (r = 0.54, P < 0.05) was positively related to, and 2-h insulin (r = 0.49, P = 0.06) tended to be positively related to, IL-6 gene expression; and glucose area (r = -0.56, P < 0.05) was negatively related to, and insulin area (r = -0.49, P = 0.06) tended to be negatively related to, adiponectin gene expression. Also, adiponectin gene expression was significantly lower in women with vs. without the metabolic syndrome (adiponectin-beta-actin ratio, 2.26 +/- 0.46 vs. 3.31 +/- 0.33, P < 0.05). We conclude that abdominal subcutaneous adipose tissue expression of inflammatory cytokines is a potential mechanism linking obesity with its metabolic comorbidities.


Early Results of Surgical Simultaneous Therapy for Significant Carotid Artery Stenosis and Heart Disease.
The purpose of this single-center, retrospective study was to review the early outcome with defined endpoints of myocardial infarction, brain injury, and death after coronary bypass grafting and simultaneous carotid endarterectomy with or without combined valve replacement. During the preoperative investigation 52 (44 males, 8 females) patients were examined by cardiac catheterization within the scope of their coronary status. In addition, all patients underwent echocardiography, CT angiography of the supra-aortic vessels, and full description of their neurologic status. Included were all patients with the need of coronary revascularization, valve disease, and either symptomatic or asymptomatic carotid artery stenosis with a maximal narrowing of >70%. There was one (2%) patient with a perioperative stroke and paresis of the upper limb. One (2%) patient experienced PRIND (2%). Eighteen (35%) patients suffered symptomatic transitory psychotic syndrome that fully recovered in all patients. One (2%) patient incurred a perioperative myocardial infarction that could be cured by percutaneous coronary stent implantation. Overall mortality was 4%. Simultaneous coronary bypass grafting as a single procedure or in combination with valve replacement and endarterectomy of severe carotid lesions with or without patients' history of previous stroke can be performed with a calculated low surgical risk. The complication rate for neurologic and myocardial events is low compared with the hazard of a single surgical repair. The in-hospital mortality is not significantly different to isolated procedures. The concomitant appearance of coronary stenosis and carotid artery disease is reasonable due to the nature of arteriosclerosis. In our opinion these patients benefit from a combined surgical approach.


NK-1 receptor antagonists: a new paradigm in pharmacological therapy.
The neuropeptide substance P (SP) shows a widespread distribution in both the central and peripheral nervous systems and it is known that after binding to the neurokinin-1 (NK-1) receptors, SP regulates many biological functions in the central nervous system such as emotional behaviour, stress, depression, anxiety, emesis, migraine, alcohol addiction and neurodegeneration. SP has been also implicated in pain, inflammation, hepatotoxicity and in virus proliferation, and it plays an important role in cancer (e.g., tumour cell proliferation, angiogenesis, and the migration of tumour cells for invasion and metastasis). By contrast, it is known that after binding to NK-1 receptors, NK-1 receptor antagonists specifically inhibit the above-mentioned biological functions mediated by SP. Thus, these antagonists exert an anxyolitic, antidepressant, antiemetic, antimigraine, antialcohol addiction or neuroprotector effect in the central nervous system, and they play a role in analgesic, antiinflammatory, hepatoprotector processes and in antivirus proliferation. Regarding cancer, NK-1 receptor antagonists exert an antitumour action (inducing tumour cell death by apoptosis), and induce antiangiogenesis and inhibit the migration of tumour cells. It is also known that NK-1 receptors have a widespread distribution and that they are overexpressed in tumour cells. Thus, NK-1 receptor antagonists are molecularly targeted agents. In general, current drugs have a single therapeutic effect, although less commonly they may exert several. However, the data reported above indicate that NK-1 receptor antagonists are promising drugs, exerting many therapeutic effects (the action of such antagonists is dose-dependent and, depending on the concentration, has more positive effects). In this review, we update the multiple therapeutic effects exerted by NK-1 receptor antagonists.


Risk factors for low colloid osmotic pressure during infant cardiopulmonary bypass with a colloidal prime.
Extensive variations of colloid osmotic pressure (COP) measured in the priming as well as during infant cardiopulmonary bypass motivated us to audit clinical and laboratory data to identify the risk factors for low COP at the end of bypass. Data of 73 consecutive infant patients with body weight <10 kg, who underwent elective, first time open-heart surgery between March 2005 and December 2006 were examined. The following variables were analyzed: COP, blood loss, transfusion requirements and hematological data. Univariate and multivariate analysis of risk factors for low COP (<15 mmHg) was performed. Forty-eight percent of patients had COP <15 mmHg at the end of bypass. Those patients had significantly lower COP before start of bypass, during, and at the end of the operation. Significant univariate predictors of low COP at the end of bypass were: lower patient weight; lower COP before start of bypass, lower priming COP and larger volume of cardioplegia received into the circulation. After multivariable analysis, lower patient COP before bypass remained the only significant predictor for low COP at the end of bypass. Pre-bypass crystalloid dilution during induction should be avoided, as this is the most important cause of low COP during the bypass. Priming COP and COP management strategy should be adapted to the individual patient demand.


Rapid Reductions in Breast Density following Tamoxifen Therapy as Evaluated by Whole-Breast Ultrasound Tomography.
Women whose mammographic breast density declines within 12-18 months of initiating tamoxifen for chemoprevention or adjuvant treatment show improved therapeutic responses compared with those whose density is unchanged. We tested whether measuring changes in sound speed (a surrogate of breast density) using ultrasound tomography (UST) could enable rapid identification of favorable responses to tamoxifen. We evaluated serial density measures at baseline and at 1 to 3, 4 to 6, and 12+ months among 74 women (aged 30-70 years) following initiation of tamoxifen for clinical indications, including an elevated risk of breast cancer (20%) and diagnoses of in situ (39%) or invasive (40%) breast carcinoma, enrolled at Karmanos Cancer Institute and Henry Ford Health System (Detroit, MI, USA). For comparison, we evaluated an untreated group with screen negative mammography and frequency-matched on age, race, and menopausal status (n = 150), at baseline and 12 months. Paired t-tests were used to assess differences in UST sound speed over time and between tamoxifen-treated and untreated patients. Sound speed declined steadily over the 12 month period among patients receiving tamoxifen (mean (SD): -3.0 (8.2) m/s; p = 0.001), whereas density remained unchanged in the untreated group (mean (SD): 0.4 (7.1) m/s; p = 0.75 (relative change between groups: p = 0.0009)). In the tamoxifen group, we observed significant sound speed reductions as early as 4-6 months after tamoxifen initiation (mean (SD): -2.1 (6.8) m/s; p = 0.008). Sound speed reductions were greatest among premenopausal patients (P-interaction = 0.0002) and those in the middle and upper tertiles of baseline sound speed (P-interaction = 0.002). UST can image rapid declines in sound speed following initiation of tamoxifen. Given that sound speed and mammographic density are correlated, we propose that UST breast imaging may capture early responses to tamoxifen, which in turn may have utility in predicting therapeutic efficacy.


Nonadherence in outpatient thromboprophylaxis after major orthopedic surgery: a systematic review.
The necessity for extended medication-based thromboprophylaxis after hip/knee-replacement surgery (major orthopedic surgery) has been acknowledged in international guidelines. In this article, we review and critically appraise the literature regarding patients' nonadherence (NA) in outpatient thromboprophylaxis after major orthopedic surgery. We conducted a systematic literature review. All studies published since 1990 and that were found to report research about NA in outpatient thrombosis prophylaxis after major orthopedic surgery were included. Only six relevant contributions could be identified. All these studies dealt with parenteral low-molecular-weight heparins or fondaparinux prophylaxis. The extent of NA (defined as existing when a patient fails to take the prescribed medication on at least 1 day) ranged from 13 to 37%. In one large German survey, patients who were nonadherent missed between 38 and 43% of their outpatient low-molecular-weight heparin injections. Subjective factors can play a role in increasing NA; such factors include a lack of knowledge of or having no fear concerning thrombosis in general and a lack of specific knowledge regarding measures to prevent it, as well as a negative evaluation of injections as the form of therapy application. Waiting times between acute in-hospital treatment and admission to rehabilitation clinics, as well as abstention from stationary rehabilitation programs, form objective adherence barriers. Therefore, NA is a phenomenon influenced by subjective patient-related factors as well as objective, care-provision structural factors. Current trends in patient care (e.g., shorter hospital stays and lengthened ambulant care) are likely to increase both the number of nonadherent patients and the extent of NA, if the current state of knowledge proves an accurate predictor of the future. At present, it appears that between one and two of every five patients are not adherent when parenteral prophylaxis is used. Whether or not new oral anticoagulation alternatives will be capable of improving the situation remains open for future research.


Evidence for the extracellular reduction of ferricyanide by rat liver. A trans-plasma membrane redox system.
1. Reduction of ferricyanide by the isolated perfused rat liver and by isolated rat hepatocytes was studied. 2. Ferricyanide was reduced to ferrocyanide by the perfused liver at a linear rate of 0.22mumol/min per g of liver. Ferricyanide was not taken up by the liver and the perfusate concentration of ferricyanide+ferrocyanide remained constant throughout the perfusion. Perfusate samples from livers perfused without ferricyanide did not reduce ferricyanide. 3. Isolated hepatocytes reduced ferricyanide in a biphasic manner. The initial rate of 2.3mumol/min per g of cells proceeded for approx. 3min and derived from low-affinity sites (apparent K(m)>1.3mm). The secondary rate of 0.29mumol/min per g of cells was maintained for the remainder of the incubation and derived from higher affinity sites (apparent K(m)0.13mm). Disruption of the cells resulted in an increase in the low-affinity rate and a decrease in the high-affinity rate. 4. Ferrocyanide was oxidized by isolated hepatocytes but not by perfused liver. The apparent K(m) for ferrocyanide oxidation by hepatocytes was 1.3mm. 5. Oxidized cytochrome c was reduced by isolated hepatocytes in the presence of 1mm-KCN but at a rate less than that of the reduction of ferricyanide. 6. Properties of the ferricyanide-reducing activities of intact hepatocytes and the perfused liver were examined. The low-affinity rate, present only in cell and broken cell preparations, was inhibited by 1mum-rotenone and 0.5mm-ferrocyanide, and stimulated by 0.1mm-KCN. The mitochondrial substrate, succinate, also stimulated this rate. The perfused liver showed only a high-affinity activity for ferricyanide reduction. This activity was also present in liver cells and was unaffected by rotenone, antimycin A, KCN, NaN(3), or p-hydroxymercuribenzoate but was inhibited by 2.6mm-CaCl(2), 2-heptyl-4-hydroxyquinoline-N-oxide and ferrocyanide. Overall, these results are consistent with the occurrence of a trans-plasma membrane redox system of liver that reduces extracellular ferricyanide to ferrocyanide. The reduction process shows properties which are similar to that of the NADH:ferricyanide oxidoreductase found in isolated liver plasma membranes but different from that of mitochondria.


Incidence of respiratory syncytial virus infection in infants and young children referred to the emergency departments for lower respiratory tract diseases in Italy.
Respiratory Syncytial Virus (RSV) is the leading cause of emergency visits and hospitalization for acute lower respiratory tract infections (LRTI) in infants and young children worldwide. To collect specific epidemiological data on the incidence of RSV infection among infants referred to Emergency Departments (ED) for LRTI in a Mediterranean country, an Italian multicenter epidemiological surveillance program was established. Eight pediatric centers throughout Italy participated in this study. The study population included 272 children < or =4 years of age, admitted to the ED between October 2000 and April 2001 for respiratory problems that might be possibly related to LRTI. 152 children were <1 year of age, 50 between 1 and 2 years, and 70 >2 years of age. Data regarding medical history and physical examination were recorded for each child, whereas an immunoenzymatic RSV test (TestPack RSV, Abbott) was performed on nasal and pharyngeal secretions. Out of 272 tested children, 85 were positive for RSV. The peak of the RSV epidemic occurred in February, with an earlier start and end of the RSV season in the northern and central regions, compared to the southern regions. Major risk factors for RSV infection were younger age (p<0.05) and low weight at birth (p<0.05). Among children positive for RSV infection, 55.2% were <1 year of age, 18.3% were between 1 and 2 years, and 25.7% were > 2 years of age. RSV positivity was associated with a higher rate of hospitalization in the whole study population (p<0.01) and especially in the children < or =12 months of age (p<0.01). Clinical evidence of lower respiratory tract involvement, was also more frequently observed in RSV positive than in RSV negative children, both in the whole study population (p<0.01) and in the < or =12 months of age subgroup (p<0.01). These data confirm that the patterns of RSV infection in Italy are similar to those reported for other countries in the northern hemisphere: RSV is associated with a higher risk of hospitalization and clinically evident LRTI involvement than respiratory infections of other etiologies, especially in infants.


CD25 and CD69 induction by α4β1 outside-in signalling requires TCR early signalling complex proteins.
Distinct signalling pathways producing diverse cellular outcomes can utilize similar subsets of proteins. For example, proteins from the TCR (T-cell receptor) ESC (early signalling complex) are also involved in interferon-α receptor signalling. Defining the mechanism for how these proteins function within a given pathway is important in understanding the integration and communication of signalling networks with one another. We investigated the contributions of the TCR ESC proteins Lck (lymphocyte-specific kinase), ZAP-70 (ζ-chain-associated protein of 70 kDa), Vav1, SLP-76 [SH2 (Src homology 2)-domain-containing leukocyte protein of 76 kDa] and LAT (linker for activation of T-cells) to integrin outside-in signalling in human T-cells. Lck, ZAP-70, SLP-76, Vav1 and LAT were activated by α4β1 outside-in signalling, but in a manner different from TCR signalling. TCR stimulation recruits ESC proteins to activate the mitogen-activated protein kinase ERK (extracellular-signal-regulated kinase). α4β1 outside-in-mediated ERK activation did not require TCR ESC proteins. However, α4β1 outside-in signalling induced CD25 and co-stimulated CD69 and this was dependent on TCR ESC proteins. TCR and α4β1 outside-in signalling are integrated through the common use of TCR ESC proteins; however, these proteins display functionally distinct roles in these pathways. These novel insights into the cross-talk between integrin outside-in and TCR signalling pathways are highly relevant to the development of therapeutic strategies to overcome disease associated with T-cell deregulation.


[Intraorganic and intratissue mechanisms of protective action of activated Kupffer cells on hepatocytes].
Endotoxine activated Kupfer cells release into the intercellular space several mediators which act directly on hepatocytes as well as via stellet cells. In both cases Kupfer cells downregulate hepatocytes as a part of paracrine system. However, downregulated part of liver parenchyma might be extended by several mechanisms. The first one is release of vasoconstrictors from activated Kupfer cells which stimulate stellet cells contraction. This effect may also be achieved by formation of hypermetabolicfocuses by Kupffer cells mediators with further activation of hepatocyte-hepatocyte interactions based on the principle of cell competition for oxygen in the intercellular space. Regulatory influence of activated Kupfer cells may be spread in liver parenchyma with participation of the mechanism of intratissue hepatocyte-hepatocyte interactions which also realize tissue stress reaction.


Effective control of Schistosoma haematobium infection in a Ghanaian community following installation of a water recreation area.
Urogenital schistosomiasis caused by Schistosoma haematobium was endemic in Adasawase, Ghana in 2007. Transmission was reported to be primarily through recreational water contact. We designed a water recreation area (WRA) to prevent transmission to school-aged children. The WRA features a concrete pool supplied by a borehole well and a gravity-driven rainwater collection system; it is 30 m(2) and is split into shallow and deep sections to accommodate a variety of age groups. The WRA opened in 2009 and children were encouraged to use it for recreation as opposed to the local river. We screened children annually for S. haematobium eggs in their urine in 2008, 2009, and 2010 and established differences in infection rates before (2008-09) and after (2009-10) installation of the WRA. After each annual screening, children were treated with praziquantel and rescreened to confirm parasite clearance. Initial baseline testing in 2008 established that 105 of 247 (42.5%) children were egg-positive. In 2009, with drug treatment alone, the pre-WRA annual cumulative incidence of infection was 29 of 216 (13.4%). In 2010, this incidence rate fell significantly (p<0.001, chi-squared) to 9 of 245 (3.7%) children after installation of the WRA. Logistic regression analysis was used to determine correlates of infection among the variables age, sex, distance between home and river, minutes observed at the river, low height-for-age, low weight-for-age, low Body Mass Index (BMI)-for-age, and previous infection status. The installation and use of a WRA is a feasible and highly effective means to reduce the incidence of schistosomiasis in school-aged children in a rural Ghanaian community. In conjunction with drug treatment and education, such an intervention can represent a significant step towards the control of schistosomiasis. The WRA should be tested in other water-rich endemic areas to determine whether infection prevalence can be substantially reduced.


Feeding and attraction of non-target flies to spinosad-based fruit fly bait.
A spinosad-based fruit fly bait, GF-120, has recently become a primary tool for area-wide suppression or eradication of pest tephritid fruit flies. The present study assessed the attraction and feeding of five non-target fly species to GF-120 in Hawaii. These non-target flies include three beneficial tephritid species [Eutreta xanthochaeta (Aldrich), Tetreuaresta obscuriventris (Loew), Ensina sonchi (L.)] introduced for weed biological control, an endemic Hawaiian tephritid [Trupanea dubautiae (Bryan)] (all Diptera: Tephritidae) and the cosmopolitan Drosophila melanogaster Meigen (Diptera: Drosophilidae). All five non-target fly species were susceptible to GF-120, as was the target pest Mediterranean fruit fly Ceratitis capitata (Wiedemann). Feeding on, or even brief tasting of, GF-120 killed all fly species within 2 h. When individual flies were provided with a choice of GF-120 or honey solution, there was no difference in the frequency of first food encounter by E. xanthochaeta, D. melanogaster or C. capitata. The other three non-target species approached honey more often than GF-120 in their first food encounter. Feeding times on GF-120 and honey were not significantly different for D. melanogaster and C. capitata, while the other four non-target species fed longer on honey than on GF-120. There was no significant difference in feeding time on honey versus GF-120 between males and females of each species. These results suggest that area-wide treatment using GF-120 for the purpose of eradication of pest fruit flies has potential negative impacts on these and other non-target fly species in Hawaii.


Unraveling the terpene synthase family and characterization of BsTPS2 contributing to (S)-( +)-linalool biosynthesis in Boswellia.
Boswellia tree bark exudes oleo-gum resin in response to wounding, which is rich in terpene volatiles. But, the molecular and biochemical basis of wound-induced formation of resin volatiles remains poorly understood. Here, we combined RNA-sequencing (RNA-seq) and metabolite analysis to unravel the terpene synthase (TPS) family contributing to wound-induced biosynthesis of resin volatiles in B. serrata, an economically-important Boswellia species. The analysis of large-scale RNA-seq data of bark and leaf samples representing more than 600 million sequencing reads led to the identification of 32 TPSs, which were classified based on phylogenetic relationship into various TPSs families found in angiosperm species such as TPS-a, b, c, e/f, and g. Moreover, RNA-seq analysis of bark samples collected at 0-24 h post-wounding shortlisted 14 BsTPSs that showed wound-induced transcriptional upregulation in bark, suggesting their important role in wound-induced biosynthesis of resin volatiles. Biochemical characterization of a bark preferentially-expressed and wound-inducible TPS (BsTPS2) in vitro and in planta assays revealed its involvement in resin terpene biosynthesis. Bacterially-expressed recombinant BsTPS2 catalyzed the conversion of GPP and FPP into (S)-( +)-linalool and (E)-(-)-nerolidol, respectively, in vitro assays. However, BsTPS2 expression in Nicotiana benthamiana found that BsTPS2 is a plastidial linalool synthase. In contrast, cytosolic expression of BsTPS2 did not form any product. Overall, the present work unraveled a suite of TPSs that potentially contributed to the biosynthesis of resin volatiles in Boswellia and biochemically characterized BsTPS2, which is involved in wound-induced biosynthesis of (S)-( +)-linalool, a monoterpene resin volatile with a known role in plant defense.


Preserved Analgesia With Reduction in Opioids Through the Use of an Acute Pain Protocol in Enhanced Recovery After Surgery for Open Hepatectomy.
Enhanced recovery after surgery (ERAS) pathways are designed to restore baseline physiology, mitigate surgical stressors, and hasten recovery. Paramount to this approach is optimal pain control through multimodal analgesia and limiting reliance on opioid-based medications. Recent studies have fostered growing controversy surrounding the use of epidural analgesia in the ERAS setting, especially for higher-risk procedures. We examine the analgesic end points associated with the use of epidural within the ERAS framework for open hepatectomy. From November 2013 through March 2016, postoperative analgesic end points including daily morphine equivalent administration and self-reported pain scores were prospectively collected and analyzed for 180 consecutive patients scheduled for open hepatectomy. Patients whose surgeries performed prior to July 2014 were managed using traditional strategy (pre-ERAS, n = 60), and those after July 1 underwent a comprehensive perioperative ERAS pathway (ERAS, n = 120). Patients managed using the ERAS pathway had a significant reduction in morphine equivalent requirements at 24 hours (median, 10.0 vs 116.0 mg; P < 0.001), 48 hours (median, 10.1 vs 85.4 mg; P < 0.001), and 72 hours (median, 2.5 vs 60.0 mg; P < 0.001) compared with pre-ERAS counterparts with a reduction in average pain scores at 24 hours (numeric pain rating scale, 4.1 ± 1.6 vs 5.1 ± 1.8) and similar scores at other time points. Within ERAS, patients who received epidural (n = 87) required significantly less morphine equivalents at 24 hours (median, 2.7 vs 65.0 mg; P < 0.001) and 48 hours (median, 8.0 vs 50.0 mg; P < 0.001) but not at 72 hours (median, 1.3 vs 4.5 mg; P = 0.56), as well as improved pain scores at 24 hours (visual analog scale score, 3.8 ± 1.3 vs 5.0 ± 1.8; P < 0.001) and 48 hours (3.4 ± 1.8 vs 4.7 ± 1.9; P = 0.001) compared with those who did not receive epidural (n = 33). Other associated postoperative end points including provision of fluids, rates of clinically significant hypotension, and lengths of stay between epidural and nonepidural groups were similar. A novel ERAS protocol for open hepatectomy successfully reduced reliance on perioperative opioids without expensing adequate analgesia compared with traditional care. Patients within ERAS benefitted from application of epidural, which further reduced opioid requirements and optimized pain control without increasing complication rates. Epidurals should remain an integral part of ERAS protocols for liver resection surgery.


Simultaneous determination of oridonin, ponicidin and rosmarinic acid from Herba Isodi Rubescentis extract by LC-MS-MS in rat plasma.
An analytical method was developed for the simultaneous quantification of oridonin, ponicidin and rosmarinic acid in rat plasma using liquid chromatography-tandem mass spectrometry with electrospray ionization in positive and negative ion modes of operation. Multiple reaction monitoring was used for quantification using the precursor to product ion pairs of m/z 359.100 → 160.9 (rosmarinic acid), m/z 150.1 → 106.9 (acetaminophen as internal standard 1), m/z 365.1 → 347.3 (oridonin), m/z 363.3 → 345.2 (ponicidin) and m/z 258.3 → 201.0 (dextrorphan as internal standard 2). Optimum chromatographic separation was achieved with a BDS Hypersil C18 column (100 × 2.1 mm, 2.4 µm) and a mobile phase consisting of acetonitrile-5mM ammonium acetate containing 0.1% acetic acid (25:75, v/v). The analytes were extracted from rat plasma using a single liquid-liquid extraction technique and the calibration curve was linear within the concentration range of 2-1,000.0 ng/mL for three analytes. The extraction recovery was above 70%, the intra-day and inter-day precision of the quality control (QC) samples was less than 12.45% and the accuracy of the QC samples was 96.99-105.79% of the nominal values. The method was sensitive and reliable and was successfully applied to a pharmacokinetic and bioavailability study of Herba Isodi Rubescentis extract in rats.


Who is the key worker in palliative home care?
Palliative home care involves coordination of care between the professionals involved. The NICE guideline on supportive and palliative care (UK) recommends that teams, regardless of their base, should promote continuity for patients. This may involve nomination of a coordinating "key worker". This study aimed to explore who acts as key worker and who ought to take on this role in the views of patients, relatives, and primary care professionals. Furthermore, it aimed to explore the level of agreement on this issue between study participants. Interview and questionnaire study. Former County of Aarhus, Denmark (2008-2009). Ninety-six terminally ill cancer patients, their relatives, general practitioners (GPs), and community nurses (CNs). Actual key worker as valued by patients, relatives, and primary care professionals; ideal key worker as valued by patients and relatives. RESULTS. Patients, relatives, GPs, and CNs most often saw themselves as having been the key worker. When asked about the ideal key worker, most patients (29%; 95%CI: 18;42) and relatives (32%; 95%CI: 22;45) pointed to the GP. Using patients' views as reference, we found very limited agreement with relatives (47.7%; k = 0.05), with GPs (30.4%; k = 0.01) and with CNs (25.0%; k = 0.04). Agreement between patients and relatives on the identity of the ideal key worker was of a similar dimension (29.6%; k = 0.11). Poor agreement between patients, relatives, and professionals on actual and ideal key worker emphasizes the need for matching expectations and clear communication about task distribution in palliative home care.


Earth's earliest non-marine eukaryotes.
The existence of a terrestrial Precambrian (more than 542 Myr ago) biota has been largely inferred from indirect chemical and geological evidence associated with palaeosols, the weathering of clay minerals and microbially induced sedimentary structures in siliciclastic sediments. Direct evidence of fossils within rocks of non-marine origin in the Precambrian is exceedingly rare. The most widely cited example comprises a single report of morphologically simple mineralized tubes and spheres interpreted as cyanobacteria, obtained from 1,200-Myr-old palaeokarst in Arizona. Organic-walled microfossils were first described from the non-marine Torridonian (1.2-1.0 Gyr ago) sequence of northwest Scotland in 1907. Subsequent studies found few distinctive taxa-a century later, the Torridonian microflora is still being characterized as primarily nondescript "leiospheres". We have comprehensively sampled grey shales and phosphatic nodules throughout the Torridonian sequence. Here we report the recovery of large populations of diverse organic-walled microfossils extracted by acid maceration, complemented by studies using thin sections of phosphatic nodules that yield exceptionally detailed three-dimensional preservation. These assemblages contain multicellular structures, complex-walled cysts, asymmetric organic structures, and dorsiventral, compressed organic thalli, some approaching one millimetre in diameter. They offer direct evidence of eukaryotes living in freshwater aquatic and subaerially exposed habitats during the Proterozoic era. The apparent dominance of eukaryotes in non-marine settings by 1 Gyr ago indicates that eukaryotic evolution on land may have commenced far earlier than previously thought.


The Role of Fluorodeoxy-D-glucose Positron Emission Tomography/Computed Tomography in Nodal Staging of Nonsmall Cell Lung Cancer in Sequential Surgical Algorithm.
With nonsmall cell lung cancer (NSCLC), accurate mediastinal nodal staging is crucial to determine whether a patient is or is not a surgical candidate. Traditionally, computed tomography (CT) and fluorodeoxy-D-glucose (FDG) positron emission tomography (PET)/CT are the initial steps followed by tissue sampling through mediastinoscopy and/or thoracotomy, which are invasive procedures. There is controversy regarding the possibility of omission of the invasive diagnostic procedures and solely relying on noninvasive presurgical staging CT and FDG PET/CT results. Eighty-three patients who had PET/CT, mediastinoscopy, and thoracotomy for NSCLC were analyzed. For all lymph nodes that may be sampled by mediastinoscopy, PET/CT sensitivity was 80%, specificity was 86%, positive predictive value was 47%, and negative predictive value (NPV) was 97%; and for those in this group whose clinical stage was T1/T2 M0, sensitivity was 100% and specificity was 84%. For lymph nodes accessible only at thoracotomy, sensitivity was 42% and specificity was 88%. FDG PET/CT is accurate in assessing stations 2R/L, 4R/L, and 7 nodes and has the potential to replace mediastinoscopy in the treatment algorithm of T1/T2 M0 disease. A negative PET/CT may potentially prevent the patient from invasive mediastinoscopy given its high NPV. However, a patient with positive PET/CT should undergo tissue biopsy with pathology confirmation.


[Use of catabolic repression in the selection of the glucoamylase-producer Aspergillus niger].
The effect of various carbon sources and cAMP on the glucoamylase synthesis in Aspergillus niger was studied to find carbon sources repressed the enzyme synthesis and conditions for the selection of catabolite stable mutants. Maltose at a concentration of 0.5% stimulated the glucoamylase synthesis, but at a concentration of 4% it repressed not only the enzyme synthesis but the growth of the parental strain on the agar medium. The more active mutant 66 was obtained as a result of treatment of Asp. niger st 6 with NG. This mutant is able to grow on the Czapek's medium containing maltose at concentrations 4 or 6%. The mutant 66 produced about 2.9 times more glucoamylase than its parent when maltose was added at 0.5% concentration to the medium. The glucoamylase synthesis in the parental strain was completely repressed under repressing conditions, while the level of the mutant strain activity was 35% from the level of enzyme activity on the medium without the repressor. The addition of cAMP (5.10(-5] resulted in a partial release of maltose (4%) repression of the glucoamylase synthesis in both strains. The results obtained indicate a possibility to select Asp niger mutants with the partially derepressed glucoamylase synthesis. Other regulation mechanisms in addition to catabolite repression may be involved in the regulation of the glucoamylase synthesis.


[Teamwork and team success in multi- and interdisciplinary teams in medical rehabilitation].
Multidisciplinary or interdisciplinary team models are often used to describe teams in medical rehabilitation. There is only little evidence whether these models are used in the daily routine of medical rehabilitation and to what extent the quality (team work) and effectiveness (team success) differ between the models. For the present study an interview guide was designed to assess the team models. In each of the six clinics phone interviews were conducted with one member of the management. Data on teamwork and team success were collected with a staff questionnaire. The questionnaires were distributed to all employees (n = 266; response rate n = 127, 48%) of the six teams. Two teams were classified as interdisciplinary teams and four teams as multidisciplinary. The comparison between the two models shows better results for the interdisciplinary team model. Several methodical restrictions (e. g. unconsidered moderator variables, representativeness of the sample) need to be considered. Systematic analyses of teamwork as presented are essential to identify the conditions of successful teamwork and also form the basis for implementation of team development measures in medical rehabilitation.


Electrostatic charge on a plastic spacer device influences the delivery of salbutamol.
The aim of this study was to determine whether electrostatic charge on a plastic spacer decreases the delivery of salbutamol from a pressurized metered-dose inhaler (pMDI) and, if so, to find an optimal and practical treatment to remove the charge. Ten single actuations from a salbutamol pMDI were drawn through different Volumatic spacers at a constant flow of 60 L.min-1. The efficacies of different methods of removing charge were tested, including detergent coating of the spacers. A multistage liquid impinger was used to determine the particle size distribution of the output of the pMDI through the Volumatic spacers. The electrostatic charge on the inner surface of the spacers was measured both quantitatively with an electrometer, and qualitatively by the attraction of a thin strip of cellulose membrane to the wall of the spacer. Each experiment was repeated four times. Ionic detergent coating of the spacers removed the charge for at least 24 h. This resulted in an increase of 55-70% in small particle (< 6.8 microns) delivery compared to delivery from new spacers with high charge. We have demonstrated that electrostatic charge plays a major role in the delivery of salbutamol through plastic spacers. Adequate treatment with ionic detergent removes the charge and improves drug delivery.


TP53 codon 72 polymorphism and P53 protein expression in colorectal cancer specimens in Isfahan.
The TP53 tumor suppressor gene plays important roles in genomic stability. A common polymorphism at codon 72 of TP53 gene has been associated with increased risk for many human cancers. The p53 protein is expressed in colorectal cancer, but the reported prevalence of its expression varies widely. In the present study, the p53 protein expression in different genotypes of its codon 72 , was investigated. We undertook a case-control study on 250 controls and 250 paraffin block specimens of sporadic colorectal adenocarcinomas from the city of Isfahan. PCR amplification of TP53 codon 72 polymorphism: TP53 codon 72 genotypes were detected by PCR using specific primer pairs for amplifying the proline or the arginine Alleles. The PCR reaction was done separately for each of the two polymorphic variants. The amplified products were subjected to electrophoresis on 1% agarose gel in 1× TBE buffer and visualized on a transilluminator using ethidium bromide. Immunohistochemical Staining: We evaluated the expression patterns of p53 protein, as potential prognostic marker in colorectal cancer specimens by immunohistochemical staining. The χ2-test was used to assess the significance of any difference in the prevalence of TP53 codon 72 polymorphism between colorectal cancer patients and controls. The odds ratio and 95% CI (confidence intervals) was used as a measure of the strength of the association. Statistical significance level was set to P≤0.05. In control samples, the genotype distribution for TP53 polymorphism showed 30.4%, 45.2% and 24.4% for the arginine/arginine, arginine/proline and proline/proline genotypes, respectively. Allelic frequencies corresponded to 0.663 for the arginine allele and 0.338 for the proline allele. In the cancer group 38.8% of the cases were arginine/arginine, 40.4% were arginine/proline and 20.8% were proline/proline. The corresponding frequencies were 0.590 for the arginine allele and 0.410 for the proline allele. A significant difference between cases and controls was found for the arginine/arginine genotype compared with (grouped) arginine/proline and proline/proline genotypes (Odds Ratio = 1.451 (1.002-2.103), P=0.048). Overexpression of p53 was observed in 50.8 percent of cancer specimens which most of them were arginine/arginine genotype (P<0.001). TP53 polymorphism and arginine/arginine genotype may be correlated with overexpression of p53 and increased risk for colorectal cancer in city of Isfahan.


Consideration of stiffness and mass effects of relatively thicker electrodes with Mindlin plate theory.
Mindlin plate theory has been widely used in the high-frequency vibrations of piezoelectric crystal plates with emphasis on its applications in crystal resonator analysis and design. The plate equations were derived without considering the effect of electrodes from the beginning. But continuing efforts have been made to include the mechanical effect, or the mass loading, through the consideration of the mass ratio of the electrodes and crystal blank. Such a consideration has been effective for relatively thin electrodes before, but the ever-increasing mass ratio has been pressing further improvement to take into account relatively thicker electrodes. To extend Mindlin plate equations for these applications, we derive the plate equations systematically with the approximation of displacements in electrodes with those in the crystal blank. As a result, both mass and stiffness effects of electrodes are considered through ratios of the thickness, density, and elastic constants of the electrodes to those of the crystal blank, respectively, and the plate equations are modified accordingly. A practical design of the electrodes and crystal blank are analyzed to demonstrate the necessity of such modifications to Mindlin plate equations.


Technique for interaction of optical fields with turbulent medium containing particles.
We develop a method for transmission of stochastic fields through turbulent media (atmosphere, ocean, biotissues) containing randomly distributed particles. The method is based on the angular spectrum representation of stochastic, statistically stationary, scalar fields, the Rytov perturbation series for propagation in weakly fluctuating media, and the first Born approximation for weak scattering from particulate media. The results for transmission of the deterministic (laser) field may be obtained from our general results as a limiting case.


Inhibitor of DNA binding 1 (Id1) mediates stemness of colorectal cancer cells through the Id1-c-Myc-PLAC8 axis via the Wnt/β-catenin and Shh signaling pathways.
Inhibitor of DNA binding 1 (Id1) is upregulated in multiple cancers, and Id1overexpression correlates with cancer aggressiveness and poor clinical outcomes in cancer patients. However, its roles in cancer stem-like cells (CSCs) and epithelial-mesenchymal transition (EMT) are still elusive. This study aimed to examine the role of Id1 on the mediation of CRC stemness and explore the underlying mechanisms. Id1 and CD133 expression was detected by qPCR assay and immunohistochemistry (IHC) in normal mucosal and primary colorectal cancer (CRC) specimens. Id1 was stably knocked down (KD) in human CRC cell lines. Spheres forming assay and tumorigenic assay were performed to evaluate self-renewal capacity and tumor initiation. Expression of CSC- and EMT-related markers and TCF/LEF activity were assessed in HCT116 cells after Id1 KD. qPCR assay showed higher Id1 and CD133 expression in CRC specimens than in normal mucosal specimens (P<0.05). IHC detected high cytoplasmic Id1 expression in 35 CRC specimens (46.7%), and high CD133 expression in 22 CRC specimens (29.3%) and negative expression in 18 normal mucosal specimens. High Id1 expression positively correlated with poor differentiation (P=0.034), and CD133 expression correlated with T category in CRC patients (P=0.002). Spearman correlation analysis revealed a positive correlation between Id1 and CD133 expression in CRC patients (P<0.05). Id1 KD resulted in suppression of proliferation, cell-colony formation, self-renewal capability and CSC-like features in HCT116 cells, and impaired the tumor-initiating capability in CRC cells. In addition, Id1 maintained the stemness of CRC cells via the Id1-c-Myc-PLAC8 axis through activating the Wnt/β-catenin and Shh signaling pathways. Id1 expression significantly correlates with CD133 expression in CRC patients, and Id1 KD impairs CSC-like capacity and reverses EMT traits, partially via the Wnt/β-catenin signaling. Id1 may be a promising therapeutic target against colon CSCs.


Rationale and design of the ADVOR (Acetazolamide in Decompensated Heart Failure with Volume Overload) trial.
Decisive evidence on the optimal diuretic agent, dosing schedule, and administration route is lacking in acute heart failure (AHF) with congestion. The Acetazolamide in Decompensated heart failure with Volume OveRload (ADVOR) trial is designed to test the hypothesis that the carbonic anhydrase inhibitor acetazolamide, a potent inhibitor of proximal tubular sodium reabsorption, improves decongestion when combined with loop diuretic therapy in AHF, potentially leading to better clinical outcomes. The ADVOR trial is set up as a multicentre, randomized, double-blind, placebo-controlled study, aiming to recruit 519 patients with AHF and clinically evident volume overload. All study participants receive high-dose intravenous loop diuretics as background therapy and are randomized towards intravenous acetazolamide at a dose of 500 mg once daily vs. placebo, stratified according to including study centre and ejection fraction (< 40% vs. ≥ 40%). The primary endpoint is successful decongestion with no more than trace oedema assessed on the third morning after hospital admission, with good diuretic efficacy defined as a urine output > 3.5 L during the first 30-48 h of decongestive treatment. Secondary endpoints include all-cause mortality or heart failure readmission after 3 months, length of hospital stay for the index admission, and longitudinal changes in the EuroQol-5 dimensions questionnaire. ADVOR will investigate if acetazolamide combined with loop diuretic therapy improves decongestion in AHF with volume overload.


Impact of environmental change and schistosomiasis transmission in the middle reaches of the Yangtze River following the Three Gorges construction project.
With the construction of the Three Gorges high dam on the Yangtze River in China in mind, a serious of ecological environmental factors that might affect the transmission of Schistosoma japonicum in Jian Han Plain were investigated by means of data collection, field surveys and observation in Hubei Province. Several ecological factors including water level of the Yangtze River; riparian water table, annual rainfall and yearly evaporation were investigated in relation to the prevalence of schistosomiasis. The results suggest that after the dam construction, middle water level flows (ie between flood flows and dry-weather flows) will persist in the flood season due to a rise in the water table. The investigation indicated that snail distribution and human schistosomiasis prevalence differed significantly between years which had typically high, middle and low typical water levels in the Yangzte. Moreover, the prevalence of the disease showed a significant linear regression relationship with density of snail intermediate hosts, water table, annual rainfall, yearly evaporation and ground altitude in the survey area. Systematic and careful monitoring and surveillance is necessary to investigate the impact of the environmental changes brought about by the dam construction on schistosomiasis transmission.


Pulse broadening caused by deviations from the optimal index profile.
The excess pulse broadening caused by deviations from the optimal index profile is analyzed for multimode optical fibers. First-order perturbation theory is used to evaluate the effects of arbitrary index perturbations having longitudinal spatial frequencies which are too low to cause mode coupling. A simplified expression is found for the excess pulse broadening in terms of a double integral over a weighting function. Weighting functions are evaluated and used to calculate the effects of two kinds of circularly symmetric perturbations, a Gaussian-shaped bump or dip in the index profile and an oscillating perturbation. These perturbations are found to affect the pulse broadening significantly according to the position and width of the bump and the transverse spatial frequency of the oscillating perturbation. The more narrow bumps and those located at larger core radii are found to have the greatest effect. For a typical multimode fiber, perturbations making 2-18 oscillations along the radius are found to cause significant increases in pulse broadening, while more rapid oscillations are found to have negligible effect. These results indicate the type and degree of index profile control required to keep the pulse broadening below a specified level.


A pathway EM-algorithm for estimating vaccine efficacy with a non-monotone validation set.
Here, we consider time-to-event data where individuals can experience two or more types of events that are not distinguishable from one another without further confirmation, perhaps by laboratory test. The event type of primary interest can occur only once. The other types of events can recur. If the type of a portion of the events is identified, this forms a validation set. However, even if a random sample of events are tested, confirmations can be missing nonmonotonically, creating uncertainty about whether an individual is still at risk for the event of interest. For example, in a study to estimate efficacy of an influenza vaccine, an individual may experience a sequence of symptomatic respiratory illnesses caused by various pathogens over the season. Often only a limited number of these episodes are confirmed in the laboratory to be influenza-related or not. We propose two novel methods to estimate covariate effects in this survival setting, and subsequently vaccine efficacy. The first is a pathway expectation-maximization (EM) algorithm that takes into account all pathways of event types in an individual compatible with that individual's test outcomes. The pathway EM iteratively estimates baseline hazards that are used to weight possible event types. The second method is a non-iterative pathway piecewise validation method that does not estimate the baseline hazards. These methods are compared with a previous simpler method. Simulation studies suggest mean squared error is lower in the efficacy estimates when the baseline hazards are estimated, especially at higher hazard rates. We use the pathway EM-algorithm to reevaluate the efficacy of a trivalent live-attenuated influenza vaccine during the 2003-2004 influenza season in Temple-Belton, Texas, and compare our results with a previously published analysis.


Three different scenarios for epileptic spasms.
Epileptic Spasms (ES) is a type of seizure usually occurring in the context of a severe childhood epileptic syndrome associated to significant Electroencephalogram (EEG) abnormalities. There are three scenarios in which ES may occur. The first one is represented by West Syndrome (WS): ES occur in a previously non encephalopathic infant in association with the development of a hypsarrhythmic EEG pattern. In most cases, standard treatment with Adrenocorticotropic Hormone (ACTH), steroids or vigabatrin leads to a reversal of the electroclinical picture. The second scenario is represented by Developmental and Epileptic Encephalopathies (DEEs): ES are documented, often along other seizures types, in an infant who often shows developmental delay since birth; the EEG pattern is pathological both in wakefulness and in sleep, without typical features of hypsarrhythmia; therapies (with the exception of few potentially treatable syndromes) are poorly effective. The last scenario is represented by ES in the context of Focal Epilepsies (FEs): ES, sometimes showing focal signs or closely related to focal seizures, are associated with focal brain lesions. Treatment with ACTH, steroids or vigabatrin may not be effective as well as antiepileptic drugs for focal epilepsies. In drug-resistant patients, surgery should be considered. Although there are some gaps in our current scientific knowledge concerning the peculiar electroclinical and physiopathological features of ES, we nowadays possess the necessary tools to correctly frame this unique seizure type into one of these scenarios and therefore properly manage the diagnostic and therapeutic workup.


Electrical properties of motor units in Parkinsonism and a possible relationship with bradykinesia.
The electrical activity of single motor units was recorded from the first dorsal interosseous muscles of nine patients with Parkinson's disease. Six of these patients had a combination of the following abnormal motor unit properies: (1) a variable delay period of 20 seconds to 3 minutes between the initiation of voluntary effort and the recruitment of the first group of motor units; (2) after recruitment, some of the motor units stopped firing for durations of 10s, 40s, 75s... 3 min.; (3) some of the motor units fired at abnormally low frequencies of 2-3 per second. All these six patients had slowed finger movement, and five of the six were studied while off levodopa for two to seven days. One of these patients, reinvestigated after levodopa therapy had been restarted, demonstrated improvement in motor unit control. The three remaining patients who were studied while on uninterrupted levodopa therapy could make rapid finger movements, could recruit motor units without delay, and could fire recruited motor units continuously at normal frequencies of 6-14 per second. These results suggest that levodopa therapy is effective in Parkinson's disease at least partly because of its ability to correct abnormalities in the recruitment of motor units. Levodopa also corrects the abnormal motor unit firing pattern. The abnormal motor unit properties found in these patients could account for some aspects of bradykinesia.


A unified framework for chaotic neural-network approaches to combinatorial optimization.
As an attempt to provide an organized way to study the chaotic structures and their effects in solving combinatorial optimization with chaotic neural networks (CNN's), a unifying framework is proposed to serve as a basis where the existing CNN models can be placed and compared. The key of this proposed framework is the introduction of an extra energy term into the computational energy of the Hopfield model, which takes on different forms for different CNN models, and modifies the original Hopfield energy landscape in various manners. Three CNN models, namely the Chen and Aihara model with self-feedback chaotic simulated annealing (CSA), the Wang and Smith model with timestep CSA, and the chaotic noise model, are chosen as examples to show how they can be classified and compared within the proposed framework.


Identification of a Marine Cyanophage in a Protist Single-cell Metagenome Assembly.
Analysis of microbial biodiversity is hampered by a lack of reference genomes from most bacteria, viruses, and algae. This necessitates either the cultivation of a restricted number of species for standard sequencing projects or the analysis of highly complex environmental DNA metagenome data. Single-cell genomics (SCG) offers a solution to this problem by constraining the studied DNA sample to an individual cell and its associated symbionts, prey, and pathogens. We used SCG to study marine heterotrophic amoebae related to Paulinella ovalis (A. Wulff) P.W. Johnson, P.E. Hargraves & J.M. Sieburth (Rhizaria). The genus Paulinella is best known for its photosynthetic members such as P. chromatophora Lauterborn that is the only case of plastid primary endosymbiosis known outside of algae and plants. Here, we studied the phagotrophic sister taxa of P. chromatophora that are related to P. ovalis and found one SCG assembly to contain α-cyanobacterial DNA. These cyanobacterial contigs are presumably derived from prey. We also uncovered an associated cyanophage lineage (provisionally named phage PoL_MC2). Phylogenomic analysis of the fragmented genome assembly suggested a minimum genome size of 200 Kbp for phage PoL_MC2 that encodes 179 proteins and is most closely related to Synechococcus phage S-SM2. For this phage, gene network analysis demonstrates a highly modular genome structure typical of other cyanophages. Our work demonstrates that SCG is a powerful approach for discovering algal and protist biodiversity and for elucidating biotic interactions in natural samples.


Three Months of Strength Training Changes the Gene Expression of Inflammation-Related Genes in PBMC of Older Women: A Randomized Controlled Trial.
Here, we investigate changes in inflammation-related gene-expression in peripheral mononuclear blood cells (PBMC) by strength training. A total of 14 women aged ≥65 years were randomized into 3 months of either 3×/week intensive strength training (IST: 3×10 rep at 80% 1RM), strength endurance training (SET: 2×30 reps at 40% 1RM) or control (CON: 3×30 sec stretching). Differentially expressed genes (fold change ≤0.67 or ≥1.5) were identified by targeted RNA-sequencing of 407 inflammation-related genes. A total of 98 genes (n = 61 pro-inflammatory) were significantly affected. IST and SET altered 14 genes in a similar direction and 19 genes in the opposite direction. Compared to CON, IST changed the expression of 6 genes in the same direction, and 17 genes in the SET. Likewise, 18 and 13 genes were oppositely expressed for, respectively, IST and SET compared to CON. Changes in gene expression affected 33 canonical pathways related to chronic inflammation. None of the altered pathways overlapped between IST and SET. Liver X Receptor/Retinoid X Receptor Activation (LXR/RXR) and Triggering Receptor Expressed On Myeloid Cells 1 (TREM1) pathways were enriched oppositely in both training groups. We conclude that three months IST and SET can induce changes in CLIP-related gene expression in PBMC, but by affecting different genes and related pathways.


[A bird's eye view of the algorithms and software packages for reconstructing phylogenetic trees].
The prototype phylogenetic tree, i.e., evolutionary "tree" or "tree of life", was first conceived by Charles Darwin in his seminal book "The Origin of Species", and its reconstructions have been approached by generations of biologists ever since. In this article, we briefly reviewed the major algorithms and software packages for reconstructing phylogenetic trees. Specifically we discuss four categories of phylogeny algorithms including distance-matrix, maximum parsimony, maximum likelihood, and Bayesian framework, as well as software packages (PHYLIP, MEGA, MrBayes) based on them.


Strategies and endpoints of antifibrotic drug trials: Summary and recommendations from the AASLD Emerging Trends Conference, Chicago, June 2014.
There is an urgent need to develop antifibrotic therapies for chronic liver disease, and clarify which endpoints in antifibrotic trials will be acceptable to regulatory agencies. The American Association for the Study of Liver Diseases sponsored an endpoints conference to help accelerate the efficient testing of antifibrotic agents and develop recommendations on clinical trial design for liver fibrosis. In this review, we summarize the salient and novel elements of this conference and provide directions for future clinical trial design. The article follows the structure of the conference and is organized into five areas: (1) antifibrotic trial design; (2) preclinical proof-of-concept studies; (3) pharmacological targets, including rationale and lessons to learn; (4) rational drug design and development; and (5) consensus and recommendations on design of clinical trials in liver fibrosis. Expert overviews and collaborative discussions helped to summarize the key unmet needs and directions for the future, including: (1) greater clarification of at-risk populations and study groups; (2) standardization of all elements of drug discovery and testing; (3) standardization of clinical trial approaches; (4) accelerated development of improved noninvasive markers; and (5) need for exploration of potential off-target toxicities of future antifibrotic drugs.


Elevation of micronuclei frequency in mouse bone marrow treated with various doses of teniposide (VM-26).
The effect of various doses (0-10 mg/kg body wt.) of teniposide (VM-26) was studied on the induction of micronuclei at 12, 24 and 36 h post-treatment. The frequency of micronuclei (MPCE and MNCE) increased in a dose-dependent manner up to a dose of 0.3125 mg/kg VM-26, where a peak frequency of micronuclei was observed. A further increase in the drug dose resulted in the reduction in micronuclei frequency in comparison with 0.3125 mg/kg drug dose reaching a nadir at 10 mg/kg. However, it was significantly higher than DDW (double distilled water) treated controls. The pattern of micronuclei induction was similar for all the post-treatment time periods. The frequency of micronuclei also increased with scoring time and the highest frequency of micronuclei was observed at 24 h post-treatment, which declined thereafter without restoration to DDW treated control level. Conversely, the PCE/NCE ratio registered a dose-dependent decline after treatment of mice with various doses of VM-26. A peak decline was observed at a dose of 0.3125 mg/kg, thereafter the decline became consistently less resulting in an elevation in the PCE/NCE ratio in comparison with 0.3125 mg/kg VM-26.


Intramuscular Electrical Stimulation for the Treatment of Trigger Points in Patients with Chronic Migraine: A Protocol for a Pilot Study Using a Single-Case Experimental Design.
Background and Objectives: Trigger points (TrPs) are prevalent in patients with migraine headaches. Needling interventions targeting TrPs in migraine patients may reduce the intensity and frequency of headaches, yet systematic reviews reveal a lack of robust evidence. Intramuscular electrical stimulation (IMES) is a modality that delivers electrical current into muscles and TrPs, with recent studies suggesting it may amplify the therapeutic effects of dry needling peripherally and centrally. This could be advantageous for patients with migraine and symptomatic TrPs. Materials and Methods: This study will implement a multiple baseline single-case experimental design (SCED). In a clinical setting, a SCED study lends itself to conducting research with only a few patients that each serve as their own controls. In this SCED study, four participants with chronic migraine will be enrolled in a non-concurrent manner and randomized to one of four baseline measurement periods (4, 5, 6 or 7 weeks), leading to four potentially different start dates for each participant in the intervention phase. During the intervention phase, patients will receive five sessions of dry needling with IMES, one session per week for five weeks. The primary outcome measure will be headache frequency, i.e., the reduction in the number of headache days over a one-month period using electronic headache diary data from the Migraine Buddy smartphone application. Secondary outcome measures will be changes in mean migraine pain intensity using a numeric pain rating scale (NPRS), migraine disability using the Migraine Disability Assessment Test (MIDAS), the Headache Impact Test (HIT-6), and changes in selected cervical musculoskeletal impairments including pressure pain thresholds (PPTs) over TrPs, the craniocervical flexion test (CCFT), and cervical active range of motion (AROM). Primary and secondary outcome measures will be analyzed separately using both visual and statistical analyses. Results: Actively recruiting participants. This project was approved by the Mass General Brigham Institutional Review Board (protocol #2023P000931) and is registered with ClinicalTrials.gov (NCT05893914). Conclusions: This study will seek to determine the effects of a five-week intervention period of IMES to TrPs in the posterior cervical muscles of subjects with chronic migraine.


Detailed molecular fingerprinting of four new anaplastic thyroid carcinoma cell lines and their use for verification of RhoB as a molecular therapeutic target.
Anaplastic thyroid carcinoma (ATC) is a highly aggressive carcinoma in need of therapeutic options. One critical component of drug discovery is the availability of well-characterized cell lines for identification of molecular mechanisms related to tumor biology and drug responsiveness. Up to 42% of human thyroid cancer cell lines are redundant or not of correct tissue origin, and a comprehensive analysis is currently nonexistent. Mechanistically, RhoB has been identified as a novel molecular target for ATC therapy. The aim was to develop four ATC cell lines detailing genetic, molecular, and phenotypic characteristics and to test five classes of drugs on the cell lines to determine whether they inhibited cell proliferation in a RhoB-dependent fashion. Four cell lines were derived from ATC tumors. Short tandem DNA repeat and mutational status of the originating tumors and cell lines were performed along with molecular and phenotypic characterizations. Compounds were tested for growth inhibition and ability to up-regulate RhoB. Cell line authenticity was confirmed by DNA short tandem repeat analysis. Each proved unique regarding expression of thyroid markers, oncogene status, amplified and deleted genes, and proliferative growth rates. FTI-277, GGTI-286, lovastatin, romidepsin, and UCN-01 up-regulated RhoB and inhibited cell proliferation in a dose-responsive fashion with only romidepsin and FTI-277 being RhoB dependent. Molecular descriptions of thyroid lines were matched to the originating tumors, setting a new standard for cell line characterization. Furthermore, suppressed RhoB is implicated as a molecular target for therapy against ATC because five classes of drugs up-regulate RhoB and inhibit growth dose-responsively.


Denaturing gradient electrophoresis (DGE) and single-strand conformation polymorphism (SSCP) molecular fingerprintings revisited by simulation and used as a tool to measure microbial diversity.
The exact extent of microbial diversity remains unknowable. Nevertheless, fingerprinting patterns [denaturing gradient electrophoresis (DGE), single-strand conformation polymorphism (SSCP)] provide an image of a microbial ecosystem and contain diversity data. We generated numerical simulation fingerprinting patterns based on three types of distribution (uniform, geometric and lognormal) with a range of units from 10 to 500,000. First, simulated patterns containing a diversity of around 1000 units or more gave patterns similar to those obtained in experiments. Second, the number of bands or peaks saturated quickly to about 35 and were unrelated to the degree of diversity. Finally, assuming lognormal distribution, we used an estimator of diversity on in silico and experimental fingerprinting patterns. Results on in silico patterns corresponded to the simulation inputs. Diversity results in experimental patterns were in the same range as those obtained from the same DNA sample in molecular inventories. Thus, fingerprinting patterns contain extractable data about diversity although not on the basis of a number of bands or peaks, as is generally assumed to be the case.


Pre- and post-diagnostic beta-blocker use and prognosis after colorectal cancer: Results from a population-based study.
Recent experimental and epidemiological studies have suggested that beta-blocker use might be associated with better cancer prognosis, but results were inconclusive and only few studies have investigated the association specifically for colorectal cancer (CRC) patients. We investigated this hypothesis using a linked dataset of the Eindhoven area of the Netherlands Cancer Registry and the PHARMO record linkage, including patients diagnosed with CRC between 1998 and 2011. Eligible patients were matched on propensity scores to control for potential confounders such as socio-demographic factors, comorbidity, cancer treatment and use of other medications. Controls were subsequently restricted to active comparators. The association between pre-diagnostic and time-dependent post-diagnostic beta-blocker use and overall survival was estimated using Cox proportional hazard regression models. Subgroup analyses by cancer site and stage and by beta-blocker type were conducted. Of 8,100 CRC patients with a median follow-up of 6.6 years, 1,813 (22%) used beta-blockers prior to diagnosis. In multivariate analysis, we observed no significant association in overall mortality for pre-diagnostic [hazard ratio 1.07, 95% confidence interval (0.96-1.19)] and post-diagnostic [1.10 (0.98-1.23)] beta-blocker use, respectively. Analyses by beta-blocker type, by cancer site, cancer stage and by cumulative dose showed no significant survival improvements for beta-blocker users. However, there was a significant association between cumulative duration of use of 1-12 months and increased overall mortality [1.20 (1.03-1.39)]. Thus, our results do not support the hypothesis of a beneficial effect of pre- or post-diagnostic beta-blocker intake on CRC prognosis, neither for specific patient subgroups nor for specific types of beta-blockers.


Anisotropic dynamics of the JE-2147-HIV protease complex: drug resistance and thermodynamic binding mode examined in a 1.09 A structure.
The structure of HIV protease (HIV Pr) bound to JE-2147 (also named AG1776 or KNI-764) is determined here to 1.09 A resolution. This highest-resolution structure for HIV Pr allows refinement of anisotropic displacement parameters (ADPs) for all atoms. Clustering based on the directional information in ADPs defines two sets of subdomains such that within each set, subdomains undergo similar anisotropic motion. These sets are (a) the core of monomer A grouped with both substrate-binding flaps and (b) the core of monomer B coupled to both catalytic aspartates (25A/B). The four-stranded beta-sheet (1-4 A/B and 95-99 A/B) that forms a significant part of the dimer interface exhibits large anisotropic amplitudes that differ from those of the other sets of subdomains. JE-2147 is shown here to be a picomolar inhibitor (K(i) = 41 +/- 18 pM). The structure is used to interpret the mechanism of association of JE-2147, a second-generation inhibitor for which binding is enthalpically driven, with respect to first-generation inhibitors for which binding is predominantly entropically driven [Velazquez-Campoy, A., et al. (2001) Arch. Biochem. Biophys. 390, 169-175]. Relative to the entropically driven inhibitor complexes, the JE-2147-HIV Pr complex exhibits an approximately 0.5 A movement of the substrate flaps in toward the substrate, suggesting a more compatible enthalpically driven association. Domains of the protease identified by clustering of ADPs also suggest a model of enthalpy-entropy compensation for all HIV Pr inhibitors in which dynamic coupling of the flaps is offset by an increased level of motion of the beta-sheet domain of the dimer interface (1-4 A/B and 95-99 A/B).


Manipulation of Ploidy in Caenorhabditis elegans.
Mechanisms that involve whole genome polyploidy play important roles in development and evolution; also, an abnormal generation of tetraploid cells has been associated with both the progression of cancer and the development of drug resistance. Until now, it has not been feasible to easily manipulate the ploidy of a multicellular animal without generating mostly sterile progeny. Presented here is a simple and rapid protocol for generating tetraploid Caenorhabditis elegans animals from any diploid strain. This method allows the user to create a bias in chromosome segregation during meiosis, ultimately increasing ploidy in C. elegans. This strategy relies on the transient reduction of expression of the rec-8 gene to generate diploid gametes. A rec-8 mutant produces diploid gametes that can potentially produce tetraploids upon fertilization. This tractable scheme has been used to generate tetraploid strains carrying mutations and chromosome rearrangements to gain insight into chromosomal dynamics and interactions during pairing and synapsis in meiosis. This method is efficient for generating stable tetraploid strains without genetic markers, can be applied to any diploid strain, and can be used to derive triploid C. elegans. This straightforward method is useful for investigating other fundamental biological questions relevant to genome instability, gene dosage, biological scaling, extracellular signaling, adaptation to stress, development of resistance to drugs, and mechanisms of speciation.


Automatic processes moderate the association between reflective processes and movement behavior: A cross-sectional study.
Dual-process theories postulate that both reflective and automatic processes regulate health behavior. Further research is required to test the basic postulates of dual-process theories. We investigated the direct associations and moderating effect between automatic processes and multiple indicators of reflective processes on various levels of physical activity and sedentary behavior assessed using accelerometry in adults who were not participating in regular physical exercise programs. This cross-sectional study included 257 adults. Each participant completed a computerized test for automatic associations, a set of questions assessing reflective processes, a seven-day assessment using accelerometers to determine their levels of physical activity and sedentary behavior. The results showed a direct relationship between perceived benefits (B = 15.90, p = 0.043), perceived cons (B = -12.81, p = 0.034), decisional intention (B = -0.07, p = 0.049) with light physical activity, and self-efficacy with daily steps (B = 485.71, p = 0.008). There was a positive association between intention strength and daily steps when implicit associations favored physical activity (b = 623.36, LLCI = 79.09, ULCI = 1167.62, p = 0.025); a negative association between self-efficacy and sedentary behavior when implicit associations favored sedentary behavior (b = -25.73, LLCI = -49.77, ULCI = -1.70, p = 0.035); and a positive association between intention strength and sedentary behavior when implicit associations favored physical activity (b = 34.18, LLCI = 8.81, ULCI = 59.56, p = 0.008). These findings underscore the importance of considering the interplay between reflective and automatic processes in shaping movement behavior.


Identification of cell surface receptors for murine macrophage inflammatory protein-1 alpha.
We have produced recombinant proteins for a cytokine, L2G25BP (macrophage inflammatory protein-1 alpha) (MIP-1 alpha). By using the recombinant protein (rMIP-1 alpha), receptors for MIP-1 alpha were identified on Con A-stimulated and unstimulated CTLL-R8, a T cell line, and LPS-stimulated RAW 264.7, a macrophage cell line. The 125I-rMIP-1 alpha binds to the receptor in a specific and saturable manner. Scatchard analysis indicated a single class of high affinity receptor, with a Kd of approximately 1.5 x 10(-9) M and approximately 1200 binding sites/Con A-stimulated CTLL-R8 cell and a Kd of 0.9 x 10(-9) M and approximately 380 binding sites/RAW 264.7 cell. 125I-rMIP-1 alpha binding was inhibited by unlabeled rMIP-1 alpha in a dose-dependent manner, but not by IL-1 alpha or IL-2. rMIP-1 alpha inhibited the proliferation of unstimulated CTLL-R8 cells. Rabbit anti-rMIP-1 alpha antibodies blocked the growth-inhibitory effect of the rMIP-1 alpha on CTLL-R8 cells.


The Army study to assess risk and resilience in servicemembers (Army STARRS).
IMPORTANCE/OBJECTIVE: Although the suicide rate in the U.S. Army has traditionally been below age-gender matched civilian rates, it has climbed steadily since the beginning of the Iraq and Afghanistan conflicts and since 2008 has exceeded the demographically matched civilian rate. The Army Study to Assess Risk and Resilience in Servicemembers (Army STARRS) is a multicomponent epidemiological and neurobiological study designed to generate actionable evidence-based recommendations to reduce Army suicides and increase knowledge about risk and resilience factors for suicidality and its psychopathological correlates. This paper presents an overview of the Army STARRS component study designs and of recent findings. DESIGN/SETTING/PARTICIPANTS/INTERVENTION: Army STARRS includes six main component studies: (1) the Historical Administrative Data Study (HADS) of Army and Department of Defense (DoD) administrative data systems (including records of suicidal behaviors) for all soldiers on active duty 2004-2009 aimed at finding administrative record predictors of suicides; (2) retrospective case-control studies of fatal and nonfatal suicidal behaviors (each planned to have n = 150 cases and n = 300 controls); (3) a study of new soldiers (n = 50,765 completed surveys) assessed just before beginning basic combat training (BCT) with self-administered questionnaires (SAQ), neurocognitive tests, and blood samples; (4) a cross-sectional study of approximately 35,000 (completed SAQs) soldiers representative of all other (i.e., exclusive of BCT) active duty soldiers; (5) a pre-post deployment study (with blood samples) of soldiers in brigade combat teams about to deploy to Afghanistan (n = 9,421 completed baseline surveys), with sub-samples assessed again one, three, and nine months after returning from deployment; and (6) a pilot study to follow-up SAQ respondents transitioning to civilian life. Army/DoD administrative data are being linked prospectively to the large-scale survey samples to examine predictors of subsequent suicidality and related mental health outcomes. Measures (self-report and administratively recorded) of suicidal behaviors and their psychopathological correlates. Component study cooperation rates are comparatively high. Sample biases are relatively small. Inefficiencies introduced into parameter estimates by using nonresponse adjustment weights and time-space clustering are small. Initial findings show that the suicide death rate, which rose over 2004-2009, increased for those deployed, those never deployed, and those previously deployed. Analyses of administrative records show that those deployed or previously deployed were at greater suicide risk. Receiving a waiver to enter the Army was not associated with increased risk. However, being demoted in the past two years was associated with increased risk. Time in current deployment, length of time since return from most recent deployment, total number of deployments, and time interval between most recent deployments (known as dwell time) were not associated with suicide risk. Initial analyses of survey data show that 13.9% of currently active non-deployed regular Army soldiers considered suicide at some point in their lifetime, while 5.3% had made a suicide plan, and 2.4% had attempted suicide. Importantly, 47-60% of these outcomes first occurred prior to enlistment. Prior mental disorders, in particular major depression and intermittent explosive disorder, were the strongest predictors of these self-reported suicidal behaviors. Most onsets of plans-attempts among ideators (58.3-63.3%) occurred within the year of onset of ideation. About 25.1% of non-deployed U.S. Army personnel met 30-day criteria for a DSM-IV anxiety, mood, disruptive behavior, or substance disorder (15.0% an internalizing disorder; 18.4% an externalizing disorder) and 11.1% for multiple disorders. Importantly, three-fourths of these disorders had pre-enlistment onsets. Integration across component studies creates strengths going well beyond those in conventional applications of the same individual study designs. These design features create a strong methodological foundation from which Army STARRS can pursue its substantive research goals. The early findings reported here illustrate the importance of the study and its approach as a model of studying rare events particularly of national security concern. Continuing analyses of the data will inform suicide prevention for the U.S. Army.


Ample use of physician-prescribed medications in Finnish elite athletes.
The present study aimed at determining the use of physician-prescribed medication in a large number of elite athletes compared with a representative control sample of the general population. Of all the athletes (N = 494) financially supported by the National Olympic Committee, 446 completed a structured questionnaire (response rate 90.3 %) in 2002. A control group (N = 1503, response rate 80.1 %) comprised an age-matched sample from the population-based study collected by the National Public Health Institute. Any prescribed medication was used by 34.5 % of the athletes and 24.9 % of the controls during the past seven days. The most frequently reported physician-prescribed medications among athletes during the previous seven days were anti-allergic medicines (12.6 % of the respondents), non-steroidal anti-inflammatory drugs (NSAIDs; 8.1 %), anti-asthmatic medicines (7.0 %), and oral antibiotics (2.7 %). The adjusted odds ratios (95 % CI) for the physician-prescribed medications used during the previous seven days was 2.42 (1.69 - 3.46), 3.63 (2.25 - 5.84), 3.42 (2.05 - 5.70), and 2.15 (1.03 - 4.45) for use of anti-allergic medication, NSAIDs, anti-asthmatic medication, and oral antibiotics, respectively, in the athletes compared with controls. Every fifth athlete reported some NSAID-related adverse effect. In conclusion, the athletes used NSAIDs, antibiotics, anti-asthmatic and anti-allergic medication significantly more often than a representative sample of age-matched controls. All these medicines have potential adverse effects that may have a deleterious impact on the maximum exercise performance of elite athletes. Adverse effects were commonly reported in connection with NSAID use.


Pharmacokinetics and In Vitro Blood-Brain Barrier Screening of the Plant-Derived Alkaloid Tryptanthrin.
The indolo[2,1-b]quinazoline alkaloid tryptanthrin was previously identified as a potent anti-inflammatory compound with a unique pharmacological profile. It is a potent inhibitor of cyclooxygenase-2, 5-lipooxygenase-catalyzed leukotriene synthesis, and nitric oxide production catalyzed by the inducible nitric oxide synthase. To characterize the pharmacokinetic properties of tryptanthrin, we performed a pilot in vivo study in male Sprague-Dawley rats (2 mg/kg bw i. v.). Moreover, the ability of tryptanthrin to cross the blood-brain barrier was evaluated in three in vitro human and animal blood-brain barrier models. Bioanalytical UPLC-MS/MS methods used were validated according to current international guidelines. A half-life of 40.63 ± 6.66 min and a clearance of 1.00 ± 0.36 L/h/kg were found in the in vivo pharmacokinetic study. In vitro data obtained with the two primary animal blood-brain barrier models showed a good correlation with an immortalized human monoculture blood-brain barrier model (hBMEC cell line), and were indicative of a high blood-brain barrier permeation potential of tryptanthrin. These findings were corroborated by the in silico prediction of blood-brain barrier penetration. P-glycoprotein interaction of tryptanthrin was assessed by calculation of the efflux ratio in bidirectional permeability assays. An efflux ratio below 2 indicated that tryptanthrin is not subjected to active efflux.


Host Specificity and Ecology of Digenean Parasites of Nassariid Gastropods in Central Queensland, Australia, with Comments on Host-Parasite Associations of the Nassariidae.
Nassariid gastropods are well represented in Australia, and Nassarius dorsatus (Röding), Nassarius olivaceus (Bruguière), and Nassarius pullus (Linnaeus) are common in the intertidal mangroves of the Capricornia region in Central Queensland. A survey of the digenean trematode fauna of nassariids of the region revealed cercariae of 12 species from six trematode families associated with these gastropods. Six species of Acanthocolpidae were found, including three putative species of the genus Stephanostomum. Additionally, two species of Himasthlidae and one species each of Microphallidae, Opecoelidae, Zoogonidae, and Lepocreadiidae were identified. The majority of the trematode species identified were found in only one host species; only four infected more than one species of nassariid. Sixteen new host-parasite combinations were added to the fauna known to infect the Nassariidae. Broader examination of host-parasite associations shows that families using nassariids commonly as first intermediate host are the Acanthocolpidae, Lepocreadiidae, and Zoogonidae, as well as Himasthla spp. from the Himasthlidae. Nassariid gastropods are also the dominant marine first intermediate hosts for the Acanthocolpidae, Lepocreadiidae, and Zoogonidae and one of the three most common first intermediate host families for marine Himasthla spp.


SARS-CoV-2 pathophysiology and its clinical implications: An integrative overview of the pharmacotherapeutic management of COVID-19.
Common manifestations of COVID-19 are respiratory and can extend from mild symptoms to severe acute respiratory distress. The severity of the illness can also extend from mild disease to life-threatening acute respiratory distress syndrome (ARDS). SARS-CoV-2 infection can also affect the gastrointestinal tract, liver and pancreatic functions, leading to gastrointestinal symptoms. Moreover, SARS-CoV-2 can cause central and peripheral neurological manifestations, affect the cardiovascular system and promote renal dysfunction. Epidemiological data have indicated that cancer patients are at a higher risk of contracting the SARS-CoV-2 virus. Considering the multitude of clinical symptoms of COVID-19, the objective of the present review was to summarize their pathophysiology in previously healthy patients, as well as in those with comorbidities. The present review summarizes the current, though admittedly fluid knowledge on the pathophysiology and symptoms of COVID-19 infection. Although unclear issues still remain, the present study contributes to a more complete understanding of the disease, and may drive the direction of new research. The recognition of the severity of the clinical symptoms of COVID-19 is crucial for the specific therapeutic management of affected patients.


Metoprolol for intermittent explosive disorder.
Metoprolol, a selective beta 1-adrenoreceptor blocker, was administered to two patients with intermittent explosive disorder who had not done well with previous medications, including propranolol and carbamazepine. Both patients improved dramatically, suggesting clinical and theoretical relevance.


Molecular mechanisms of the biological clock in cultured fibroblasts.
In mammals, the central circadian pacemaker resides in the hypothalamic suprachiasmatic nucleus (SCN), but circadian oscillators also exist in peripheral tissues. Here, using wild-type and cryptochrome (mCry)-deficient cell lines derived from mCry mutant mice, we show that the peripheral oscillator in cultured fibroblasts is identical to the oscillator in the SCN in (i) temporal expression profiles of all known clock genes, (ii) the phase of the various mRNA rhythms (i.e., antiphase oscillation of Bmal1 and mPer genes), (iii) the delay between maximum mRNA levels and appearance of nuclear mPER1 and mPER2 protein, (iv) the inability to produce oscillations in the absence of functional mCry genes, and (v) the control of period length by mCRY proteins.


Vitamin D deficiency in children with renal disease.
The purpose of this study was to assess the vitamin D status of children with renal disease attending the outpatient clinics of our tertiary nephrology centre, allowing us to determine the prevalence of vitamin D deficiency and study its relationship with glomerular filtration rate (GFR) and serum parathyroid hormone (PTH) concentration. 90 boys and 53 girls (99 white Caucasians, 38 of South Asian origin and six from other ethnic groups) were enrolled into the study. 18 were on dialysis (15 peritoneal dialysis and three haemodialysis), 61 had a functioning renal transplant (19 with reduced GFR), 18 had chronic renal failure and 46 had a variety of renal disorders with normal renal function. Serum/plasma concentrations of 25(OH)D, 1,25(OH)2D, PTH, creatinine, calcium, phosphate and alkaline phosphatase were measured. Patients with serum 25(OH)D concentrations of less than 25 nmol/l were considered to be deficient and those with levels of 25-50 nmol/l considered insufficient. 26% of all patients were 25(OH)D deficient and a further 32% were insufficient. The prevalence in South Asians was higher (87% deficient/insufficient) than in white Caucasians (46% deficient/insufficient, p<0.001). In children with reduced GFR who were not on dialysis, 25(OH)D levels of less than 50 nmol/l were associated with a high PTH in 90% of cases compared to only 50% of those with normal vitamin D concentrations (p = 0.013). A high proportion of renal patients were vitamin D deficient/insufficient, particularly children of South Asian origin. High PTH values in the setting of reduced GFR might be due to vitamin D deficiency and should lead to estimation of serum 25(OH)D concentration.


[Hydrogen peroxide inhibits acetylcholinesterase of myometrium sarcolemma].
The action of hydrogen peroxide on acetylcholinesterase enzymatic activity in myometrium sarcolemma fraction is investigated. Hydrogen peroxide (0.1-26 microM), depending on the concentration, suppressed the activity. Acetylcholinesterase proved to be highly sensitive to the action of H2O2, making Ki = 2.4 +/- 0.4 microM, nH = 0.65 +/- 0.08 (n = 4-5). It is established, that hydrogen peroxide in the range of 1.6 - 6.4 microM essentially reduce V(0,max) and K(M). In the presence of dithiothreitole (a reducer of SH-groups of the membrane surface) the investigated substance effect considerably decreased.


The role of D-serine as co-agonist of NMDA receptors in the nucleus accumbens: relevance to cocaine addiction.
Cocaine addiction is characterized by compulsive drug use despite adverse consequences and high rate of relapse during periods of abstinence. Increasing consensus suggests that addiction to drugs of abuse usurps learning and memory mechanisms normally related to natural rewards, ultimately producing long-lasting neuroadaptations in the mesocorticolimbic system. This system, formed in part by the ventral tegmental area and nucleus accumbens (NAc), has a central role in the development and expression of addictive behaviors. In addition to a broad spectrum of changes that affect morphology and function of NAc excitatory circuits in cocaine-treated animals, impaired N-methyl-D-aspartate receptor (NMDAR)-dependent synaptic plasticity is a typical feature. D-serine, a D-amino acid that has been found at high levels in mammalian brain, binds with high affinity the co-agonist site of NMDAR and mediates, along with glutamate, several important processes including synaptic plasticity. Here we review recent literature focusing on cocaine-induced impairment in synaptic plasticity mechanisms in the NAc and on the fundamental role of D-serine as co-agonist of NMDAR in functional and dysfunctional synaptic plasticity within this nucleus. The emerging picture is that reduced D-serine levels play a crucial role in synaptic plasticity relevant to cocaine addiction. This finding opens new perspectives for therapeutic approaches to treat this addictive state.


The application of gamma stereotactic body radiation therapy in stage I/II non-small-cell lung cancer. Promising and encouraging outcome.
To further evaluate the efficacy and toxicity of the gamma-ray stereotactic body radiation therapy (SBRT) in patients with stage I/II non-small-cell lung cancer (NSCLC). Twenty-nine newly diagnosed patients with stage I/II NSCLC who had no previous treatments, underwent OUR-QGD type of the gamma-SBRT at the Radiation Oncology Department, People's Liberation Army Airforce General Hospital, Beijing, China from January 2007 to July 2010. All patients were immobilized by vacuum bag, and then a slow CT scan was performed without any respiration gating. The total radiation dose of 50%, 60%, and 70% isodose line were prescribed in 50, 60, and 70 Grey (Gy) correspondingly, covering 100% of the planning target volume (PTV), 90% of the clinical target volume (CTV), and 80% of the gross target volume (GTV) in 10 fractions. The CT scans of the chest were required at one, 3, 6, 12, 18, and 24 months to evaluate the efficacy of the treatment. The median follow-up duration was 24 months, and the final follow-up rate is 96.6%. Local control rates of one and 2 years were all 93.1%. The progression-free survival rates versus overall survival rate of one year was 89.7% versus 96.6%, and 2 years was 86.1% versus 89.4%. Acute radiation reactions was diagnosed in 34.5%, and late radiation reactions in 37.9% of patients. The gamma-SBRT results in a good curative effects, and minimal toxicity in the treatment of stage I/II NSCLC.


Nrs1, a repetitive element linked to pisatin demethylase genes on a dispensable chromosome of Nectria haematococca.
We have identified a repetitive DNA element in Nectria haematococca mating population VI, isolate T-2. This repetitive sequence has been called Nrs1. DNA hybridization analysis indicates the sequence is found in several isolates of the fungus pathogenic to Pisum sativum. A 2,027-bp clone containing the Nrs1-2 allele contains a long polyA sequence, imperfect RNA polymerase III promoter sequences, multiple inverted repeats, and the potential for extensive secondary structure similar to known RNA polymerase III transcripts and related retroelements. Ten of the 11 HindIII restriction fragments from isolate T-2 DNA that hybridize to Nrs1-2 segregate in a manner consistent with a 1:1 ratio for random ascospore progeny. The 10 restriction fragment length polymorphism (RFLP) loci define three linkage groups and correspond to three chromosome-sized DNAs from T-2 separated by pulsed field gel electrophoresis. Three RFLP loci defined by hybridization to the gene for pisatin demethylase and localized on the 1.6 million base pair (Mb) chromosome were genetically linked to each other and to several Nrs1 loci. These sequences recombined despite the fact that no obvious homolog exists for the 1.6-Mb chromosome in one parent strain. Allelic RFLPs corresponding to the gene sequence of cutinase were unlinked to Nrs1 loci.


The Montreal Neurological Institute.
In this centenary year of Wilder Penfield, his contribution to the creation of the Montreal Neurological Institute is reviewed. In the early 1930's, a confluence of favorable circumstances at McGill University and the Royal Victoria Hospital, Montreal, made it possible for Wilder Penfield, with his partner William Cone, to realize his dream of a combined neurological hospital and research institute. Endowed by the Rockefeller Foundation and with initial and ongoing support from community and governments, the Montreal Neurological Institute has continued its exponential growth over the past half-century as a world center for the study and treatment of disorders of the brain and nerves.


Long-term Outcomes After Flap Reconstruction in Pediatric Pressure Ulcers.
Pressure ulcers refractory to nonoperative management may undergo flap reconstruction. This study aims to evaluate the long-term outcomes and recurrence rates of flap reconstruction for pediatric pressure ulcers. We reviewed the records of patients who underwent flap reconstruction for pressure ulcer(s) from 1995 to 2013. Twenty-four patients with 30 pressure ulcers, requiring 52 flaps were included. Ulcers were stages III and IV and mostly involved either the ischia (15/30) or sacrum (8/30). Flaps were followed for a median of 4.9 years. Twenty-three patients were wheelchair dependent, and 20 had sensory impairment at their ulcer site(s). Ten patients had a history of noncompliance with preoperative management, 8 of whom experienced ulcer recurrence. Twenty-one ulcers had underlying osteomyelitis, associated with increased admissions (P = 0.019) and cumulative length of stay (P = 0.031). Overall, there was a 42% recurrence rate in ulceration after flap reconstruction. Recurrence was associated with a preoperative history of noncompliance with nonoperative therapy (P = 0.030), but not with flap type or location, age, sex, body mass index, osteomyelitis, or urinary/fecal incontinence (P > 0.05, all). Flap reconstruction can be beneficial in the management of pediatric pressure ulcers. Although high rates of long-term success with this intervention have been reported in children, we found rates of ulcer recurrence similar to that seen in adults. Poor compliance with nonoperative care and failure to modify the biopsychosocial perpetuators of pressure ulcers will likely eventuate in postoperative recurrence. Despite the many comorbidities observed in our patient sample, compliance was the best indicator of long-term skin integrity and flap success.


Severe personality disorder in the secure estate: continuity and change.
The Response to Offender Personality Disorder Consultation was released in October 2011. For some this is a welcome step in the right direction due to its therapeutic optimism, however for practitioners operating in the secure estate there are significant challenges ahead. This aim of this article is to discuss the increasing convergence of health and criminal justice and their inherent ideological and practical difficulties. It does so with reference to the consultation on offender personality disorder pathways and in particular the implications regarding multi-disciplinary and cross agency approaches to risk, public protection and personality disorder respectfully. It concludes that before embarking on a new wave of determining and responding to those with personality disorder, offender or otherwise, a more in-depth and empirically informed critical reflection is warranted.