A novel shape complementarity scoring function for protein-protein docking.
Shape complementarity is the most basic ingredient of the scoring functions for protein-protein docking. Most grid-based docking algorithms use the total number of grid points at the binding interface to quantify shape complementarity. We have developed a novel Pairwise Shape Complementarity (PSC) function that is conceptually simple and rapid to compute. The favorable component of PSC is the total number of atom pairs between the receptor and the ligand within a distance cutoff. When applied to a benchmark of 49 test cases, PSC consistently ranks near-native structures higher and produces more near-native structures than the traditional grid-based function, and the improvement was seen across all prediction levels and in all categories of the benchmark. Without any post-processing or biological information about the binding site except the complementarity-determining region of antibodies, PSC predicts the complex structure correctly for 6 test cases, and ranks at least one near-native structure in the top 20 predictions for 18 test cases. Our docking program ZDOCK has been parallelized and the average computing time is 4 minutes using sixteen IBM SP3 processors. Both ZDOCK and the benchmark are freely available to academic users (http://zlab.bu.edu/~ rong/dock).


[Effectiveness of platinum-based chemotherapy in ovarian cancer patients with BRCA1/2 mutations].
The purpose of this study was to examine the clinical significance of mutations in BRCA1/2 in the formation of response to neoadjuvant platinum-based chemotherapy for ovarian cancer (OC). All patients who had had neoadjuvant chemotherapy (NCT) in our Institute from January 2000 till January 2013 were tested for carrier of mutations in BRCA1/2. In accordance with the BRCA-status we formed two groups--a group with hereditary advanced OC and a group with non-hereditary advanced OC. In the formed groups there was studied the effectiveness of chemotherapy. Patients carriers of mutations in BRCA1/2 showed a complete clinical response in 34% of cases, compared to 4% in the non-hereditary OC. Analysis of the results of cytoreductive surgery showed that in the group of hereditary cancer it was significantly higher the percentage of performing optimal cytoreductive operations (71% vs 48%). We analyzed the cases of complete pathologic response in all patients NCT and found that full pathomorphosis significantly associated with BRCA-status and the type of ongoing chemotherapy. It was important to note that all carriers of mutations in BRCA1/2 responded to cisplatin chemotherapy.


Ability of Lactobacillus kefiri LKF01 (DSM32079) to colonize the intestinal environment and modify the gut microbiota composition of healthy individuals.
Probiotics have been observed to positively influence the host's health, but to date few data about the ability of probiotics to modify the gut microbiota composition exist. To evaluate the ability of Lactobacillus kefiri LKF01 DSM32079 (LKEF) to colonize the intestinal environment of healthy subjects and modify the gut microbiota composition. Twenty Italian healthy volunteers were randomized in pre-prandial and post-prandial groups. Changes in the gut microbiota composition were detected by using a Next Generation Sequencing technology (Ion Torrent Personal Genome Machine). L. kefiri was recovered in the feces of all volunteers after one month of probiotic administration, while it was detected only in three subjects belonging to the pre-prandial group and in two subjects belonging to the post-prandial group one month after the end of probiotic consumption. After one month of probiotic oral intake we observed a reduction of Bilophila, Butyricicomonas, Flavonifractor, Oscillibacter and Prevotella. Interestingly, after the end of probiotic administration Bacteroides, Barnesiella, Butyricicomonas, Clostridium, Haemophilus, Oscillibacter, Salmonella, Streptococcus, Subdoligranolum, and Veillonella were significantly reduced if compared to baseline samples. L. kefiri LKF01 showed a strong ability to modulate the gut microbiota composition, leading to a significant reduction of several bacterial genera directly involved in the onset of pro-inflammatory response and gastrointestinal diseases.


Visualizing Rank Time Series of Wikipedia Top-Viewed Pages.
Visual clutter is a common challenge when visualizing large rank time series data. WikiTopReader, a reader of Wikipedia page rank, lets users explore connections among top-viewed pages by connecting page-rank behaviors with page-link relations. Such a combination enhances the unweighted Wikipedia page-link network and focuses attention on the page of interest. A set of user evaluations shows that the system effectively represents evolving ranking patterns and page-wise correlation.


Effects of cytosine arabinoside, 6-aminonicotinamide, and 6-mercaptopurine riboside on ectoderm and mesoderm of mouse limb buds.
The effects of cytosine arabinoside, 6-aminonicotinamide, and 6-mercaptopurine riboside on the incorporation of [14C] glucose moieties and [32P] phosphate into acid-soluble material and lipids, RNA, DNA, and protein were measured in the dissected mesoderm and ectoderm of mouse limb buds at the 42-45 (day 11) somite stage. Due to the different proliferative capacities of the two tissues the incorporation of the precursors into mesodermal cells was considerably higher the than into ectodermal ones. Cytosine arabinoside inhibited the incorporation of the precursor moieties only into DNA, but very early after its application. This effect was more obvious in mesoderm than ectoderm. 6-Aminonicotinamide interfered only with glucose metabolism, whereas the incorporation of phosphate was not affected. 14C radioactivity in the various cell components was similarly reduced in mesoderm and ectoderm. 6-mercaptopurine riboside caused an increased incorporation of precursor material in all fractions studied in the mesoderm as well as in the ectoderm during the first 12 hours. This was succeeded by a dramatic decrease of incorporated 14C and 32P radioactivity. Differences of response in the tissues could not be detected with this drug. It is suggested that the malformations of the extrmities caused by these antimetabolites may be predominantly attributed to changes in the cell function rather than to gross effects on cell metabolism.


[The efficacy of botulinum therapy in treatment of delayed facial palsy after resection of vestibular schwannoma].
Delayed facial palsy is a complication developing 3 or more days after surgery. The etiology and pathogenesis of this condition has not been fully explored, and there are no treatment standards for it. As in the case of Bell's paralysis, glucocorticosteroids (GCSs) are currently used to treat delayed facial palsy. However, patients with contraindications to GCSs need new therapy modalities. We aimed to evaluate the efficacy and safety of botulinum therapy in patients with delayed facial palsy after neurosurgical interventions. We examined 33 patients with delayed facial palsy developed 3 or more days after resection of vestibular schwannoma. The main group included 18 patients with contraindications to GCSs who received injections of botulinum toxin A (BTA) into the facial muscles of the healthy side for muscle relaxation. The comparison group consisted of 15 patients who received a course of prednisolone (1 mg/kg/day) for 5-7 days. The efficacy of treatment was assessed using the House-Brackmann scale and Clinical Global Impression Scale. The follow-up period after therapy was 3 months. Delayed facial palsy was more common in the following cases: the facial nerve was located near the antero-inferior tumor pole; the tumor was adherent to the facial nerve; the tumor extended in the oral direction; the tumor had with unclear borders and was 11 to 30 mm in size. In most patients of both groups, facial muscle palsy developed more than 11 days after surgery. Treatment both in the main and control groups resulted in a significant improvement: complete regression of the facial asymmetry in patients of the main group and comparison groups 3 months after treatment onset was 83.3 and 93.3% (House-Brackmann scale), respectively. Botulinum therapy may be recommended for patients with delayed facial palsy developed after vestibular schwannoma resection, who have contraindications to GCSs.


The politics of 'branding' in policy transfer: the case of DOTS for tuberculosis control.
How and why policies are transferred between countries has attracted considerable interest from scholars of public policy over the last decade. This paper, based on a larger study, sets out to explore the processes involved in policy transfer between international and national levels. These processes are illustrated by looking at a particular public health policy--DOTS for the control and treatment of tuberculosis. The paper demonstrates how, after a long period of neglect, resources were mobilised to put tuberculosis back on international and national public policy agendas, and then how the policy was 'branded' and marketed as DOTS, and transferred to low and middle income countries. It focuses specifically on international agenda setting and policy formulation, and the role played by international organisations in those processes. It shows that policy communities, and particular individuals within them, may take political rather than technical positions in these processes, which can result in considerable contestation. The paper ends by suggesting that while it is possible to raise the profile of a policy dramatically through branding and marketing, success also depends on external events providing windows of opportunity for action. Second, it warns that simplifying policy approaches to 'one-size-fits-all' carries inherent risks, and can be perceived to harm locally appropriate programmes. Third, top-down internationally driven policy changes may lead to apparent policy transfer, but not necessarily to successfully implemented programmes.


Mandibular advancement splints for obstructive sleep apnoea - a cautionary tale.
Obstructive sleep apnoea is a serious medical condition of increasing prevalence that has a multitude of treatment modalities. The aim of this study was to review the results of patients treated with mandibular advancement splints in the Oral and Maxillofacial Surgery Unit of South Australia to inform ongoing practice in this area. The casenotes of patients who received a splint for obstructive sleep apnoea between January 2008 and June 2014 were audited. Data collection included referral and demographic details, sleep study results, splint type and follow-up details. Three hundred and four patients were identified as having been provided a mandibular advancement splint for obstructive sleep apnoea, of which 82 had diagnostic and splint-in-situ sleep studies available for comparison. It was observed that 27 had clinically significant objective improvement, 40 had no clinically significant change and 15 had clinically worse OSA following MAS therapy. Mean reductions in Apnoea-Hypopnoea Index and Respiratory Disturbance Index were eight events (27%) and 13 events (33%) per hour respectively with treatment. Based on this study, mandibular advancement splint therapy is a viable treatment for a subset of patients with obstructive sleep apnoea. Protocol driven, multi-disciplinary care with auditing of results is recommended.


Mast cell-nerve interactions in children with functional dyspepsia.
Functional dyspepsia in childhood is commonly triggered by food allergen in sensitised individuals. We investigated the topography of eosinophils and mast cells in gastric antral lamina propria, the interaction of mast cell products with mucosal nerve fibres, and changes in gastric antral muscle slow wave activity in children with atopy and non-atopy-related functional dyspepsia. Open label study of gastric mucosal cow's milk challenge in 10 atopic and 6 nonatopic children (ages 2-12 years) investigated consecutively with gastroscopy for functional dyspepsia. Simultaneous surface electrogastrography and milk challenge were undertaken and laser scanning fluorescence microscopy used to examine the association of mast cell tryptase with mucosal nerves in the gastric mucosa before and after challenge. Eosinophils and mast cells within the lamina propria were increased in number in children with atopic functional dyspepsia and degranulated rapidly after cow's milk challenge in the atopic group. For degranulating eosinophils, median = 13.0% (interquartile range = 3.7-31.0) premilk versus 32.0% (12.0-42.0) after milk biopsies (P < 0.05); for degranulating mast cells, 5.35% (2.7-10.9) premilk biopsies versus 18.75% (12.9-22.1) after milk biopsies (P < 0.05). No such differences were seen in nonatopic patients. Mast cells were closely associated with mucosal nerve fibres and released tryptase, which colocalised with proteinase-activated receptors on mucosal nerve fibres. The gastric antral slow wave became abnormal within 2 minutes of antigen challenge in atopics with an increase in dominant frequency instability coefficient (P < 0.005), decrease in 3 cycles per minute myoelectrical activity (P < 0.01), and increase in bradygastria (P < 0.01). Early-onset neuroimmune interactions induced by cow's milk in the gastric mucosa of atopic children are associated with rapid disturbance of gastric myoelectrical activity and dyspeptic symptoms.


Biomathematical analysis of the neuronal loss in the aging human brain of both sexes, demonstrated in pigment preparations of the pars cerebellaris loci coerulei.
The pars cerebellaris loci coerulei is a flat accumulation of perikarya representing a subnucleus of the locus coeruleus in the roof of the fourth ventricle of the brain in man. A quantitative study was done on this subnucleus; it comprised 67 specimens from individuals between 11 and 90 years. Nerve cell counts were performed on cleared mounted block preparations. Mathematical analysis of the data was performed using logistic growth functions. The results showed that with advancing age there is a loss of neurons in both sexes. This begins in the female approximately after age 40; in the male already at age 20 (reaching a lower value in old age). The final loss of neurons is more pronounced in the male than in the female. Male and female specimens did not differ in the total number of neurons. In both sexes the left side of the pars cerebellaris loci coerulei contained more neurons than the right.


MiR-454 prompts cell proliferation of human colorectal cancer cells by repressing CYLD expression.
Previous studies have shown that miR-454 plays an important role in a variety of biological processes in various human cancer cells. However, the underlying mechanisms of this microRNA in colorectal cancer (CRC) cells remain largely unknown. In the present study, we investigated the miR-454 role in CRC cell proliferation. We found that miR-454 expression is markedly upregulated in CRC tissues and CRC cells compared with the matched tumor adjacent tissues and the FHC normal colonic cell line. Ectopic expression of miR-454 promoted the proliferation and anchorage-independent growth of CRC cells, whereas inhibition of miR-454 reduced this effect. Bioinformatics analysis further revealed cylindromatosis (CYLD), a putative tumor suppressor as a potential target of miR-454. Data from luciferase reporter assays showed that miR-454 directly binds to the 3'-untranslated region (3'-UTR) of CYLD mRNA and repressed expression at both transcriptional and translational levels. In functional assays, CYLD-silenced in miR-454-in-transfected SW480 cells have positive effect to promote cell proliferation, suggesting that direct CYLD downregulation is required for miR-454-induced CRC cell proliferation. In sum, our data provide compelling evidence that miR-454 functions as an onco-miRNA, playing a crucial role in the promoting cell proliferation in CRC, and its oncogenic effect is mediated chiefly through direct suppression of CYLD expression.


Kinetic selectivity of complementary nucleic acids: bcr-abl-directed antisense RNA and ribozymes.
Efficacy and sequence specificity are two major requirements in the use of antisense nucleic acids and ribozymes. For long-chain complementary RNA sequences (>30 nt), effects in living cells are correlated with the association rate of the complementary RNA in vitro, but not with the stability of the formed double strand. Thus, sequence selectivity of complementary RNA has to be defined as fast versus slow annealing with the appropriate target or non-target sequences, respectively. In this work, we performed a systematic kinetic analysis to evaluate the selectivity of bcr-abl-directed antisense RNA and hammerhead ribozymes with a length of the complementary sequences of between 20 and 80 bases. By kinetic in vitro selection, we identified oligomeric as well as long-chain complementary RNA that annealed at least tenfold faster with the bcr-abl sequence in comparison with either of the wild-type sequences bcr or abl, respectively. In the presence of selected oligodeoxynucleotide sequences and RNase H, the bcr-abl transcript was specifically hydrolysed out of a mixture containing abl and bcr sequences as well. Hammerhead ribozymes were designed such that binding with their target was facilitated either via helix I or helix III-forming antisense arms but not both. Further, cleavage and binding occurred on opposite sides of the bcr-abl fusion point. Target selectivity was found for a ribozyme that annealed fast via abl sequences and cleaved within the bcr portion of bcr-abl RNA. Kinetic probing and calculations of the local folding potential indicate that the bcr-abl fusion point sequences are not easily accessible for complementary nucleic acids. This study supports the need for more detailed structural investigations of the bcr-abl fusion sequence and forms a more rational basis for the therapeutic use of nucleic acid inhibitors of the aberrant bcr-abl gene expression in Philadelphia chromosome-positive cells.


Cannabis Use and Outcomes in Patients With Aneurysmal Subarachnoid Hemorrhage.
The incidence of cannabis use in patients with aneurysmal subarachnoid hemorrhage (aSAH) and its impact on morbidity, mortality, and outcomes are unknown. Our objective was to evaluate the relationship between cannabis use and outcomes in patients with aSAH. Records of consecutive patients admitted with aSAH between 2010 and 2015 were reviewed. Clinical features and outcomes of aSAH patients with negative urine drug screen and cannabinoids-positive (CB+) were compared. Regression analyses were used to assess for associations. The study group consisted of 108 patients; 25.9% with CB+. Delayed cerebral ischemia was diagnosed in 50% of CB+ and 23.8% of urine drug screen negative patients (P=0.01). CB+ was independently associated with development of delayed cerebral ischemia (odds ratio, 2.68; 95% confidence interval, 1.03-6.99; P=0.01). A significantly higher number of CB+ than urine drug screen negative patients had poor outcome (35.7% versus 13.8%; P=0.01). In univariate analysis, CB+ was associated with the composite end point of hospital mortality/severe disability (odds ratio, 2.93; 95% confidence interval, 1.07-8.01; P=0.04). However, after adjusting for other predictors, this effect was no longer significant. We offer preliminary data that CB+ is independently associated with delayed cerebral ischemia and possibly poor outcome in patients with aSAH. Our findings add to the growing evidence on the association of cannabis with cerebrovascular risk.


Immediate compared with on-demand maternal full feeding after planned caesarean delivery: a randomised trial.
To evaluate immediate compared with on-demand full maternal oral feeding after caesarean delivery STUDY DESIGN: A randomised trial. Obstetric unit of a university hospital in Kuala Lumpur, Malaysia. Women admitted for a planned caesarean under spinal anaesthesia. Participants were randomised to a sandwich meal served immediately on return to the ward or on-demand. Primary outcomes were patient satisfaction VAS (visual analog scale of 100 mm) on the feeding regimen and vomiting at 24 hours. 453 women were initially enrolled, 395 were randomised and available for analysis. Median (full range) patient satisfaction VAS scores were 82 (15-100) versus 84 (0-100) mm, P = 0.88 and vomiting rates were 1/197 (0.5%) versus 2/198 (1.0%), P > 0.99 for immediate compared with on-demand feeding, respectively. The immediate versus on-demand arms first ate at a median of 105 (35-210) versus 165 (45-385) minutes, P < 0.001, had second meal at 5.3 (1.2-15.5) versus 5.8 (2.2-29.7), P < 0.001, flatus passage at 9.5 (3.1-29.0) versus 10.3 (2.8-24.6), P = 0.023 hours post-caesarean and opiate analgesia use was 10/197 (5.1%) versus 23/198 (11.6%), P = 0.028, RR 0.4 (95% CI 0.2-0.9), NNTb 16 (95% CI 8-89). The median visual numerical rating scale (0-10 scale) for nausea and bloating at 8, 16 and 24 hours was similarly scored at zero in both arms. Other outcomes were similar. Immediate full feeding has some advantage over on-demand feeding. Both regimens are tolerated well. Patients probably should be fed as soon as practicable after a caesarean. Full maternal oral feeding should commence as soon as practicable after an uncomplicated caesarean section.


Policing Diversity: Examining Police Resistance to Training Reforms for Transgender People in Australia.
Using field notes collected from participant observation of Australian police officers training to work with the transgender community, the current research builds on previous work examining social identity theory (Tajfel, 2010) to explain how one training program implemented to educate police about transgender people challenges police culture. This research determines that police culture, training procedures, and stereotypes of gender are equally influential on police perceptions of all transgender people. Overall, the results indicate that negative police perceptions toward police training reforms strengthen in-group identity of police, and negative out-group perceptions of transgender people.


Unraveling the Amine-Induced Disproportionation Reaction of Perchlorinated Silanes-A DFT Study.
A detailed quantum-chemical study on the amine-induced disproportionation reaction of perchlorinated silanes to neo-Si5 Cl12 is reported. The key intermediate in the resulting mechanistic scenario is a dichlorosilylene amine adduct, which is in tune with recent experimental findings. Yet, at variance with the generally accepted notion of silicon-chain growth by concerted silylene insertion into Si-Cl bonds of lower silanes, the formation of neo-Si5 Cl12 follows more complex pathways. The reactivity is dominated by the Lewis-base character of the dichlorosilylene amine adduct and characterized by three elementary steps that bear close resemblance to the key elementary steps identified earlier for the chloride-induced disproportionation of Si2 Cl6 . NBO and QTAIM analyses of the key reactive species SiCl2 ⋅NMe3 and SiCl3 (-) provide a rationale for these striking similarities.


Treatment of traumatic brain injury in female rats with intravenous administration of bone marrow stromal cells.
To study the effect of bone marrow stromal cells administered intravenously to female rats subjected to traumatic brain injury. We injected marrow stromal cells harvested from male rat bone marrow (n = 24) into the tail vein of the female rat (n = 8) 24 hours after traumatic brain injury; the rats were killed at Day 7 or 14 after treatment. The neurological function of the rats was evaluated using the rotarod test and the neurological severity score. The distribution of the male donor cells in brain, heart, lung, kidney, liver, muscle, spleen, and bone marrow of the female recipient rats was measured by identifying Y chromosome-positive cells using fluorescent in situ hybridization. We found that marrow stromal cells injected intravenously significantly reduced motor and neurological deficits compared with control groups by Day 15 after traumatic brain injury (P < 0.05, analysis of covariance for repeated measures). The transplanted cells preferentially engrafted into the parenchyma of the injured brain and expressed the neuronal marker NeuN and the astrocytic marker glial fibrillary acidic protein. Marrow stromal cells were also found in other organs in female rats subjected to traumatic brain injury without any obvious adverse effects. These data suggest that the intravenous administration of marrow stromal cells may be a promising therapeutic strategy that warrants further investigation for patients with traumatic brain injury.


Afterloading increases the left ventricular end-systolic force-length relation slope in dogs.
We evaluated the effects of pressure loading produced by gradual aortic occlusion on left ventricular (LV) myocardial contractility by assessing changes in the slope of the LV end-systolic force-diameter (Fes-Des) relation. Eleven adult mongrel dogs were prepared with ultrasonic crystals for measuring LV diameter and a micromanometer for measuring LV pressure. Preload was decreased by vena caval occlusion (VCO), and afterload was increased by aortic occlusion (AOO). The slopes (Ec) and extrapolated diameter intercepts (Do) of the LV Fes-Des relation were determined for each dog from the end-systolic data obtained during VCO and AOO. During VCO and also AOO, the heart rate showed little change (134 +/- 18 vs. 134 +/- 16 bpm in VCO, 135 +/- 16 vs. 132 +/- 17 bpm in AOO). The values of Ec and Do during VCO were 62.8 +/- 15.6 g/cm and 1.20 +/- 0.36 cm, respectively, while during AOO the respective values were 122.6 +/- 18.6 g/cm and 1.66 +/- 0.38 cm. Ec and Do were both significantly increased during AOO in comparison with VCO (p less than 0.001). These results suggest that gradual aortic occlusion increases LV myocardial contractility in anesthetized, open-chest dogs under autonomic blockade. This phenomenon might be related to length-dependent calcium activation.


Inhibitory effects of Montelukast on mediator release by nasal epithelial cells from asthmatic subjects with or without allergic rhinitis.
This study tested inhibitory effects of in vitro Montelukast treatment on nasal airway epithelial cells (AEC) cultured from asthmatic patients treated with Montelukast with and without concomitant allergic rhinitis. We further examined the effect of Montelukast withdrawal in these patients on cytokine release from cultured nasal AEC. Nasal AEC were collected by brushings from subjects with a history of stable (no exacerbations or change in medication for ≥ 1 month) physician confirmed mild/moderate asthma whose asthma symptoms were documented to benefit from Montelukast treatment (NCT01230437). Release of the following mediators by nasal AEC were measured: IL-8, IL-6, IL-10, GM-CSF, RANTES, eotaxin and IFN-γ. Nasal AEC were cultured before and one week after withdrawal of their Montelukast treatment. Forty two asthmatics were recruited. Nasal AEC were successfully cultured in 17 at the first assessment, 14 at the second assessment and in 10 individuals at both assessments. Nasal AEC release was no different between asthmatics with or without allergic rhinitis. Montelukast significantly suppressed the release of IL-8 (p = 0.016), IL-6 (p = 0.006), RANTES (p = 0.002) and IFN-γ (p = 0.046), in a dose dependent manner in unstimulated cultures but not in those stimulated with IL-1/TNF. Withdrawal of Montelukast treatment, was associated with increased IL-8 and RANTES secretion in unstimulated nasal AEC cultured from subjects with asthma and allergic rhinitis but not with asthma alone. Montelukast treatment for asthma symptoms reversibly suppresses nasal AEC release of pro-inflammatory mediators (i.e. IL-8 and RANTES) but only in those cells cultured from subjects with concomitant allergic rhinitis.


Drug-nutrient interactions: a case and clinical guide.
Advances in pharmacokinetics and pharmacodynamics require new competencies related to pharmaceutical prescribing. First, both physicians and pharmacists need to recognize the potential negative impact of nutrients and dietary supplements on the absorption, metabolism, and utilization of prescription drugs. Second, physicians, even more than pharmacists, need to recognize the potential negative effects of pharmaceuticals on the absorption, metabolism, and utilization of nutrients. This article discusses common drug-nutrient interactions and presents a case that illustrates how unrecognized nutrient disruption may negatively affect a patient's health and potentially result in unnecessary prescribing of medications. In presenting the case, we also provide a conceptual framework for assessing and treating this patient and a summary of current knowledge regarding drug-nutrient interactions.


Four-year lower extremity disability trajectories among African American men and women.
We examined 4-year lower extremity disability trajectories. Nine hundred ninety-eight African American men and women 49-65 years old were evaluated at baseline and at four annual follow-ups. Lower extremity disability was the number of difficulties with nine standard activities of daily living (ADL), instrumental ADLs (IADL), and lower body function items. Mixed-effect models were used. The 9-item lower extremity disability measure had factorial validity and high reliability (alpha > 0.88). The mean baseline lower extremity disability score was 2.43, and at the subsequent follow-ups it was 2.23, 2.35, 2.60, and 2.70. The mixed-effect model included significant random intercept and aging effects. Fixed factors with the largest effect sizes (all p < or =.001) were physical performance (-0.238 lower extremity disabilities per point on the Short Physical Performance Battery [SPPB]), fear of falling (1.094), poor or fair self-rated health (0.735), self-reported arthritis (0.659), clinically relevant levels of depression symptoms (0.641), body mass index (0.047 per kg/m(2) unit), aging (0.082 per year), and asthma (0.558). To improve lower extremity disability trajectories among African Americans, interventions should focus on improving SPPB scores. In addition, fear of falling, poor or fair self-rated health, and clinically relevant levels of depression symptoms should be considered potential intervention candidates warranting further evaluation.


Polyphosphate produced in recombinant Escherichia coli confers mercury resistance.
An Escherichia coli strain was generated by fusion of a merA-deleted broad-spectrum mer operon from Pseudomonas K-62 with a bacterial polyphosphate kinase gene (ppk) from Klebsiella aerogenes in vector pUC119. A large amount of the ppk-specified polyphosphate was identified in the mercury-induced bacterium with the fusion plasmid designated pMKB18 but not in the cells without mercury induction. These results suggest that the synthesis of polyphosphate as well as the expression of the mer genes is mercury-inducible and regulated by merR. The E. coli strain with pMKB18 was more resistant to both Hg2+ and C6H5Hg+ than its isogenic strain with cloning vector pUC119. The recombinant strain accumulated more mercury from Hg2+- and C6H5Hg+-contaminated medium. Hg2+ transported into the cytoplasm appeared to be bound by chelation with the polyphosphate produced by the recombinant cells. The transported phenylmercury was degraded to Hg2+ before the chelation since polyphosphate did not directly chelate with C6H5Hg+. These results indicate that polyphosphate is capable of reducing the cytotoxicity of the transported Hg2+ probably via chelation between polyphosphate and Hg2+.


Predicting New-Onset Postoperative Atrial Fibrillation in Cardiac Surgery Patients.
To derive a simple clinical prediction rule identifying patients at high risk of developing new-onset postoperative atrial fibrillation (POAF) after cardiac surgery. Retrospective analysis on prospectively collected observational data. A university-affiliated cardiac hospital. Adult patients undergoing coronary artery bypass grafting and/or valve surgery. Observation for the occurrence of new-onset postoperative atrial fibrillation. Details on 28 preoperative variables from 999 patients were collected and significant predictors (p<0.2) were inserted into multivariable logistic regression and reconfirmed with recursive partitioning. A total of 305 (30.5%) patients developed new-onset POAF. Eleven variables were associated significantly with atrial fibrillation. A multivariable logistic regression model included left atrial dilatation, mitral valve disease, and age. Coefficients from the model were converted into a simple 7-point predictive score. The risk of POAF per score is: 15.0%, if 0; 20%, if 1; 27%, if 2; 35%, if 3; 44%, if 4; 53%, if 5; 62%, if 6; and 70%, if 7. A score of 4 has a sensitivity of 44% and a specificity of 82% for POAF. A score of 6 has a sensitivity of 11% and a specificity of 97%. Bootstrapping with 5,000 samples confirmed the final model provided consistent predictions. This study proposed a simple predictive score incorporating three risk variables to identify cardiac surgical patients at high risk of developing new-onset POAF. Preventive treatment should target patients ≥ 65 years with left atrial dilatation and mitral valve disease.


Ca2+ signaling in HEK-293 and skeletal muscle cells expressing recombinant ryanodine receptors harboring malignant hyperthermia and central core disease mutations.
Malignant hyperthermia (MH) and central core disease (CCD) are caused by mutations in the RYR1 gene encoding the skeletal muscle isoform of the ryanodine receptor (RyR1), a homotetrameric Ca(2+) release channel. Rabbit RyR1 mutant cDNAs carrying mutations corresponding to those in human RyR1 that cause MH and CCD were expressed in HEK-293 cells, which do not have endogenous RyR, and in primary cultures of rat skeletal muscle, which express rat RyR1. Analysis of intracellular Ca(2+) pools was performed using aequorin probes targeted to the lumen of the endo/sarcoplasmic reticulum (ER/SR), to the mitochondrial matrix, or to the cytosol. Mutations associated with MH caused alterations in intracellular Ca(2+) homeostasis different from those associated with CCD. Measurements of luminal ER/SR Ca(2+) revealed that the mutations generated leaky channels in all cases, but the leak was particularly pronounced in CCD mutants. Cytosolic and mitochondrial Ca(2+) transients induced by caffeine stimulation were drastically augmented in the MH mutant, slightly reduced in one CCD mutant (Y523S) and completely abolished in another (I4898T). The results suggest that local Ca(2+) derangements of different degrees account for the specific cellular phenotypes of the two disorders.


Keeping up with evolution. Choosing strategies as group purchasing gets more complex.
The next stage of group purchasing contracts has arrived. Increasing demands of alliances and groups require a more sophisticated response from manufacturers than in the past. Innovative programs, contracts and a thorough knowledge of the group are now required to remain ahead of the competition.


Inhibitory effects of tumor necrosis factor-alpha on migration of human periodontal ligament cells.
Tumor necrosis factor-alpha (TNF-alpha) is associated with chronic gingival inflammation and is suspected to influence periodontal destruction. However, the exact roles of TNF-alpha in wound healing and periodontal tissue regeneration are largely unknown. In the present study, we examined the effects of TNF-alpha on migration and proliferation of human periodontal ligament (PDL) cells. PDL cells were cultured in the presence of TNF-alpha to determine its effects on cellular migration and proliferation. The protein expression profiles of alpha5 and beta1 integrin subunits and their related molecules, paxillin and focal adhesion kinases (FAK), were investigated. Gene expression of fibronectin also was assayed. Further, the activation of Rho-family small guanosine triphosphate (GTP)-binding protein (RhoA) was evaluated using a GTP-loading pull-down assay, and focal adhesion formation by PDL cells after transfection with the expression vector of paxillin-fused green fluorescent protein (GFP) also was observed with confocal microscopy. Cellular migration was impaired by TNF-alpha and recovered following the addition of anti-TNF-alpha antibodies. In contrast, PDL cell proliferation was not affected by TNF-alpha. TNF-alpha upregulated the expression of the alpha5 and beta1 integrin subunits, whereas fibronectin was not overexpressed. Phosphorylation of paxillin and FAK by PDL cells was induced, and RhoA activation also was induced. Confocal microscopic analysis revealed that TNF-alpha induced focal adhesion and stress fiber formation in all parts of the cells. Our results suggested that TNF-alpha impairs cellular migration by enhancing cellular adhesive ability following significant focal adhesion and stress fiber formation.


Isolated leptomeningeal carcinomatosis (carcinomatous meningitis) after taxane-induced major remission in patients with advanced breast cancer.
To identify the incidence of leptomeningeal carcinomatosis (LMC), as the first site of systemic progression, in breast cancer patients after having obtained a major response (CR or near CR) to first-line taxane-based chemotherapy and compare these findings in retrospect with a matched-pair group of historical control patients from our database treated with nontaxane regimens. Patients with histologically proven breast cancer having either metastatic disease or high-risk locoregional disease that were entered into treatment protocols with first-line taxane (paclitaxel or docetaxel) plus anthracyclines or mitoxantrone combinations and developed LMC as the first evidence of progression after major response (CR or >80% PR) were analyzed in the present study (n = 155), and compared, as regards the incidence of LMC, to a matched-pair retrospective group of 155 patients treated with nontaxane regimens in our unit. Seven patients with a median age of 54 years (range 40-70) developed LMC as their first evidence of progression after taxane-based regimens with a median interval of 6 months (range 2-18) from start of treatment to diagnosis of LMC. Five patients received intrathecal (i.t.) methotrexate treatment and whole brain radiotherapy (RT), while 1 patient received i.t. methotrexate and RT to the lumbar spine. Two patients responded to treatment for LMC, while 2 achieved stable disease and 3 progressed. Two patients had elevated cerebrospinal fluid tumor markers (more than serum marker levels) that proved useful in monitoring response to treatment. Median survival after LMC was 3.6 months (range 1-17+) and correlated positively to the interval from the initiation of taxane-based therapy to LMC (r = 0.84, p = 0.019). Seven out of 86 responders (8.13%) in the taxane group versus 1 out of 72 responders (1.4%) in the non-taxane-treated group developed LMC as the first sign of progression after a major response to first-line chemotherapy (p < 0.1). LMC after a major response to front-line taxane-based regimens represents a grave disease manifestation and its incidence appears increased, but not significantly so, when compared retrospectively to non-taxane-treated patients. Prospective evaluation of the incidence of LMC after taxane versus non-taxane-based treatment from large randomized multi-institutional trials is warranted and identification of potential prognostic factors might help to identify patients requiring appropriate prophylactic therapy.


Degenerative adult onset scoliosis.
There are people who have no history of scoliosis who develop spinal deformity of a progressive nature as adults, associated with severe degenerative disc disease. The clinical syndrome associated with this deformity is not well documented. In an attempt to describe this clinical syndrome more precisely, 21 patients with the diagnosis of degenerative scoliosis were identified and reviewed. Review included history with pain drawings when available, physical examination, bone densities, and standing spinal roentgenograms. Patients with spinal compression fractures, spondylolyses, prior history of scoliosis or radiographic findings consistent with an idiopathic scoliosis were excluded. Our review shows that these patients can develop, along with progressive scoliosis, loss of lumbar lordosis with a resulting flat back deformity. These patients commonly present in the sixth decade with a predominantly stenotic symptom complex, but often lack the classic feature of relief in a sitting posture. The number of male and female patients was approximately equal. Roentgenogram findings show a high angle deformity over a short number of spinal segments and an absence of bony features associated with idiopathic scoliosis such as lateral vertebral wedging and alterations of the lamina. The incidence of this condition remains to be established.


Validation of the cephalosporin intradermal skin test for predicting immediate hypersensitivity: a prospective study with drug challenge.
Cephalosporin is a major offending agent in terms of drug hypersensitivity along with penicillin. Cephalosporin intradermal skin tests (IDTs) have been widely used; however, their validity for predicting immediate hypersensitivity has not been studied. This study aimed to determine the predictive value of cephalosporin intradermal skin testing before administration of the drug. We prospectively conducted IDTs with four cephalosporins, one each of selected first-, second-, third-, or fourth-generation cephalosporins: ceftezol; cefotetan or cefamandole; ceftriaxone or cefotaxime; and flomoxef, respectively, as well as with penicillin G. After the skin test, whatever the result, one of the tested cephalosporins was administered intravenously and the patient was carefully observed. We recruited 1421 patients who required preoperative cephalosporins. Seventy-four patients (74/1421, 5.2%) were positive to at least one cephalosporin. However, none of responders had immediate hypersensitivity reactions after a challenge dose of the same or different cephalosporin, which were positive in the skin test. Four patients who suffered generalized urticaria and itching after challenge gave negative skin tests for the corresponding drug. The IDT for cephalosporin had a sensitivity of 0%, a specificity of 97.5%, a negative predictive value of 99.7%, and a positive predictive value (PPV) of 0%, when challenged with the same drugs that were positive in the skin test. Routine skin testing with a cephalosporin before its administration is not useful for predicting immediate hypersensitivity because of the extremely low sensitivity and PPV of the skin test (CRIS registration no. KCT0000455).


Prevalence of Lyme borrelia in Ixodes persulcatus ticks from an area with a confirmed case of Lyme disease.
In this study, the prevalence of Borrelia infections in Ixodes ticks from a site in Hokkaido, Japan, with confirmed cases of Lyme disease was determined by a PCR method capable of detecting and differentiating between strains of pathogenic Borrelia, with particular emphasis on Borrelia garinii (B. garinii) and Borrelia afzelli (B. afzelli), using tick-derived DNA extracts as template. A total of 338 ticks, inclusive of 284 Ixodes persulcatus (I. persulcatus), were collected by flagging vegetation in mid-spring. Ninety-eight (34.5%) of I. persulcatus tested positive for Borrelia species DNA, whereas the overall prevalence of Borrelia species in Ixodes ovatus and Haemaphysalis longicornis ticks was 19.5 and 7.7%, respectively. PCR-RFLP and sequence analysis of Borrelia rrf(5S)-rrl(23S) intergenic spacer DNA amplicons indicated that they originated from three different Borrelia species namely, B. garinii, B. afzelii and B. japonica. Among the I. persulcatus species, which is a known vector of human borreliosis, 86 were mono-infected with B. garinii, 2 ticks were mono-infected with B. afzelii and whereas 12 ticks had dual infections. Most significant, 11 of the I. persulcatus ticks were coinfected with Anaplasma phagocytophilum and B. garinii. The difference between the number of obtained and expected co-infections was significant (χ(2)=4.32, P=0.038).


Retinal tubulin binds macular carotenoids.
To investigate the biochemical mechanisms responsible for the specific uptake, concentration, and stabilization of the carotenoids lutein and zeaxanthin in the macula. Soluble extracts of bovine retina mixed with radioactive carotenoids were purified by hydrophobic interaction, ion exchange, and gel filtration chromatography. Carotenoid-associated proteins in these purified preparations were identified through photoaffinity labeling and protein microsequencing. Similar purifications on human macular tissue without the addition of exogenous carotenoids also were performed. Experiments on bovine retinal tissue demonstrated that tubulin is the major soluble carotenoid-binding protein. When soluble extracts of human macular protein were examined, the endogenous carotenoids lutein and zeaxanthin were found to copurify with tubulin. Tubulin is found in abundance in the receptor axon layer of the fovea, where it can serve as a locus for the deposition of the high concentrations of macular carotenoids found there. The binding interaction of carotenoids and tubulin in the Henle's fiber layer could play an important role in the photoprotective effects of the macular carotenoids against the progression of age-related macular degeneration. The association of carotenoids with tubulin, a protein that can form highly ordered linear arrays, may provide an explanation for the dichroic phenomenon of Haidinger's brushes.


Intravenous self-administration of heroin, cocaine, and the combination in Balb/c mice.
Polydrug abuse, including the abuse of cocaine + heroin combinations (or 'speedballs') is an increasingly significant problem. The use of genetically defined populations of mice has the potential to add considerably to the study of polydrug abuse. Balb/cByJ (Balb/c) mice have been shown to self-administer opiates, but not cocaine, therefore these mice were chosen for the initial characterization of intravenous self-administration of cocaine + heroin combinations. Mice were implanted with chronic indwelling jugular catheters and given the opportunity to self-administer heroin, cocaine or heroin + cocaine combinations. Heroin was self-administered, while, under the same conditions, none of the mice tested acquired cocaine self-administration. However, heroin + cocaine combinations were self-administered in naive mice as well as in mice that had failed to self-administer cocaine alone. The heroin + cocaine combination dose-effect curve resembled the heroin dose-effect curve. It is hypothesized that heroin may interact with effects of cocaine that function to limit self-administration in Balb/c mice, facilitating the acquisition and maintenance of self-administration of cocaine + heroin combinations.


[The second phase clinical observation of anti-radiation effect by superoxide dismutase].
Multiple center randomized controlled double blind clinical trait was conducted to evaluate the anti-radiotherapy effect by SOD (produced by Hunan Biochemical Work) in 159 patients. Injection of 4000U SOD immediately after receiving radiotherapy significantly reduced the occurrence rate of skin, oral mucosal, pelvic visceral and systematic adverse reaction, only the reduction of leukopenia did not reach the statistical significant level. No adverse effect of SOD injection was observed. The results suggest that SOD is a safe and effective agent to attenuate the radiotherapy reactions.


Atrial natriuretic factor attenuates prostaglandin E2-stimulated plasma renin activity in humans.
The effect of atrial natriuretic factor (ANF) 99-126 3 or 10 pmol/kg/min on increase in plasma renin activity (PRA) stimulated by a 30-min infusion of prostaglandin E2 (PGE2) 80 ng/kg/min was studied in healthy salt-replete male volunteers. PGE2 increased PRA to approximately 230% of basal levels (p less than 0.001). Concomitant infusion of ANF 3 pmol/kg/min significantly attenuated this rise in PRA to approximately 130% of baseline values (p = 0.02, n = 10). A quantitatively similar effect was observed with ANF 10 pmol/kg/min (n = 5). We suggest, in light of previous findings, that this inhibitory effect of ANF probably represents a nonspecific action rather than a specific effect of ANF on PGE2-mediated increases in PRA.


Bioelectric toxicity caused by chlorpromazine in human lung epithelial cells.
Endogeneous and exogeneous amine-containing substances possess pneumophilic properties. Among them, tricyclic amphiphilic amine drugs like neuroleptics intensively accumulate in the lung cell membrane and occasionally cause severe respiratory disorders. In the present study, we examined the bioelectric toxicity of chlorpromazine (CPZ), a commonly used neuroleptic, in human lung epithelial cells. CPZ concentration-dependently inhibited the isoproterenol (ISO)-generated short-circuit current (I(sc)) sensitive to a nonselective K(+) channel blocker, clotrimazole (30 microM), but insensitive to a selective Ca(2+)-activated K(+) (K(Ca)) channel blocker, charybdotoxin (ChTx, 100 nM). The effects of apical CPZ on the ISO-induced responses were greater than those of basolateral CPZ. Forskolin- and 8-bromo-cyclic AMP-induced I(sc) were partially prevented by CPZ. Nystatin permeabilization of the monolayers revealed that CPZ attenuated the basolateral K(+) current elicited by ISO more than that elicited by forskolin and that the apical Cl(-) current elicited by forskolin was instead potentiated by CPZ, although it inhibited the ISO-induced Cl(-) current. 1-Ethyl-2-benzimdazolinone (1-EBIO, a K(Ca) channel opener, 500 microM)- and ionomycin (Ca(2+) ionophore, 1 microM)-evoked Cl(-) secretions were also sensitive to CPZ. These results indicate that CPZ inhibits transepithelial Cl(-) transport, affecting at least two different targets: the beta-adrenergic receptor and the basolateral K(+) channels (especially the K(Ca) channel). Electrostatic interactions at the inner surface of the membrane between the protonated amines of CPZ and negatively charged portions of the plasma membrane may be involved in the mechanisms.


Individual differences in flow proneness are linked to a dopamine D2 receptor gene variant.
Flow is a special mental state characterized by deep concentration that occurs during the performance of optimally challenging tasks. In prior studies, proneness to experience flow has been found to be moderately heritable. In the present study, we investigated whether individual differences in flow proneness are related to a polymorphism of the dopamine D2 receptor coding gene (DRD2 C957T rs6277). This polymorphism affects striatal D2 receptor availability, a factor that has been shown to be related to flow proneness. To our knowledge, this is the first study to investigate the association between this trait and a specific gene variant. In a sample of 236 healthy Hungarian adults, we found that CC homozygotes report higher flow proneness than do T allele carriers, but only during mandatory activities (i.e., studying and working), not during leisure time. We discuss implications of this result, e.g., the potential mediators of the relationship.


Regulation of RAW264.7 macrophage polarization on smooth and rough surface topographies by galectin-3.
Recognition of topographical features induces phenotypic changes in macrophages although the receptors and signaling pathways are not completely characterized. As integrin molecules in focal adhesions/podosomes are in intimate contact with topography and topography modulates the NFkB pathway through cholesterol enriched raft-associated adhesive signaling structures we hypothesized that a cell-surface signaling complex comprised of galectin-3 together with its ligand CD98 and integrinβ1 is important for topography-directed lineage determination. This study used polished, sand blasted and acid etched (SLA) surfaces and two novel grooved topographies (G1 and G2) produced by anisotropic etching of Si <1 1 0> to evaluate the role of galectin-3 in macrophage polarization in RAW 264.7 macrophages, as determined by gene expression and morphology. In the presence of the galectin-3 inhibitor, lactose, the M2 marker (mannose receptor) was down-regulated while the M1 marker (iNOS) was up-regulated on smooth and rough surfaces. This skewing of phenotype suggests a role for galectin-3 in macrophage polarization towards the M2 phenotype. Additionally, we evaluated the role of PI3K on polarization using PI3K inhibitor LY294002. We found that the M2 marker was down-regulated on both PO (surface polished) and G1 surfaces implicating PI3K in lineage determination. We also found that surface topography altered cell morphology; macrophages had a larger area on G2 surfaces. Lactose treatment significantly reduced the cell area on all topographies suggesting that the galectin-3 is also involved in signaling complexes triggering the rearrangement of the actin cytoskeleton. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2499-2509, 2017.


Diffusion-weighted MR imaging of benign and malignant breast lesions before and after contrast enhancement.
Many publications describe the use of diffusion-weighted imaging (DWI) in breast MRI. This article addresses the question of when to apply the DWI sequence in the course of the scan protocol. The effect of T1-shortening contrast media (CM) on the ADC values of breast lesions is investigated. Data were acquired on a 1.5 T scanner. 60 patients with 79 lesions (20 benign, 59 malignant) were included. The DWI sequence (4 mm slice thickness, b-values: 50, 400, 800) was applied before and after CM administration. Before calculating the ADC map, the b50, b400 and b800 series were analyzed concerning lesion displacement. ADC values before and after CM application were compared. The mean lesion size was 1.5 ± 0.8 cm. On the basis of the b50 and b400 measurements, the mean ADC value of benign lesions was 1.89 ± 0.30 × 10-3 mm2/s before and 1.85 ± 0.28 ×10-3 mm2/s after CM administration. The consecutive values for two pure mucinous carcinomas were 1.88 × 10-3 mm2/s and 1.81 × 103 mm2/s and for the remaining malignant lesions 1.00 ± 0.18 × 10-3 mm2/s and 0.88 ± 0.21 × 10-3 mm2. On the basis of the b50, b400 and b800 measurements, the mean ADC value of benign lesions was 1.99 ± 0.37 × 10-3 mm2/s before and 1.97 ± 0.30 × 10-3 mm2/s after CM application, whereas the mean ADC value of the malignant lesions was 0.90 ± 0.14 × 10-3 mm2/s before and 0.80 ± 0.14 × 10-3 mm2/s after CM application. While there was no significant change for benign lesions, the ADC value decrease in post-contrast malignant lesions was highly significant. DWI after CM is possible and even leads to slightly better lesion discrimination between benign and malignant. However, further studies need to be performed to verify this. Citation Format: • Janka R, Hammon M, Geppert C et al. Diffusion-Weighted MR Imaging of Benign and Malignant Breast Lesions Before and After Contrast Enhancement. Fortschr Röntgenstr 2014; 186: 130 - 135.


Differentiation pathways in duodenal and ampullary carcinomas: a comparative study on mucin and trefoil peptide expression, including gastric and colon carcinomas.
Duodenal carcinomas, such as ampullary tumors, may be a heterogeneous group of neoplasms that share differentiation features with gastric or colorectal carcinomas. Because of the cell- and tissue-specific expression patterns of mucins and trefoil peptides, these markers were used to investigate the differentiation status of duodenal and ampullary carcinomas in comparison with gastric and colorectal carcinomas. Adenocarcinomas (14 duodenal, 10 gastric, 11 ampullary and 10 colorectal) were examined immunohistochemically for the mucin gene products MUC1, MUC2, MUC5AC, MUC6 and the trefoil peptides TFF1 and TFF2. The tumors' expression profile for MUC5AC, MUC6 and TFF1 was used to distinguish between gastric- and intestinal-directed differentiation. The mucins that were most often expressed in the individual tumor types were MUC1 (duodenal and ampullary carcinomas), MUC2 (colorectal carcinomas) and MUC5AC (gastric carcinomas). Further classification focusing on the expression profile for MUC5AC, MUC6 and TFF1 revealed that 21% of the duodenal and 45% of the ampullary carcinomas demonstrated mainly gastric differentiation (positivity for all three markers or only two of them). The remaining duodenal and ampullary carcinomas showed nongastric, i.e., intestinal differentiation (all three markers negative or only one marker positive). The gastric differentiation pattern characterized 60% of gastric carcinomas. Colorectal carcinomas showed intestinal differentiation in 100% of cases. Duodenal carcinomas have a heterogeneous mucin expression pattern that is mainly related to either gastric differentiation or intestinal differentiation. This also holds for ampullary carcinomas. Among the markers used, MUC5AC, MUC6 and TFF1 are most useful for revealing differentiation pathways in duodenal and ampullary carcinoma.


Autocrine secretion of GM-CSF in acute myeloblastic leukemia.
Three cases of acute myeloblastic leukemia (AML) were identified in which clonogenic cells proliferated autonomously in vitro. Cells from two of these cases were found to secrete a colony-stimulating factor (CSF) that was immunologically and molecularly related to GM-CSF. Growth of AML-CFU could be blocked by the addition of a neutralizing antiserum to GM-CSF. Northern blot hybridization of leukemic cell mRNA with a cDNA probe for the GM-CSF gene revealed a 1-kb message identical in size to the normal GM-CSF message in stimulated T cells. No GM-CSF message was detected in the third case. These results indicate that constitutive expression of the GM-CSF gene, apparently by leukemic cells, can result in autonomous in vitro proliferation of AML-CFU in some cases of AML.


Gelatin density gradient for isolation of islets of Langerhans.
Isolation of islets of Langerhans (islets) has been performed by means of collagenase digestion of the pancreatic tissue combined with density gradient separation of islets from unwanted exocrine tissues. An enormous number of islets are necessary for clinical islet transplantation. The density gradient used for isolation of a large number of islets should satisfy several requirements in addition to those for the conventional density gradients, such as high viscosity for creating fine interfaces with a large area, easy sterilization, and low cost. This study is concerned with the development of a new density gradient made of low-molecular-weight gelatin. We isolated islets from the hamster pancreatic tissue using the gelatin density gradients. The yield and purity of islet and its insulin release function were compared with those of islets isolated using Ficoll and Ficoll-Conray density gradients that have been conventionally used. The new gelatin density gradient can separate islets from the unwanted exocrine tissue as effectively as the Ficoll density gradient and more effectively than the Ficoll-Conray density gradients. The islets collected using the gelatin gradient retain ability of insulin release increase in response to glucose stimulation, similar to those isolated by the Ficoll-Conray gradient and more than those collected by the Ficoll gradient. In addition, the gelatin effectively inhibited enzyme activities, that is, collagenase and proteolytic enzymes released from the exocrine tissue, and thus it can inhibit overdigestion of islets during their density gradient isolation. The gelatin gradient satisfies most of the additional requirements for islet isolation from the pancreatic tissue of large animals mentioned above. Although several factors, such as molecular weight of gelatin, osmolality of the gradient, and centrifugal conditions, still remain to be optimized, our results suggest that the gelatin gradient has potentiality to isolate islets from the pancreatic tissue of a large animal.


Associations among circulating branched-chain amino acids and tyrosine with muscle volume and glucose metabolism in individuals without diabetes.
Amino acid metabolites, including branched-chain amino acids (BCAA) and tyrosine (Tyr), affect glucose metabolism. The effects of BCAA on insulin resistance in patients with diabetes seem to conflict with mechanisms determined in animal models and cultured cells. The aim of this study was to clarify the controversy surrounding the effects of BCAA by investigating the physiological effects of BCAA and Tyr on glucose metabolism in healthy community dwellers. We investigated associations among BCAA and Tyr and metabolic parameters in 78 residents (median age, 52 y) of Mie, Japan, who did not have prediabetes, diabetes, or a body mass index >30 kg/m(2). Muscle volume, serum BCAA, and Tyr levels were higher in men than in women (n = 32 and 46, respectively; all P < 0.0001). Stepwise multiple regression analysis associated BCAA positively with muscle volume (regression coefficient/t/p/95% confidence interval, 281.8/3.7/0.0004/129.7-433.8), fasting blood glucose (FBG; 12699.4/3.22/0.0020/4830.9-20567.8), fasting immunoreactive insulin (IRI; 8505.1/2.75/0.0078/2322.5-14687.6), and homeostasis model assessment of β-cell function (HOMA-β; 893.6/2.58/0.0122/201.8-1585.5), and negatively with the HOMA-insulin resistance (HOMA-IR; -9294.1/-2.89/0.0052/-15711.0 to -2877.1). Tyr positively correlated with fasting IRI (26/2.77/0.0072/7.3-44.7). Insulin sensitivity and muscle volume are positively associated with BCAA in individuals without diabetes. In turn, BCAA correlate with increased FBG and fasting IRI levels. Tyr correlated with fasting IRI, but not with insulin sensitivity.


Latitudinal variation in speciation mechanisms in frogs.
Speciation often has a strong geographical and environmental component, but the ecological factors that potentially underlie allopatric and parapatric speciation remain understudied. Two ecological mechanisms by which speciation may occur on geographic scales are allopatric speciation through niche conservatism and parapatric or allopatric speciation through niche divergence. A previous study on salamanders found a strong latitudinal pattern in the prevalence of these mechanisms, with niche conservatism dominating in temperate regions and niche divergence dominating in the tropics, and related this pattern to Janzen's hypothesis of greater climatic zonation between different elevations in the tropics. Here, we test for latitudinal patterns in speciation in a related but more diverse group of amphibians, the anurans. Using data from up to 79 sister-species pairs, we test for latitudinal variation in elevational and climatic overlap between sister species, and evaluate the frequency of speciation via niche conservatism versus niche divergence in relation to latitude. In contrast to salamanders, we find no tendency for greater niche divergence in the tropics or for greater niche conservatism in temperate regions. Although our results support the idea of greater climatic zonation in tropical regions, they show that this climatic pattern does not lead to straightforward relationships between speciation, latitude, and niche evolution.


Brief report: "Quick and (not so) dirty" assessment of change in autism: cross-cultural reliability of the Developmental Disabilities CGAS and the OSU autism CGI.
There are few evaluated economic tools to assess change in autism. This study examined the inter-rater reliability of the Developmental Disabilities Children's Global Assessment Scale (DD-CGAS), and the OSU Autism Clinical Global Impression (OSU Autism CGI) in a European setting. Using these scales, 16 clinicians with multidisciplinary background and varying experience independently rated eight vignettes of autism spectrum disorder for severity and general psychosocial functioning at referral and discharge. Intraclass correlation coefficient (ICCs) for experienced clinicians were .75 for the DD-CGAS and .72 for the OSU Autism CGI. In inexperienced clinicians these ICCs were .58 and .59. Results confirm previous North American studies, and further extents the reliability of the instruments to untrained, less experienced clinicians with different professions.


Design and validation of a new immunoassay for soluble forms of thrombomodulin and studies on plasma.
Thrombomodulin (TM), purified from human placental homogeneate by affinity chromatography on DIP-Thrombin agarose, was used to develop monoclonal antibodies (MAbs). Two of them, 3E2 and 24FM (both IgG1, K), which were not calcium-dependent, were found convenient for developing a two-site enzyme immunoassay. Testing of recombinant and truncated forms of TM26 demonstrated that the species containing the amino terminus including the lectin-like domain and the epidermal growth factor (EGF)-like domains 1-4 were fully measured. The working range was from 2 to 100 ng/ml with a detection threshold of 2 ng/ml. Intraassay and interassay reproducibilities were, respectively, below 7.4% and 8.6%, whereas recovery of purified TM was between 88 and 114% in plasma. Mean plasma concentration was 42.1 (+/- 11.3) ng/ml (males 51.8 +/- 7.9 ng/ml, females 34.8 +/- 7.8 ng/ml) and it was established on 62 normal individuals between the ages of 21 and 55 (28 males and 34 females). This new assay is a convenient tool for measuring plasma TM and establishing its diagnostic and predictive value in diseases associated to endothelial damage.


Psychophysical determination of load carrying capacity for a 1-h work period by Chinese males.
This study used the psychophysical approach to examine the effects of container width, the presence or absence of container handles, and different load-carrying frequencies and distances on the maximum acceptable weight carried and the resulting response (heart rate and rating of perceived exertion) by well-conditioned males for a 1-h work period. After training, 12 male subjects performed a load-carrying task at knuckle height. Each subject performed 30 different carrying combinations. The conditions examined were container width, from 15.2 to 55.9 cm; carrying frequency, from 1 carry to 5 carries/min; and carrying distance from 1 to 6 m. The results were compared with prior studies and led to the following conclusions: (1) the use of container handles leads to the subjects carrying a significantly higher maximum acceptable weight than when containers do not have handles, which differs from the results of a previous study by Morrissey and Liou; (2) there were significant reductions in the maximum acceptable carrying weight with increases in container width, frequency and distance; (3) the presence or absence of container handles, different frequencies and load-carrying distances had significant effects on heart rate, although the effect of container width was not significant. In addition, the various frequencies and distances for load carrying had significant interaction effects on heart rate; (4) the effects of various frequencies and load-carrying distances on the rating of perceived exertion were statistically significant. The most stressed body parts were the wrists and arms.


Expression of acquired immunity to a local isolate of Haemonchus contortus by the Nigerian West African Dwarf goat.
The capacity of young Nigerian West African Dwarf (WAD) goats to express good acquired immunity to their native geographic strain of Haemonchus contortus and the correlates of this responsiveness were studied in a laboratory experiment involving forty 7-8 month old kids. A primary immunising infection with 2000 L3 (equivalent to 260-450 L3/kg body weight) with or without challenge on D42 with 2000 L3 resulted in a mild chronic infection with a pre-patent period of 18-20 days and little or no reduction in worm burden between D14 and D56. In contrast, another group (D) of kids, whose immunising infection had been truncated with fenbendazole on D35 and later received similar challenge infection, developed good protection against challenge. Thus, worm burdens were largest in group E (challenge control), larger in group C (primary+challenge) and least in group D. Of the measures of infection used, namely faecal worm egg counts (FECs), circulating eosinophil (EOS) responses, packed cell volume (PCV) and body weight, FEC and EOS responses exhibited marked individual variability, but only FEC (geometric mean of transformed counts) and PCV showed strong correlation with worm burden. There was also a significant negative correlation between FEC and PCV. The size of inoculum used was well tolerated by the kids, as it induced only mild changes in PCV in some goats and no effect at all on body weights. This suggests that the WAD goat may possess a good measure of resistance to the pathogenic effects of its native strain of H. contortus. The wide individual variability in FEC and its strong relationships to worm burden and PCV are pointers to its likely genetic basis. There are, therefore, good prospects for further studies to identify H. contortus resistant genotypes among the WAD goat population.


Pyroelectric effect induced by the built-in field of the p-n junction in the quantum paraelectric PbTe: experimental study.
We report here the first observation of a pyroelectric effect in a nonpolar semiconductor. This effect originates in the temperature-dependent electric dipole of the PbTe p-n junction. The junction was illuminated by a chopped CO(2) laser beam, and periodic and single-pulse pyroelectric signals were observed and measured as a function of temperature, reverse bias voltage, and chopper frequency. The measured pyroelectric coefficient is approximately 10(-3) microC/cm(2) K in the region of 40-80 K. The theoretical model describes quantitatively all experimental features. The time evolution of the temperature inside the junction region was reconstructed.


Case of chondroid syringoma mimicking a nodular hidradenoma: A diagnostic pitfall on cytopathology.
Limited literature is available on the use of fine needle aspiration (FNA) for skin tumors. Awareness of cytological features of these lesions is important to prevent misdiagnosis. A 45-year-old male, presented with slowly growing swelling over the thigh since 4 years. FNA from multiple sites yielded highly cellular smears with a dual cell population consisting of cells with small round nucleus scanty cytoplasm and polygonal cells with moderate to abundant cytoplasm. Extracellular basement membrane like material was also noted. A diagnosis of benign adnexal neoplasm, suggestive of nodular hidradenoma was made. Paraffin sections showed an encapsulated dermal tumor composed of ductal and myoepithelial cells in ducts, sheets, strands with chondromyxoid stroma. A final diagnosis of chondroid syringoma was given following evaluation of histopathology slides and Immunohistochemistry. FNA interpretation of skin tumors can be challenging since literature is limited. The placement of tumors into benign or malignant category is realistic and can direct management guidelines. Greater awareness of these lesions and following stringent diagnostic criteria can help in establishing a more accurate diagnosis.


CD4+ T cell epitope discovery and rational vaccine design.
T cell epitope-driven vaccine design employs bioinformatic algorithms to identify potential targets of vaccines against infectious diseases or cancer. Potential epitopes can be identified with major histocompatibility complex (MHC)-binding algorithms, and the ability to bind to MHC class I or class II indicates a predominantly CD4(+) or CD8(+) T cell response. Furthermore, an epitope-based vaccine can circumvent evolutionary events favoring immune escape present in native proteins from pathogens. It can also focus on only the most relevant epitopes (i.e. conserved and promiscuous) recognized by the majority of the target population. Mounting evidence points to the critical role of CD4(+) T cells in natural antigen encounter and active immunization. In this paper the need for CD4(+) T cell help in vaccine development, the selection of CD4(+) T cell epitopes for an epitope-based vaccine, and how the approach can be used to induce a protective effect are reviewed.


Characterization of an anti-beta-galactosidase antibody recognizing a precursor but not a mature enzyme in solution.
A monospecific antibody was raised against a recombinant human 84-kDa beta-galactosidase precursor protein, which had been produced with a baculovirus gene expression system and affinity-purified. It bound to the 64-kDa mature enzyme protein extracted from human fibroblasts and the antigenic recombinant precursor on immunoblotting, but neither the fibroblast-derived mature enzyme nor the 64-kDa mature protein-like tryptic product of the recombinant precursor protein was immunoprecipitated. We conclude that it specifically recognized the precursor but not the mature protein in solution. Immunoprecipitation with this anti-precursor antibody revealed that the precursor protein mainly accounted for the residual enzyme activity in fibroblasts from an adult GM1-gangliosidosis patient, and the mature protein accounted for the activity in fibroblasts from a juvenile GM1-gangliosidosis patient.


A disposable end-tidal CO2 detector to verify endotracheal intubation.
We compared the performance of the Fenem FEF end-tidal CO2 detector with the TRIMED capnometer to verify endotracheal intubation. The FEF indicates the presence of CO2 by the color change of a chemically treated indicator; the TRIMED uses infrared technology. Both devices were used during 60 intubations. Intubations during in-hospital emergency situations outside of the operating room were studied. Adult patients undergoing intubation for respiratory failure, CPR, and other airway protection situations were enrolled in the study. The TRIMED monitor and FEF detector were placed in series between the manual resuscitator and the patient's endotracheal tube adapter after endotracheal tube placement. We defined the acceptable criterion for detection of CO2 as production of a positive signal within six manual resuscitator bag breaths. The TRIMED met this criterion in 58 of 60 patients (sensitivity, 0.97) and the FEF met this criterion in 59 of 60 patients (sensitivity, 0.98). A paired t test showed no statistically significant difference in performance between the two devices. In five of nine cases of intubation during CPR, the color change of the FEF was described as "subtle." In one CPR case, a positive signal was not obtained by either device. We conclude that the performance of the FEF CO2 detector is equal to that of the TRIMED monitor for verification of endotracheal intubation in nonCPR situations. Interpretation of FEF color changes during CPR should be approached with caution until further studies using the FEF during CPR are completed.


Hypoxic-ischemic encephalopathy: a review for the clinician.
Hypoxic-ischemic encephalopathy (HIE) occurs in 1 to 8 per 1000 live births in developed countries. Historically, the clinician has had little to offer neonates with HIE other than systemic supportive care. Recently, the neuroprotective therapy of hypothermia has emerged as the standard of care, and other complementary therapies are rapidly transitioning from the basic science to clinical care. To examine the pathophysiology of HIE and the state of the art for the clinical care of neonates with HIE. We performed a literature review using the PubMed database. Results focused on reviews and articles published from January 1, 2004, through December 31, 2014. Articles published earlier than 2004 were included when appropriate for historical perspective. Our review emphasized evidence-based management practices for the clinician. A total of 102 articles for critical review were selected based on their relevance to the incidence of HIE, pathophysiology, neuroimaging, placental pathology, biomarkers, current systemic supportive care, hypothermia, and emerging therapies for HIE and were reviewed by both of us. Seventy-five publications were selected for inclusion in this article based on their relevance to these topics. The publications highlight the emergence of serum-based biomarkers, placental pathology, and magnetic resonance imaging as useful tools to predict long-term outcomes. Hypothermia and systemic supportive care form the cornerstone of therapy for HIE. The pathophysiology of HIE is now better understood, and treatment with hypothermia has become the foundation of therapy. Several neuroprotective agents offer promise when combined with hypothermia and are entering clinical trials.


The human basal forebrain integrates the old and the new.
Acquisition of new learning is challenged by the phenomenon of proactive interference (PI), which occurs when previous learning disrupts later learning. Whereas human neuroimaging studies have focused on the cortical contributions to interference resolution, animal studies demonstrate that efficient resolution of PI depends on cholinergic modulation from basal forebrain (BF). Whether the BF promotes PI resolution in humans is unknown. Here, we adapted a PI paradigm from animal studies for use in a functional MRI experiment. During PI resolution, neurologically intact subjects recruited a BF network that included afferent anterior and posterior cortical sites associated with efficient memory acquisition and perceptual processing. Despite normal performance, nonamnesic patients with alcoholism, which is known to disrupt BF function, did not activate a BF network but instead invoked anterior cortical sites traditionally associated with executive function. These results provide evidence for parallel neural systems, each with the potential to resolve interference in the face of competing information.


Planar biaxial testing of soft biological tissue using rakes: A critical analysis of protocol and fitting process.
Mechanical characterization of soft biological tissue is becoming more and more prevalent. Despite the growing use of planar biaxial testing for soft tissue characterization, testing conditions and subsequent data analysis have not been standardized and vary widely. This also influences the quality of the result of the parameter fitting. Moreover, the testing conditions and data analysis are often not or incompletely reported, which impedes the proper comparison of parameters obtained from different studies. With a focus on planar biaxial tests using rakes, this paper investigates varying testing conditions and varying data analysis methods and their effect on the quality of the parameter fitting results. By means of a series of finite element simulations, aspects such as number of rakes, rakes׳ width, loading protocol, constitutive model, material stiffness and anisotropy are evaluated based on the degree of homogeneity of the stress field, and on the correlation between the experimentally obtained stress and the stress derived from the constitutive model. When calculating the aforementioned stresses, different definitions of the section width and deformation gradient are used in literature, each of which are looked into. Apart from this degree of homogeneity and correlation, also the effect on the quality of the parameter fitting result is evaluated. The results show that inhomogeneities can be reduced to a minimum for wise choices of testing conditions and analysis methods, but never completely eliminated. Therefore, a new parameter optimization procedure is proposed that corrects for the inhomogeneities in the stress field and induces significant improvements to the fitting results. Recommendations are made for best practice in rake-based planar biaxial testing of soft biological tissues and subsequent parameter fitting, and guidelines are formulated for reporting thereof in publications.


Dopamine-functionalized upconversion nanoparticles as fluorescent sensors for organophosphorus pesticide analysis.
Organophosphorus pesticide (OP) residues in agricultural products, herbal medicines and environment have attracted increasing concerns because they cause high healthy risk. Herein, a tyrosinase-mediated photoinduced electron transfer system was constructed for OPs analysis by using dopamine-functionalized upconversion nanoparticles (UCNPs) as fluorescent (FL) sensors. Dopamine quinone was produced by tyrosinase-mediated oxidation of dopamine on the surface of UCNPs, which acted as electron accepter to quench the FL emission of UCNPs. The FL quenching was inhibited by OP since it inhibited the activity of tyrosinase. Chlorpyrifos was used as a model analyte to investigate the feasibility of the FL sensor for the analysis of OPs. Under the optimal conditions, chlorpyrifos can be analysed in a wide range of 1.0 ‒ 1000 ng mL-1, with a detection limit of 0.38 ng mL-1 (3σ). Some other groups pesticides, including organonitrogen pesticide, organochlorine pesticide and chloronicotinyl insecticide all showed negligible interference. The proposed sensor was successfully used to analyse chlorpyrifos spiked in Balloonflower and Angelica with acceptable recovery values of 95.4-120.0%, demonstrating its application potential for real samples. It exhibits some advantages like low cost, high sensitivity and free of autofluorescent interference and photobleaching.


Overexpression in Escherichia coli, purification, and characterization of recombinant 60S ribosomal acidic proteins from Saccharomyces cerevisiae.
The 60S ribosomal subunits from Saccharomyces cerevisiae contain a set of four acidic proteins named YP1alpha, YP1beta, YP2alpha, and YP2beta. The genes for each were PCR amplified from a yeast cDNA library, sequenced, and expressed in Escherichia coli cells using two expression systems. The first system, pLM1, was used for YP1beta, YP2alpha, and YP2beta. The second one, pT7-7, was used for YP1alpha. Expression in both cases was under the control of a strong inducible T7 promoter. The amount of induced recombinant proteins in the host cells was around 10 to 20% of the total soluble bacterial proteins. A new protocol for purification of all four recombinant proteins was established. The preliminary steps of purification were done by ammonium sulfate precipitation (YP1alpha, YP1beta) or NH4Cl/ethanol extraction (YP2alpha, YP2beta). The recombinant proteins were then purified to apparent homogeneity by only two steps of classical chromatographies, ion exchange (DEAE-cellulose) and gel filtration (Sephacryl S-200). Isoelectrofocusing analysis of YP2alpha and YP2beta showed the pIs of the recombinant proteins are the same as that of the native yeast ribosomal P2 proteins. The pI of YP1alpha is changed due to the addition of five amino acids attached to the N-terminus of recombinant polypeptide from the expression vector. YP1beta was obtained as a truncated form of polypeptide, similar to its ribosomal counterpart, YP1beta'. This was proved by isoelectrofocusing gel analysis.


[Urticarial vasculitis: apropos of 12 cases].
12 cases of urticarial vasculitis in 6 males and 6 females of average age are studied. Cutaneous lesions are accompanied with fever (3 cases), arthralgias (5 cases), renal manifestations (2 cases) and gastrointestinal pain (2 cases). The majority presented elevated VSG and increment of the alpha-2-globulin. Histopathologically, we find two different patterns: leukocytic-classic vasculitis and lymphocytic vasculitis of small vessels. Clinical characteristics give up with dapsone better than with doses of corticoids. We consider the influence of climatological agents (fundamentally solar exposition) and we compare our cases with the literature.


Natural history of valvular pulmonary stenosis in children below the age of two years. Long-term follow-up with serial heart catheterizations.
18 patients with mild and moderate valvular pulmonary stenosis diagnosed by heart catheterization before the age of 2 yr (mean 11.6 mth) were followed for 5 to 21 yr without operation. 15 cases had a repeat catheterization study after a mean interval of 10.7 yr. Of 7 cases with an initial right ventricular peak systolic pressure (RVSP) of 50 mm Hg or below, 6 had a decrease and 1 a moderate increase in RVSP. Of 8 cases with an initial RVSP of greater than 50 mm Hg 2 had a decrease, 2 had no change and 4 had a significant increase in RVSP; the latter subsequently underwent operation. Electrocardiography yielded some information, but neither roentgenography nor history was of any help in evaluating the severity of the stenosis at follow-up. As mild and moderate valvular pulmonary stenosis in infants and children below the age of 2 may progress and require operation a repeat heart catheterization is usually to be recommended later in childhood.


Influence of the new AJCC breast cancer staging system on sentinel lymph node positivity and false-negative rates.
The sixth and newest edition of the American Joint Committee on Cancer (AJCC) staging system for breast cancer now defines axillary sentinel lymph nodes with micrometastatic deposits 0.2 mm in diameter or smaller as node-negative. The aim of this study was to determine how this new classification scheme would affect axillary sentinel lymph node positivity, false-negative rate, and overall accuracy of an inception cohort of 205 breast cancer patients undergoing definitive surgery that included sentinel lymph node biopsy plus level I/II axillary lymphadenectomy. Based on the previous AJCC system for staging breast cancer, in which all sentinel lymph node metastases were considered positive, the rate of nodal positivity in this cohort was 47%, the overall accuracy was 99%, and the false-negative rate was 2.1%. According to the new classification system, the rate of nodal positivity in this cohort was 39.5% and the overall accuracy was 98%. The false-negative rate rose to 4.9% because two patients with micrometastatic deposits 0.2 mm or smaller, which are considered node-negative in the new system, had macroscopically positive disease in non-sentinel lymph nodes found in the completion lymphadenectomy.


Partial gonadal dysgenesis associated with a pathogenic variant of PBX1 transcription factor.
A term neonate was admitted to the Neonatal Intensive Care Unit for respiratory distress, hypotonia and atypical genitalia. Significant findings included a small phallic structure, labial folds, no palpable gonads and two perineal openings. Pelvic ultrasound showed uterine didelphys and a gonad in the right inguinal canal. The right gonad was removed during diagnostic laparoscopy with microscopic evaluation showing infantile testicular tissue and fluorescence in-situ hybridisation showed only XY signal suggesting that the removed gonad was a male-developed testis. Infant was 46,XY, SRY probe positive. The parents chose a female sex assignment prior to gonadectomy. The infant had respiratory insufficiency and central hypotonia that persisted on discharge. Whole exome sequencing showed a heterozygous pathogenic variant of the PBX1 gene. This variant encodes the pre-B-cell leukaemia homeobox PBX transcription factor and has been associated with malformations and severe hypoplasia or aplasia of multiple organs including lungs and gonads. Whole exome sequencing was crucial in providing a unifying diagnosis for this patient.


BldD is a direct regulator of key developmental genes in Streptomyces coelicolor A3(2).
BldD is a transcription factor required for aerial hyphae formation in the filamentous bacterium Streptomyces coelicolor. Three targets of BldD regulation were discovered by a number of means, including examination of bld gene interdependence, selective enrichment of chromosomal DNA fragments bound by BldD and searching the promoter regions of known developmental genes for matches to a previously characterized BldD binding site. The three BldD targets identified were the developmental sigma factor genes, whiG and bldN, and a previously uncharacterized gene, designated bdtA, encoding a putative transcription factor. In each target gene, the sequences bound by BldD were characterized by electrophoretic mobility shift and DNase I footprinting assays, and their alignment suggested AGTgA (n)m TCACc as a consensus BldD operator. The in vivo effect of mutation in bldD on the expression of these three target genes was assessed using S1 nuclease protection assays. In each case, target gene expression was upregulated during early colony development in the bldD background, suggesting that, in the wild type, BldD acts to repress premature expression of whiG, bldN and bdtA during vegetative growth.


Urethro-balanitis xerotica obliterans.
A clinical report of 5 cases of balanitis xerotica obliterans (BXO) with urethral involvement is presented. Involvement of the urethra in the BXO is emphasized and the fact that this process is often unnoticed, is stressed. The clinical and histological characteristics of this disease are described in its different localizations. Investigations are suggested to diagnose urethral involvement and the suitable therapy is outlined.


Bilateral oculomotor nerve palsy in Guillain-Barre syndrome.
Guillain-Barre syndrome (GBS) is an acquired immune-mediated inflammatory disorder of the peripheral nervous system. GBS is also called acute idiopathic polyradiculoneuritis. Cranial nerves are affected in over 50% of all cases, with the facial nerves being affected the most. Otherwise, oculomotor nerves affection is rare and might occur in about 10% of cases. In this case report we present 61 years old female with GBS (acute motor and sensory axonal neuropathy subtype) associated with bilateral oculomotor nerve palsy. At the admittance in the neurological status were flaccid paraplegia, tendon reflexes absent at legs and reduced at arms, sensory disturbances in a distal (stocking-glove) distribution and bilateral ptosis. The disease was diagnosed on clinical features, nerve conduction velocity test (NCV), electromyogram (EMG) and cerebrospinal fluid (CSF) tests. After treatment with intravenous immunoglobulins and physical treatment the patient improved. She was able to walk by her own, mild semiptosis remained and she had no paresthesia.


ACOG Technical Bulletin. Gynecologic ultrasonography. Number 215--November 1995.
Advances in ultrasound imaging equipment have allowed increased applications of pelvic imaging by the practicing clinician. The myriad clinical uses presented here should provide the reader with a basic understanding of when ultrasonography may be helpful in different clinical conditions.


Using intravital microscopy to observe bevacizumab-mediated anti-angiogenesis in human head and neck squamous cell carcinoma xenografts.
The study showed the value of using intravital microscopy (IVM) analysis for the study of neoangiogenesis. It demonstrated that the model and the analytical methodology could be used to evaluate in detail the effects of treatment strategies for solid tumours. Neoangiogenesis is a key component of tumour progression, invasion and metastasis. In clinical trials monoclonal antibodies specific for vascular endothelial growth factor - VEGF (bevacizumab) - have been shown to significantly affect tumour progression when given in combination with standard chemotherapy, and also to improve the overall survival of patients. For squamous cell carcinoma of the head and neck (HNSCC), we still await definitive evidence of the effect of such treatment. The present study was designed to investigate the anti-angiogenesis effect of beviacizumab in green fluorescent protein (GFP)-labelled HNSCC xenografts using IVM technology. We performed IVM and used image analysis for quantification of angiogenesis and of effects of bevacizumab on cell viability, combined with histochemical and immunohistochemical analysis to standardize the digital analysis of changes in tumour vascularization and cell viability. We found significant effects of bevacizumab on angiogenesis and cancer cell survival in HNSCC. Repeated injections of bevacizumab were found to provide the greatest effects.


Postprandial sleep in healthy men.
Twenty-one healthy men between 18 and 30 years of age were studied to determine the effects of midday food intake on sleep. Twelve subjects were administered liquid carbohydrate meals at lunchtime on 2 consecutive days. Subjects slept on 22 of the 24 study days for an average of 93 minutes during 3 hours of postprandial polysomnographic recording. Nine subjects were used as controls and were deprived of a lunch meal. Six of the nine subjects slept for an average of 30 minutes during the postprandial period. This time was significantly shorter than that of subjects in the meal condition (p < 0.005). There was no difference in latency to sleep onset following food intake between the two study groups. The results of this study suggest that lunchtime food intake does not promote the initiation of sleep, but that it does increase the duration of sleep episodes occurring during the postprandial period.


Lyn contributes to regulation of multiple Kit-dependent signaling pathways in murine bone marrow mast cells.
SCF induces autophosphorylation of Kit and activates a variety of signaling components including Jnks, Erks, PI 3 Kinase, the JAK-Stat pathway and members of the Src family. Previously we showed that Lyn is activated at multiple points during SCF-induced cell cycle progression and contributes to SCF-mediated growth, chemotaxis and internalization of Kit. However, the Kit-dependent biochemical events that require Lyn are unknown. In this study, we used Lyn-deficient bone marrow mast cells (BMMC) to examine the contribution of this Src family member to tyrosine phosphorylation of Kit and SCF-induced activation of Jnks, Akt, Stat3 and Erks. Although surface expression of Kit was increased in Lyn-deficient BMMC, SCF-induced phosphorylation and growth was reduced compared to wild-type BMMC. Downstream of Kit, SCF-induced activation of Jnks was markedly reduced in Lyn-deficient BMMC. Further, Lyn was required for SCF-induced tyrosine phosphorylation of Stat3. Interestingly, Kit was constitutively associated with PI 3 Kinase in Lyn-deficient BMMC and this correlated with constitutive phosphorylation of Akt. This was in marked contrast to wild-type BMMC, where both these events were induced by SCF. These data indicate that in BMMC, Lyn contributes to SCF-induced phosphorylation of Kit, as well as phosphorylation of Jnks and Stat3. In contrast, Lyn may negatively regulate the PI 3 Kinase/Akt pathway. The opposing effects of Lyn on these signaling pathways may explain the pleiotropic effects ascribed to this Src family member in the literature.


[Pharmacognostical studies on cortex Indigoferae Pseudotinctoriae].
This paper reported the studies on descriptions, microscopical characteristics of cross section and podwer, chemical examination and TLC of Cortex indigoferae pseudotinctoriae. These studies provided referential information for clinics, quality control, development and identification of this crude drug.


[Metastatic involvement of the heart and pericardium--MR case reports].
Secondary metastatic tumors of the heart and pericardium are in comparison with primary cardiac tumors more common, but their overall prognosis is very poor. Metastases can occur in every part of the heart and pericardium. The most common source are tumors--lung cancer, breast cancer, melanoma and lymphoma. In this case report we described two patients with metastatic involvement of the heart. We are pointing out the possibilities of diagnostic imaging of cardiac tumors using echocardiography and magnetic resonance.


Effect of propranolol and labetalol on renal haemodynamics at rest and during exercise in essential hypertension.
The effect of exercise on renal haemodynamics was studied in young patients with mild essential hypertension. Four groups of subjects were studied: thirteen normotensive, healthy control subjects, fifteen untreated, eleven propranolol-treated, and six labetalol-treated patients. Renal plasma flow (RPF) and glomerular filtration rate (GFR) were measured during four consecutive periods, a pre-exercise control period, two exercise periods with loads of 450 kpm/min and 600 kpm/min, respectively, and a postexercise control period. In the untreated patients RPF and GFR were lower during exercise than in the normotensive control subjects, whereas no significant differences were found at rest. In the propranolol-treated patients the reduction in RPF and GFR during exercise were more pronounced than in the untreated hypertensives. In the labetalol-treated patients however, RPF and GFR were reduced only to the same degree as in the untreated hypertensive patients. The reduced renal blood flow in propranolol-treated patients could be attributed to a compensatory increased sympathetic activity caused by an impaired cardiac response to exercise. The lack of reduction in renal blood flow during labetalol therapy could partly be related to alpha-adrenoceptor blockade in the renal vascular bed induced by labetalol, and partly to a lesser degree of reduction in cardiac output during labetalol than propranolol therapy.


Effects of Wholesale and Retail Contamination on the Case Life of Beef 1.
To determine effects of wholesale and retail contamination on steak retail case life, rib steaks were fabricated from wholesale beef ribs using laboratory-simulated extremes of retail processing sanitation. Steak retail case life was more highly correlated with the psychrotrophic bacterial load on the surface of wholesale ribs and retail steaks than with the level of retail processing sanitation. Although steak case life could be predicted as a function of both steak and rib bacterial loads or rib and retail equipment bacterial loads the, following is recommended as the most useful predictor of case life: Steak Case Life (days) = 3.97 - 0.19 (log bacteria/cm2 on wholesale ribs) -0.14 (log bacteria/cm2 on retail processing equipment). This equation could also be used by the retailer to assess the quality of wholesale product received from different suppliers.


[Clinical diagnosis of malaria and amoebiasis].
In all patients with fever after returning from tropical countries malaria must be excluded. Amebic liver abscess is an important differential diagnosis, especially in patients with pain in the right epigastrium. In all patients with diarrhea after returning from tropical countries amebic colitis is an important differential diagnosis. The diagnosis of malaria is based on microscopical demonstration of the parasite in blood slides. In patients with Plasmodium falciparum malaria the extent of organ complications has to be checked. The diagnosis of intestinal amebiasis rests on parasitological stool examination. Amebic liver abscess is diagnosed by demonstration of the liver abscess in imaging techniques and verification by finding specific antibodies.