["Expressed emotion", marital quality and risk of recurrence in depressed patients].
Long-term course of depression appears to be clearly correlated to marital quality and quality of the relationship with spouses. Aim of the present study is the replication of results, indicating that expressed emotion (EE) is an important factor in prediction of relapse in depression. Additionally, the correlation between EE shown by spouses and the severeness of depressive symptomatology as well as the importance of martial quality as seen by patients for relapse are investigated. Results of our study indicate that spouses EE does not predict relapse, and status of EE does not correlate with the extent of depressive symptoms. However, patient self ratings of satisfaction with marital relationship and of perceived criticism are related to relapse frequency. Additionally, discrepant ratings of marital satisfaction given by spouses predict relapse to some extent. Implications for research in affective disorders and clinical practice are discussed.


Recombinant expression of mouse osteocalcin protein in Escherichia coli.
Osteocalcin is the most abundant non-collagenous protein of bone. Recombinant mouse osteocalcin protein (mOC) that includes the highly conserved central domain for binding to hydroxyapatite (HA), a mineral component of bone, was expressed in Escherichia coli. Purified mOC protein exhibited a significant increase in HA adhesion and differentiation in osteoblast cells as well as binding to HA with high affinity.


Sensitivity of cultured human pancreatic carcinoma cells to dihydroxyanthracenedione.
We tested the effectiveness of dihydroxyanthracenedione (DHAD) on cell growth of two human pancreatic carcinoma cell lines MIA PaCa-2 and PANC-1. At the level of ID50, the drug was almost equally effective against both cell lines. When the time exposure of MIA PaCa-2 cells to the drug was increased from 1 h to continuous exposure for 5 days, the ID50 was decreased about three-fold only (1.4 X 10(-8)M and 4 X 10(-9)M respectively). At the level of ID50 also the difference between 6 h exposure and continuous exposure for 5 days was minimal. In equimolar concentrations and with 1 h exposure, DHAD was more effective against MIA PaCa-2 cells than other chemotherapeutic agents including adriamycin, mitomycin-C, 5-FU, vincristine, vindesine, vinblastine, VP-16-213, bleomycin, cis-platinum, asparaginase and acivicin. In concentrations of 5 X 10(-7)M, DHAD caused about 40% inhibition of 14C-thymidine incorporation of MIA PaCa-2 cells. Treatment of MIA PaCa-2 cells with the ID50 of DHAD for 1 h caused retardation of cellular traverse, with the major effect appearing to be in G2 + M phase of the cycle. From these data DHAD appears to be a potent drug against human pancreatic carcinoma in vitro.


[Wet environmental cure for Dupuytren pathology. Evaluation and follow-up].
Dupuytren disease starts with a fracture in the hand which in most cases requires surgery. In the Galdakao Hospital during 2006 and early 2007, medical personnel carried out a study to evaluate the effectiveness of a Aquacel wet cure dressing (Hydrofiber Technology, ConvaTec, Bristol-Myers Squibb) in treatment for secondary wounds after surgery to deal with Dupuytren pathology. 52 patients who underwent surgery for Dupuytren pathology were included in this study, in stages from N to IV, using surgical techniques Skoog and Skoog and McCash. Average patient age was 62.8 (range 29-79); 43 (82.7%) were male. Perilesional skin condition improved in 100% of the cases after the use of this dressing. Average time for complete cicatrisation was 2.7 weeks (DE 1.04). Initial pain sensation tied to the wound had a 4.9 average score (evaluated according to an analogical pain scale from 1 to 10). After cure sessions with this dressing, a 1.8 score was registered (for pain reduction of 63%). Adaptation and ease of application of this dressing received an excellent rate in 47 cases or 90.4% and a good rate in 4 cases, or 9.6%. Comfort sensation was rated as excellent by 48 patients, or 92.3% and good by 4 patients, or 7.7%. Removal of this dressing was rated as excellent in 33 cases, or 63.5% and good in 19 cases, or 36.5%. After this wet environment cure, 32 patients, or 61.5% showed good mobility in their hand, acceptable mobility in 14 cases, or 26.9%, and reduced mobility in 5 cases, or 9.6%. Only one case, 7.9%, did not permit an evaluation.


Differences in the efficiency and stability of gene expression after transfection and nuclear injection: a study with a chick delta-crystallin gene.
The efficiency of an exogenous gene's expression was compared after its transfection and injection into various mouse cells to systematically evaluate these two gene transfer techniques. Special attention was paid to the period of transient expression. The gene used was a derivative of chicken delta-crystallin gene with the 5' end region replaced by a promoter base sequence of a retrovirus. Nuclear injection was more efficient than transfection in several respects: it was roughly one thousand times more efficient in producing gene-expressing cells than the transfection technique; it produced positive cells in every challenged cell line in contrast to the results of some unsuccessful trials found with transfection; and the maximum expression of the exogenous gene in a gene-transferred cell was much higher after injection than after transfection. With the transfection technique, use of a DNA-calcium phosphate coprecipitate was slightly more efficient than the use of DEAE-dextran. The stability of gene expression in transfected and nuclear-injected cells differed greatly: Expression of the exogenous gene in transfected cells was transmitted to 92% of the daughter cells per division, whereas its expression in injected cells was transmitted to only 32% of the daughter cells. This great difference in stability probably reflects different states of the major fraction of the exogenous gene: integration into chromosomes in transfected cells versus extrachromosomal localization in injected cells.


Procalcitonin in patients with acute coronary syndromes and cardiogenic shock submitted to percutaneous coronary intervention.
Procalcitonin (PCT) is known to be a biological diagnostic marker for severe sepsis, or septic shock in critically ill patients. There are still contrasting data about a role of procalcitonin in patients with acute myocardial infarction or cardiogenic shock, and in those with acute coronary syndromes, that is, non-ST-elevation myocardial infarction or unstable angina. We evaluated plasma levels of procalcitonin and C-reactive protein (CRP) in 52 patients admitted to our intensive cardiac care unit (ICCU): 14 patients with cardiogenic shock (CS) following ST-elevation myocardial infarction (STEMI), 15 patients with uncomplicated ST-elevation myocardial infarction (STEMI), and 24 with non-ST-elevation myocardial infarction or unstable angina (NSTEMI/UA). In all patients, infective processes were excluded. Procalcitonin values were significantly higher in CS patients with respect to the other two subgroups (P < 0.001, P < 0.001) while CRP levels were higher than NSTEMI/UA patients (P < 0.001) but not with respect to STEMI patients (P = 0.063). No correlations were found in cardiogenic shock patients between CRP and PCT values (R = 0.02; P = 0.762, ns). Procalcitonin levels measured on ICCU admission are significantly higher in patients with cardiogenic shock following the acute myocardial infarction, and they are not correlated with those of CRP. The degree of myocardial ischemia (clinically indicated by the whole spectrum of ACS, from unstable angina to cardiogenic shock ST-elevation following myocardial infarction) and the related inflammatory-induced response are better reflected by CRP (which was positive in most acute cardiac care patients of all our subgroups), than by PCT which seems more reflective of a higher degree of inflammatory activation, being positive only in all CS patients.


Alcalilimnicola halodurans gen. nov., sp. nov., an alkaliphilic, moderately halophilic and extremely halotolerant bacterium, isolated from sediments of soda-depositing Lake Natron, East Africa Rift Valley.
An alkaliphilic, halotolerant, Gram-negative, heterotrophic, aerobic and rod-shaped organism was isolated from drying soda and at a water-covered site of Lake Natron, Tanzania, by means of the most-probable-number technique developed for anoxygenic, phototrophic sulfur bacteria. It had an absolute requirement for alkalinity, but not for salinity; growth occurred at salt concentrations of 0-28% (w/v), with optimal growth at 3-8% (w/v) NaCl. The bacterium preferentially metabolized volatile fatty acids and required vitamins for growth. The name Alcalilimnicola halodurans gen. nov., sp. nov. is proposed for the novel isolate, placed in the gamma-Proteobacteria within the family Ectothiorhodospiraceae on the basis of analysis of the 16S rDNA sequence, polar lipids, fatty acids and DNA base composition. Although Alcalilimnicola halodurans is closely related to the extreme anoxygenic, phototrophic sulfur bacteria of the genus Halorhodospira, it is not phototrophic.


Release of polycyclic aromatic hydrocarbons from Yangtze River sediment cores during periods of simulated resuspension.
The role of resuspension duration on release of 16 PAHs was measured experimentally using a particle entrainment simulator (PES). Three sediment cores were resuspended for 12h at 0.2 and 0.5 N m(-2). PAHs in water column and total suspended solids (TSS) were monitored at intervals. After 0.25 h of resuspension, PAH release was on average 42% of their concentrations after 12h of resuspension, indicating fast release of PAHs from sediments in an initial short time. Moreover, PAHs released faster at 0.5 N m(-2) than at 0.2N m(-2); low molecular weight PAHs (2-3-ring) released faster than median molecular weight (4-ring) PAHs. PAH concentrations in TSS showed generally increase with time and differences in magnitudes based on sediment type and energy. Overall, the composition of sediments is the major factor in determining the amount of released PAHs, more so than the level of resuspension energy applied.


The synthesis and antiviral properties of acyclic nucleoside analogues with a phosphonomethoxy fragment in the side chain.
Acyclic nucleoside analogues bearing phosphonomethoxy residues in the side chain (ANP) attract much attention due to a very beneficial combination of biological properties. Intensive work of organic chemists during the last two decades resulted in a large panel of new compounds that were evaluated as potential antiviral drugs. Herein, we present an overview of major chemical structures within the group of acyclic nucleoside analogues containing phosphonomethoxy side fragments and describe main aspects of their synthesis and antiviral potential. We also describe progress in "prodrug" approaches applied to this chemical group to improve pharmacokinetic profiles of the potential candidates. Chemical modifications in the molecule of parental ANP aimed at blocking of phosphonate charges resulted in a set of promising derivatives, two of which have been recently approved for treatment of hepatits B (Hepsera) and HIV (Viread). The preparation, antiviral properties and some aspects of metabolic transformations and pharmacokinetics of ANP prodrugs are discussed.


The role of CNP-mediated PKG/PKA-PLCβ pathway in diabetes-induced gastric motility disorder.
Our previous work demonstrated that the C-type natriuretic peptide (CNP)/cyclic guanosine monophosphate (cGMP)/cyclic adenosine monophosphate (cAMP) pathway in gastric antrum smooth muscle of rats with diabetes was upregulated and played an important role in the development of diabetic gastric dysmotility. Our goal for this study was to explore the downstream signaling pathways of CNP. We found that the expressions of protein kinase G (PKG) and protein kinase A (PKA) in gastric smooth muscle tissue of rats with diabetes were significantly upregulated. The expressions of β-type phospholipase C 3(PLCβ3) and β-type phospholipase C 1(PLCβ1) protein were reduced, whereas Phosphor-PLCβ3Ser1105 (P-PLCβ3Ser1105) was increased. The inhibitory effect of CNP on gastric antral smooth muscle in diabetic rats was significantly greater than in the normal group. The content of trisphosphate inositol (IP3) in the gastric antral smooth muscle of rats with diabetes was significantly lower than that of the normal group. After blocking PKA with N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H-89, a blockage PKA), the effect of CNP on the production of IP3 was decreased, while blocking PKG with KT5823 (a blockage PKG) simultaneously, and CNP can no longer reduce the IP3 production. CNP promoted the phosphorylation of PLCβ3Ser1105, thereby inhibiting the activity of PLCβ3 in gastric smooth muscle tissue of rats with diabetes; this effect can be abolished by blocking PKA and PKG. These results suggested that CNP can decrease IP3 level in gastric smooth muscle cells and thus inhibit gastric smooth muscle contraction through PKG/PKA-PLCβ pathway, which may play an important role in the development of diabetic gastroparesis.


Adult Dental Trauma: What Should the Dental Practitioner Know?
The management of adult dental trauma can be a daunting challenge for practitioners at any level. Like medical emergencies, initial management can have a large influence on prognosis. It is important that practitioners understand the basic principles of managing the acute presentations of dental trauma. This article aims to illustrate a step-by-step approach in order to improve the management within general dental practice for better outcomes for patients.


Serum protein pattern in sarcoidosis analysed by a proteomics approach.
An altered protein expression in bronchoalveolar lavage fluid (BALF) of sarcoidosis patients has previously been reported. As this disease is systemic, involving not only the lungs, a change in the serum components is also to be expected. In the present study we therefore analysed the total serum protein profile of the patients earlier reported, with active sarcoidosis. We used a proteomics approach to assess overall changes in the protein content of serum in patients with acute sarcoidosis (n = 6) compared to healthy controls (n = 4). Our results show a significantly higher number of protein-spots in serum of the patients compared to the healthy individuals in the pH range 4.5-6.7 (median 886 vs. 742, p < 0.05). The total protein concentrations of the patients' sera were also significantly higher compared to the controls (median 62 vs. 56 mg/mL, p < 0.05). Measurement of the optical densities of the protein-spots from two-dimensional electrophoresis gels, covering pH interval 4.5-6.7, showed varying levels of expression of 22 different serum proteins in the patients compared to the controls. These proteins are involved in immune responses and some are known markers of inflammation. We found three proteins, which were changed concomitantly in the BALF as described in a previous report, and sera of the same patients. Our results are of importance in order to identify biochemical markers in blood of sarcoidosis patients. Ideally, a combination of markers in BALF and serum may turn out to be disease specific.


[CANCER PATIENTS FOLLOW-UP – CROATIAN SOCIETY OF MEDICAL ONCOLOGY CLINICAL GUIDELINES Part I: breast cancer, uterine cancer, cervical cancer, ovarian cancer].
Treatment of oncological patients must be based upon multidisciplinary approach, and takes place in specialized oncological centers. By the end of a specific oncological treatment further follow-up is managed mostly by the oncologists, but the role of the general practitioners becomes more important every day and therefore should be precisely defined. Nowadays, most of the existing follow-up guidelines are not based on prospective studies, but on the experts opinion of a particular oncological center or specialists. The aim of the Croatian Society of Medical Oncology (CSMO) with these recommendations is to standardise and rationalise the diagnostic procedures algorithm in the follow-up of oncological patients after primary treatment.


[Homeopathy from the viewpoint of the clinical pharmacologist].
The homeopathy is a therapeutic line developed about 170 years ago which is based on the sciences of the late 18th century. The homeopathy is founded (1) on the vitalism of Aristoteles, (2) on the examination of remedy reactions in healthy subjects as basis of the therapy of patients, (3) on the "simile" rule, that means the treatment of symptoms with remedies which produce similar symptoms of a mild intoxication in healthy subjects, (4) on the principle of potentiation with the opinion that low doses provoke stronger therapeutic effects, (5) on the individual finding of a remedy due to an extensive anamnesis and (6) on the principle of nosodes, that means the use of diluted body secretion due to infectious diseases for therapy. The therapeutic principles of homeopathy are based on insecure hypotheses and on the patient's information of improved behaviour. Thus, homeopathy does not agree with the present natural science.


Changes in thrombin generation, fibrinolysis, platelet and endothelial cell activity, and inflammation following endovascular abdominal aortic aneurysm repair.
Abdominal aortic aneurysm (AAA) is a chronic inflammatory condition associated with a prothrombotic, hypofibrinolytic diathesis that may increase the risk of cardiovascular events. The effect of endovascular aneurysm repair (EVAR) on this prothrombotic diathesis is not fully understood, especially over the medium and long term. A better understanding of these postintervention changes may improve the risk of cardiovascular complications in the long term. The purpose of this study was to examine thrombin generation, fibrinolysis, platelet and endothelial activation, and the inflammatory response during the 12 months following EVAR. Twenty-nine patients (mean age, 76.9 years) undergoing EVAR for AAA (mean diameter 6.9 cm) had prothrombin fragment (PF) 1 + 2, thrombin-antithrombin complex (TAT), plasminogen activator inhibitor (PAI) activity, tissue plasminogen activator (t-PA) activity and antigen, soluble P- and E-selectin, and highly sensitive C-reactive protein (hsCRP) measured before and at 24 hours, and 1, 6, and 12 months after surgery. PF1 + 2 were markedly elevated prior to EVAR and remained so at 24 hours and 1 month, but had decreased significantly at 6 and 12 months. TAT was also elevated prior to EVAR and increased still further by 24 hours, but fell to below baseline levels thereafter. PAI activity and t-PA antigen were normal prior to EVAR, increased significantly at 24 hours, and then fell to baseline levels. t-PA activity was only detectable at 1 and 6 months; there was a significant rise in soluble P- and E-selectin after EVAR, which was sustained for 12 months. hsCRP increased transiently in response to EVAR but returned to preoperative levels by 1 month. The prothrombotic, hypofibrinolytic diathesis associated with AAA is normalized 12 months after EVAR. This beneficial systemic effect of EVAR for AAA disease may help protect patients against future thromboembolic cardiovascular events.


How specific are periodic complexes in the diagnosis of herpes simplex encephalitis?
The specificity of localized periodic complexes (PC) in the EEG is discussed by means of a case. This patient presented rapidly evolving focal symptoms of the CNS together with fever. The EEG showed localized PCs and in the CT scan there was a hyperdense lesion in a region corresponding to the EEG finding. Postmortem examination revealed a subcortical bleeding in the temporal region. Possible mechanisms for the origin of these PCs and their diagnostic value are discussed.


[Primary hyperparathyroidism disclosed by heart arrhythmia].
The authors report a case of primary hyperparathyroidism in a 47 year old woman presenting with cardiac arrhythmias. She had paraoxystic supraventricular tachycardias and rate-dependent ventricular hyperexcitability suggestive of a catecholamine-induced phenomenon which were resistant to antiarrhythmic therapy over a 3 year period; the plasma calcium concentration was 3.30 mmol/l. An ectopic parathyroid adenoma was removed surgically, normalising the biological changes, and no further arrhythmias were detected by Holter monitoring during the 2 year follow-up period. The association of atrial and ventricular arrhythmias seems to be relatively rate in hyperparathyroidism; patients usually present with atrioventricular block and ventricular hyperexcitability. The other cardiovascular abnormalities observed in hyperparathyroidism are hypertension and myocardial hypertrophy. The electrophysiological mechanisms underlying the clinical manifestations remain obscure.


Cancer Therapeutics, Molecular Targets, Pharmacology and Clinical Applications--First International Conference 19-21 February 2004, Florence, Italy.
The treatment of cancer is currently undergoing a conceptual revolution due to the discovery of cancer pathways and mechanisms of oncogenesis. There are presently two complementary approaches in the field of cancer chemotherapy. Firstly, the development of "classical" anticancer agents aimed at the inhibition of cellular proliferation as a whole and bearing global cytotoxic properties that are aimed not only at cancer cells but also at all actively proliferative cells of the organism. Secondly, the development of "targeted" agents, which inhibit specific cancer targets including cyclin-dependent kinases, tyrosine kinase receptors and metalloproteases involved in cell invasion. These new molecularly targeted therapies are still experimental, with the exception of two or three compounds that have been marketed recently; a number of molecules, however, appear promising. It is generally believed that the "cytostatic" properties of classical compounds (in contrast to the cytotoxic properties) will avoid the general toxicity symptoms that usually accompany cancer chemotherapy, and that many of them will find specific indications in the cancers bearing the alteration they target. This renewal in the concept of anticancer drug discovery and development explains the emergence of new research groups involved in this field, and the continued interest in organizing meetings devoted to this rapidly expanding field. In addition, identification of the molecular targets of classical as well as novel agents, together with the completion of genome sequencing, has allowed the emergence of the science of pharmacogenomics, which tries to relate gene structure and/or expression to the activity of anticancer drugs. It is anticipated that in a few years from now cancer chemotherapy will no longer be prescribed as a function of the localization of the tumor (breast or colon, for example), but as a function of the oncogenic alteration (mutated Ras tumor, ErbB-2 amplifying tumor, etc), wherever it is localized. The paradigm of antibiotics and infectious diseases may well serve as a guide for this evolution.


Adjuvant carboplatin-based chemotherapy in resected stage IIIA-N2 non-small cell lung cancer.
We determined whether adjuvant vinorelbine/paclitaxel plus carboplatin prolonged overall survival among patients with completely resected stage IIIA-N2 non-small cell lung cancer (NSCLC). We randomly assigned patients with completely resected stage IIIA-N2 NSCLC to a control group or to a treatment group with vinorelbine/carboplatin or paclitaxel/carboplatin doublet adjuvant chemotherapy. The primary endpoint was overall survival; secondary endpoints were disease-free survival and the toxicity and safety of the regimen. This trial was terminated before accumulation of the planned numbers for registration because of the results of bigger clinical trial. Finally, 150 patients underwent randomization to vinorelbine/paclitaxel plus carboplatin (79 patients) or observation. In the chemotherapy group, 38 patients received vinorelbine plus carboplatin and 41 patients received paclitaxel plus carboplatin. In both groups, the median age was 57 years, 73% were men, and 28% had squamous carcinoma. Chemotherapy caused neutropenia in 82% of the patients (including grade 3 and 4 neutropenia in 42%), and there were no treatment-related deaths in this trial. After a median follow-up of 29 months (range, 1-110 months), the overall survival was significantly prolonged in the chemotherapy group, when compared with the observation group (33 months [95% confidence interval {CI}, 27.4-38.6] versus 24 months [95% CI, 15.8-32.2], p = 0.037), as was disease-free survival (32 months [95% CI, 21.3-42.7] versus 20 months (95% CI, 13.1-26.9), p = 0.020). The 5-year overall survival rates were 31.1% and 19.1%, respectively. Although with limitations, this clinical trial showed that adjuvant vinorelbine/paclitaxel plus carboplatin has an acceptable level of toxicity and prolongs disease-free and overall survival among patients with completely resected stage IIIA-N2 NSCLC.


Identification of two polyketide synthase gene clusters on the linear plasmid pSLA2-L in Streptomyces rochei.
The 200kb linear plasmid pSLA2-L was suggested to be involved in the production of two macrolide antibiotics, lankamycin (Lm) and lankacidin (Lc), in Streptomyces rochei 7434AN4. Hybridization experiments with the polyketide synthase (PKS) genes for erythromycin and actinorhodin identified two eryAI-homologous regions and an actI-homologous region on pSLA2-L. The nucleotide sequence of a 3.6kb SacI fragment carrying one of the eryAI-homologs revealed that it codes for part of a large protein with four domains for ketoreductase, acyl carrier protein, ketosynthase, and acyltransferase. Gene disruption confirmed that the two eryAI-homologs are parts of a large type-I PKS gene cluster for Lm. A 4.8kb DNA carrying the actI-homologous region contains four open reading frames (ORF1-ORF4) as well as an additional ORF, i.e. ORF5, which might code for a thioesterase. Deletion of the ORF2-ORF4 region showed that it is not involved in the synthesis of Lm or Lc. Thus, it was confirmed that pSLA2-L contains two PKS gene clusters for Lm and an unknown type-II polyketide.


The palpation of pulses.
In 554 anaesthetised patients, the times taken to separately palpate and identify each of the carotid, radial, brachial and femoral pulses were recorded. The patients were divided into three groups based on the form of airway management chosen (tracheal tube, facemask or laryngeal mask airway). Our results demonstrate that in the operating theatre environment the identification of the radial pulse is the most rapid and reliable; by 5 s, 98% and by 10 s, more than 99% of radial pulses were identified. The carotid pulse was not so easily identified, requiring 10 s to enable an identification rate of greater than 95%. The presence of a laryngeal mask airway or a tracheal tube did not hinder the identification of carotid pulse.


Structural basis of effector regulation and signal termination in heterotrimeric Galpha proteins.
This chapter addresses, from a molecular structural perspective gained from examination of x-ray crystallographic and biochemical data, the mechanisms by which GTP-bound Galpha subunits of heterotrimeric G proteins recognize and regulate effectors. The mechanism of GTP hydrolysis by Galpha and rate acceleration by GAPs are also considered. The effector recognition site in all Galpha homologues is formed almost entirely of the residues extending from the C-terminal half of alpha2 (Switch II) together with the alpha3 helix and its junction with the beta5 strand. Effector binding does not induce substantial changes in the structure of Galpha*GTP. Effectors are structurally diverse. Different effectors may recognize distinct subsets of effector-binding residues of the same Galpha protein. Specificity may also be conferred by differences in the main chain conformation of effector-binding regions of Galpha subunits. Several Galpha regulatory mechanisms are operative. In the regulation of GMP phospodiesterase, Galphat sequesters an inhibitory subunit. Galphas is an allosteric activator and inhibitor of adenylyl cyclase, and Galphai is an allosteric inhibitor. Galphaq does not appear to regulate GRK, but is rather sequestered by it. GTP hydrolysis terminates the signaling state of Galpha. The binding energy of GTP that is used to stabilize the Galpha:effector complex is dissipated in this reaction. Chemical steps of GTP hydrolysis, specifically, formation of a dissociative transition state, is rate limiting in Ras, a model G protein GTPase, even in the presence of a GAP; however, the energy of enzyme reorganization to produce a catalytically active conformation appears to be substantial. It is possible that the collapse of the switch regions, associated with Galpha deactivation, also encounters a kinetic barrier, and is coupled to product (Pi) release or an event preceding formation of the GDP*Pi complex. Evidence for a catalytic intermediate, possibly metaphosphate, is discussed. Galpha GAPs, whether exogenous proteins or effector-linked domains, bind to a discrete locus of Galpha that is composed of Switch I and the N-terminus of Switch II. This site is immediately adjacent to, but does not substantially overlap, the Galpha effector binding site. Interactions of effectors and exogenous GAPs with Galpha proteins can be synergistic or antagonistic, mediated by allosteric interactions among the three molecules. Unlike GAPs for small GTPases, Galpha GAPs supply no catalytic residues, but rather appear to reduce the activation energy for catalytic activation of the Galpha catalytic site.


The influence of aging on poststroke depression using a rat model via middle cerebral artery occlusion.
Poststroke depression (PSD) is the most frequent psychological sequela following stroke. While previous studies describe the impact of age on brain infarct volume, brain edema, and blood-brain barrier (BBB) breakdown following ischemia, the role of age on PSD has yet to be described. Here, we examine the influence of age on PSD progression in a rat model of PSD by middle cerebral artery occlusion (MCAO). One hundred forty-three rats were divided into three groups. 48 rats 20 weeks of age underwent a sham procedure, 51 rats 20 weeks of age had MCAO, and 44 rats 22-26 months of age had MCAO. Groups were further divided into two subgroups. The first subgroup was used to measure infarct lesion volume, brain edema, and BBB breakdown at 24 h. In the second subgroup at 3 weeks after MCAO, rats were subjected to a sucrose preference test, two-way shuttle avoidance task, forced swimming test, and a brain-derived neurotrophic factor (BDNF) protein level measurement. Total and striatal infarct volume, brain edema, and BBB breakdown in the striatum were increased in older rats, as compared with younger rats. While both old and young rats exhibited depressive-like behaviors on each of the behavioral tests and lower BDNF levels post-MCAO, as compared with control rats, there were no differences between old and young rats. Although older rats suffered from larger infarct volumes, increased brain edema and more BBB disruption following MCAO, the lack of behavioral differences between young and old rats suggests that there was no effect of rat age on the incidence of PSD.


Quality of life and functional status of terminally ill head and neck cancer patients: a nation-wide, prospective observational study at tertiary cancer centers in Japan.
Little is known about quality of life and functional status of patients with terminally ill head and neck cancers. We conducted a multicenter, prospective, observational study to examine quality of life and functional status in terminally ill head and neck cancer patients. Of the 100 patients meeting inclusion criteria, 72 were observed until death. There was no significant difference in the quality of life score between baseline and Week 3. Forty patients (54.9%) could speak and 22 patients (30.5%) could have oral intake upon study entry. Fifty-three patients (74.6%) received enteral nutrition. Twenty-six patients (36.6%) required dressing changes for fungating tumors. The route of nutritional intake (nasogastric tube vs. percutaneous gastric tube) might be predictive for the duration of hospital stay (64 vs. 21 days, P = 0.0372). There was no significant relationship between quality of life and functional status seen in this study. Feeding tube type could have the most impact on quality of life.


The multilayer flow modulator stent for the treatment of arterial aneurysms.
The purpose of this manuscript was to: 1) report our experience with the Multilayer Flow Modulator (MFM) for the treatment of peripheral, visceral and aortic aneurysms; 2) review the published literature regarding the outcomes of patients treated with the MFM; and 3) develop initial treatment guidelines regarding the MFM. We reviewed our clinical experience with the MFM in 58 high surgical risk patients. Thirty-one peripheral (PAAs), 9 visceral (VAAs) and 18 aortic aneurysms (10 thoracoabdominal [TAAA]; 8 abdominal) were treated. In addition, the PubMed database through April 2013, along with relevant websites and scientific presentations at international meetings, were quered regarding the MFM. Seventeen articles and 3 presentations were identified. Data regarding 178 patients treated with the MFM were included for analysis including 57 PAAs, 31 VAAs and 90 complex degenerative aortic lesions. Outcomes including technical success, 30-day mortality, endoleak rate and aneurysm-related survival were studied. In our experience, there were 47 males and the mean age was 62 years (16-80). In patients with PAAs and VAAs, technical success with the MFM was 100%. At 30 days, there were no deaths. Initial MFM patency was 97.5% (39/40) with patency of the thrombosed MFM successfully restored. Longer-term follow-up (mean 16 ± 8 months) demonstrated progressive thrombosis and shrinkage of the aneurysm sacs and all side-branches were patent. In patients with aortic aneurysms, technical success was 100%, with no complications and no deaths at 30 days. Longer-term follow-up (8 ± 7 months) demonstrated aneurysm-related survival of 100%, all-cause survival of 83.3%, intervention-free survival of 100% and 100% patency of the side branches. The longest duration for aneurysm sac thrombosis was 18 months. A significant mean diameter reduction was observed at 6 months (17.3 mm for the transversal maximal diameter and 13.83 mm for the antero-posterior diameter) in the TAAA group. In the literature review, there was 100% technical success and a 97.7% 30-day survival rate in patients with PAAs and VAAs treated with the MFM. In follow-up (range 5-26 months), there were no aneurysm-related deaths or aneurysm ruptures and the overall survival was 95.5%. Complete aneurysm exclusion was observed in 94.3% of the patients with significant aneurysm shrinkage in 83% of the patients. Nine (10%) MFMs occluded with most occlusions resulting from pre-existing conditions. Patency of 5 occluded MFMs were restored and 4 occluded MFMs were not treated and were asymptomatic. All covered side branches were patent except in a patient with thrombophilia who also had an occluded MFM. The treatment of complex aortic degenerative lesions with MFM demonstrates a 95.5% 30-day survival with 2 aneurysm ruptures for contrindicated use (previously ruptured aneurysm; mycotic aneurysm). Over the follow-up (range 3-28 months), all-cause survival was 87.8% and aneurysm-related survival was 96.7% (1 late rupture due to a type 1 endoleak). Side branches were patent for 97.7% of the treated cases and a 13.3% endoleak rate was reported. There were no neurological, renal or respiratory complications. Complete exclusion and size stability were achieved for most of the cases. Clinical experience with the MFM is increasing. The MFM has been used to treat many types of aneurysms including peripheral, visceral and aortic. Early results suggest that use of the MFM can help prevent aneurysm-related mortalities while maintining branch vessel patency. Additional study and investigation is needed.


Dynamic behavior of the cadherin-based cell-cell adhesion system during Drosophila gastrulation.
During Drosophila gastrulation, morphogenesis occurs as a series of cell shape changes and cell movements which probably involve adhesive interactions between cells. In the present study, we examined the dynamic aspects of cadherin-based cell-cell adhesion in the morphogenetic events to assess its contribution to morphogenesis. DE- and DN-cadherin show complementary expression patterns in the presumptive ectoderm and mesoderm at the mRNA level. We found that switching of cadherin expression from the DE- to the DN-type in the mesodermal germ layer occurred downstream of the mesoderm-determination genes twist and snail. However, examination of their protein expression patterns showed that considerable amounts of DE-cadherin remained on the surfaces of mesodermal cells during invagination, while DN-cadherin did not appear on the cell surfaces at this stage. Further immunocytochemical analysis of the localizations of DE-cadherin and its associated proteins Armadillo (beta-catenin) and Dalpha-catenin revealed dynamic changes in their distributions which were accompanied by changes in cell morphology in the neuroectoderm and mesoderm. Simultaneously, adherens junctions (AJs), based on the cadherin-catenin system, were shown to change their location, size, and morphology. These dynamic aspects of cadherin-based cell-cell adhesion appeared to be associated with the following: (1) initial establishment of the blastoderm epithelium, (2) acquisition of cell motility in the neuroectoderm, (3) cell sheet folding, and (4) epithelial to mesenchymal conversion of the mesoderm. These observations suggest that the behavior of the DE-cadherin-catenin adhesion system may be regulated in a stepwise manner during gastrulation to perform successive cell-morphology conversions. Moreover, the processes responsible for loss of epithelial cell polarity and elimination of preexisting DE-cadherin-based epithelial junctions during early mesodermal morphogenesis are discussed.


Electron microscopical study of initial and final stages of fowl plague virus-replication in chick embryo cells.
Cellular uptake of fowl plague virus occurs 10-30 minutes after inoculation of chick embryo cells. The penetration of the virions is by pinocytosis (viropexis); fusion with the cellular membrane has not been observed. After pinocytosis the virions become gradually disintegrated. Budding of newly formed virions from the cellular membrane starts 3 hours post inoculation (p.i.) and reaches its maximum 8 hours p.i. At the same time budding takes place into electron microscopically empty and autophagic vacuoles. Eight hours p.i. about 3 per cent of the infected cells show budding of virions from the surface and into cytoplasmic vacuoles. Labelling of the cellular membrane with ruthenium red demonstrated that these cytoplasmic vacuoles are not simple cross-sections of invaginations of the cellular membrane. Cluster-like structures were found at 6 hours p.i. in the nuclei of infected cells; however, the suggestion that the clusters develop from nucleoli could not be confirmed.


Evaluation of polyanion-coated biodegradable polymeric micelles as drug delivery vehicles.
Polymeric micelles, as drug delivery vehicles, must achieve specific targeting and high stability in the body for efficient drug delivery. We recently reported the preparation of polyanion-coated biodegradable polymeric micelles by coating positively charged polymeric micelles consisting of poly(L-lysine)-block-poly(L-lactide) (PLys-b-PLLA) AB diblock copolymers with anionic hyaluronic acid (HA) by polyion complex (PIC) formation. The obtained HA-coated micelles showed significantly higher stability in aqueous solution. In this study, to evaluate the HA-coated polymeric micelles as a drug carrier, model drug release from the micelles and cytotoxicity of the micelles were investigated. The HA-coated micelles showed sustained release of model drugs and low cytotoxicity. It is known that there are receptors for HA on liver sinusoidal endothelial cells (LSEC). Specific interactions of HA-coated micelles with LSECs and Kupffer cells were investigated and compared with polymeric micelles coated with other polyanionic polysaccharides, i.e., heparin (Hep) and carboxymethyl-dextran (CMDex). Although Hep-coated micelles and CMDex-coated micelles were incorporated into both Kupffer cells and LSECs, HA-coated micelles were taken up only into LSECs. These results suggest HA-coated micelles have potential utility as drug delivery vehicles exhibiting specific accumulation into LSECs.


A prospective phase II trial of concomitant chemotherapy and radiotherapy with delayed accelerated fractionation in unresectable tumors of the head and neck.
Our study is a prospective evaluation of unresectable malignant cancers of the head and neck treated with concomitant chemotherapy and radiotherapy (RT) using delayed accelerated fractionation (concomitant boost). Between January 1988 and March 1995, 82 patients with unresectable cancers of the head and neck were enrolled in this phase II study. Of these, 52 patients were treated and followed for a minimum of 3 years and are the subject of this analysis. All patients had T4 lesions and were stage IV according to the American Joint Committee on Staging Criteria (AJCC). Patients received RT with accelerated fractionation to a total of 70 Gy in 6 weeks using a concomitant-boost technique. Concomitant cis platin (100 mg/M2) was given on days 1 and 22 of RT. Twenty-seven patients received mitomycin-C (7.5 mg/M2) on days 1 and 22, and 1 patient received mitomycin-C on day 1. In addition, 27 patients received adjuvant chemotherapy with cis platin and vinblastine. The mean follow-up was 45 months (range, 36-72 months). The minimum follow-up for surviving patients in 3 years. At 3 years, the local control rate was 58%. Three-year local control rates for paranasal sinus, nasopharynx, oropharynx, and larynx/hypopharynx were 78%, 78%, 64%, and 100%, respectively. For all patients, the distant-metastasis-free survival was 56%, and the overall survival rate was 36%. Patients with oral cavity cancers experienced worse overall survival versus other sites, 0% versus 47% (p = .03). Salivary cancers also showed worse survival versus other sites, 0% versus 47%, but was not statistically significant. Severe acute complications occurred in 34% of patients. Treatment-related toxicity also resulted in the death of 2 patients. Severe late complications occurred in 7% of patients. Treatment of this poor prognostic group of patients with aggressive chemotherapy and RT produced surprisingly good local control and survival.


Dietary fiber, source foods and colorectal cancer risk: the Fukuoka Colorectal Cancer Study.
Despite much evidence from laboratory work, epidemiological evidence remains elusive regarding the role of dietary fiber in colorectal carcinogenesis. We investigated associations of dietary fiber and source foods with colorectal cancer risk in the Fukuoka Colorectal Cancer Study, a community-based case-control study. The study subjects were 816 incident cases of colorectal cancer and 815 community controls. Nutrient and food intakes were estimated on the basis of a computer-assisted interview regarding 148 dietary items. Odds ratios of colorectal cancer according to quintile categories of energy-adjusted intakes of dietary fiber and food groups were obtained with adjustment for non-dietary factors and dietary intakes of calcium and n-3 fatty acids. Total, soluble and insoluble dietary fibers were not measurably associated with overall risk or subsite-specific risk of colorectal cancer. By contrast, rice consumption was associated with a decreased risk of colorectal cancer (trend p = 0.03), particularly of distal colon and rectal cancer (trend p = 0.02), and high intake of non-rice cereals tended to be related to an increased risk of colon cancer (trend p = 0.07). There was no association between vegetable consumption and colorectal cancer, whereas individuals with the lowest intake of fruits tended to have an increased risk of colorectal cancer. The present study did not corroborate a protective association between dietary fiber and colorectal cancer, but suggested a decreased risk of distal colorectal cancer associated with rice consumption.


[Overlap of irritable bowel syndrome and functional dyspepsia based on Rome III criteria in medical students from a private university in Lima, Peru].
To determine the prevalence of irritable bowel syndrome and functional dyspepsia in medical students from a private university in Lima, Peru. Furthermore, to determine the associated factors with these diseases. Observational descriptive and retrospective cross sectional study. Medical students from a private university in Lima were surveyed using a Rome III questionnaire for functional disorders and the STEPwise auto survey for defining the variables of alcohol and tobacco. For data analysis the Stata 11.0 program was used. Of the 608 students, 543 answered the questionnaire. The prevalence of Irritable Bowel Syndrome was 12.4%, 16.9% of dyspepsia, and of both diseases simultaneously, a prevalence of 7.1% was found. Alcohol consumption in the total population was 89.4% and 29.0% of tobacco. Association between both disorders was found (OR 10.47, 95% CI 5.08 to 21.55; p < 0.001), dyspepsia was associated with sex (OR: 0.16, 95% CI 0.07 to 0.36 p < 0.001), with alcohol consumption (OR: 5.22, 95% CI 23.99 1,13- p = 0.034) and with irritable bowel syndrome (OR: 9.88, 95% CI 4.78 to 20.46 p <0.001). Both conditions together were associated with sex (OR: 0.20, 95% CI from 0.06 to 0.60 p = 0.004) and daily consumption of tobacco (OR: 3.23, 95% CI 1.17 to 8.89 p = 0.023). A prevalence of 12.4% of Irritable Bowel Syndrome and 16.9% of dyspepsia was determined. An overlap of 7.1% of these diseases was reported.


[Radioimmunoassay for plasma betamethasone and its application for clinical study (author's transl)].
A sensitive and specific radioimmunoassay for plasma betamethasone has been developed. The antiserum used was obtained by immunizing rabbits with betamethasone-3-oxime-human serum albumin (BM-3-oxime-HSA) conjugate. The sensitivity was 20 pg and a standard curve was established with a useful range from 20 pg to 4 ng. The cross reactivity of all tested endogenous steroids was less than 0.2%. Cortisol with a cross reactivity of 0.14% caused slight interference at very high concentrations, but this factor is negligible when plasma cortisol level is less than 30 microgram/dl. The calculated interference by cortisol was less than 10%, and plasma not receiving BM consistently gave blanks which were less than 20 pg/tube. Reliability criteria were satisfactory. By this method plasma BM could be measured directly in dichloromethane extract of plasma. The plasma concentrations of BM were measured in normal subjects and patients with liver diseases following oral administration of BM. The peaks of the plasma concentrations for 1.0 mg and 1.5 mg of BM were 345 +/- 40 (n = 3) and 710 +/- 200 ng/dl (n = 5) respectively within 2 hours after administration in normal subjects. After the peak level, plasma BM rapidly fell and disappeared 24 hours after administration in all examined normal subjects. In patients with chronic active hepatitis, the peak levels for 1.0 mg and 1.5 mg of BM were 428 +/- 48 (n = 4) and 837 +/- 83 ng/dl (n = 7) respectively within 2 hours after administration. However, the peak levels of plasma BM were higher than those of normal subjects, and the disappearance of BM from the blood was markedly delayed, reaching a level of 318 +/- 88 nad 622 +/- 148 ng/dl respectively for 1.0 mg and 1.5 mg of BM at 5 hours. The relatively high plasma concentration of BM, ranging from 135 to 170 ng/dl was maintained even 24 hours after administration in all patients with chronic active hepatitis. The disappearance of cortisol from the blood also rapidly fell in normal subjects, but was markedly delayed in patients with chronic active hepatitis. There was good correlation between the severity of the liver disease as measured by the ICG retention at fifteen minutes and removals of BM and cortisol from the blood.


Unilateral pulmonary oedema/atelectasis in the lateral decubitus position.
An obese male patient developed hypoxia, hyercarbia and radiological signs of pulmonary oedema/atelectasis in the dependent lung after surgery in the lateral decubitus position. This appears to have been due to ventilation-perfusion mismatch, although other factors were considered. The patient recovered following 36 hours of intermittent positive pressure ventilation of the lungs.


[Drug therapy of urge incontinence in the female].
In this study we urodynamically examined 105 women suffering from urge incontinence before and 4 weeks after treatment with various drugs. The subjective and objective findings in the 3 patients' groups are compared. The results showed that the patients treated by spasmolytics (Oxyphenonium bromatum/Hexasonium jodatum/Papaverin) had the lowest rate of side effects in comparison with the women treated by calcium channel blockers and spasmolytics (Diltiazem/Oxyphenonium bromatum) and those with imipramine therapy. But the success rate of the first group was the lowest, too.


C.T. findings in the Hajdu-Cheney syndrome.
We describe a patient with the typical features of Hajdu-Cheney Syndrome. The previously undescribed cranial computerised tomographic (C.T.) findings in this condition are discussed. In addition we describe a possible association with bowel malrotation.


Ellagic acid: a potent naturally occurring inhibitor of benzo[a]pyrene metabolism and its subsequent glucuronidation, sulfation and covalent binding to DNA in cultured BALB/C mouse keratinocytes.
The metabolism of [3H]benzo[a]pyrene (BP) by cultured primary keratinocytes prepared from BALB/C mouse epidermis was found to be largely inhibited by the dietary plant phenol, ellagic acid. Varying concentrations of ellagic acid added to the keratinocyte cultures resulted in a dose-dependent inhibition of the cytochrome P-450-dependent monooxygenases aryl hydrocarbon hydroxylase (AHH) and 7-ethoxycoumarin-O-deethylase (ECD). The major organic solvent-extractable metabolites found intracellularly in the cultured cells were trans-7,8-dihydro-7,8-dihydroxybenzo[a]-pyrene (BP-7,8-diol) and 3-hydroxybenzo[a]pyrene (3-OH-BP), although small amounts of 9-hydroxybenzo[a]pyrene, quinones and trans-9,10-dihydro-9,10-dihydroxybenzo[a]-pyrene (BP-9,10-diol) were also present. The major organic solvent-extractable metabolites found in the extracellular culture medium were BP-7,8-diol and BP-9,10-diol, with smaller quantities of unconjugated phenols and quinones. The major intracellular and extracellular water-soluble metabolites of BP were conjugated with glucuronide (primarily 3-OH-BP and several BP-quinones), and to a lesser extent with sulfate (primarily BP-7,8-diol). Both intracellular and extracellular metabolism of organic solvent-extractable and water-soluble conjugates was significantly inhibited by ellagic acid in a dose-dependent manner. The intracellular enzyme-mediated binding of BP to mouse keratinocyte DNA was also largely inhibited in a dose-dependent fashion by ellagic acid. Our results indicate that cultured primary mouse keratinocytes offer a useful model system for studying factors affecting the metabolic activation and detoxification of polycyclic aromatic hydrocarbon carcinogens in the epidermis, and that polyphenolic compounds such as ellagic acid may prove useful in modulating the risk of cutaneous cancer that results from exposure to these environmental chemicals.


Thymic hormone modulation of leukemogenic virus replication.
The effect of purified calf thymus extract, thymosin, on infection with murine leukemia virus (MuLV) (Rauscher) was studied in adult thymectomized BALB/c mice. The course of infection was determined by virus titer in the blood plasma, by enumeration of spleen cells replicating MuLV (i.e., infectious centers), and by the examination of cells for expression of virus-induced cell membrane antigen. Thymectomy (performed 1 week prior to the virus injection) decreased MuLV infection by all three parameters. However, administration of thymosin (500 mug, given in equal doses 2 days before and at the time of injection of virus) caused a marked increase of plasma titer of virus, as well as an increased number of spleen cells generating MuLV as compared to controls treated either with purified spleen fraction or with diluent (buffered salt solution). More than two doses of thymic extract (up to nine, given every 2 days after infection) had no further effect on plasma virus titer or number of splenic infectious centers. Furthermore, treatment of mice with either thymic or splenic extracts increased the incidence of virus related membrane antigen appearing on the surface of spleen cells, as compared to diluent-treated control animals. These results indicate that thymosin may enhance infection by or replication of murine oncornaviruses.


Alpha-adrenergic signal transduction in renin transgenic rats.
The alpha1-adrenoceptor-G protein-phosphoinositide-specific phospholipase C (PLC) signal transduction pathway is assumed to play an important role in the regulation of contractile force and in the pathophysiology of myocardial hypertrophy. In the present study, the components of this pathway were investigated in left ventricles of hearts from hypertensive transgenic rats overexpressing the mouse renin gene [TG(mREN2)27] in comparison to age- and weight-matched Sprague-Dawley control rats. Contractile force was assessed in isolated electrically driven left ventricular papillary muscle strips. Alpha1-adrenoceptor density was measured by radioligand binding using [3H]prazosin, steady state levels of alpha q/11, and G protein beta-subunits by Western blotting. PLC activity was determined by a cell-free assay using exogenous phospholipid vesicles containing [3H]phosphatidylinositol (4,5)-bisphosphate as a substrate. Alpha1-adrenoceptor density was significantly increased (by 80%) in transgenic rats compared with control rats, while the positive inotropic response to the alpha1-adrenoceptor agonist phenylephrine was significantly reduced, suggesting a postreceptor defect in TG(mREN2)27. The expression of alpha q and alpha11 was verified by reverse transcription-polymerase chain reaction, and alpha q/11 steady state protein levels were shown to be similar in transgenic and control rats. Western blotting using a beta-common antibody revealed two bands at approximately 35 and 36 kD. The quantities of both were similar in TG(mREN2)27 compared with those in control rats. In contrast, PLC activity was significantly reduced (by 32%) in transgenic rats. In conclusion, our findings are consistent with a desensitization of the alpha1-adrenergic signal transduction pathway at the level of the effector.


Interferon and the treatment of polycythemia vera, essential thrombocythemia and myelofibrosis.
Recombinant IFN-α (rIFN-α) induces complete hematologic remissions in patients with myeloproliferative neoplasms (MPNs), but its use has been limited by side effects owing to the relatively high doses used. Now, low-dose rIFN-α is stressed, starting relatively early in the course of the MPNs. In polycythemia vera, this has resulted in a significant clinical, hematologic, morphologic and molecular response manifested by reduction in the JAK2(V617F) allele burden, sustained even after discontinuation of recombinant IFN. In essential thrombocythemia, platelet count reduction is prompt and durable without treatment for varying periods. In hypercellular primary myelofibrosis, rIFN-α has restored normal blood counts, reduced splenomegaly and induced morphologic marrow remissions. This article highlights our current use of rIFN-α in MPNs.


Integration of cell cycle control with transcriptional regulation by the retinoblastoma protein.
Rapid progress in several areas of molecular biology has led to the realization that the retinoblastoma protein may play a pivotal role in the coordination between cell cycle control and regulation of gene expression. This role is a subtle one, and is important in only certain mammalian cell types. Exploring the details of these connections, and why only some cells rely on them, is already beginning to shed light on the regulation of cell multiplication.


Duplication of the extrahepatic bile duct.
Duplication of the extrahepatic bile duct is an extremely rare anomaly. There are five types of this anomaly, with type V being the least common variant. Only two cases of type V (one each of type Va and type Vb) have been reported. Here we report the second case of type Va duplication of the common bile duct presenting with choledocholithiasis. The patient underwent open choledocholithotomy and T-tube choledochostomy after failed endoscopic stone clearance. Duplication of the bile duct was detected on T-tube cholangiogram, which was confirmed by magnetic resonance cholangiopancreaticography (MRCP). No associated pancreaticobiliary maljunction was found on the imaging.


Kinetics of benzene adsorption onto activated carbon.
An activated carbon bed adsorption process is influenced by the adsorbents' characteristics, volatile organic compound (VOC) characteristics, and process conditions. In the literatures, the adsorption processes of the adsorbents and VOCs were usually considered to be in equilibrium. In this study, the VOC adsorption processes by activated carbon were considered to be a kinetic process, i.e. they are not in equilibrium. Then, isothermal adsorption curves and a small column experiment were simulated.


Social well-being and its measurement in the nursing home, the SWON-scale.
The aim of this study was to develop an observational scale to measure the social well-being of nursing home residents, by assessing not only the social behaviour of the resident towards others, but also the behaviour of others towards the resident. Traditionally, aspects of the social well-being of nursing home residents are assessed according to the social activities and interactions where they engage. Although these are important indicators of social well-being, other important indicators may include the positive social behaviour of others towards the resident (e.g. confirming the resident's behaviour or showing affection). A cross-sectional descriptive survey design. From the perspective of human social needs, items relating to fulfillment of the needs for affection, behavioural confirmation and status were formulated and tested. This took place in three nursing homes in the Netherlands that provide somatic and psycho-geriatric care. The study (sample n = 306) yielded a short and reliable scale, the Social Well-being Of Nursing home residents-scale, with separate sub-scales (three items each) for fulfillment of the three social needs. These first results indicate that overall social well-being and its sub-dimensions can be measured with this new observational scale, although its validity needs to be confirmed. Including the social behaviour of others towards the resident may have provided a more comprehensive measure of the social well-being of nursing home residents. This measure may help to underscore the importance of the social behaviour of others (e.g. caregivers) for the overall social well-being of residents and with that assist care-providers in nursing homes to improve the social well-being of the residents.


The risk of acute myocardial infarction and arterial hypertension in a cohort of male employees of a Siberian Group of Chemical Enterprises exposed to long-term irradiation.
During the period from 1998 to 2007, a prospective cohort study of acute myocardial infarction morbidity cases as well as a "case-control" study of arterial hypertension was carried out. The risk of acute myocardial infarction was assessed as well as arterial hypertension; the dose-response relationship and the role of radiation in the mechanism of acute myocardial infarction as well as arterial hypertension development were studied. As a result of this study, a statistically significant increased risk of acute myocardial infarction among the male staff at the Siberian Group of Chemical Enterprises [standardized relative risk = 1.16 (1.04; 1.29)] exposed to external irradiation in comparison with employees unexposed to ionizing radiation was observed. A significant increase in the risk of acute myocardial infarction was observed at external radiation dose accumulation of more than 300 mSv [standardized relative risk = 1.46 (1.09; 1.91)]. The increase in arterial hypertension risk has been established among the analyzed group of employees exposed to long-term irradiation in the absence of the linear dependence of risk, based on cumulative dose of external γ-irradiation [risk due to external radiation dose in the range of 7.3-21.3 mSv = 1.6 (0.96; 2.51) and in the range of external radiation dose 21.4-68.5 mSv = 1.7 (1.04; 2.67) for 68.6-864 mSv = 1.6 (1.01; 2.57)]. This led to the conclusion that radiation can act also as a factor that might potentiate the negative effects of the "traditional" risk factors in the pathogenesis of acute myocardial infarction and hypertension.


Heterogeneity in TV fast food advertisement exposure in South Korea.
To assess TV fast food ad exposure in South Korea. We assessed time trends of targeted ratings (licensed from Nielsen Media Research Korea) by household income and education during 2004-2010. Lower income groups saw more fast food TV ads during the study period. Exposure decreased in all income groups with a bigger income gap in 2010 than in 2004. The relative exposure to local fried chicken franchise TV ads surged from one fifth in 2004 to half of all TV fast food ads seen in 2010 in all socioeconomic status (SES). Future studies should assess the link between TV fast ood ad exposure, an important contextual factor for individual food choices, and actual consumption.


E2F4 is required for early eye patterning.
Increasingly, studies reveal novel functions for cell cycle proteins during development. Here, we investigated the role of E2F4 in eye development. E2F4-deficient mouse embryos exhibit severe early eye patterning defects, which are evident from embryonic day 11.5 and characterized by aberrant shape of the optic cup, coloboma as well as abnormal eye pigmentation. Loss of E2F4 is associated with proximal-distal patterning defects in the optic vesicle. These defects are characterized by the expansion of optic stalk marker gene expression to the optic cup and reduced expression of ventral optic cup markers. These defects are associated with a split of Shh expression domain at the ventral midline of the forebrain and expansion of the Shh activity into the ventral optic cup. Despite these patterning defects, early neuronal differentiation and Shh expression in the retina are not affected by E2F4 deletion. Overall, the results of our studies show a novel role of E2F4 in the early eye development.


Stratification and weighting via the propensity score in estimation of causal treatment effects: a comparative study.
Estimation of treatment effects with causal interpretation from observational data is complicated because exposure to treatment may be confounded with subject characteristics. The propensity score, the probability of treatment exposure conditional on covariates, is the basis for two approaches to adjusting for confounding: methods based on stratification of observations by quantiles of estimated propensity scores and methods based on weighting observations by the inverse of estimated propensity scores. We review popular versions of these approaches and related methods offering improved precision, describe theoretical properties and highlight their implications for practice, and present extensive comparisons of performance that provide guidance for practical use.


Surface-functionalized nanoparticles for controlled drug delivery.
Nanoparticles have been extensively investigated in drug-delivery systems. Especially, the effectiveness of the surface-functionalized nanoparticles, which consist of copolymers with functional molecules, is well demonstrated. This chapter describes the complete technique for the preparation of surface-functionalized nanoparticles. Tetracycline with an affinity to bone was chosen as a model material for surface functionalization. There are two steps for the preparation of tetracycline-modified nanoparticles. The first step is the conjugation of poly(D,L-lactide-co- glycolic acid) with tetracycline via carbodiimide chemistry and is the most often employed. Three kinds of techniques--the emulsification-diffusion method, nanoprecipitation, and the dialysis method are used for nanoparticle formation of the resulting copolymer. Prepared nanoparticles having a size <200 nm and a hydrophilic surface layer can be applied for bone-specific drug delivery.


[Algorithm for the molecular analysis of Bardet-Biedl syndrome in Spain].
Bardet-Biedl syndrome (BBS) is a multisystemic genetic disorder, which is not widespread among the Caucasian population, characterized by a highly variable phenotype and great genetic heterogeneity. BBS belongs to a group of diseases called ciliopathies, caused by defects in the structure and/or function of cilia. Due to the diagnostic complexity of the syndrome, the objective of this study was to analyse our whole group of patients in order to create an algorithm to facilitate the routine molecular diagnosis of BBS. We also calculated several epidemiological parameters in our cohort. We analysed 116 BBS patients belonging to 89 families from the whole Spanish geography. All probands fulfilled diagnosis criteria established for BBS. For this, we used: genotyping microarray, direct sequencing and homozygosis mapping (in consanguineous families). By means of the different approaches, it was possible to diagnose 47% of families (21% by genotyping microarray, 18% by direct sequencing of predominant BBS genes, and 8% by homozygosis mapping). With regard to epidemiological data, a prevalence value of 1:407,000 was obtained for BBS in Spain, and a sex ratio of 1.4:1 (men:women). The proposed algorithm, based on the analysis of predominant BBS genes combined with homozygosis mapping, allowed us to confirm the molecular diagnosis in a significant percentage of families with clinically suspected BBS. This diagnostic algorithm will be useful for the improvement of the efficiency of molecular analysis in BBS.


Lipase and pancreatic amylase versus total amylase as biomarkers of pancreatitis: an analytical investigation.
To evaluate the biomarkers of pancreatitis Colorimetric Lipase, Total Amylase and Pancreatic Amylase (immunoinhibition) assays on the Roche COBAS INTEGRA 700. Pancreatic and Total Amylase assays and Colorimetric Lipase showed excellent imprecision of 1.6 to 2.3% and linearity (slope = 0.94-0.99, y-intercepts-1 to +3 U/L, r = 0.999) over the range of 17 to 900, 35 to 880, and 21 to 150 U/L, respectively. There was an excellent correlation between Pancreatic and Total Amylase: Pancreatic Amylase = 0.99 (+/- 0.02) x Total Amylase-36(+/- 8) (n = 106, r = 0.97, p < 1 x 10(-5), y intercept p < 1 x 10(-5)). Colorimetric Lipase showed some correlation to Total and Pancreatic Amylase results: Colorimetric Lipase = 1.54 (+/- 0.16) x Total Amylase-81(+/- 37) (n = 100, r = 0.70, p < 1 x 10(-6), y intercept p = 0.03), and Colorimetric Lipase = 1.78 (+/- 0.15) x Pancreatic Amylase-50(+/- 29) (n = 99, r = 0.78, p < 1 x 10(-6), y intercept p = 0.09). We recommend running the more specific Pancreatic Amylase as biomarker of pancreatitis on the Roche COBAS INTEGRA.


Electronic cigarette: use and perceptions among French military nurses in 2013.
Paramedical personnel are exposed to tobacco smoking. Electronic cigarettes (e-cigarettes) may be considered as a lower-risk substitute for cigarettes. The aim of the study was to estimate the prevalence of e-cigarette use, the motives for use and the perceptions among French military nurses. A cross-sectional survey, using self-administered questionnaires, was conducted in 2013 among 300 students and instructors of the French school of military paramedical personnel. Prevalences of e-cigarette use among smokers and nonsmokers were compared using logistic regressions adjusted on age and gender. The prevalence of smoking was 40% among the 200 responders. E-cigarette current use prevalence was 25% (6% daily users), without significant difference according to gender and age. Tobacco smokers reported significantly more e-cigarette current use (51% vs7%). Motives for e-cigarette use reported by smokers were curiosity (48%), intention to reduce tobacco consumption (43%) or to quit smoking (8%). Among users of both tobacco and e-cigarettes, 48% reported a significant decrease in tobacco consumption following e-cigarette initiation (average decrease of 5-10 cigarettes smoked per day; p <0.001). Both tobacco smokers and nonsmokers (88%) estimated that e-cigarette use was potentially harmful for health, but it was perceived as less harmful than tobacco by 46%. E-cigarette use among military nurses follows the trends observed in the general population in terms of prevalence and motives. E-cigarettes, which are seen as an attractive alternative to cigarettes, may contribute to a reduction in tobacco use among healthcare workers.


Relationship of perceived parenting practices to anger regulation and coping strategies in African-American adolescents.
This study examined the relationship of perceived parenting practices to anger and coping in a sample of 70 African-American adolescents. Regression analyses reveled that perceptions of parental strictness were negatively related to anger temperament, anger-out, and avoidance (all p<0.05), and was positively associated with engaging in demanding activities (p<0.02). Additionally, perceptions of parental involvement were positively related to seeking diversions (p<0.04), and perceptions of parental autonomy granting were negatively related to anger-out (p<0.02). These findings highlight the potential importance of the perceived parent-adolescent relationship for African-American adolescents.


Effect of different postharvest drying temperatures on Aspergillus flavus survival and aflatoxin content in five maize hybrids.
After harvest, maize is dried artificially to halt fungal growth and mycotoxin production while in postharvest storage. The process often limits harvest capacity and has been a frequent cause of seed injury. Higher drying temperatures could lead to shorter drying periods and faster turnover; however, there is often a deterioration of the physical grain quality, including increased breakage susceptibility and loss of viability. The goals of this study were to determine the effect of different postharvest drying temperatures on Aspergillus filavus and Fusarium verticillioides survival and aflatoxin content in maize and to determine the viability of the seed. Five corn hybrids varying in resistance to A. flavus were side needle-inoculated with A. flavus, harvested at physiological maturity, and dried at temperatures ranging from 40 to 70 degrees C. Kernels were evaluated for aflatoxin, stress cracks, germination, and kernel infection by A. flavus and a natural infestation of F. verticillioides. Drying temperature had no effects on aflatoxin concentration given the heat stability of the toxin. With increased temperatures from 40 to 70 degrees C, germination decreased significantly, from 96 to 27%, and stress cracks increased significantly (1.4 up to 18.7). At temperatures above 60 degrees C, F. verticillioides kernel infection was significantly reduced to less than 18%. At 70 degrees C, there was a significant reduction in A. flavus kernel infection, from 11 to 3%. This information is useful in determining a range of temperatures that can be used for drying seed when fungal infection, stress cracks, and seed viability are of interest.


Rapid analysis of avenacosides in grain and husks of oats by UPLC-TQ-MS.
Avenacosides A (AA) and B (AB) as well as 26-desglucoavenacoside A (26dAA) were quantified in oats using rapid and sensitive method utilising UPLC-TQ-MS. In the grain, AA and AB were revealed as the primary saponins, whereas in the husks, dAA was predominant. Inconsistent with the published data, observed concentrations of AA and AB in the grain were very similar. Presumably, this is due to higher sensitivity and better selectivity of the mass spectrometry-based quantification method, thus allowing for more precise measurements. Elevated level of an active fungicidal form of saponin, dAA in the husks possibly indicates they are more prone to fungal attacks.


Wastewater reuse in on-site wastewater treatment: bacteria and virus movement in unsaturated flow through sand filter.
In on-site wastewater treatment plants, effluents are pre-treated by septic tank and treated by soil infiltration or sand filtration systems, with unsaturated flow conditions. These systems remove efficiently carbon, nitrogen and suspended solids. But for microbial pollution, the treatment efficiency depends on the hydrodynamic behaviour and filtering media characteristics. Contamination of superficial water and groundwater due to pathogenic viruses and pathogenic bacteria is responsible for many diseases. The objective of this study is to approach the mechanisms and operating conditions to control bacteria and virus release in the environment. Experiments were carried out on reactors of different length packed with sand. Hydraulic load of 90 cm x d(-1) with a pulse periodic flow was used. The influence of chemical composition of the solution on the treatment efficiency has also been studied. For the first time, the residence time distribution (RTD) has been studied using a conservative tracer (KI), to determine the main hydrodynamic parameters. For the second time, the RTD with bacterial and viral tracers (E. coli, bacteriophage MS2) was applied, with the aim to define microbial behaviour in filtering media, including adsorption and filtration phenomena. This work allowed us to determine retardation factors according to the hydraulic loads and chemical composition.


Influence of clonogenic assay methodology on measurement of drug sensitivities in vitro.
To assess the influence of clonogenic assay methodology on measurement of reproductive cell kill, the in vitro sensitivities of a human bladder cancer cell line, RT112, to methotrexate and adriamycin were determined using ten different procedures. Marked differences in dose-response to methotrexate, but not adriamycin, were observed.


State-dependent alterations in the perception of life events in menstrual-related mood disorders.
The authors examined the reporting of daily life events by women with prospectively confirmed menstrual-related mood disorder (N = 40) and asymptomatic control subjects (N = 20). During the follicular and late luteal phases of the menstrual cycle, subjects completed a schedule of life events that monitors an individual's perception of 1) the frequency of occurrence of life events and 2) the degree of associated distress or pleasure. The patient group reported significantly more negative life events than the control group. Further, the patients with menstrual-related mood disorder showed significantly more distress associated with the same event when it occurred in the premenstrual phase than when it occurred in the post-menstrual phase.


Reduction potential tuning of the blue copper center in Pseudomonas aeruginosa azurin by the axial methionine as probed by unnatural amino acids.
The conserved axial ligand methionine 121 from Pseudomonas aeruginosa azurin (Az) has been replaced by isostructural unnatural amino acid analogues, oxomethionine (OxM), difluoromethionine (DFM), trifluoromethionine (TFM), selenomethionine (SeM), and norleucine (Nle) using expressed protein ligation. The replacements resulted in < 6 nm shifts in the S(Cys)-Cu charge transfer (CT) band in the electronic absorption spectra and < 8 gauss changes in the copper hyperfine coupling constants (AII) in the X-band electron paramagnetic resonance spectra, suggesting that isostructural replacement of Met resulted in minimal structural perturbation of the copper center. The slight blue shifts of the CT band follow the trend of stronger electronegativity of the ligands. This trend is supported by 19F NMR studies of the fluorinated methionine analogues. However, the order of AII differs, suggesting additional factors influencing AII. In contrast to the small changes in the UV-vis and EPR spectra, a large variation of > 227 mV in reduction potential was observed for the series of variants reported here. Additionally, a linear correlation was established between the reduction potentials and hydrophobicity of the variants. Extension of this analysis to other type 1 copper-containing proteins reveals a linear correlation between change in hydrophobicity and change in reduction potential, independent of the protein scaffold, experimental conditions, measurement techniques, and steric modifications. This analysis has also revealed for the first time high and low potential states for type 1 centers, and the difference may be attributable to destabilization of the protein fold by disruption of hydrophobic or hydrogen bonding interactions that stabilize the type 1 center.


[Paul-Bunnell negative glandular fever (author's transl)].
Cytomegalovirus infections may occur either as congenital and neonatal diseases respectively or postnatally in adults in connection with conditions of antibody or immunologic deficiencies. Reports of postnatally acquired cytomegalovirus infections in apparently healthy adults are scarce. A case of an acquired cytomegalo-virus infection in a 23-year old male patient who never was ill before is reported. Clinical and laboratory features showed a typical course of a Paul-Bunnell negative glandular fever. Diagnosis could be established by detection of cytomegalovirus IgM serum levels by means of immunofluorescence technique.


Effect of melatonin on the peripheral concentrations of LH and progesterone after oestrus, and on conception rate in ewes.
It has previously been shown that administration of the indoleamine melatonin to advance the breeding season of ewes is also associated with an increase in ovulation rate and subsequent litter size. Experiment 1 assessed whether, in ewes receiving melatonin to advance the breeding season, the indoleamine acts directly on the corpus luteum to enhance progesterone secretion or indirectly through increased activity of the hypothalamic pulse generator. Six ewes received 3 mg melatonin orally at 15.00 h daily from 22 March onwards, six were induced to ovulate during mid-anoestrus following withdrawal of a progestagen pessary and injection of exogenous gonadotrophin and six acted as naturally ovulating controls. First overt oestrus occurred between 17 May and 8 July in melatonin-treated ewes, between 21 October and 3 January in control ewes and on 8 July in all induced ewes. On days 2 and 10 after the first overt oestrus, melatonin-treated ewes had pulsatile LH activity characteristic of that measured in control ewes ovulating naturally during the breeding season. There was an absence of any pulsatile LH activity in the induced ewes. Progesterone concentrations between days 7 and 12 following oestrus were significantly higher in melatonin-treated than in control and induced ewes, suggesting a luteotrophic role for melatonin. Experiment 2 was carried out to determine whether administration of melatonin commencing after induced ovulation and insemination would alter the endocrine status of the ewe and thereby influence the establishment of pregnancy and embryo survival.(ABSTRACT TRUNCATED AT 250 WORDS)


Globus sensation after laser-assisted uvulopalatoplasty.
To determine the cause of globus sensation following laser-assisted uvulopalatoplasty (LAUP). Sixty consecutive LAUP patients were questioned regarding globus sensation. Palate sensation on globus patients was mapped and compared with asymptomatic LAUP control patients. Symptomatic patients were tested again 2 months later. There was a 25% incidence of globus with 8% rated severe and 16% rated mild. The globus sensation corresponds to an insensate area of palate (P < .01). Resolution of the symptoms coincides with reemergence of sensation on the palate. Sensory dennervation of the central palate is a likely source of post-LAUP globus sensation. As sensation returns, symptoms abate. Palate surgery should be designed with this in mind. It is possible that undiagnosed globus patients may have areas of anesthetic pharynx.


Recurrence Patterns and Long-term Results After Induction Chemotherapy, Chemoradiotherapy, and Curative Surgery in Patients With Locally Advanced Esophageal Cancer.
The long-term follow up data of 2 prospective phase II trials is reported (NCT00072033, NCT00445861), which investigated neoadjuvant chemoradiation followed by surgery in patients with esophageal carcinoma. Postoperative complications as well as prognostic factors and patterns of relapse during long-term observation are shown. Long-term follow-up is often missing in the complex setting of multimodal treatments of esophageal carcinoma; this leads to rather undifferentiated follow-up guidelines for this tumor entity. In the first trial, patients received induction chemotherapy followed by chemoradiation and surgery. In the second trial, cetuximab was added to the same neoadjuvant treatment concomitant with induction chemotherapy and chemoradiation. Eighty-two patients underwent surgery; the median follow-up time was 6.8 and 6.4 years, respectively. Fifty-five percent were diagnosed with adenocarcinoma, 80% clinically node-positive, 68% received transthoracic esophagectomy, and 32% transhiatal or transmediastinal resection. Five patients died postoperatively in-hospital due to complications (6%). The median overall survival was 4.3 years, and the median event-free survival was 2.7 years. Patients with adenocarcinoma rarely relapsed after a 3-year event-free survival. Whereas patients with residual tumor cells after neoadjuvant therapy primarily experienced relapse within the first 2 postoperative years, this in contrast to several patients with complete remission who also experienced late relapses 4 years after surgery. After curative surgery in a multimodal setting, the histological type and the response to neoadjuvant therapy predicted the time frame of relapse; this knowledge may influence further follow-up guidelines for esophageal carcinoma.


[Investigating patients with differentiated thyroid carcinoma and elevated serum thyroglobulin but negative whole-body scan].
Findings of elevated thyroglobulin (Tg) and a negative whole-body scan (WBS) are not uncommon during the follow-up of differentiated thyroid carcinoma. In 12% of our patients submitted to thyroidectomy and radioiodine with Tg >10 ng/ml during hypothyroidism had a negative diagnostic WBS. This finding generally corresponds to a false-negative WBS. Inadequate preparation in terms of iodine exposure and insufficient elevation of TSH should be excluded. Micrometastases which do not accumulate sufficient iodine to be detected by low radioiodine activity and the loss of the capacity to express the sodium/iodine symporter explain many cases. In patients with elevated Tg, metastases can be identified after the administration of a therapeutic radioiodine dose, with this procedure being indicated in cases with Tg >10 ng/ml during hypothyroidism or >5 ng/ml after recombinant TSH, after exclusion of lung and cervical macrometastases. In the present study, 5 of 7 patients with these criteria showed ectopic uptake on post-therapy WBS. If the post-therapy scan is negative or reveals discrete uptake in the thyroid bed, other methods (e.g. FDG PET) can be performed, and the physician should not insist on radioiodine therapy. If WBS detect lymph node metastases, surgery is indicated, while in cases of diffuse lung metastases radioiodine is indicated until the occurrence of a negative WBS or normalization of stimulated Tg levels. Patients with a positive post-therapy scan may show a significant reduction in Tg, with even complete remission in some cases after radioiodine, but the impact of this treatment on mortality remains controversial.


Insulin-dependent adipogenesis in stromal ST2 cells derived from murine bone marrow.
ST2 cells, a cloned stromal-cell line from mouse bone marrow have the phenotypes of osteoblasts and hematopoietic supporting cells, but little is known about the adipogenesis in this cell line. We found that treatment of ST2 cells with a cocktail, a combination of insulin, dexamethasone (DEX) and 3-isobutyl-1-methylxanthine (IBMX), induced adipocyte differentiation. The cells were adipocytic, as seen by the accumulation of triglycerides and with lipid droplets stained with Oil Red O. Expressions of the adipogenic transcriptional factors peroxisome-proliferator-activated receptor gamma and CCAAT-enhancer binding protein alpha were stimulated, triggering the expression of the late adipocyte-specific marker alpha-glycerophosphate-3-dehydrogenase. Unlike another bone marrow stromal cell line PA6, ST2 cells responded to the adipogenic effects of insulin, as do the extramedullary preadipocytes 3T3-L1. Like PA6 and 3T3-L1 cells, adipogenesis in ST2 cells was inhibited by 1,25-dihydroxyvitamin D3, retinoic acid, tumour necrosis factor alpha, and transforming growth factor beta. Therefore, ST2 cells can differentiate into osteoblasts, adipocytes, and hematopoietic supporting cells, in which the process of adipogenesis differs from that of bone marrow preadipocytes PA6, and resembles that of the extramedullary preadipocytes 3T3-L1.


Direct evidence for the cell surface location of a protease-inhibitor complex on intact leukaemia cells.
The interaction of a protease with two fluorescent inhibitors has been studied using intact fixed leukaemia cells as the source of the membrane bound enzyme. Fresh rat leukaemia cells were disrupted and the cytosol collected; this extract was known to contain a protein inhibitor of guanidinobenzoatase (GB) associated with leukaemia cells. All the cytosolic proteins were derivatised with Texas red acid chloride. Leukaemia cells with latent GB failed to bind the Texas red inhibitor protein but did so after activation of GB. Competition experiments with 9-amino acridine (a fluorescent marker for the active site of GB) demonstrated that the Texas red-inhibitor protein could only bind to intact leukaemia cells when the active centre of GB was not already occupied by 9-amino acridine. This competition between these two fluorescent inhibitors demonstrated their specificity for GB. The use of intact leukaemia cells and the high molecular weight of the inhibitor protein precludes the possibility of any interaction between GB and inhibitor within the cells. It is concluded that GB and the GB-inhibitor complex of latent GB are located on the external surface of intact leukaemia cells.


Lower sensitivity of nasal polyp fibroblasts to glucocorticoid anti-proliferative effects.
Treatment with glucocorticoids (GCs) is the cornerstone of nasal polyp (NP) therapy, but some patients respond poorly to them. Fibroblasts are involved in both inflammation and remodelling of NP. We aimed to evaluate whether NP fibroblasts are less sensitive to GCs' anti-proliferative and anti-inflammatory effects, compared to nasal mucosa (NM) fibroblasts. Fibroblasts were obtained from NP (n = 8) from asthmatic patients undergoing endoscopic surgery and NM (n = 8) from patients undergoing nasal corrective surgery. Fibroblasts were stimulated with DMEM at 0.5% or 5% FBS, or TGF-β (5 ng/ml), with or without dexamethasone (10(-11) to 10(-5)M) for different times. Cell proliferation, collagen mRNA expression and IL-6 and IL-8 release were measured. After 3-days, dexamethasone dose-dependently inhibited proliferation of NM (p < 0.001) but not that of NP fibroblasts. Dexamethasone (10(-6)M) reduced by 25% the proliferation of NM fibroblasts. Dexamethasone also inhibited proliferation of NM (p < 0.01) but not that of NP fibroblasts at 5-days. TGF-β induced collagen-1α1, -1α2, and -3α1 mRNA levels in both NM and NP fibroblasts (p < 0.05), and dexamethasone did not alter TGF-β-induced collagen mRNA levels in either fibroblast type at 24 h. Dexamethasone dose-dependently decreased (p < 0.05) FBS-induced IL-6 and IL-8 release in both NM and NP fibroblasts at 4 h, although at 10(-8)M, dexamethasone inhibited cytokine production in NM (p < 0.05) but not in NP fibroblasts. This impaired sensitivity of nasal polyp fibroblasts to in vitro glucocorticoid effects concurs in part with the poor clinical response that these nasal polyp patients show to glucocorticoid treatment.


Cell proliferation after flap surgery, root conditioning and fibronectin application.
This study evaluated the effects of citric acid demineralization and autologous fibronectin application on cell proliferation after mucoperiosteal flap surgery. Three adult rhesus monkeys were used. After flaps were raised, the roots were surgically exposed and planed. Surfaces on the experimental sides were decalcified with citric acid, and after thorough rinsing, the inner aspect of the flaps and the roots were bathed with 1 ml of autologous plasma fibronectin in normal saline (400 micrograms m/ml) and the flaps sutured. Contralateral teeth, acting as controls, were treated only with the surgical procedure. One hour prior to sacrifice, the animals were injected with an intravenous injection of tritiated thymidine (1 microCi/gm body weight). Surgeries were staggered to produce the following time periods: 3, 7, 15, 21 and 28 days. After processing, autoradiographs were obtained for evaluation, and labeled cells were counted in five compartments at 400 x: (1) oral epithelium, (2) crevicular area, (3) supracrestal connective tissue, (4) coronal periodontal membrane and (5) coronal bone marrow. Forty tissue sections per procedure (20 slides per tooth) were counted and means obtained for the three monkeys. Differences between experimental and control values were statistically evaluated for each component, at each time interval, using pairwise t tests. Fibronectin-treated areas showed significantly increased cellular proliferation (P less than 0.01) during the first 2 weeks, affecting mainly all the supracrestal tissues. Histologically, the establishment of a well-organized fibrinous clot at 3 days was noted in these areas. Results show a faster healing after surgery with the use of citric acid and fibronectin. It was concluded that citric acid followed by fibronectin enhanced cellular proliferation.


Recent advances in the research and development of marine antimicrobial peptides.
Antimicrobial peptides are a group of natural or semi-synthetic molecules possessing antimicrobial activities against bacteria, fungi, viruses, parasites, etc. They are considered as promising candidates for treatment of microbial infections and suppression of microbial resistance. The increasing emergence of bacterial resistance has required development of new efficient antibiotics that can be added to the antibacterial armamentarium. The marine world provides a rich source of antimicrobial peptides. That world has not been highly explored yet, and much more effort is required in order to discover new efficient antimicrobial peptides. In the present article, we have reviewed the recent progress in the field of marine antimicrobial peptides from 2009 until the mid of 2012.


Hormone therapy and age-related macular degeneration: the Women's Health Initiative Sight Exam Study.
To determine the effectiveness of treatment with conjugated equine estrogens (CEE) or with CEE combined with progestin (CEE + P) on age-related macular degeneration (AMD). In an ancillary study to the Women's Health Initiative clinical trial of hormone therapy, 4262 women 65 years and older underwent fundus photography for the determination of AMD. Participants were recruited from April 2000 to June 2002 at 21 clinical sites an average of 5 years after randomization. Participants were randomized to treatment with CEE, CEE + P, or placebo. Participants had been treated for an average of 5 years at the ophthalmic evaluation for AMD. The overall prevalence of any AMD was 21.0%. No association was found between CEE + P (odds ratio [OR], 0.91; 95% confidence interval [CI], 0.75-1.11) or CEE alone (OR, 0.98; 95% CI, 0.78-1.25) and early-stage AMD. The CEE + P was associated with a reduced risk of soft drusen (OR, 0.83; 95% CI, 0.68-1.00) after adjustment for covariates and with a reduced risk of neovascular AMD (OR, 0.29; 95% CI, 0.09-0.92). Treatment with CEE alone or CEE + P does not affect early- or late-stage AMD. Treatment with CEE + P may reduce the risk of soft drusen or neovascular AMD.


Development of cell lines stably expressing human immunodeficiency virus type 1 proteins for studies in encapsidation and gene transfer.
Experiments were done to test cell lines for their capacity to express human immunodeficiency virus type 1 (HIV-1) proteins in a stable manner. Marked differences were seen in the ability to stably express and export viral Gag and Pol proteins. Two cell lines, one suspension (MDS) and one monolayer (SW480), were established which exported these proteins at high level. Two other cell lines, HeLa and THP-1, showed poorer expression and very limited particle release. Single cell cloning was used to select the optimal producing clones from the lines. These produced large quantities of viral core particles pelletable from the supernatants. Cell lines were constructed from these clones which stably expressed in addition either the HIV-1 Envelope or a packageable HIV-based vector. The vector was shown to be packaged within the viral core particles. Transient transfection of envelope expressing constructs into a gag-pol plus vector cell line, or the vector into a gag-pol plus envelope expressing cell line resulted in gene transfer to CD4+ target cells. These cell lines provide useful tools with which to study the assembly and export of viral proteins and RNA, for assay of alternative envelope proteins to pseudotype HIV cores, for assessment of antiviral drugs and as a source of correctly processed proteins for immunological studies.


Current status of cardiovascular surgery in Japan 2013 and 2014: A report based on the Japan Cardiovascular Surgery Database. 2: Congenital heart surgery.
We analyzed the mortality and morbidity of congenital heart surgery in Japan using the Japan Cardiovascular Surgery Database (JCVSD). Data regarding congenital heart surgery performed between January 2013 and December 2014 were obtained from JCVSD. The 20 most frequent procedures were selected and the mortality rates and major morbidities were analyzed. The mortality rates of atrial septal defect repair and ventricular septal defect repair were less than 1%, and the mortality rates of tetralogy of Fallot repair, complete atrioventricular septal defect repair, bidirectional Glenn, and total cavopulmonary connection were less than 2%. The mortality rates of the Norwood procedure and total anomalous pulmonary venous connection repair were more than 10%. The rates of unplanned reoperation, pacemaker implantation, chylothorax, deep sternal infection, phrenic nerve injury, and neurological deficit were shown for each procedure. Using JCVSD, the national data for congenital heart surgery, including postoperative complications, were analyzed. Further improvements of the database and feedback for clinical practice are required.


[Typical skeletal changes in metastatic neuroblastoma].
Compared with other solid tumours in childhood, neuroblastomas show a marked tendency to metastasise to the skeleton. The differentiation of these lesions from inflammatory and other malignant bone lesions in this age group is often difficult. The radiological findings in ten patients with metastasising and histologically confirmed neuroblastomas have been reviewed and the typical appearances in the skeleton are described. The most important features in the differential diagnosis are discussed and the significance of bone changes in the diagnosis of neuroblastoma have been evaluated.


Early pottery at 20,000 years ago in Xianrendong Cave, China.
The invention of pottery introduced fundamental shifts in human subsistence practices and sociosymbolic behaviors. Here, we describe the dating of the early pottery from Xianrendong Cave, Jiangxi Province, China, and the micromorphology of the stratigraphic contexts of the pottery sherds and radiocarbon samples. The radiocarbon ages of the archaeological contexts of the earliest sherds are 20,000 to 19,000 calendar years before the present, 2000 to 3000 years older than other pottery found in East Asia and elsewhere. The occupations in the cave demonstrate that pottery was produced by mobile foragers who hunted and gathered during the Late Glacial Maximum. These vessels may have served as cooking devices. The early date shows that pottery was first made and used 10 millennia or more before the emergence of agriculture.


Solid phase synthesis without repetitive acidolysis. Preparation of leucyl-alanyl-glycyl-valine using 9-fluorenylmethyloxycarbonylamino acids.
The utility of repetitive nonhydrolytic base cleavage of alpha-amino protective groups in solid phase peptide synthesis is shown by a preparation of the model tetrapeptide leucyl-alanyl-glycyl-valine on a p-benzyloxybenzyl ester polystyrene--1% divinylbenzene resin support. Nalpha-9-Fluorenylmethyloxycarbonyl (Fmoc: Carpino & Han, 1970, 1972) amino acids were coupled by the symmetrical anhydride procedure, followed by Fmoc group cleavage using 50% piperidine in methylene chloride. Quantitative removal of the Fmoc-tetrapeptide from the solid support was effected by treatment with 55% trifluoroacetic acid in methylene chloride. Homogeneous free tetrapeptide was obtained in 87% overall yield. The procedure is proposed to offer advantages over present solid phase methods which use acidolysis for repetitive alpha-amino group deblocking.