Control of bleeding by silk ligation and diathermy coagulation during tonsillectomy: A comparison of efficacy of the two techniques in the first 24 hours after surgery.
To assess and compare the relative efficacy of silk ligation and diathermy coagulation techniques in controlling bleeding during tonsillectomy in the first 24 hours. This prospective study was conducted at the Department of ENT, Khalifa Gul Nawaz Teaching Hospital, Bannu and this department related consultants' private clinics from January 1, 2012 to December 31, 2014. The study included 180 cases. All patients included were having history of recurrent, acute tonsillitis, with more than 6-7 episodes in one year, five episodes per year for two years, or three episodes per year for three years. All the surgeries were performed by dissection method. Haemostasis during the procedure was secured by either ligation with silk 1 or using diathermy. The results were analyzed using SPSS 16.0 for windows. A total of180 cases were included in the study. The ages of the patients ranged from 5 to 40 years with the mean age of 15.56 years and a std.deviation of +/- 8.24. The male to female ratio was 1.25:1. The number of hemorrhages occurring was greater in the 'diathermy coagulation' group as compared to the 'silk ligation' group. However, the observed difference was statistically insignificant (p >.05). Primary haemorrhage occurring during tonsillectomy is a serious threat and control of bleeding during the procedure should therefore be meticulous. Both suture ligation and coagulation diathermy for control of bleeders during the procedure by dissection method are equally effective.


Derepression of prophage P2 by satellite phage P4: cloning of the P4 epsilon gene and identification of its product.
Escherichia coli phage P4 lacks all of the genetic information necessary for capsid, tail, and lysis functions. P4 is therefore dependent on a helper phage, such as P2, for lytic propagation. During P4 superinfection of a P2 lysogen, the P2 prophage is derepressed by the action of the P4-encoded epsilon gene. We have cloned the epsilon gene and identified the 10-kDa E protein. The epsilon gene product is the only P4 protein required to derepress prophage P2, which leads to in situ P2 DNA replication. A two-plasmid derepression assay system has been developed to examine the derepression activity of E. The reporter plasmid contains the two face-to-face promoters, Pe and Pc, involved in the lysis-lysogeny transcriptional switch of phage P2 and the immunity repressor C. The Pe promoter is coupled to a cat reporter gene. In the construct, the C repressor is transcribed from the Pc promoter and represses the Pe promoter, which mimics the in situ-repressed P2 prophage. The E protein is supplied in trans from a compatible plasmid in which the epsilon gene is under the control of the T7 promoter. We show here that in the two-plasmid assay system, induction of the E protein derepresses the Pe promoter. The ash9 mutation, which is located upstream of the epsilon gene, enhances the E-mediated derepression of the Pe promoter. The purified E protein shows no specific DNA binding activity, and the implications of this are discussed.


Characterization of Arabidopsis secretory phospholipase A2-gamma cDNA and its enzymatic properties.
Plant secretory phospholipases A(2) (sPLA(2)s) probably play important roles in phospholipid signaling based on the data reported from other organisms, but their functions are poorly understood because of the lack of cloned sPLA(2) genes. In this study, we cloned and characterized an Arabidopsis secretory phospholipase A(2)-gamma (AtsPLA(2)-gamma) cDNA, and examined its enzymatic properties. The recombinant protein of AtsPLA(2)-gamma showed maximal enzyme activity at pH 8.0, and required Ca(2+) for activity. Moreover, AtsPLA(2)-gamma showed sn-2 position specificity but no prominent acyl preference, though it showed head group specificity to phosphatidylethanolamine rather than to phosphatidylcholine. AtsPLA(2)-gamma was found to predominate in the mature flower rather than in other tissues, and subcellular localization analysis confirmed that AtsPLA(2)-gamma is secreted into the intercellular space.


Acquired ABC-transporter overexpression in cancer cells: transcriptional induction or Darwinian selection?
Acquired multidrug resistance (MDR) in tumor diseases has repeatedly been associated with overexpression of ATP-binding cassette transporters (ABC-transporters) such as P-glycoprotein. Both in vitro and in vivo data suggest that these efflux transporters can cause MDR, albeit its actual relevance for clinical chemotherapy unresponsiveness remains uncertain. The overexpression can experimentally be achieved by exposure of tumor cells to cytotoxic drugs. For simplification, the drug-mediated transporter overexpression can be attributed to two opposite mechanisms: First, increased transcription of ABC-transporter genes mediated by nuclear receptors sensing the respective compound. Second, Darwinian selection of sub-clones intrinsically overexpressing drug transporters being capable of extruding the respective drug. To date, there is no definite data indicating which mechanism truly applies or whether there are circumstances promoting either mode of action. This review summarizes experimental evidence for both theories, suggests an algorithm discriminating between these two modes, and finally points out future experimental approaches of research to answer this basic question in cancer pharmacology.


[Enzyme activities and structural features of mitochondria and expression of mRNA of complex I].
The functional ability of the muscle is closely related to the activities of mitochondria, which are the energy producing organelles in the muscle cell. Complex I, III and IV exist as respiratory chain enzymes in the inner membrane of mitochondria and are responsible for transporting electrons from NADH to O2. In this paper, rat masseter muscle, rat and frog cerebral cortex, rat tongue, and frog tibialis anterior muscle mitochondria were described in terms of their correlation with enzyme activity of mitochondrial respiratory chain, the structure and number of mitochondria, and transcription levels of ND1 (mitochondrial) and 51K (nuclear) subunits of Complex I, especially regarding the change in feeding behavior in rat and hibernation in frog. The cristae in mitochondria gradually developed during the postnatal period. The changes in feeding behavior were closely correlated with the level of mitochondrial enzyme activity and were also supported by the structural changes in mitochondria. Based on results of this study, the expression of ND1 and 51K is thought to be involved in the rate-limiting step in the enzyme production. The results also suggest that there is a correlation between biochemical properties and structural features in mitochondria during development, effects of a soft diet, and hibernation.


The design of automated analysis machines.
Since the introduction of the first automatic chemical analysis machine 25 years ago, development has progressed rapidly and a wide range of machines are now available. The original machines were constrained by the available analytical techniques, but later developments in methodology have permitted much more latitude. There are four main groups of machines and these differ markedly in operating principles. In addition there are many unclassifiable devices which have been developed for special purposes. The principles underlying the design of the machines are discussed, and as far as possible an account is given of the steps in development which led to their modern form. The principles of fluid flow, which govern the operation of many of these machines, are considered in some detail. Proposals are made regarding the areas in which new development is possible and desirable in the hope of encouraging further work in this field.


FoxO1 negatively regulates leptin-induced POMC transcription through its direct interaction with STAT3.
FoxO1, which is up-regulated during early stages of diet-induced leptin resistance, directly interacts with STAT3 and prevents STAT3 from binding to specificity protein 1 (SP1)-pro-opiomelanocortin (POMC) promoter complex, and thereby inhibits STAT3-mediated regulation of POMC transcription. FoxO1 also binds directly to the POMC promoter and negatively regulates its transcription. The present study aims to understand the relative contribution of the two interactions in regulating POMC expression. We studied the structural requirement of FoxO1 for its interaction with STAT3 and POMC promoter, and tested the inhibitory action of FoxO1 mutants by using biochemical assays, molecular biology and computer modelling. FoxO1 mutant with deletion of residues Ala137-Leu160 failed to bind to STAT3 or inhibit STAT3-mediated POMC activation, although its binding to the POMC promoter was unaffected. Further analysis mapped Gly140-Leu160 to be critical for STAT3 binding. The identified region Gly140-Leu160 was conserved among mammalian FoxO1 proteins, and showed a high degree of sequence identity with FoxO3, but not FoxO4. Consistently, FoxO3 could interact with STAT3 and inhibit POMC promoter activity, whereas FoxO4 could not bind to STAT3 or affect POMC promoter activity. We further identified that five residues (Gln145, Arg147, Lys148, Arg153 and Arg154) in FoxO1 were necessary in FoxO1-STAT3 interaction, and mutation of these residues abolished its interaction with STAT3 and inhibition of POMC promoter activity. Finally, a FoxO1-STAT3 interaction interface model generated by computational docking simulations confirmed that the identified residues of FoxO1 were in close proximity to STAT3. These results show that FoxO1 inhibits STAT3-mediated leptin signalling through direct interaction with STAT3.


Integration of NIRS and PCA techniques for the process monitoring of a sewage sludge anaerobic digester.
This study investigates the use of Hotelling's T(2) control charts as the basis of a process monitor for sewage sludge anaerobic digestion. Fourier transform near infrared spectroscopy was used to produce partial least squares regression models of volatile fatty acids, bicarbonate alkalinity and volatile solids. These were utilised in a series of principle component analysis models along with spectral data from digestate and feedstock samples to produce a pseudo steady state model, which was then used with an independent test set to evaluate the system. The system was able to identify disturbances to the digester due to a temporary alteration of the type of feedstock to the digester and separately, halving of the hydraulic retention time of the digester. It could also provide advance warning of disturbances to the digester. This technique could be used to improve the performance of sewage sludge anaerobic digesters by enabling optimisation of the process.


Initial Characterization and Optimization of the Liquid Sampling-Atmospheric Pressure Glow Discharge Ionization Source Coupled to an Orbitrap Mass Spectrometer for the Determination of Plutonium.
Plutonium measurements are essential to the nuclear forensics and safeguards community. The liquid sampling-atmospheric pressure glow discharge (LS-APGD) microplasma ionization source coupled with an Orbitrap mass spectrometer is a proven platform for uranium isotope ratio determinations. This work expands the LS-APGD-Orbitrap platform capabilities by reporting the first-ever analysis of plutonium with the LS-APGD and the first-ever measurement of elemental plutonium with an Orbitrap mass spectrometer. This coupling has the potential to dramatically reduce the complex sample manipulations required for traditional analysis techniques employed for actinide isotope ratio determinations. As a first step toward the goal of simultaneous uranium and plutonium isotope ratio determinations, the initial characterization and optimization of the platform for the detection of plutonium are reported. Collision-induced dissociation modality settings were optimized to reduce water-related and other molecular clusters containing plutonium, maximizing 242Pu16O2+ responses. A design of experiments study was conducted to optimize the discharge conditions of the dual-electrode LS-APGD toward the responsivity of 242Pu16O2+. The measurement sensitivity was determined from a Pu response curve, yielding a limit of detection of 10 fg (absolute) of total analyte when data was collected and processed with a Spectroswiss FTMS Booster X2 data acquisition system. Additionally, plutonium and uranium were measured in a simultaneous acquisition, and each analyte remained unaffected by the other. It is believed that the LS-APGD-Orbitrap platform could be a valuable addition to the nuclear forensics' toolbox and, indeed, other scientific disciplines and regulatory communities in which rapid, high-resolution plutonium determinations are paramount.


2D-IR spectroscopy for oil paint conservation: Elucidating the water-sensitive structure of zinc carboxylate clusters in ionomers.
The molecular structure around metal ions in polymer materials has puzzled researchers for decades. This question has acquired new relevance with the discovery that aged oil paint binders can adopt an ionomer structure when metal ions leached from pigments bind to carboxylate groups on the polymerized oil network. The characteristics of the metal-polymer structure are expected to have important consequences for the rate of oil paint degradation reactions such as metal soap formation and oil hydrolysis. Here, we use two-dimensional infrared (2D-IR) spectroscopy to demonstrate that zinc carboxylates formed in paint films containing zinc white pigment adopt either a coordination chain- or an oxo-type cluster structure. Moreover, it was found that the presence of water governs the relative concentration of these two types of zinc carboxylate coordination. The results pave the way for a molecular approach to paintings conservation and the application of 2D-IR spectroscopy to the study of polymer structure.


Predicting deterioration in patients with lobar haemorrhages.
To study the clinical course and determine predictors of deterioration in patients with lobar haemorrhages). A comprehensive review of 61 consecutive patients with lobar haemorrhages was performed. Neurological deterioration was defined as (1) decrease in Glasgow coma sum score by 2 points, (2) new neurological deficit, or (3) clinical signs of brain herniation. A univariate logistic regression was performed and expressed in odds ratios. Sixteen of 61 (26%) patients with lobar haemorrhages deteriorated after admission. In a univariate analysis, only a Glasgow coma score <14 predicted deterioration (75% of deteriorators v 24% who did not deteriorate; p<0.0001). Initial CT characteristics predictive of deterioration included haemorrhage volume >60 ml (63% v 16%, p< 0.0001), shift of the septum pellucidum (75% v 36%, p<0.01), effacement of the contralateral ambient cistern (33% v 0%, p<0.0001), and widening of the contralateral temporal horn (50% v 0%, p<0.0001). Patients presenting and deteriorating within 12 hours of ictus declined due to enlargement of the haemorrhage. Those who deteriorated more than 12 hours after initial neurological symptoms, showed increased mass effect secondary to oedema. CONCLUSION-Patients with lobar haemorrhages presenting immediately after ictus are at risk for deterioration from enlargement of the haemorrhage and predictors of deterioration may be absent. Patients with large volume lobar haemorrhages presenting to the emergency department with decreased level of consciousness and shift on CT are at risk for further deterioration from worsening oedema. These patients require close observation and early aggressive management may be warranted.


An acute increase in skeletal muscle carnitine content alters fuel metabolism in resting human skeletal muscle.
Carnitine plays an essential role in the integration of fat and carbohydrate oxidation in skeletal muscle, which is impaired in obesity and type 2 diabetes. The aim of the present study was to investigate the effect of an increase in skeletal muscle total carnitine (TC) content on muscle fuel metabolism. A 5-h iv infusion of saline (control) or l-carnitine was administered while serum insulin was maintained at a physiologically high concentration during two randomized visits. Seven healthy, nonvegetarian young men (body mass index, 26.1 +/- 1.6 kg/m2) participated in the present study at the University of Nottingham. Skeletal muscle pyruvate dehydrogenase complex (PDC) activity and associated muscle metabolites were measured. The combination of hypercarnitinemia (600 micromol/liter) and hyperinsulinemia (160 mU/liter) increased muscle TC content by 15% (P < 0.01) and was associated with decreased pyruvate dehydrogenase complex activity (P < 0.05) and muscle lactate content (P < 0.05) by 30 and 40%, respectively, and an overnight increase in muscle glycogen (P < 0.01) and long-chain acyl-coenzyme A content (P < 0.05) by 30 and 40%, respectively, compared with control. These results suggest that an acute increase in human skeletal muscle TC content results in an inhibition of carbohydrate oxidation in conditions of high carbohydrate availability, possibly due to a carnitine-mediated increase in fat oxidation. These novel findings may have important implications for our understanding of the regulation of muscle fat oxidation, particularly during exercise, when carnitine availability may limit fat oxidation, and in obesity and type 2 diabetes where it is known to be impaired.


Advances in combination therapies based on nanoparticles for efficacious cancer treatment: an analytical report.
The main objective of nanomedicine research is the development of nanoparticles as drug delivery systems or drugs per se to tackle diseases as cancer, which are a leading cause of death with developed nations. Targeted treatments against solid tumors generally lead to dramatic regressions, but, unfortunately, the responses are often short-lived due to resistant cancer cells. In addition, one of the major challenges of combination drug therapy (called "cocktail") is the crucial optimization of different drug parameters. This issue can be solved using combination nanotherapy. Nanoparticles developed in oncology based on combination nanotherapy are either (a) those designed to combat multidrug resistance or (b) those used to circumvent resistance to clinical cancer drugs. This review provides an overview of the different nanoparticles currently used in clinical treatments in oncology. We analyze in detail the development of combinatorial nanoparticles including dendrimers for dual drug delivery via two strategic approaches: (a) use of chemotherapeutics and chemosensitizers to combat multidrug resistance and (b) use of multiple cytotoxic drugs. Finally, in this review, we discuss the challenges, clinical outlook, and perspectives of the nanoparticle-based combination therapy in cancer.


Recommendations for the use of molecular diagnostics in the diagnosis of allergic diseases.
The Study Group on Allergology of the Italian Society of Clinical Pathology and Laboratory Medicine (SIPMeL) and the Associazione Italiana degli Allergologi e Immunologi Territoriali e Ospedalieri (AAIITO) developed the present recommendations on the diagnosis of allergic diseases based on the use of molecular allergenic components, whose purpose is to provide the pathologists and the clinicians with information and algorithms enabling a proper use of this second-level diagnostics. Molecular diagnostics allows definition of the exact sensitization profile of the allergic patient. The methodology followed to develop these recommendations included an initial phase of discussion between all the components to integrate the knowledge derived from scientific evidence, a revision of the recommendations made by Italian and foreign experts, and the subsequent production of this document to be disseminated to all those who deal with allergy diagnostics.


Rapid degradation of poly(ADP-ribose) after injection into the mouse bloodstream.
Extensive DNA damage leads to the activation of poly(ADP-ribose) polymerase and subsequently to the formation of poly(ADP-ribose). When the damage is severe or leads to cell death, poly(ADP-ribose) may leak into the blood circulation. The metabolism of poly(ADP-ribose) in the bloodstream is not well understood. Thus, in the present study, the metabolism of P-labeled poly(ADP-ribose) was followed in mice after injection of this labeled compound into the tail vein. The results showed that 5 min after injection more than half of the radioactivity was concentrated in acid-soluble fractions, namely in low molecular weight compounds in the blood, liver, and kidneys. Most of this radioactivity was in the form of inorganic phosphate, detected 5 min post-injection in the blood, kidneys, and urine. By contrast, the metabolites ADP-ribose and phosphoribosyl-AMP were not detected in any of the tissues nor in blood or urine. Taken together, these findings suggest that once poly(ADP-ribose) enters the bloodstream it is rapidly degraded, thereby preventing its accumulation in the blood.


A multicentre comparison of piroxicam and indomethacin in acute soft tissue sports injuries.
One hundred and five patients suffering from sport-associated soft tissue injuries were included in a randomized controlled trial comparing the efficacy and side-effects of piroxicam and indomethacin. Patients were treated for 7 days. Both groups showed a similar degree of improvement in joint tenderness, swelling and pain. Eighty-eight per cent of patients on piroxicam showed a marked or moderate improvement in their injury at the end of the trial and 79% of indomethacin-treated patients. Approximately 50% of patients in both groups resumed full activity within 7 days. One patient, in the indomethacin group, was withdrawn from treatment due to a side-effect.


Ovine alpha-amylase genes: isolation, linkage mapping and association analysis with milk traits.
On the basis of comparisons between cattle and sheep genome mapping information the ovine alpha-amylase gene was examined as a possible genetic marker for milk traits in sheep. The objective of the present study was to isolate, map and determine whether this gene is a candidate gene for milk traits. DNA fragments (832 and 2360 bp) corresponding to two different AMY genes were isolated, and one SNP in intron 3 and one GTG deletion in exon 3 of the 2360 bp DNA fragment were found. The 2360 bp ovine AMY DNA fragment was located on chromosome 1 by linkage mapping using the International Mapping Flock. No association was found between estimated breeding values for milk yield, protein and fat contents and AMY genotypes in a daughter design comprising 13 Manchega families with an average of 29 daughters (12-62) per sire.


Post-traumatic stress disorder in chronic post-traumatic headache patients.
The prevalence of post-traumatic stress disorder (PTSD) was examined in a sample of chronic post-traumatic headache patients. All subjects had been injured in a motor vehicle accident and had developed headache or experienced marked exacerbation of an existing headache condition as a result. Nearly 30% of the sample was diagnosed with PTSD. Depression and suppressed anger were significantly higher in subjects with PTSD compared to the rest of the sample. Those with PTSD were more likely to have a history of headache prior to the accident than subjects without PTSD. Comparisons between the PTSD and non-PTSD groups on other headache and demographic variables were non-significant. The importance of evaluating post-traumatic headache patients for the presence of PTSD is discussed.


Influence of climatic and management factors on Eimeria infections in goats from semi-arid zones.
A survey of Eimeria infections was performed in dairy goats and kids (<6 months old) of six farms from a dry desert area of Gran Canaria Island (Spain). The number of oocysts per gram of faeces (OPG) was determined by a modified McMaster technique over a total of 2,616 individual faecal samples taken from the rectum in monthly intervals. Eimeria oocysts were found in 96.1% of the samples with OPG ranging from 1 x 10(2) to 1.4 x 10(6). Kid goats had significantly (P < 0.001) higher OPG counts (46,496 +/- 5,228) than dairy females (2,225 +/- 287). Eight Eimeria species were identified, with Eimeria ninakohlyakimovae (30.0%), Eimeria arloingi (28.6%) and Eimeria alijevi (20.5%) being the most frequent species followed by Eimeria caprina (9.1%), Eimeria christenseni (4.5%), Eimeria jolchijevi (3.4%), Eimeria caprovina (3.2%) and Eimeria hirci (0.7%). Although significant differences were observed among goat groups and herds, the eight species were present in the six farms in both dairy goats and kids. The intensity of oocysts shedding was related to some factors such as the size of the herd and was further influenced by the prevailing climatic conditions of the area. The highest OPG counts were recorded during the hot season in dairy goats and close to weaning time in kids reared in small farms having no prophylactic treatments against eimeriosis.


Effect of sodium hyaluronate/carboxymethyl cellulose (Guardix-sol) on retear rate and postoperative stiffness in arthroscopic rotator cuff repair patients: A prospective cohort study.
Hyaluronate-based anti-adhesive agents are expected to enhance rotator cuff healing; however, their effect on the incidence and extent of postoperative complications such as stiffness and retears has not been investigated. From July 2012 to February 2013, 80 patients undergoing arthroscopic rotator cuff repair surgery were prospectively enrolled. Forty patients were assigned to the control group, while the other 40 were assigned to the injection group and received a Guardix-sol injection immediately after surgery. Passive range of motion, pain visual analog scale, and functional score were assessed at 8 weeks, 6 months, and 24 months postoperatively. Gliding motion between the deltoid muscle and the greater tuberosity of the proximal humerus was evaluated using ultrasonography at 2 and 8 weeks postoperatively, and tendon integrity was evaluated using magnetic resonance imaging at 6 months postoperatively. We found no significant difference between the groups regarding gliding motion at 2 weeks postoperatively. However, at 8 weeks, the incidence of poor gliding motion was 2.5% and 15% for the injected patients and control group, respectively, which was statistically significant. At 6 months after surgery, the retear rate between the two groups was not statistically significant. We found no statistically significant difference between the two groups regarding retear rate and clinical score throughout the follow-up period. We noted no complications related to the use of Guardix-sol. Patients who received the Guardix-sol injection showed improved gliding motion between the deltoid muscle and the greater tuberosity in the early postoperative period.


Adenosine triphosphate-induced electron transfer in 2-hydroxyglutaryl-CoA dehydratase from Acidaminococcus fermentans.
2-hydroxyglutaryl-CoA dehydratase from Acidaminococcus fermentans catalyzes the chemical difficult elimination of water from (R)-2-hydroxyglutaryl-CoA to glutaconyl-CoA. The enzyme consists of two oxygen-sensitive protein components, the homodimeric activator (A) with one [4Fe-4S]1+/2+ cluster and the heterodimeric dehydratase (D) with one nonreducible [4Fe-4S]2+ cluster and reduced riboflavin 5'-monophosphate (FMNH2). For activation, ATP, Mg2+, and a reduced flavodoxin (16 kDa) purified from A. fermentans are required. The [4Fe-4S](1+/2+) cluster of component A is exposed to the solvent since it is accessible to iron chelators. Upon exchange of the bound ADP by ATP, the chelation rate is 8-fold enhanced, indicating a large conformational change. Oxidized component A exhibits ATPase activity of 6 s(-1), which is completely abolished upon reduction by one electron. UV-visible spectroscopy revealed a spontaneous one-electron transfer from flavodoxin hydroquinone (E(0)' = -430 mV) to oxidized component A, whereby the [4Fe-4S]2+ cluster of component A became reduced. Combined kinetic, EPR, and Mössbauer spectrocopic investigations exhibited an ATP-dependent oxidation of component A by component D. Whereas the [4Fe-4S]2+ cluster of component D remained in the oxidized state, a new EPR signal became visible attributed to a d1-metal species, probably Mo(V). Metal analysis with neutron activation and atomic absorption spectroscopy gave 0.07-0.2 Mo per component D. In summary, the data suggest that in the presence of ATP one electron is transferred from flavodoxin hydroquinone via the [4Fe-4S]1+/2+ cluster of component A to Mo(VI) of component D, which is thereby reduced to Mo(V). The latter may supply the electron necessary for transient charge reversal in the unusual dehydration.


Reversible contraceptive effect of PH-20 immunization in male guinea pigs.
Sperm proteins are currently being studied as antigens on which to base a contraceptive vaccine. Sperm plasma membrane proteins offer the theoretical possibility of immunizing either males or females and achieving a contraceptive effect. In this study, we investigated the sperm plasma membrane protein PH-20 as an antigen for inducing infertility in males. We found that infertility can reproducibly be induced in male guinea pigs immunized with purified PH-20: 100% (29 of 29) of PH-20-immunized males became infertile, whereas all 22 controls were fertile. The males were extremely responsive to PH-20 immunization: infertility could be induced with a single injection of only 5 microg PH-20. Among males that received their initial injection when they were approximately 300 g (body weight), 14 of 15 had regained fertility at about 1 yr after initial injection. Surprisingly, in another group of males that received their first injection when they were approximately 650 g (body weight), only 1 of 5 had regained fertility about 1 yr after initial injection. Anti-PH-20 titers in antisera (2 mo after initial injection) were generally in the range 1.1-4.2 x 10(4) in twice-injected males and the range 1.8-9.4 x 10(3) in once-injected males. Over the next 6-11 mo, twice-injected males' titers decreased > or = 4-fold, whereas once-injected males' titers decreased slightly (1.1 - to 1.8-fold). After 6-11 mo, anti-PH-20 titers were in the range 1.0-4.8 x 10(3), and the precise residual titer did not correlate with fertility/infertility. The results show that immunization of males with PH-20, even at low doses, results in a reproducible, completely effective contraceptive action.


Evaluation of Salmonella enteritidis in molting hens after administration of an experimental chlorate product (for nine days) in the drinking water and feeding an alfalfa molt diet.
The method most commonly used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production is to fast the hens. Unfortunately, increased risk of Salmonella enteritidis (SE) infection may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE infection are needed. Hens over 50 wk of age were divided into 12 groups of 11 hens each and placed in individual laying cages. One week prior to dietary changes, hens were placed on an 8-h light and 16-h dark photoperiod that continued for the 9-d molt. All hens were challenged orally with 10(6) cfu of SE on the fourth day of the molt. Treatments were nonfed hens with distilled water (NFD), nonfed hens with the experimental chlorate product (ECP, which provided 15 mM chlorate ion) water (NFECP), alfalfa diets with distilled water (ALD), and alfalfa diets with ECP water (ALECP). In the NFD hens, 67% (log10 2.74) of the crops and 94% (log10 5.62) of the ceca were colonized, whereas for the NFECP hens significant reductions to 22% (log10 1.05) of the crops and 61% (log10 2.44) of the ceca were observed. In the ALD hens, 61% (log10 2.52) of the crops and 94% (log10 4.06) of the ceca were colonized. In the ALECP hens, highly significant reductions to 11% (log10 1.26) of the crops and 39% (log10 1.12) of the ceca were observed. When compared with the NFD hens, significant reductions in SE invasion of the ovary, liver, and spleen occurred in all other treatments, except the ovary in the ALD hens. The low alfalfa intake is probably a factor in our lowered protection against SE when compared with previous results. For several parameters, these results suggest that ECP or the combination of ECP and alfalfa may be a useful tool to reduce the risk of SE during an induced molt.


Sweetened beverages, snacks and overweight: findings from the Young Lives cohort study in Peru.
To determine the association between consumption of snacks and sweetened beverages and risk of overweight among children. Secondary analysis of the Young Lives cohort study in Peru. Twenty sentinel sites from a total of 1818 districts available in Peru. Children in the younger cohort of the Young Lives study in Peru, specifically those included in the third (2009) and the fourth (2013) rounds. A total of 1813 children were evaluated at baseline; 49·2 % girls and mean age 8·0 (sd 0·3) years. At baseline, 3·3 (95 % CI 2·5, 4·2) % reported daily sweetened beverage consumption, while this proportion was 3·9 (95 % CI 3·1, 4·9) % for snacks. Baseline prevalence of overweight was 22·0 (95 % CI 20·1, 23·9) %. Only 1414 children were followed for 4·0 (sd 0·1) years, with an overweight incidence of 3·6 (95 % CI 3·1, 4·1) per 100 person-years. In multivariable analysis, children who consumed sweetened beverages and snacks daily had an average weight increase of 2·29 (95 % CI 0·62, 3·96) and 2·04 (95 % CI 0·48, 3·60) kg more, respectively, than those who never consumed these products, in approximately 4 years of follow-up. Moreover, there was evidence of an association between daily consumption of sweetened beverages and risk of overweight (relative risk=2·12; 95 % CI 1·05, 4·28). Daily consumption of sweetened beverages and snacks was associated with increased weight gain v. never consuming these products; and in the case of sweetened beverages, with higher risk of developing overweight.


A critical examination of distortion measurement in hearing aids.
This study was designed to examine relationships between measures of total distortion (TD) and individual 2nd and 3rd harmonic distortion in various types of hearing aids. Measures of 2nd harmonic, 3rd harmonic, and TD were determined for six instruments. Analysis of the results revealed that little additional information is obtained from a measure of TD as opposed to individual harmonic measures. Furthermore, in two instances TD measures resulted in erroneously high distortion readings because of ambient and system noise. It is recommended that in less than ideal acoustic environments, measures of 2nd and 3rd harmonic distortion be used in lieu of TD.


The effect of awarding disability benefits on opioid consumption.
Strong empirical evidence points towards a significantly higher prevalence of opioid consumption among people receiving disability benefits (DB) than in the general population of the United States. However, no previous research established a causal relationship between the decision to award DB to applicants and their subsequent opioid use. We aim to contribute towards filling this gap. There are channels through which awarding DB may both increase and depress opioid consumption, and thus, ex ante, the sign of a potential causal relationship is ambiguous. To correct for the treatment endogeneity, since an individual's age at the time of the decision on an application impacts discontinuously at certain age cutoffs the award decision, we employ a fuzzy Regression Discontinuity model with three age cutoffs used for identification. We find that awarding DB increases the likelihood of using opioids by about 27-30 percentage points. This suggests that the positive association between DB receipt and opioid consumption is likely to be causal.


Combined Directional Atherectomy and Drug-Eluting Balloon Angioplasty for Isolated Popliteal Artery Lesions in Patients With Peripheral Artery Disease.
To evaluate the midterm results of combined directional atherectomy (DA) and drug-eluting balloon (DEB) angioplasty for atherosclerotic lesions of the popliteal artery. In a single-arm, prospective study, 21 patients (mean age 63±16 years; 16 men) with isolated popliteal artery lesions were enrolled and underwent treatment with combined DA and DEB angioplasty under filter protection between October 2009 and February 2014. The majority (18, 86%) presented with lifestyle-limiting intermittent claudication and 3 with critical limb ischemia. Fifteen (71%) target sites were de novo lesions; 4 were occlusions. The main outcome was primary patency; secondary outcomes were technical success, secondary patency, and early and midterm morbidity and mortality. The TurboHawk atherectomy device was used in 15 (71%) patients and the SilverHawk peripheral plaque excision system in the remaining 6 patients. The In.Pact Admiral/Pacific DEB was used in the majority of cases (15, 71%). The technical success rate was 90% (n=19). One flow-limiting dissection was treated with bailout stenting. Complications included a perforation of the popliteal artery and 2 puncture site hematomas; there was no distal embolic event. The mean follow-up was 18±12 months. Two restenoses were retreated successfully. Kaplan-Meier estimates of primary patency at 12 and 18 months were 95% and 90%, respectively; the secondary patency was 100%. One (5%) patient died in follow-up. None of the patients had an amputation. In this prospective single-arm study, the combined therapy of DA and DEB angioplasty for popliteal artery lesions showed promising midterm performance. The combination of DA and DEB may, in highly selected patients, overcome the challenges presented by the mobility of the knee joint.


Construction of mini-chemostats for high-throughput strain characterization.
To achieve large-scale, high-throughput experiments for systems biology research of microorganisms, reliable data from robust cultivation systems are needed. Chemostats are such systems, ensuring reproducibility and quality by providing a stable, well-controlled environment for the cells. However, many of the available chemostat systems require large amounts of media and are complex to set up and expensive to purchase and maintain. To address these concerns, we developed a mini-chemostat (MC) system with 16 reactors, each at a working volume of 40 ml. Sensors measure dissolved oxygen in the reactor, while OD600 is measured in the outflow. We further developed a CO2 and pH sensor array that can be plugged into the outflow of the reactors. The system was used to characterize yeast physiology at four metabolically different conditions: limitations of glucose, both aerobic and anaerobic, nitrogen, and ethanol. The physiology of yeast cells grown at the four different conditions in the MC system was compared with the yeast cells grown in a DASGIP 1 L system using RNAseq analysis. The results show that the MC system provides the same environmental conditions as the DASGIP system and that the MC system is reproducible between different runs. The system is built to be easily scalable with more reactors and to include more sensors, if available. Our study shows that a robust, reproducible chemostat system for high-throughput and large-scale experiments can be built at low costs.


Multiple Negative Differential Resistance Device by Using the Ambipolar Behavior of Tunneling Field Effect Transistor with Fast Switching Characteristics.
We propose a novel double-peak negative differential resistance (NDR) characteristic at the conventional single-peak MOS-NDR circuit by employing ambipolar behavior of TFET. The fluctuated voltage transfer curve (VTC) from ambipolar inverter is analyzed with simple model and successfully demonstrated with TFET, as a practical example, on the device simulation. We also verified that the fluctuated VTC generates additional peak and valleys on NDR characteristics by using circuit simulations. Moreover, by adjusting the threshold voltage of conventional MOSFET, ultra-high 1st and 2nd peak-to-valley current ratio (PVCR) over 10(7) is obtained with fully suppressed valley currents. The proposed double-peak NDR circuit expected to apply on faster switching and low power multi-functional applications.


Exponential increases of the brominated flame retardants, polybrominated diphenyl ethers, in the Canadian Arctic from 1981 to 2000.
A suite of 37 polybrominated diphenyl ether (PBDE) congeners and all of the homologue groups from mono- to deca-brominated were determined in ringed seal (Phoca hispida) blubber collected from subsistence hunts in the Canadian Arctic in 1981, 1991, 1996, and 2000. Total PBDE (sum(PBDE)) concentrations have increased exponentially over this period in male ringed seals aged 0-15 years. Penta- and hexa-BDEs are increasing at approximately the same rate (t2 = 4.7 and 4.3 years, respectively) and more rapidly than tetra-BDEs (t2 = 8.6 years) and tri-BDEs (t2 = infinity) in this age/sex grouping. In contrast to declining PBDE concentrations since 1997 in human milk from Sweden, sum(PBDE) concentrations in arctic ringed seals continue to increase exponentially similar to worldwide commercial penta-BDE production. PBDE congener profiles in male ringed seals aged 0-15 years from 1991 to 2000 also differ significantly from other aquatic organisms and semipermeable membrane devices collected from temperate coastal regions of British Columbia. While PBDE concentrations are 50 times lower than those of mono-ortho and non-ortho PCBs, and approxiamately 500 times higher than PCDD/Fs, our data indicate that, at current rates of bioaccumulation, PBDEs will surpass PCBs to become the most prevalent organohalogen compound in Canadian arctic ringed seals by 2050.


[Ventricular tachycardia in bursts. Electrophysiological and therapeutical study].
22 patients with salvoes of ventricular tachycardia were investigated electrophysiologically and pharmacologically to determine the mechanism of the arrhythmia. The appearance of the ventricular complexes was suggestive of a septal origin. The arrhythmia appeared to occur and regress within precise limits of sinus rhythm. 24 hour ECG recording and endocavitary stimulation techniques showed its essentially variable character, but occasionally a fixed coupling interval with the first extrasystole was observed, suggesting the presence of an unprotected parasystolic focus. The repetition of the extrasystolic activity, which could be triggered and terminated at will, is explained by phenomena of delayed after potentials. The results of the systematic investigation of the effects of quinidine-like drugs, beta-blockers and verapamil do not invalidate these hypotheses and show a difference of degree but not of nature between the two populations A and B of patients classified according to the quantitative character of the extrasystolic activity over 24 hours (more or less than 1000/hour). The observation of intermediate forms with simple extrasystoles and other forms with sustained tachycardia extends the significance of these results to many forms of ventricular arrhythmia.


Sample size of thorough QTc clinical trial adjusted for multiple comparisons.
A thorough QT trial is typically designed to test for two sets of hypotheses. The primary set of hypotheses is for demonstrating that the test treatment will not prolong QT interval. The second set of hypotheses is to demonstrate the assay sensitivity of the positive control treatment in the study population. Both analyses require multiple comparisons by testing the treatment difference measured repeatedly at multiple selected time points. Tsong and Zhong (2010) indicated that for prolongation testing, this involves an intersection-union test that leads to the reduction of study power. It requires type II error rate adjustment in order to maintain proper sample size and power of the test. Tsong et al. (2010) indicated also that the assay sensitivity analysis is carried out using a union-intersection test that leads to the inflation of the family-wise type I error rate. Type I error rate adjustment is required to control the family-wise type I error rate. Zhang and Machado (2008) proposed the sample size calculation of test-placebo QT response difference based on simulation with a multivariate normal distribution model. Even though the results are generally used as guidance for sample size determination for balanced arm TQT trials, they are limited in generalization to various advanced and adaptive designs of TQT trials (Zhang, 2011 ; Tsong, 2013). In this article, we propose a power equation based on multivariate normal distribution of TQT trials. Sample sizes of various TQT designs can be obtained through numerical iteration of the equation.


Fluorescence in situ hybridization (FISH) in the diagnosis of bladder and upper tract urothelial carcinoma: the largest single-institution experience to date.
We evaluated the UroVysion (Abbott Molecular, IL, USA) fluorescence in situ hybridization (FISH) assay for the diagnosis of urothelial cancer in patients diagnosed with or suspected to have bladder, upper tract urothelial carcinoma (UTUC), and combined upper and lower tract urothelial carcinoma (BC). A single institution retrospective analysis comparing sensitivity, specificity, positive predictive value, and negative predictive values for FISH and urinary cytology. FISH within 6 months of endoscopic evaluation were obtained from outpatient voided urine samples. Our institutional pathology department confirmed pathologic disease from specimens obtained during endoscopic evaluations for lower tract disease. For upper tract disease, disease was confirmed by retrograde ureteroscopy, biopsies of visual lesions, and site-specific upper tract cytology. A total of 415 patients submitted FISH specimens. Overall, FISH was more sensitive than cytology 54.9% in comparison with cytology 42.2% (p = 0.01), specificity favored cytology 92.9% compared to 73.5% with FISH (p < 0.01). For BC only patients, the same significant finding of increased sensitivity and decreased specificity was identified, but for UTUC alone and combined UTUC and BC, there was no significant difference. Cytology had improved positive predictive value (PPV) over FISH, 76.9% in comparison to 64.6% (p = 0.02). Negative predictive value (NPV) also favored cytology 74.2% versus 64.9% (p = 0.02). When analyzing individual cohorts, cytology had improved PPV for BC alone patients. UTUC showed no difference for PPV and NPV. For both UTUC and BC, NPV was slightly favored for FISH over cytology 93.2% versus 91.2% (p = 0.03). Voided urine FISH testing does offer a higher detection of urothelial carcinoma for BC compared to voided cytology; however, specificity was worse. FISH does not appear to improve detection of urothelial carcinoma in patients with either UTUC only or both BC and UTUC.


Systemic mast cell disease: a review of the literature with special focus on the gastrointestinal manifestations.
Mastocytosis is a chronic disease accompanied by specific infiltration of mast cells into the skin and other tissues. In 90% of cases only the skin is involved. In systemic mastocytosis there is an abnormal proliferation of mast cells in various organs including the gastrointestinal tract and liver. Release of various mediators and infiltrative growth of the mast cells lead to the symptoms of the disease. Therefore, many patients present with abdominal symptoms. The clinical manifestations are reviewed with special focus on the gastrointestinal and hepatic involvement. The prognosis of the disease is variable but most often benign. The treatment is based on H1 and H2-blockers and disodium cromoglycate.


Maternal and placental responses before preterm birth: adaptations to increase fetal thyroid hormone availability?
During pregnancy, maternal thyroid hormone supply is crucial for fetal development. Preterm infants often present with hypothyroxinemia. Preterm birth, either spontaneous or medically indicated, is always the result of a complicated pregnancy. We hypothesized that in preterm birth, the maternal transplacental thyroid hormone supply is influenced by the pregnancy complication and we questioned whether maternal and placental compensatory mechanisms are activated to increase thyroid hormone transfer. Observational case-control study in mother-infant-dyads with complicated pregnancies ending in spontaneous preterm birth (n = 31) or indicated preterm birth due to vascular complications (n = 45) and normal pregnancies (healthy term controls; n = 41). At delivery, maternal and cord blood and placenta samples were collected. Cord and maternal plasma concentrations of thyroid stimulating hormone (TSH), total T4, fT4/FTI, total T3, and T4 binding globulin (TBG), and maternal serum concentrations of thyroid peroxidase (TPO)-antibodies were measured. Placental maturity was evaluated histologically and mRNA and/or protein levels of thyroid hormone deiodinases (DiO) 1, 2 and 3, and transporters (MCT8, MCT10, and OATP1c1) were quantified. In indicated and spontaneous preterm births, cord plasma T4 concentrations were lower than in healthy term controls (p ≤ .001), whereas T3 was only decreased in spontaneous preterm birth (p ≤ .001). Compared with spontaneous preterm births and healthy term controls, indicated preterm birth was characterized by higher maternal plasma TSH (p ≤ .05), earlier placental maturation, higher placental DiO2 gene and MCT10 protein levels and lower DiO3 gene levels (all p ≤ .01). Low T4 was observed in preterm infants irrespective of the cause of preterm birth, while maternal (TSH) and placental (DiO2, DiO3, and MCT10) compensatory responses were only activated in indicated preterm birth due to vascular complications. This may have mediated the normal fetal T3 availability in preterm infants born after indicated preterm birth but not after spontaneous preterm birth.


Adaptive dual-energy algorithm based on pre-calibrated weighting factors for chest radiography.
Objective.To develop a dual-energy (DE) algorithm with spatially varying weighting factors for material selection and noise suppression.Approach.Calibration step-phantoms, with overlapping slabs of solid water and bone with different thicknesses, were used to obtain the pre-calibrated material selection and noise reduction weighting factors. The Material selection weighting factors were calculated by finding a zero of contrast-to-noise-ratio (CNR) between regions with two overlapping materials and regions of only target material, while noise suppression weighting factors were determined by maximizing signal-to-noise ratio for overlapping regions. The pre-calibrated weighting factors were fitted with low and high energy radiograph of two Rando phantoms to create maps of material selection and noise suppression weighting factors, which used with DE algorithm and anti-correlated noise reduction (ACNR) algorithm to generate DE images. Three different implementations, including two different sizes of Rando phantoms and two different orientations (oblique and anterior-posterior), were investigated. Soft-tissue and bone only images of Rando phantoms were obtained with five combinations of DE algorithms and CNR, contrast, and noise values of selected regions of interest were compared to evaluate the performance of the novel method: simple log subtraction (SLS), SLS with uniform ACNR, adaptive DE (aDE), aDE with uniform ACNR, and aDE and adaptive ACNR (aACNR).Main results.Compared to SLS, the aDE algorithm demonstrated improved image quality in all three orientations. CNR increased with better contrast for both soft-tissue and bone images. Implementation of aACNR algorithm resulted in further reduction of image noise and improvements in CNR at the cost of contrast. However, aACNR algorithm showed better contrast compared to ACNR method.Significance.A novel DE algorithm was proposed, which showed improved material selection and noise suppression as compared to the conventional DE techniques and can be easily implemented in a clinical environment for real-time DE image generation.


Increase in complement iC3b is associated with anti-inflammatory cytokine expression during late pregnancy in mice.
Immunological tolerance between fetal allograft and mother is crucial for pregnancy establishment and maintenance; however, these mechanisms particularly those during the latter part of pregnancy have not been definitively elucidated. The aim of this study was to examine the presence and potential function of innate immunity characteristic to the middle to late pregnancy. We first characterized up-regulated proteins in decidua from day 11 pregnant (P11) mice using 2D-PAGE, followed by MALDI-TOF/MS analysis. These analyses identified increased complement component 3 (C3) and its derivatives in P11 decidua. We then found that in the decidual tissues, C3 mRNA increased on P15 and remained high on P19. C3 is converted to C3b and then iC3b by complement component factor I (Cfi) and complement receptor 1-like protein (Crry), both of which were present in P19 placentas. In addition, iC3b proteins and its receptor CR3 (Cd11b/Cd18) in decidual and placental tissues increased toward the latter phase of pregnancy. Moreover, CR3 subunit CD11b protein was predominantly localized to spongiotrophoblast layer in the P19 placenta. Because iC3b is known to induce anti-inflammatory cytokine production, the analysis was extended to examine changes in pro- and anti-inflammatory cytokines, Il12, Il10, and Tgfb1. Il12 expression decreased in P15 and P19 placenta, while high mRNA expression of Il10 and Tgfb1 was found in P19 placental tissues. Furthermore, placental Il10 and Tgfb1 mRNAs were down-regulated when pregnant mice were treated with an anti-C3 antibody, detecting C3, C3b and iC3b. These results indicated that C3 derivatives, in particular, iC3b and its receptor CR3 were up-regulated at the fetal-maternal interface, and suggest that iC3b may regulate the placental expression of anti-inflammatory cytokines, IL10 and TGFB1, during the latter phase of pregnancy.


Evolutionary dynamics of collective index insurance.
Index-based insurances offer promising opportunities for climate-risk investments in developing countries. Indeed, contracts conditional on, e.g., weather or livestock indexes can be cheaper to set up than conventional indemnity-based insurances, while offering a safety net to vulnerable households, allowing them to eventually escape poverty traps. Moreover, transaction costs by insurance companies may be additionally reduced if contracts, instead of arranged with single households, are endorsed by collectives of households that bear the responsibility of managing the division of the insurance coverage by its members whenever the index is surpassed, allowing for additional flexibility in what concerns risk-sharing and also allowing insurance companies to avoid the costs associated with moral hazard. Here we resort to a population dynamics framework to investigate under which conditions household collectives may find collective index insurances attractive, when compared with individual index insurances. We assume risk sharing among the participants of each collective, and model collective action in terms of an N-person threshold game. Compared to less affordable individual index insurances, we show how collective index insurances lead to a coordination problem in which the adoption of index insurances may become the optimal decision, spreading index insurance coverage to the entire population. We further investigate the role of risk-averse and risk-prone behaviors, as well as the role of partial correlation between insurance coverage and actual loss of crops, and in which way these affect the original coordination thresholds.


Stent graft treatment of cerebrovascular wall defects: intermediate-term clinical and angiographic results.
Endovascular stent graft (SG) deployment offers a useful vessel-preserving strategy for vascular wall lesions such as pseudoaneurysms and fistulae. Although deployment of expanded polytetrafluoro-ethylene-covered SGs within the carotid and vertebral arteries is technically feasible, data on long-term efficacy, safety, and patency rate remain sparse. Six patients with traumatic (n = 4), iatrogenic (n = 1), or spontaneous (n = 1) internal carotid and vertebral artery injuries (direct carotid-cavernous fistula, n = 2; pseudoaneurysms, n = 4) were treated with nine balloon-mounted coronary expanded polytetrafluoro-ethylene SGs. Angiographic (mean, 2.3 yr; range, 1.7-4.2 yr) and neurological follow-up (mean, 2.7 yr) was performed for all patients. Complete angiographic exclusion of the lesion was achieved by the initial procedure in five of the six patients; one ruptured cavernous carotid aneurysm leading to a direct carotid-cavernous fistula showed persistent slow shunting despite tandem deployment of two SGs. All six patients revealed complete and persistent angiographic obliteration at delayed follow-up, with minimal in-stent stenosis (<20%) seen in two instances. Difficulty with SG navigation was encountered in five patients, resulting in one instance of guide catheter-induced intimal dissection. Type I endoleak was observed in five patients, requiring secondary angioplasty in four patients and deployment of an additional tandem SG in three. Technical challenges in current-generation SG deployment include sizing, navigation, positioning, and propensity for endoleak. When managed successfully, stent grafting provides a valuable approach for the treatment of vascular wall defects for which vessel preservation is preferred. Intermediate-term safety is satisfactory, with no delayed complications and minimal in-stent stenosis in follow-up periods of more than 2 years.


A case of systemic lupus erythematosus with complex partial status epilepticus.
We describe a woman with systemic lupus erythematosus who had complex partial status epilepticus as the sole epileptic manifestation. The patient presented with a confusional state, initially considered to be psychotic in nature, due to SLE. However, the EEG demonstrated the epileptic etiology of her symptoms. In patients with lupus presenting with prolonged episodes of confusion and behavioural disturbance, an EEG should be performed, whenever possible during the episode, even in the absence of a previous history of epilepsy.


Type II interferon induction and passive transfer depress the murine cytochrome P-450 drug metabolism system.
Induction of type II interferon by sensitization of mice with Mycobacterium tuberculosis strain BCG and challenge with tuberculin resulted in a depression of the cytochrome P-450 drug metabolism system of the liver. The degree of depression was significantly greater than in mice that were only sensitized to BCG. Cytochrome b5 levels were not affected. In addition, the level of the depression of the cytochrome P-450 system correlated with the levels of type II interferon induced. Passive transfer of exogenous type II interferon preparations also significantly depressed the cytochrome P-450 system. Passive transfer of mock interferon or of normal serum had no effect.


Hyperinsulinism and hyperammonemia in infants with regulatory mutations of the glutamate dehydrogenase gene.
A new form of congenital hyperinsulinism characterized by hypoglycemia and hyperammonemia was described recently. We hypothesized that this syndrome of hyperinsulinism and hyperammonemia was caused by excessive activity of glutamate dehydrogenase, which oxidizes glutamate to alpha-ketoglutarate and which is a potential regulator of insulin secretion in pancreatic beta cells and of ureagenesis in the liver. We measured glutamate dehydrogenase activity in lymphoblasts from eight unrelated children with the hyperinsulinism-hyperammonemia syndrome: six with sporadic cases and two with familial cases. We identified mutations in the glutamate dehydrogenase gene by sequencing glutamate dehydrogenase complementary DNA prepared from lymphoblast messenger RNA. Site-directed mutagenesis was used to express the mutations in COS-7 cells. The sensitivity of glutamate dehydrogenase to inhibition by guanosine 5'-triphosphate was a quarter of the normal level in the patients with sporadic hyperinsulinism-hyperammonemia syndrome and half the normal level in patients with familial cases and their affected relatives, findings consistent with overactivity of the enzyme. These differences in enzyme insensitivity correlated with differences in the severity of hypoglycemia in the two groups. All eight children were heterozygous for the wild-type allele and had a mutation in the proposed allosteric domain of the enzyme. Four different mutations were identified in the six patients with sporadic cases; the two patients with familial cases shared a fifth mutation. In two clones of COS-7 cells transfected with the mutant sequence from one patient, the sensitivity of the enzyme to guanosine 5'-triphosphate was reduced, findings similar to those in the child's lymphoblasts. The hyperinsulinism-hyperammonemia syndrome is caused by mutations in the glutamate dehydrogenase gene that impair the control of enzyme activity.


Designing Sustainable Polymer Blends: Tailoring Mechanical Properties and Degradation Behaviour in PHB/PLA/PCL Blends in a Seawater Environment.
Biodegradable polyesters are a popular choice for both packaging and medical device manufacture owing to their ability to break down into harmless components once they have completed their function. However, commonly used polyesters such as poly(hydroxybutyrate) (PHB), poly(lactic acid) (PLA), and polycaprolactone (PCL), while readily available and have a relatively low price compared to other biodegradable polyesters, do not meet the degradation profiles required for many applications. As such, this study aimed to determine if the mechanical and degradation properties of biodegradable polymers could be tailored by blending different polymers. The seawater degradation mechanisms were evaluated, revealing surface erosion and bulk degradation in the blends. The extent of degradation was found to be dependent on the specific chemical composition of the polymer and the blend ratio, with degradation occurring via hydrolytic, enzymatic, oxidative, or physical pathways. PLA presents the highest tensile strength (67 MPa); the addition of PHB and PCL increased the flexibility of the samples; however, the tensile strength reduced to 25.5 and 18 MPa for the blends 30/50/20 and 50/25/25, respectively. Additionally, PCL presented weight loss of up to 10 wt.% and PHB of up to 6 wt.%; the seawater degradation in the blends occurs by bulk and surface erosion. The blending process facilitated the flexibility of the blends, enabling their use in diverse industrial applications such as medical devices and packaging. The proposed methodology produced biodegradable blends with tailored properties within a seawater environment. Additionally, further tests that fully track the biodegradation process should be put in place; incorporating compatibilizers might promote the miscibility of different polymers, improving their mechanical properties and biodegradability.


The physiological role of ADP and Mg2+ in maintaining a stable beat cycle in bull sperm.
Sperm flagella derive their motive power from the motor protein dynein. In this study, we show that maintenance of the flagellar beat cycle in detergent-extracted bull sperm models is highly dependent on the ratio of Mg(2+) to adenosine triphosphate (ATP). An excess of either ATP un-complexed with Mg(2+) , or an excess of Mg(2+) without an equivalent concentration of ATP, results in the loss of beat amplitude and a reduced curvature development in the beat cycle. In addition, we find that adenosine diphosphate (ADP) can stabilize the beat cycle and permit rhythmic beating across a broader range of ATP and Mg(2+) concentrations. We provide evidence that suggests that when ATP is un-complexed with Mg(2+) , it disrupts the beat cycle by reducing dynein adhesion and thereby, reduces the transmission of dynein-generated force between the doublets. Excess Mg(2+) does not act by the same mechanism and induces a condition where the flagellum is more resistant to bending. This is consistent with the idea that high Mg(2+) stabilizes rigor bridges, and ATP reduces the microtubule binding affinity of dynein. Our results may explain how intact sperm are able to sustain coordinated flagellar beating under a wide range of metabolic conditions, as intact sperm produce ADP in direct proportion to their consumption of ATP.


Is Intravenous Heparin a Contraindication for TPA in Ischemic Stroke?
There are approximately 2 million cardiac catheterizations that occur every year in the United States and with an aging population this number continues to rise. Adverse events due to this procedure occur at low rates and include stroke, arrhythmia, and myocardial infarctions. Due to the high volume of procedures there are a growing number of adverse events. Stroke after cardiac catheterization (SCC) has an incidence between 0.27 and 0.5% and is one of the most debilitating complications leading to high rates of mortality and morbidity. Given the relatively uncommon clinical setting of stroke after cardiac catheterization, treatment protocols regarding the use of IV or IA thrombolysis have not been adequately developed. Herein, we describe a case of a 39-year-old male who developed a stroke following a cardiac catheterization where IV thrombolysis was utilized although the patient was on heparin prior to cardiac catheterization.


Rho-associated kinase directly induces smooth muscle contraction through myosin light chain phosphorylation.
Small GTPase Rho plays pivotal roles in the Ca2+ sensitization of smooth muscle. However, the GTP-bound active form of Rho failed to exert Ca2+-sensitizing effects in extensively Triton X-100-permeabilized smooth muscle preparations, due to the loss of the important diffusible cofactor (Gong, M. C., Iizuka, K., Nixon, G. , Browne, J. P., Hall, A., Eccleston, J. F., Sugai, M., Kobayashi, S. , Somlyo, A. V., and Somlyo, A. P. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 1340-1345). Here we demonstrate the contractile effects of Rho-associated kinase (Rho-kinase), recently identified as a putative target of Rho, on the Triton X-100-permeabilized smooth muscle of rabbit portal vein. Introduction of the constitutively active form of Rho-kinase into the cytosol of Triton X-100-permeabilized smooth muscle provoked a contraction and a proportional increase in levels of monophosphorylation of myosin light chain in both the presence and the absence of cytosolic Ca2+. These effects of constitutively active Rho-kinase were wortmannin (a potent myosin light chain kinase inhibitor)-insensitive. Immunoblot analysis revealed that the amount of native Rho-kinase was markedly lower in Triton X-100-permeabilized tissue than in intact tissue. Our results demonstrate that Rho-kinase directly modulates smooth muscle contraction through myosin light chain phosphorylation, independently of the Ca2+-calmodulin-dependent myosin light chain kinase pathway.


Modified low density lipoproteins in the pathogenesis of atherosclerosis.
A fatty streak, the earliest atherosclerotic lesion consists mostly of lipid-laden macrophages. The mechanism leading to accumulation of cholestrol ester in macrophages in vivo is unknown, but it may result from the uptake of modified low density lipoproteins that enter from blood or are modified within the arterial wall. In this review I discuss the major types of modified low density lipoproteins with a special focus on their chemical nature.


The cell surface of Trypanosoma musculi bloodstream forms. II. Lectin and immunologic studies.
Living Trypanosoma musculi bloodstream trypomastigotes were agglutinated specifically with concanavalin A (ConA), wheat germ agglutinin (WGA), soybean agglutinin (SBA), and fucose-binding protein (FBP). The agglutination with these lectins of living cells from which the coat was removed by trypsinization was the same as with intact trypanosomes. Glutaraldehyde or formalin fixation did not affect the results with regard to agglutination with WGA, SBA, and FBP, but lower agglutination with ConA was observed upon fixation. By using a dense iron-dextran marker many fewer ConA marker particles were localized at the fine structural level in the intact than in trypsin-treated trypanosomes. On the basis of the results obtained by agglutination and electron microscopy, it is likely that fixation cross-links intact surface-coat components associated with the ConA binding sites. It is evident from the studies in which lectins were employed that ligands containing alpha-D-mannose, N-acetylglucosamine, N-acetylgalactosamine, and alpha-L-fucose are randomly distributed in the outer surface of the pellicular and flagellar membranes of T. musculi trypomastigotes. Results obtained with alpha-amylase- and dextranase-treated trypanosomes suggested that lectin-binding sugar ligands in the cell surface were not directly associated with alpha-1,4 or repetitive alpha-1,6 glucan-bonded polysaccharide moieties. Similar conclusions can be drawn on the basis of neuraminidase treatment with regard to N-acetylated neuraminic acids. After thorough washing, intact, but not trypsin-treated trypomastigotes were agglutinated specifically with antisera against whole mouse serum and against mouse IgG. Evidently, adsorbed constituents of mouse serum are regular components of the T. musculi surface coat. After incubation in dilute whole mouse serum or in mouse IgG solutions, also the trypsinized cells were agglutinated by the 2 antisera. No such results were obtained with trypsinized cells incubated in serum-free buffers. It was concluded that mouse serum proteins were readily readsorbed on, and firmly bound to the trypsinized cells' surfaces. Specific agglutinations were obtained with trypsinized cells after incubation in dilute rat, rabbit, bovine, and human sera and in solutions of rat and rabbit IgG in reactions with the corresponding antisera. It seems, therefore, that the host serum proteins are adsorbed nonspecifically to the cell surface of trypsinized T. musculi bloodstream forms. When examined by electron microscopy, the intact trypomastigotes were covered by an ununiform, slightly granular, fibrillar extracellular coat, applied to the entire outer lamina of the pellicular and flagellar membranes. No indication of such a coat was noted in the trypsinized organisms. Flocculent surface coat-like matrix could, however, be discerned in cells which, after trypsinization, were incubated in various sera.


Retinol-binding protein-4 in experimental and clinical metabolic disease.
Retinol-binding protein-4 (RBP4), a 21-kDa protein synthesized in the liver and adipose tissue, has recently been described as a murine adipokine involved in the development of insulin resistance. The expression of the gene encoding RBP4 was increased in the adipose tissue, but not in the liver, of insulin-resistant adipose GLUT4(-/-) mice and five other mouse models of obesity and insulin resistance. In addition, intraperitoneal injection or transgenic overexpression of RBP4 in mice induced insulin resistance. While experimental clinical approaches (mostly applying clamp techniques) in humans confirmed correlations of RBP4 with insulin resistance, studies in larger groups out of clinical routine failed to demonstrate a connection with alternative measures of insulin sensitivity. Yet, significant associations of RBP4 with atherogenic lipids were found and a focus of future studies should be the influence on atherosclerosis and related complications. Based on current data, the function of RBP4 as an adipokine exerting metabolic effects in glucose metabolism in humans remains uncertain and might be restricted to rodent models.


Central alpha-adrenoceptors during the development of hypertension in rats on high and low salt intake.
Our purpose was to investigate the binding characteristics of central alpha-adrenoceptors during the early stages of the development of hypertension in rats on high and low salt (NaCl) intake. We measured alpha 1-[( 3H]prazosin) and alpha 2-[( 3H]rauwolscine) binding in membranes of the hypothalamus and medulla oblongata of six groups of young Wistar-Kyoto (WKY) rats, spontaneously hypertensive rats (SHR) and subtotally nephrectomized WKY (SN) rats with mean arterial blood pressure (MAP) ranging from normotensive to hypertensive levels after 1 week of salt restriction or loading. In the hypothalamus the SN-high salt rats and both SHR groups had elevated alpha 1-number but there was no change in alpha 2-number. Moreover, MAP was positively correlated with mean hypothalamic alpha 1-number in the six groups. In the medulla oblongata alpha 1-number was unaffected. However, high salt diet influenced medullary alpha 2-binding in the opposite manner in WKY rats versus SN rats and SHR. In these latter groups the affinity was increased and the number decreased in response to high salt intake. Furthermore, a positive correlation between MAP and mean alpha 1:alpha 2 ratio existed in both the hypothalamus and the medulla of the six groups. The data suggest that hypothalamic alpha 1-binding capacity was increased in SHR due principally to a genetic condition which is mimicked by salt loading in the SN rats. Medullary alpha 2-adrenoceptors of WKY, which remained normotensive despite salt loading, responded differently to high salt intake than those of the SN and SHR, whose blood pressure rose significantly.(ABSTRACT TRUNCATED AT 250 WORDS)


The effects of physiotherapy on osteoarthritic knees of females.
The purpose of this study was to investigate the effect of ultrasound and shortwave diathermy, with and without therapeutic exercise on osteoarthritic (OA) knees. From February 1987 to January 1989, we collected 94 OA knees from 61 female patients and divided them into 4 groups. Group 1 received ultrasound therapy, group 2 had shortwave diathermy, group 3 received ultrasound and an exercise program, and group 4 received shortwave diathermy and exercise therapy. All 61 patients (94 knees) were assessed by functional incapacity scoring and isokinetic testing before and after treatment. At the termination of treatment, all patients had significant improvement in both functional capacity and peak torque. This study failed to prove any significant difference in treatment effect between ultrasound and shortwave diathermy for chronic OA knees. However, the exercise program in addition to the diathermy did promote the treatment effect in both function and muscle torque.


Impact of salinity on the aerobic metabolism of phosphate-accumulating organisms.
The use of saline water in urban areas for non-potable purposes to cope with fresh water scarcity, intrusion of saline water, and disposal of industrial saline wastewater into the sewerage lead to elevated salinity levels in wastewaters. Consequently, saline wastewater is generated, which needs to be treated before its discharge into surface water bodies. The objective of this research was to study the effects of salinity on the aerobic metabolism of phosphate-accumulating organisms (PAO), which belong to the microbial populations responsible for enhanced biological phosphorus removal (EBPR) in activated sludge systems. In this study, the short-term impact (hours) of salinity (as NaCl) was assessed on the aerobic metabolism of a PAO culture, enriched in a sequencing batch reactor (SBR). All aerobic PAO metabolic processes were drastically affected by elevated salinity concentrations. The aerobic maintenance energy requirement increased, when the salinity concentration rose up to a threshold concentration of 2 % salinity (on a W/V basis as NaCl), while above this concentration, the maintenance energy requirements seemed to decrease. All initial rates were affected by salinity, with the NH4- and PO4-uptake rates being the most sensitive. A salinity increase from 0 to 0.18 % caused a 25, 46, and 63 % inhibition of the O2, PO4, and NH4-uptake rates. The stoichiometric ratios of the aerobic conversions confirmed that growth was the process with the highest inhibition, followed by poly-P and glycogen formation. The study indicates that shock loads of 0.18 % salt, which corresponds to the use or intrusion of about 5 % seawater may severely affect the EBPR process already in wastewater treatment plants not exposed regularly to high salinity concentrations.


Movement-related cortical activation in familial Parkinson disease.
We sought to determine whether or not first-degree relatives of patients with familial Parkinson disease (FDRs) present impaired movement-related cortical activity. We studied 10 familial Parkinson disease subjects, 10 FDRs, and 10 controls and analyzed event-related mu desynchronization (ERD) and beta synchronization. Forty percent FDRs presented reduced premovement mu ERD latency, suggesting that premovement cortical activation is impaired in FDRs.


Status Update on the Sheehan-Suicidality Tracking Scale (S-STS) 2014.
There is a need for a choice of scales to evaluate the full range of suicidal phenomena. Such scales must be capable of use as both safety and efficacy outcome measures in research and in clinical settings. Central to the success in finding and developing effective anti-suicidal medications is having a sensitive suicidality scale that can detect an efficacy signal in conventional sample sizes used in clinical trials. The Sheehan-Suicidality Tracking Scale was developed for these purposes. This article provides a 2014 status update on the scale's progress, its use, and its properties. The authors review why and how the scale was developed; the scale structure, versions, and properties; the trials in which it was used; the time frames accommodated; its validation and reliability studies; its utility in screening and assessment; its utility in assessing treatment-emergent suicidal adverse events; its use as an efficacy outcome measure; its availability in self-rated and clinician-rated forms; the availability and linguistic validation of pediatric versions; linguistic validation in other languages; how it compares with global ratings of suicidality; and its possible utility and applications.


The optimal method for improving postural balance in healthy young and older people: specific training for postural tasks encountered in personal physical practice.
It is well known that regular exercise or physical activity (training) improves postural balance in healthy young and older subjects, but the optimal exercise or physical activity (i.e., likely to induce the greatest postural improvements) and the context in which it is carried out remain to be explored and determined for each population. The most beneficial adaptations would depend, in particular, on gestural conditions (body position, movement and gesture practiced) and material conditions (nature of the ground surface, sports equipment used, type of environment - stable or changing). In fact, the global postural adaptations induced by training do not result from the transfer between different trained and untrained postural tasks, but are the sum of the adaptations related to each trained postural task in healthy young and older subjects. Based on current knowledge, optimal training programs should include the full range of postural tasks encountered in personal physical practice for each population. To date, the method of implementing progressive postural balance tasks with different degrees of difficulty and instability has been used as the effective method to improve postural balance, but it should not be considered as the reference method. Instead, it should be considered as a complementary method to the one based on specific postural tasks. An intervention strategy is proposed for young and older adults consisting of three different steps (general, oriented and specific/ecologic training). However, some parameters still need to be explored and possibly reconsidered in future studies to improve postural balance in an optimal way.


Inhibitory effects of salvianolic acid B on the high glucose-induced mesangial proliferation via NF-kappaB-dependent pathway.
Salvianolic acid B (Sal B) is one of the major water-soluble compounds isolated from Radix Salviae Miltiorrhizae (Danshen in Chinese) that has been reported to be beneficial to treatment of diabetic complications. However, the mechanisms involved in these effects are not discussed in relation to mesangial proliferation via modulation of NF-kappaB. To explain this, human mesangial cells were pretreated with or without Sal B (0.1, 1, 10 microM) for 24 h and stimulated with high glucose (30 mM). Then the effects of Sal B on mesangial cells proliferation, extracellular matrix production and the possible mechanisms were evaluated by methylthiazoletetrazolium assay, flow cytometry assay, enzyme-linked immunosorbent assay, gelatin zymography assay and western blot assay. These results indicated that Sal B could inhibit high glucose-induced mesangial cells proliferation and extracellular matrix production in a dose-dependent manner, partially through modulating the cell-cycle progress and MMP-2 and MMP-9 activities via suppressing NF-kappaB activation, suggesting that Sal B may be a promising agent for treating diabetic nephropathy.


Effect of enalapril on the skin response to bradykinin in man.
We tested the effect of oral enalapril on intradermal bradykinin to determine if kininase II inhibition occurs with therapeutic doses in vivo. Six normal male volunteers took either 5 mg enalapril orally or placebo on 2 days. Three hours later bradykinin was injected into the skin of the back in doses increasing from 10(-11) to 10(-9) M. Enalapril increased the bradykinin-induced wheal. Inhibition of kininase II may cause accumulation of endogenous bradykinin. This could be an important mechanism in the occasionally reported side effect of angioedema with the angiotensin converting enzyme (ACE) inhibiting group of drugs.


Phase inversion in polylactide/soybean oil blends compatibilized by poly(isoprene-b-lactide) block copolymers.
Renewable composites were prepared by melt blending of polylactide and soybean oil. The blend morphology was tuned by the addition of poly(isoprene-b-lactide) block copolymers. Due to the extreme difference in the viscosities of soybean oil and polylactide, a critical block copolymer composition was found to induce a phase inversion point at which the minor soybean oil phase became the matrix surrounding polylactide particles. This transition was due to the thermodynamic interactions between the block copolymer and the two phases and shear forces acting on the mixture during blending. The size of the soybean oil droplets in the polylactide matrix was also highly dependent on the block copolymer composition. In binary polylactide/soybean oil blends, there was a limiting concentration of soybean oil that could be incorporated into the polylactide matrix (6% of the total blend weight), which could be increased up to 20% by the addition of block copolymers.


Detection of the mecA gene and identification of Staphylococcus directly from blood culture bottles by multiplex polymerase chain reaction.
Staphylococcus spp. - both S. aureus, including methicillin-resistant strains (MRSA) and coagulase negative staphylococci (CoNS) - are relevant agents of healthcare-associated infections. Therefore, the rapid recognition of MRSA and methicillin-resistant CoNS from blood stream infections is critically important for patient management. It is worth noting that inappropriate empiric therapy has been associated with higher in-hospital mortality. In this study we evaluated a multiplex polymerase chain reaction (multiplex PCR) standardized to detect Staphylococcus spp., S. aureus, and mecA gene-encoded oxacillin resistance directly from blood culture bottles. A total of 371 blood cultures with Gram-positive microorganisms confirmed by Gram-stain were analyzed. Results from multiplex PCR were compared to phenotypic characterization of isolates. Staphylococcus aureus was detected in 85 (23.0%) blood cultures and CoNS in 286 (77.0%). There was 100% agreement between phenotypic and multiplex PCR identification. Forty-three (50.6%) of the 85 S. aureus carried the mecA gene and among the 286 CoNS, 225 (78.7%) were positive for the mecA gene. The multiplex PCR assay developed here was found to be sensitive, specific, rapid, and showed good agreement with the phenotypic results besides being less expensive. This PCR method could be used in clinical laboratories for rapid identification and initiation of specific and effective treatment, reducing patient mortality and morbidity. Furthermore, this method may reduce misuse of antimicrobial classes that are more expensive and toxic, thus contributing to the selection of antibiotic-resistant Staphylococcus spp.


Influence of dual-task constraints on the interaction between posture and movement during a lower limb pointing task.
One of the challenges regarding human motor control is making the movement fluid and at a limited cognitive cost. The coordination between posture and movement is a necessary requirement to perform daily life tasks. The present experiment investigated this interaction in 20 adult men, aged 18-30 years. The cognitive costs associated to postural and movement control when kicking towards a target was estimated using a dual-task paradigm (secondary auditory task). Results showed that addition of the attentional demanding cognitive task yielded a decreased kicking accuracy and an increased timing to perform the movement, mainly during the backswing motion. In addition, significant differences between conditions were found for COP and COM displacement (increased amplitude, mean speed) on the anteroposterior axis. However, no significant differences between conditions were found on the mediolateral axis. Finally, EMG analysis showed that dual-task condition modified the way anticipatory postural adjustments (APAs) were generated. More specifically, we observed an increase of the peroneus longus activity, whereas the temporal EMG showed a decrease of its latency with respect to movement onset. These results suggested a functional adaptation resulting in an invariance of overall APAs, emphasizing that cognitive, postural, and motor processes worked dependently.


Nonmelanoma skin cancer mortality (1988-2000): the Rhode Island follow-back study.
To estimate (1) the magnitude of and the components and factors associated with nonmelanoma skin cancer (NMSC) mortality and (2) the proportion of deaths misclassified as NMSC. Population-based follow-back study. All Rhode Island residents whose deaths between 1988 and 2000 were attributed to NMSC. Distribution of diagnoses, verification of the causes of death, and characterization of associated factors. The proportion of misclassified deaths was significantly higher for nongenital NMSC (57%) than for genital NMSC (18%; P<.001). Most of the deaths misclassified as nongenital NMSC were caused by squamous cell carcinoma of mucosal surfaces. The age-adjusted NMSC mortality rate was 0.91 (per 100 000 persons per year), of which almost half (0.45) were due to genital carcinoma. Nonmelanoma skin cancer mortality increased sharply with age. The mortality rate from nongenital NMSC in men was more than twice that in women, but for genital NMSC this ratio was reversed. Skin cancers originating on the ear were responsible for more than a quarter of all deaths caused by nongenital NMSC. No cases of NMSC mortality occurred in organ transplant recipients. Many individuals had comorbid psychiatric conditions or evidence of unreasonable delay in seeking medical care for their lesions. Misclassifying the cause of death as nongenital NMSC accounts for a large source of error on death certificates in Rhode Island. Overall, nongenital squamous cell carcinoma and basal cell carcinoma death rates have declined, and mortality due to genital carcinoma was about half of total NMSC deaths. The dermatology community should emphasize prevention of mortality from genital skin cancer, while continuing to stress the importance of reducing excessive exposure to UV light and prompt treatment of NMSC.


Tryptophan mutants of troponin C from skeletal muscle--an optical probe of the regulatory domain.
We have generated a series of chicken skeletal muscle troponin C mutants to study the conformation of the regulatory domain in the N-terminal half of the molecule. These mutants each contained a single Trp at position 22 (helix A), 52 (linker of helices B and C), or 90 (central helix). Some of these mutants also contained additional mutations to introduce a single Cys at a desired position. The mutants were characterized by molecular graphics and CD and found to have a minimum of structural perturbations when compared with the native structure. They also retained the ability to regulate myofibrillar ATPase activity. The fluorescence of Trp22 was sensitive to Ca2+ binding only to the regulatory sites, whereas Trp52 and Trp90 responded to Ca2+ binding to both the regulatory and the Ca2+/Mg2+ sites. The tryptophan quantum yield (Q) of all Trp22-containing mutants was very high (0.33) in the absence of bound Ca2+, compared to that of L-tryptophan in aqueous solution (0.14). Q decreased 25% upon binding of Ca2+ to the regulatory sites. The quantum yields of Trp52 and Trp90 in apo mutants were close to 0.14. In the presence of bound Ca2+ at the regulatory sites, the quantum yield of Trp52 decreased 16%, whereas that of Trp90 increased 25%. Results from acrylamide quenching of the fluorescence of the three Trp residues indicated that Trp22 was the least exposed and Trp52 was the most exposed, consistent with other spectral data that Trp22 was in a relatively nonpolar environment and Trp52 was in a highly polar environment. The ability of Trp52 and Trp90 to sense Ca2+ binding to sites located at both domains suggests inter-domain communication in the protein. These single Trp TnC mutants provide specific signals for probing Ca2+-induced conformational changes in the regulatory domain.


Solution structure of a late embryogenesis abundant protein (LEA14) from Arabidopsis thaliana, a cellular stress-related protein.
We report the three-dimensional structure of a late embryogenesis abundant (LEA) protein from Arabidopsis thaliana gene At1g01470.1. This protein is a member of Pfam cluster PF03168, and has been classified as a LEA14 protein. LEA proteins are expressed under conditions of cellular stress, such as desiccation, cold, osmotic stress, and heat. The structure, which was determined by NMR spectroscopy, revealed that the At1g01470.1 protein has an alphabeta-fold consisting of one alpha-helix and seven beta-strands that form two antiparallel beta-sheets. The closest structural homologs were discovered to be fibronectin Type III domains, which have <7% sequence identity. Because fibronectins from animal cells have been shown to be involved in cell adhesion, cell motility, wound healing, and maintenance of cell shape, it is interesting to note that in plants wounding or stress results in the overexpression of a protein with fibronectin Type III structural features.


Mandibular Osteonecrosis due to the Pulpal-Periodontal Syndrome: a Case Report and Review of the Literature.
Ishemic bone disease has multifactorial etiologies. Cronic dental infections should be eliminated to prevent osteonecrosis of the jaw. We report an unusual case of osteonecrosis due to the pulpal-peridontal syndrome and subsequent pulp necrosis. A case of 38 year old woman who presented with exposed bone, 8 mm in diameter, in the lingual area of the right lower third molar. The patient was otherwise healthy and was not taking any medications. A detailed medical history showed no previous diseases. Patient denied any type of local trauma. A complete blood count showed no abnormalities. The panoramic radiograph revealed a deep periodontal pocket between teeth 47 and 48. The CBCT revealed a deep periodontal pocket between molars and bone sequestrum of the lingual plate. Topical treatment consisted of adhesive periodontal dressing based on the cellulose and bethamethasone oitnment together with orabase, without improvement. Therefore, peroral amoxycillin was prescribed for a week. Since there was no improvement, the third molar was removed as well as necrotic bone; the alveolar bone was remodelled and sutures were placed. After suturing, the whole area was covered using intraoral resorbable bandage. Microbial swab of the wound aspirate did not reveal polymorphonuclears or the presence of microorganisms. Microbial swab of the biopsy specimen of the necrotic bone particle and sequestrum showed a large amount of gram-positive coccae, however, polymorphonuclears were not found. Histopathological analysis revealed acute chronic inflammation. One week after the surgery, the area healed completely. This case highlights the fact that in some patients bone exposure might develop due to the pulpal-periodontal syndrome i.e. pulp necrosis.


[Histamine as the principal pathogenetic factor in gastroduodenal ulcer. Research priorities].
A research group led by L. Businco between 1937 and 1942 was able, by applying new measuring methods in a variety of experimental situations, to demonstrate for the first time a concrete pathogenetic link between histamine and gastroduodenal ulcer. These personal researches produced the following objective data: increase in blood histamine in more than 70% of patients with ulcers; presence of histamine in the gastric juice of ulcer patients; diminution of blood histamine after gastric resection; increase in blood histamine in patients with postoperative peptic ulcer; increase in tissue histamine in ulcerous zones; diminution in blood histamine, parallel clinical improvement and contemporaneous diminution in total and hydrochloric acidity in patients treated with histaminase. The discover by Black et al of cimetidine as an antagonist of H2 receptors to which is added the gastric secretory stimulus of histamine, underscores the role played by this substance in determining gastroduodenal ulcer.


Flexible robotic catheters in the visceral segment of the aorta: advantages and limitations.
Flexible robotic catheters are an emerging technology which provide an elegant solution to the challenges of conventional endovascular intervention. Originally developed for interventional cardiology and electrophysiology procedures, remotely steerable robotic catheters such as the Magellan system enable greater precision and enhanced stability during target vessel navigation. These technical advantages facilitate improved treatment of disease in the arterial tree, as well as allowing execution of otherwise unfeasible procedures. Occupational radiation exposure is an emerging concern with the use of increasingly complex endovascular interventions. The robotic systems offer an added benefit of radiation reduction, as the operator is seated away from the radiation source during manipulation of the catheter. Pre-clinical studies have demonstrated reduction in force and frequency of vessel wall contact, resulting in reduced tissue trauma, as well as improved procedural times. Both safety and feasibility have been demonstrated in early clinical reports, with the first robot-assisted fenestrated endovascular aortic repair in 2013. Following from this, the Magellan system has been used to successfully undertake a variety of complex aortic procedures, including fenestrated/branched endovascular aortic repair, embolization, and angioplasty.


Proteomics in Inflammatory Bowel Disease: Approach Using Animal Models.
Recently, proteomics studies have provided important information on the role of proteins in health and disease. In the domain of inflammatory bowel disease, proteomics has shed important light on the pathogenesis and pathophysiology of inflammation and has contributed to the discovery of some putative clinical biomarkers of disease activity. By being able to obtain a large number of specimens from multiple sites and control for confounding environmental, genetic, and metabolic factors, proteomics studies using animal models of colitis offered an alternative approach to human studies. Our aim is to review the information and lessons acquired so far from the use of proteomics in animal models of colitis. These studies helped understand the importance of different proteins at different stages of the disease and unraveled the different pathways that are activated or inhibited during the inflammatory process. Expressed proteins related to inflammation, cellular structure, endoplasmic reticulum stress, and energy depletion advanced the knowledge about the reaction of intestinal cells to inflammation and repair. The role of mesenteric lymphocytes, exosomes, and the intestinal mucosal barrier was emphasized in the inflammatory process. In addition, studies in animal models revealed mechanisms of the beneficial effects of some therapeutic interventions and foods or food components on intestinal inflammation by monitoring changes in protein expression and paved the way for some new possible inflammatory pathways to target in the future. Advances in proteomics technology will further clarify the interaction between intestinal microbiota and IBD pathogenesis and investigate the gene-environmental axis of IBD etiology.


Death due to coronary artery anomaly with coexistence of Chiari network.
Coronary artery anomalies rarely detected in autopsy series and angiograms can be a component of complex malformations, besides, can be also associated with sudden cardiac death. Presented case was 22-year-old male, who had suddenly fainted during a football match played on artificial turf, he was transferred into the hospital, however had died during intensive care therapy. He had been evaluated by local prosecutor, and sent to our center for autopsy. At autopsy, internal macroscopic examination revealed absence of the right coronary artery. A total of two coronary artery ostia were observed. One of them originated from the left aortic sinus, and the other one stemmed from 8 mm above the sinotubular line. Besides, Chiari network formation was seen in the right atrium. This case with coronary artery anomaly associated with formation of Chiari network was discussed from the perspective of forensic medicine in the light of the literature information.