Oxygen chiral phosphate esters.
During the past four years, methods have been reported for the syntheses and configurational analyses of virtually any phosphate mono- and diester chiral by virtue of oxygen isotope substitution, and these techniques have already been applied to an impressive number of enzymic and chemical reactions. At present, no experimental information is available that contradicts the simplest interpretations that have been applied to the results obtained for enzymic reactions: inversion of configuration indicating a direct displacement of the leaving group by the attacking group, and retention of configuration implying the formation of a phosphorylated or nucleotidylated enzyme intermediate. However, it does seem necessary to further investigate the mechanisms of at least some of the reactions discussed in this review to ensure that the simplest interpretation is correct. For example, the caveat we have raised about the interpretation of inversions of configurations in phosphohydrolase reactions is chemically reasonable, and these reactions should be reexamined to evaluate the importance of covalent catalysis by carboxylate groups. However, for the vast majority of the enzymic reactions that have been investigated, the stereochemical approach to ascertaining whether catalysis involves the formation of covalent intermediates remains the simplest and most direct method.


Prevalence and correlates of REM sleep behaviour disorder in patients with major depressive disorder: a two-phase study.
To investigate the prevalence and clinical correlates of video polysomnography (vPSG)-confirmed rapid eye movement sleep behaviour disorder (RBD) in patients with major depressive disorder (MDD). This is a clinic-based two-phase epidemiological study. In phase 1, patients with MDD were screened by a validated questionnaire, RBD Questionnaire-Hong Kong (RBDQ-HK). In phase 2, a subsample of both the screen-positive (RBDQ-HK >20) and screen-negative patients with MDD underwent further clinical and sleep assessment (vPSG) to confirm the diagnosis of RBD (MDD+RBD). Poststratification weighting method was used to estimate the prevalence of MDD+RBD. The total likelihood ratio and the probability of prodromal Parkinson's disease (PD) were calculated from prodromal markers and risk factors, as per the Movement Disorder Society research criteria. A total of 455 patients with MDD were screened (median age (IQR)=52.66 (15.35) years, 77.58% woman, 43.74% positive). Eighty-one patients underwent vPSG and 12 of them were confirmed MDD+RBD. The prevalence of MDD+RBD was estimated to be 8.77% (95% CI: 4.33% to 16.93%), with possibly male predominance. MDD+RBD were associated with colour vision and olfaction deficit and a higher probability for prodromal PD. Almost 9% of patients with MDD in the psychiatric outpatient clinic has vPSG-confirmed RBD. Comorbid MDD+RBD may represent a subtype of MDD with underlying α-synucleinopathy neurodegeneration. Systematic screening of RBD symptoms and necessity of vPSG confirmation should be highlighted for capturing this MDD subtype with a view to enhance personalised treatment and future neuroprotection to prevent neurodegeneration.


Consumption and acceptability of whole grain staples for lowering markers of diabetes risk among overweight and obese Tanzanian adults.
Dietary changes characterized by a reduction in carbohydrate quality are occurring in developing countries and may be associated with a higher prevalence of obesity and chronic diseases such as type 2 diabetes mellitus. We assessed the preferences and acceptability of unrefined whole grain carbohydrate staples (i.e., brown rice, unrefined maize and unrefined sorghum ugali) as substitutes for commonly consumed refined carbohydrates in Tanzania. A questionnaire was used to collect sociodemographic information and dietary habits, and pre-and post-tasting questionnaires were administered for test foods. A 10-point LIKERT scale was used to rate attributes of the three test foods. White rice and refined maize ugali were the most commonly consumed carbohydrate staples in this population; 98% and 91%, respectively. Occasional consumption of unrefined maize and sorghum ugali was reported by 32% and 23% of the participants, respectively. All of the test foods were highly rated for smell, taste, color, appearance and texture. Taste was rated highest for unrefined maize ugali. Almost all of the participants were willing to participate in a future dietary intervention involving regular consumption of these unrefined carbohydrates for at least six months duration. These findings suggest that whole grain carbohydrates are highly acceptable, and that there is a promising potential for their use in future dietary intervention studies in Tanzania.


Assessment of the microbiological quality of groundwater in three regions of the Valencian Community (Spain).
Urban groundwater development was traditionally constrained by concerns about its quality. This study was conducted in the regions of La Ribera Alta and Ribera Baja and La Plana de Requena-Utiel of the Valencian Community (Valencia, Spain) where population density, demand for drinking water and agricultural activities are high. Groundwater bodies (GWBs) are regarded as management areas within each territory, and were used to establish protection policies. This study analyzed eleven GWBs. We used two databases with microbiological measurements from 154 wells over a 7-year period (2004-2011), risk factors and groundwater information. Wells were grouped according to frequency of microbiological contamination using E. coli measurements, category <1, or wells with low-frequency microbiological contamination and high-frequency wells or category 1-100, according to World Health Organization (WHO) quality criteria of drinking water. Of all wells, 18.12% showed high-frequency microbiological contamination with a majority distribution in the Ribera Alta region (26.98%, p < 0.001). No significant differences were found between the two risk categories for flow, static level, well depth and distance from population centres. This paper reveals that the vulnerability classes established by the Geological and Mining Institute of Spain (IGME) do not match the microbiological results, and that only eight wells with high-frequency contamination coincide with the high vulnerability areas.


Multiple lipid binding sites determine the affinity of PH domains for phosphoinositide-containing membranes.
Association of peripheral proteins with lipid bilayers regulates membrane signaling and dynamics. Pleckstrin homology (PH) domains bind to phosphatidylinositol phosphate (PIP) molecules in membranes. The effects of local PIP enrichment on the interaction of PH domains with membranes is unclear. Molecular dynamics simulations allow estimation of the binding energy of GRP1 PH domain to PIP3-containing membranes. The free energy of interaction of the PH domain with more than two PIP3 molecules is comparable to experimental values, suggesting that PH domain binding involves local clustering of PIP molecules within membranes. We describe a mechanism of PH binding proceeding via an encounter state to two bound states which differ in the orientation of the protein relative to the membrane, these orientations depending on the local PIP concentration. These results suggest that nanoscale clustering of PIP molecules can control the strength and orientation of PH domain interaction in a concentration-dependent manner.


The development of the method for the determination of terbinafine in cat's plasma and hair.
Clinical investigations of terbinafine indicate its high treatment activity against infections by several dermatophytes. Its efficiency was tested also in the treatment of microsporosis in cats. The distribution of terbinafine in cat's plasma and hair is important for the identification of the drug efficiency. A fast and reliable reversed-phase high performance liquid chromatographic method with appropriate sample preparation has been developed. Reliability, good reproducibility and low detection limit (LOD 0.25 ng/ml) of the method enable determination of terbinafine in hair and also in plasma of cats infected with Microsporum canis treated by Lamisil tablets.


Blood pressure profile in Nigerian children.
To observe blood pressure (BP) pattern and its correlates in primary school children of northern Nigeria. Sitting BP and pulse were measured in quadruplicate, then repeated after four weeks in 1,721 healthy children aged five to 16 years. Body weight and height were also measured in their school environment. Primary schools located in three communities in Zaria Local Government Area (LGA) of Kaduna State, Nigeria. The communities were Tudun Wada (University community of migrants with some indigenous Hausa settlers), Zaria City (traditional Hausa community) and Zaria Kewaye (a rural Hausa settlement). BP rose with age. However, BP levels particularly systolic was highest in children from Tudun Wada (TW) (urban), followed by those from Zaria city (ZC) (semi urban), and Zaria Kewaye (ZK) (rural). The mean systolic/diastolic BP (mmHg) were: 99/61, 94/62 and 89/60 in children aged five to ten years; and 112/69, 109/68 and 107/68 in those older than 10 years respectively. The differences in BP levels were evident even as early as the age of five years and appears largely independent of physical stature and gender. These observations suggest that place of residence and ethnicity may be important factors in the progression of BP with age in some children in northern Nigeria.


[Characteristics of the use of laparoscopic cholecystectomy in middle-aged and aged patients].
Under analysis are results of using laparoscopic cholecystectomy (LChE) in 43 elderly and senile patients (from 60 to 88 years of age). The control group consisted of 67 patients of young and middle-aged patients (from 25 to 59 years of age) operated upon by the same method. Based on a comparative analysis of 2 groups of patients the authors have established age-dependent specificity of using LChE at all stages of treatment from the preoperative preparation and choice of indications for operation till discharge from the hospital. The materials presented show that most of non-standard difficult and complicated LChE take place just in the age older than 60. When performing LChE in such patients additional intraoperative technical means should be used and the indications for laparotomy must be wider. A conclusion is made that when estimating results of LChE it is expedient to analyze results of treatment of elderly and senile patients by this method.


The Important Role of Hemispherotomy for Rasmussen Encephalitis: Clinical and Functional Outcomes.
Rasmussen encephalitis (RE) is characterized by pharmacoresistant epilepsy and progressive neurological deficits concurrent with unilateral hemispheric atrophy. Evidence of an inflammatory autoimmune process has been extensively described in the literature; however, the precise etiology of RE is still unknown. Despite data supporting a beneficial effect of early immunosuppressive and immunomodulatory interventions, surgical disconnection of the affected hemisphere is considered the treatment of choice for these patients. The aim of this study was to report a series of children and adolescents who underwent hemispheric surgery (HS) for the treatment of RE, analyzing their clinical, electrographic, and neuroimaging features pre-operatively, as well as their postoperative status, including seizure and functional outcomes. All patients with RE who underwent HS in the Epilepsy Surgery Center (CIREP) of the University Hospital of Ribeirão Preto Medical School, between 1995 and 2020 were retrospectively reviewed. Preoperative and postoperative analyses included gender; age at epilepsy onset; seizure semiology; seizure frequency; interictal and ictal electroencephalographic (EEG) findings; age at surgery; duration of epilepsy; surgical complications; duration of follow-up; histopathological findings; and postoperative seizure, cognitive, and functional outcomes. Forty-four patients were evaluated. Mean age at seizure onset and epilepsy duration was 6 years and 2.5 years, respectively. Mean age at surgery was 9 years, with an average follow-up ranging from 3 months to 23 years. All patients presented with severe epilepsy and distinct neurological abnormalities on MRI. Before HS, different degrees of abnormal intellectual performance as well as hemiparesis were seen in 86% and 90%, respectively. Histopathology examination confirmed this diagnosis in 95% patients. At the last follow-up, 68% of patients were seizure free, and 70% were classified as Engel Class I or II. Postoperatively, the cognitive status remained unchanged in 64% of patients. Likewise, the gross motor function remained unchanged in 54% of patients and 74% had functional hand ability after HS. Considering the progressive damage course of RE, hemispheric surgery should be offered to pediatric patients. It has manageable risks and results in good seizure outcome, and the preoperative functional status of these children is often preserved (even when the left hemisphere is involved), thus improving their quality of life.


cis-Diamminedichloroplatinum(II) induces c-jun expression in human myeloid leukemia cells: potential involvement of a protein kinase C-dependent signaling pathway.
cis-Diamminedichloroplatinum(II) (CDDP) is a chemotherapeutic agent known to inhibit DNA, RNA, and protein synthesis. The cytotoxicity of this drug is thought to result from the formation of DNA intrastrand cross-links. The present work demonstrates that treatment of human myeloid leukemia cells (HL-60, U-937, and KG-1) with CDDP is associated with increased expression of the c-jun gene and that this effect is related to activation by a transcriptional mechanism. The results also demonstrate that treatment with CDDP is associated with increases in protein kinase C (PKC) activity. Furthermore, the finding that pretreatment with H7, an inhibitor of PKC, abrogates the effect of CDDP on c-jun expression suggested the involvement of PKC in this process. Down-regulation of PKC by prolonged pretreatment with 12-O-tetradecanoylphorbol-13-acetate was also associated with inhibition of CDDP-induced c-jun expression. The results further demonstrate that there is a temporal relationship between the CDDP-induced increase in c-jun expression and the occurrence of internucleosomal DNA cleavage characteristic of programmed cell death. These findings suggest that c-jun may be involved in the cellular response to DNA-damaging agents, such as CDDP, and that this effect may be mediated by a PKC-dependent pathway.


Optimized purification of a heterodimeric ABC transporter in a highly stable form amenable to 2-D crystallization.
Optimized protocols for achieving high-yield expression, purification and reconstitution of membrane proteins are required to study their structure and function. We previously reported high-level expression in Escherichia coli of active BmrC and BmrD proteins from Bacillus subtilis, previously named YheI and YheH. These proteins are half-transporters which belong to the ABC (ATP-Binding Cassette) superfamily and associate in vivo to form a functional transporter able to efflux drugs. In this report, high-yield purification and functional reconstitution were achieved for the heterodimer BmrC/BmrD. In contrast to other detergents more efficient for solubilizing the transporter, dodecyl-ß-D-maltoside (DDM) maintained it in a drug-sensitive and vanadate-sensitive ATPase-competent state after purification by affinity chromatography. High amounts of pure proteins were obtained which were shown either by analytical ultracentrifugation or gel filtration to form a monodisperse heterodimer in solution, which was notably stable for more than one month at 4°C. Functional reconstitution using different lipid compositions induced an 8-fold increase of the ATPase activity (k(cat)∼5 s(-1)). We further validated that the quality of the purified BmrC/BmrD heterodimer is suitable for structural analyses, as its reconstitution at high protein densities led to the formation of 2-D crystals. Electron microscopy of negatively stained crystals allowed the calculation of a projection map at 20 Å resolution revealing that BmrC/BmrD might assemble into oligomers in a lipidic environment.


Management of postoperative introital and vaginal stenosis.
Five patients with symptomatic postoperative introital or vaginal stenosis were evaluated and treated. In each case simple operative correction, utilizing basic plastic surgery principles, was successful. The five operative techniques as used alone and in combination are described. Though postoperative stenosis of the vagina and/or introitus is relatively uncommon, the resulting disability is such that every gynecologic surgeon should be familiar with the problem and solution. In most cases simple surgical correction can be successful if the surgeon is familiar with basic techniques and principles.


Porcine skin inguinal hernioplasty.
Sixty inguinal hernias were repaired using a porcine dermis to reinforce the posterior wall of the inguinal canal. This study mentions the criteria for the use of this xenograft and the technique used. A follow-up of two years is reported without any recurrences so far. An average patient has a 10 to 15 per cent risk of recurrence after an operation for indirect hernia and about 20 per cent after direct hernioplasty. It has been reported that 50 per cent of the recurrences appear during the first year post-operative period, and most of these in the first three to six months. Although inguinal hernia is one of the most common conditions encountered by a general surgeon, no prospective randomised study has yet been undertaken to evaluate the most widely used techniques. Many operative procedures for the treatment of inguinal hernias have been reported since Bassini and Halstead first reported upon this 90 years ago. While the late results of many methods have been presented, there is no unanimity concerning the procedure of choice. Recurrence rates differ according to the technique used and the expertise of the operating surgeon. Porcine dermis was used as it is the best available biological material. It does not have the problem of rejection and fragmentation like other prostheses.


Axillary nerve repair in 99 patients with 101 stretch injuries.
The purpose of this paper was to analyze outcomes in patients at the Louisiana State University Health Sciences Center (LSUHSC) who presented with contusion-stretch injuries to the axillary nerve. These injuries resulted from shoulder injury either with or without fracture/dislocation. Although recovery of deltoid function can occur spontaneously, this was not always the case. Severe deficits persisting for several months led the patients to undergo surgery. Operative categories included isolated axillary palsy (56 procedures), combined axillary and suprascapular palsies (11 procedures), axillary and radial palsies (14 procedures), and axillary palsy with another deficit, usually infraclavicular plexus loss (20 procedures). Deltoid function was evaluated pre- and postoperatively by applying the LSUHSC grading system. An anterior infraclavicular approach was usually followed during surgery, but in three patients an additional posterior approach was used. Axillary lesions usually began in the proximal portion of the posterior cord. Although several patients had distraction of the nerve, lesions in continuity were found in more than 90% of cases. Intraoperative nerve action potential (NAP) recordings were performed to determine the need for resection. Most repairs were made using grafts, although in three patients with relatively focal lesions suture was used. When an NAP was recorded across the lesion and neurolysis was performed, recovery was judged to be a mean Grade 4 according to the LSUHSC in 30 cases. Recovery following suture repairs was a mean Grade 3.8, whereas recovery after 66 graft repairs was a mean Grade 3.7. In cases in which suprascapular palsies were associated with axillary injuries, the former recovered but the latter did not necessarily do so without surgery. If the radial nerve was also injured, recovery of the triceps and brachioradialis muscles and wrist extension was usually obtained, but it was far more difficult to reverse the loss of finger and thumb extension. Although few in number, complications did occur and they are important. Operative exploration of axillary contusion-stretch lesions is worthwhile in carefully selected cases. If indicated by inspection and intraoperative electrical studies, nerve repair can lead to useful function.


Actinomycin D for methotrexate-failed low-risk gestational trophoblastic neoplasia.
To determine outcomes and factors associated with failure of 5-day actinomycin D for treatment of methotrexate-failed low-risk gestational trophoblastic neoplasia (GTN). We reviewed the records of 358 patients treated with methotrexate 0.4 mg/kg (max 25 mg) IV push q.d. x 5 d every 14 d for FIGO-defined, low-risk GTN between 1979 and 2009. Actinomycin D 0.5 mg IV push q.d. x 5 d every 14 d was given to 64 of 68 patients (18%) who failed methotrexate: 48 (75%) for resistance and 16 (25%) for toxicity. Adjuvant surgery was used in selected patients. Clinical response and survival as well as factors affecting outcomes were analyzed retrospectively. The complete response rate to secondary chemotherapy with actinomycin D for failed methotrexate treatment of low-risk GTN was 75% (48/64), including 71% (34/48) for methotrexate resistance and 88% (14/16) for methotrexate toxicity. All 20 patients (6%) who failed sequential single-agent chemotherapy with methotrexate and actinomycin D were placed into permanent remission with the use of multiagent chemotherapy with or without surgery. The only factor significantly associated with resistance to secondary actinomycin D chemotherapy was clinicopathologic diagnosis of choriocarcinoma versus postmolar GTN (56% versus 20%, p = 0.025). Actinomycin D 0.5 mg IV q.d. x 5 d every 14 d used as secondary therapy in methotrexate-failed low-risk GTN resulted in a 75% complete response rate and eventual 100% cure with subsequent multiagent chemotherapy with or without surgery. Resistance to sequential methotrexate and actinomycin D chemotherapy was significantly associated with original FIGO score > or = 3 and clinicopathologic diagnosis of choriocarcinoma.


Mast cells induce upregulation of P-selectin and intercellular adhesion molecule 1 on carotid endothelial cells in a new in vitro model of mast cell to endothelial cell communication.
It is suggested that mast cells contribute to cell recruitment in inflammation through the upregulation of endothelial adhesion molecules. P-selectin and intercellular adhesion molecule(ICAM)-1 are two key adhesion molecules that have been associated indirectly with mast cell activity. The canine C2 mastocytoma cell line and primary cultures of canine carotid endothelial cells were used to establish a new in vitro model to help study the interaction between mast cells and endothelial cells. Carotid endothelial cells were incubated with mast cell mediators to uncover their effect on endothelial ICAM-1 and P-selectin expression. To assess the relative contributions of tumour necrosis factor (TNF)-alpha and histamine to such effect, an H1 antihistamine and a TNF-alpha blocking antibody were used. Prior to activation by mast cell mediators, P-selectin was expressed only within the cytoplasm, and ICAM-1 was constitutively expressed on the surface of the canine carotid endothelial cells. Both adhesion molecules were enhanced significantly and strongly upon mast cell activation at various time points. Unstored TNF-alpha was fully responsible for ICAM-1 upregulation. P-selectin was up-regulated by both preformed and newly synthesized mast cell mediators, but neither histamine nor TNF-alpha accounted for such an effect. Therefore,a new model is proposed in which the pro-inflammatory effect of mast cells on endothelial cells can be studied in vitro. In this model, it has been demonstrated that only TNF-alpha accounts for the overexpression of ICAM-1 induced by mast cells, and that mast cells up-regulate P-selectin expression through a histamine-independent mechanism.


Proteomic analysis of the cerebrospinal fluid of patients with restless legs syndrome/Willis-Ekbom disease.
Restless Legs Syndrome/Willis-Ekbom Disease (RLS/WED) is a sensorimotor disorder that causes patients to experience overwhelming and distressing sensations in the legs compelling the patient to move their legs to provide relief. The purpose of this study was to determine if biomarkers in the cerebrospinal fluid can distinguish RLS/WED patients from neurological controls. We obtained CSF samples by lumbar puncture from 5 early-onset RLS/WED patients and 5 controls. We performed 2-dimensional difference in-gel electrophoresis (2D-DIGE). Proteins that were significantly altered were identified by Student's t-test. Protein spots that were differentially expressed (p ≤ 0.05, Av. Ratio ≥ 2.0) between RLS/WED and control CSF samples were identified using MALDI-TOF-MS. Statistical analyses of the validation immunoblot assays were performed using Student's t-test. In this discovery study we identified 6 candidate CSF protein markers for early-onset RLS/WED. Four proteins (Cystatin C, Lipocalin-type Prostaglandin D2 Synthase, Vitamin D binding Protein, and β-Hemoglobin) were increased and 2 proteins (Apolipoprotein A1 and α-1-acid Glycoprotein) were decreased in RLS/WED patients. Our results reveal a protein profile in the RLS/WED CSF that is consistent with clinical findings of disruptive sleep, cardiovascular dysfunction and painful symptoms. Moreover, protein profiles are consistent with neuropathological findings of activation of hypoxia inducible factor (HIF) pathways and alterations in dopaminergic systems. These data indicate the CSF of RLS/WED patients may provide information relevant to biological basis for RLS/WED, treatment strategies and potential new treatment targets.


Livestock abundance predicts vampire bat demography, immune profiles and bacterial infection risk.
Human activities create novel food resources that can alter wildlife-pathogen interactions. If resources amplify or dampen, pathogen transmission probably depends on both host ecology and pathogen biology, but studies that measure responses to provisioning across both scales are rare. We tested these relationships with a 4-year study of 369 common vampire bats across 10 sites in Peru and Belize that differ in the abundance of livestock, an important anthropogenic food source. We quantified innate and adaptive immunity from bats and assessed infection with two common bacteria. We predicted that abundant livestock could reduce starvation and foraging effort, allowing for greater investments in immunity. Bats from high-livestock sites had higher microbicidal activity and proportions of neutrophils but lower immunoglobulin G and proportions of lymphocytes, suggesting more investment in innate relative to adaptive immunity and either greater chronic stress or pathogen exposure. This relationship was most pronounced in reproductive bats, which were also more common in high-livestock sites, suggesting feedbacks between demographic correlates of provisioning and immunity. Infection with both Bartonella and haemoplasmas were correlated with similar immune profiles, and both pathogens tended to be less prevalent in high-livestock sites, although effects were weaker for haemoplasmas. These differing responses to provisioning might therefore reflect distinct transmission processes. Predicting how provisioning alters host-pathogen interactions requires considering how both within-host processes and transmission modes respond to resource shifts.This article is part of the theme issue 'Anthropogenic resource subsidies and host-parasite dynamics in wildlife'.


Molecular dynamics simulations of amino acid adsorption and transport at the acetonitrile-water-silica interface: the role of side chains.
The solvation and transport of amino acid residues at liquid-solid interfaces have great importance for understanding the mechanism of separation of biomolecules in liquid chromatography. This study uses umbrella sampling molecular dynamics simulations to study the adsorption and transport of three amino acid molecules with different side chains (phenylalanine (Phe), leucine (Leu) and glutamine (Gln)) at the silica-water-acetonitrile interface in liquid chromatography. Free energy analysis shows that the Gln molecule has stronger binding affinity than the other two molecules, indicating the side chain polarity may play a primary role in adsorption at the liquid-solid interface. The Phe molecule with a phenyl side chain exhibits stronger adsorption free energy than Leu with a non-polar side chain, which can be ascribed to the better solvated configuration of Phe. Further analysis of molecular orientations found that the amino acid molecules with apolar side chains (Phe and Leu) have 'standing up' configurations at their stable adsorption state, where the polar functional groups are close to the interface and the side chain is far from the interface, whereas the amino acid molecule with a polar side chain (Gln) chooses the 'lying' configuration, and undergoes a sharp orientation transition when the molecule moves away from the silica surface. Extending our simulation studies to systems with different solute concentrations reveals that there is a decrease in the adsorption free energy as well as surface diffusion as the solute concentration increases, which is related to the crowding in the interfacial layers. This simulation study gives a detailed microscopic description of amino acid molecule solvation and transport at the acetonitrile-water-silica interface in liquid chromatography and will be helpful for understanding the retention mechanism for amino acid separation.


Partial purification of the Epstein-Barr virus nuclear antigen(s).
The Epstein-Barr virus nuclear antigen (EBNA) is speculated to be involved in cell transformation by the virus. Studies on the molecular properties of EBNA, however, have yielded conflicting results. In this study, three Epstein-Barr virus(EBV)-induced antigens were isolated and purified from extracts prepared from Raji cells. These antigens were able to block the anticomplement immunofluorescence reaction, indicating that all three were related to EBNA. The soluble antigen was found wholly in the cytosol fraction. An EBV-induced nuclear antigen I was found both in the cytosol and the nucleus. The EBV-induced nuclear antigen II was found associated with the chromatin. The soluble antigen and the nuclear antigen I were separated and partially purified using phosphocellulose chromatography. Each was further purified 1,400-fold with respect to the whole cell state by chromatography on CL-Sepharose 6B followed by blue dextran-Sepharose. subunit molecular weights of 70,000 were determined for each of these antigens, both in the crude and purified state, by radioimmunoelectrophoresis and gel filtration. The nuclear antigen II was purified 2,500-fold using hydroxylapatite, CL-Sepharose 6B, and blue dextran-Sepharose chromatographies. This antigen displayed two subunits by radioimmunoelectrophoresis with molecular weights of 65,000 and 70,000. Although all antigens shared similar molecular weights, the extent of their homology remains to be determined.


Double-stranded RNA adenosine deaminase ADAR-1-induced hypermutated genomes among inactivated seasonal influenza and live attenuated measles virus vaccines.
We sought to examine ADAR-1 editing of measles and influenza virus genomes derived from inactivated seasonal influenza and live attenuated measles virus vaccines grown on chicken cells as the culture substrate. Using highly sensitive 3DI-PCR (R. Suspène et al., Nucleic Acids Res. 36:e72, 2008), it was possible to show that ADAR-1 could hyperdeaminate adenosine residues in both measles virus and influenza virus A genomes. Detailed analysis of the dinucleotide editing context showed preferences for 5'ArA and 5'UrA, which is typical of editing in mammalian cells. The hyperedited mutant frequency, including genomes and antigenomes, was a log greater for influenza virus compared to measles virus, suggesting a greater sensitivity to restriction by ADAR-1.


[The effect of thyroid function on the increase of plasma cyclic AMP following glucagon injection (author's transl)].
The increase in plasma cyclic adenosine-3':5'-monophosphate (cAMP) was measured after intravenous injection of 1 mg of glucagon in 26 normal subjects, 36 patients with hyperthyroidism, 35 patients with hypothyroidism and 24 patients with euthyroid goitre. While patients with euthyroid goitre responded normally, the plasma cyclic AMP response in patients with hyperthyroidism was considerably increased and in those with hypothyroidism decreased. 4 patients with cirrhosis of the liver had reduced responses and 1 patient with extrahepatic obstructive jaundice an enhanced response. This test seems to be a valuable additional parameter for the description of the thyroid-dependent metabolic situation. However, because of its unspecificity it cannot replace the measurement of serum T3, T4 and thyrotropin (TSH) response to thyroliberin (TRH).


Caveats and Pitfalls of SOX1 Autoantibody Testing With a Commercial Line Blot Assay in Paraneoplastic Neurological Investigations.
SOX1 autoantibodies are considered markers of small cell lung cancer (SCLC) and paraneoplastic neurological syndromes (PNS) and are usually determined by commercial line blot in many clinical services. Recent studies suggested that SOX1 autoantibodies also occur in patients with neuropathies unrelated to SCLC, questioning the value of SOX1 autoantibodies as paraneoplastic biomarkers. Here, we compared the specificity and sensitivity of a commercial line blot (Euroimmun, Lübeck, Germany) with those of an in house cell-based assay (CBA) with HEK293 cells transfected with SOX1. Overall, 210 patients were included in the study, 139 patients with polyneuropathies without SCLC, and 71 with disorders associated with SOX1 autoantibodies detected with the in-house CBA. Forty one of these 71 cases had been referred to our laboratory for onconeuronal antibody assessment and 30/71 were patients with known PNS and SCLC. None of the patients with polyneuropathies had SOX1 autoantibodies by either line blot or CBA (specificity of the immunoblot: 100%; 95%C.I.: 97.8-100). Among the 71 patients with CBA SOX1 autoantibodies, only 53 were positive by line blot (sensitivity: 74.6%; 95%C.I.: 62.9-84.2). Lung cancer was detected in 37/41 (90%; 34 with SCLC) patients referred for onconeuronal antibody assessment and 34 of them also had a PNS. Our study confirms the association of SOX1 autoantibodies with SCLC and PNS. The line blot test misses 25% of the cases; therefore, to minimize the frequency of false negative results we recommend the use of a confirmatory test, such as CBA, in patients suspected to have a SCLC-related PNS.


Effect of Pseudomonas sp. MT5 baths on Flavobacterium columnare infection of rainbow trout and on microbial diversity on fish skin and gills.
Use of Pseudomonas sp. strain MT5 to prevent and treat Flavobacterium columnare infection was studied in 2 experiments with fingerling rainbow trout Oncorhynchus mykiss. In the first experiment, length heterogeneity analysis of PCR-amplified DNA fragments (LH-PCR) was used to assess the effect of antagonistic baths on the microbial diversity of healthy and experimentally infected fish. In the 148 samples studied, no difference was found between bathed and unbathed fish, and 3 fragment lengths were detected most frequently: 500 (in 75.7% of the samples), 523 (62.2%) and 517 bp (40.5%). The species contributing to these fragment sizes were Pseudomonas sp., Rhodococcus sp. and F. columnare, respectively. A specific PCR for detection of Pseudomonas sp. MT5 was designed, but none of the tissue samples were found to be positive, most likely indicating poor adhesion of the strain during bathing. LH-PCR was found to be a more powerful tool for detecting F. columnare in fish tissue than traditional culture methods (chi2 = 3.9, df = 1, p < 0.05). Antagonistic baths had no effect on the outbreak of infection or on fish mortality. F. columnare was also detected in healthy fish prior to and after experimental infection, indicating that these fish were carriers of the disease. In the second experiment, intensive Pseudomonas sp. MT5 antagonistic baths were given daily to rainbow trout suffering from a natural columnaris infection. Again, the antagonistic bacteria had no effect on fish mortality, which reached 95 % in both control and antagonist-treated groups in 7 d.


Deubiquitinating enzyme regulation of the p53 pathway: A lesson from Otub1.
Deubiquitination has emerged as an important mechanism of p53 regulation. A number of deubiquitinating enzymes (DUBs) from the ubiquitin-specific protease family have been shown to regulate the p53-MDM2-MDMX networks. We recently reported that Otub1, a DUB from the OTU-domain containing protease family, is a novel p53 regulator. Interestingly, Otub1 abrogates p53 ubiquitination and stabilizes and activates p53 in cells independently of its deubiquitinating enzyme activity. Instead, it does so by inhibiting the MDM2 cognate ubiquitin-conjugating enzyme (E2) UbcH5. Otub1 also regulates other biological signaling through this non-canonical mechanism, suppression of E2, including the inhibition of DNA-damage-induced chromatin ubiquitination. Thus, Otub1 evolves as a unique DUB that mainly suppresses E2 to regulate substrates. Here we review the current progress made towards the understanding of the complex regulation of the p53 tumor suppressor pathway by DUBs, the biological function of Otub1 including its positive regulation of p53, and the mechanistic insights into how Otub1 suppresses E2.


BRCA1 and BRCA2 mutation prevalence and clinical characteristics of a population-based series of ovarian cancer cases from Denmark.
To evaluate the prevalence of BRCA1 and BRCA2 mutations and associations with clinical correlates of disease in a population-based series of ovarian cancer cases from Denmark. DNA sequencing and multiplex ligation-dependent probe amplification analysis were used to analyze the BRCA1 and BRCA2 genes for coding sequence mutations and large genomic rearrangements in 445 confirmed cases of ovarian cancer. We evaluated associations between mutation status and clinical characteristics, including cancer risks for first-degree relatives and clinicopathologic features of tumors. Deleterious BRCA1 or BRCA2 mutations were identified in 26 cases; thus, mutations in these genes are responsible for at least 5.8% of ovarian cancer cases in this population. Five different mutations were identified in more than one individual, suggesting that they may be founder mutations in Denmark. We identified several differences between mutation carriers and noncarriers: mutation carriers were diagnosed at a significantly early age (median, 49 and 61 years, respectively; P = 0.0001); the frequency of BRCA1 mutation carriers was 23% for women diagnosed <40 years, 15% for 40 to 49 years, 4% for 50 to 59 years, and 2% for > or =60 years (P = 0.00002); ovarian cancer in carriers was diagnosed at a later stage (P = 0.002) and tumors were of poorer grade (P = 0.0001); and first-degree relatives of mutation carriers had greater relative risks of both ovarian cancer [10.6 (95% confidence interval, 4.2-26.6); P < 0.0001] and breast cancer <60 years [8.7 (95% confidence interval, 3.0-25.0); P < 0.0001]. These data may have a significant effect on risk assessment and clinical management of individuals from Denmark who are predisposed to ovarian cancer because they carry a BRCA1 or BRCA2 mutation.


Clinicopathologic analysis of 124 biopsy-proven peripheral nerve diseases.
We reviewed dinical, histological and ultrastructural findings of 124 cases of sural nerve biopsy specimens to delineate the trends of peripheral nerve diseases in our institute. Eighty-one were men and 43 were women. We categorized them into five groups: specific diagnosis (66 cases, 53.2%), axonal degeneration type (47 cases, 37.9%), demyelinating type (4 cases, 3.2%), mixed axonal degeneration-demyelinating type (6 cases, 4.8%) and normal (1 case, 0.9%). Cases with specific diagnosis included 21 inflammatory demyelinating polyneuropathy (15 chronic inflammatory demyelinating polyradiculoneuropathy, 6 Guillain-Barre disease), 13 hereditary motor and sensory neuropathy (7 Charcot-Marie-Tooth type I, 6 Charcot-Marie-Tooth type II), 10 vasculitis, 6 toxic neuropathy, 4 leprosy, 3 diabetic neuropathy, 2 alcoholic neuropathy, 1 Fabry's disease and other specific diseases (5 cases). In our cases, the proportion of specific diagnoses was higher, while the proportion of demyelinating peripheral neuropathies and normal were lower than those of Western series. The results of this study indicate that 1) a dose clinicopathologic correlation is important to make a precise diagnosis of peripheral nerve biopsy, 2) Biopsy under strict indication may reduce unnecessary histologic examination, 3) There is no difference in disease pattern of peripheral neuropathy between Western people and Koreans.


Seasonal Changes in the Incidence of Transient Global Amnesia.
Transient global amnesia (TGA) is a stereotypic condition characterized by anterograde and retrograde amnesia that typically resolves within 24 hours. The pathophysiology of TGA is still unclear. We noted that patients hospitalized with TGA tend to appear in seasonal clusters, and decided to investigate this phenomenon. Every patient with acute presentation of amnesia at our medical center is hospitalized for observation and evaluation. We reviewed the monthly occurrence of TGA in our patient population between 2000 and 2014, and compared this to non-TGA hospitalizations during the same time period. During the analysis period, 154 patients who met the criteria for TGA were hospitalized, as well as 259,007 non-TGA hospitalizations. The annual occurrence of TGA ranged from 5 to 16 hospitalizations. There were 91 TGA events in women and 63 in men, in subjects aged 62.8±10.6 years (mean±SD). The incidence was maximal during December [odds ratio (OR)=2.83, 95% confidence interval (CI)=1.20-6.67] and March (OR=2.77, 95% CI=1.17-6.56), and minimal from April to August. The incidence exhibited an increase followed by a decrease from October to February. A seasonal trend was observed as well, with incidence peaks occurring in winter (OR=1.82, 95% CI=1.12-2.96) and spring (OR=1.80, 95% CI=1.10-2.94). Our findings suggest that the incidence of TGA exhibits seasonal variations. This observation may help to improve the understanding of the pathophysiology underlying TGA.


Rate of Viral Re-Suppression and Retention to Care Among PLHIV on Second-Line Antiretroviral Therapy at Dessie Comprehensive Specialized Hospital, Northeast Ethiopia: A Retrospective Cohort Study.
In Ethiopia, first-line antiretroviral therapy failure is growing rapidly. However, unlike first-line therapy, to date, very little is known about the outcomes of second-line therapy. Thus, this study assessed the rate of viral re-suppression and attrition to care and their predictors among people living with HIV on second-line therapy. A retrospective cohort study was conducted on 642 people living with HIV at Dessie Comprehensive Specialized Hospital from October 2016 to November 2019. A proportional Cox regression model was computed to explore predictors of viral re-suppression (viral load less than 1000 copies/mL) and attrition to care. Out of 642 subjects, 19 (3%), 44 (6.9%), 70 (10.9%), and 509 (79.3%) patients were lost to follow up, died, transferred out, and alive on care, respectively. Similarly, 82.39% (95% CI: 79.24-85.16%) of patients had achieved viral re-suppression, with 96 per 100 person-year rate of re-suppression. Patients who switched timely to second-line therapy were at a higher rate of viral re-suppression than delayed patients [adjusted hazard rate, AHR = 1.43 (95% CI: 1.17-1.74)]. Not having drug substitution history [AHR = 1.25 (95% CI: 1.02-1.52)] was positively associated with viral re-suppression. In contrast, being on anti-TB treatment [AHR = 0.67 (95% CI: 0.49-0.91)] had lower likelihood with viral re-suppression. In the current study, attrition to care was 11% (95% CI: 8.7-13.9%). Ambulatory or bedridden patients were more at risk of attrition to care as compared with workable patients [AHR = 2.61 (95% CI: 1.40-4.87)]. Similarly, being not virally re-suppressed [AHR = 6.87 (95% CI: 3.86-12.23)] and CD4 count ≤450 cells/mm3 [AHR = 2.61 (95% CI: 1.40-4.87)] were also positively associated with attrition to care. A significant number of patients failed to achieve viral re-suppression and attrition from care. Most identified factors related to patient monitoring. Hence, patient-centered intervention should be strengthened, besides treatment switch.


The gel electrophoresis markup language (GelML) from the Proteomics Standards Initiative.
The Human Proteome Organisation's Proteomics Standards Initiative has developed the GelML (gel electrophoresis markup language) data exchange format for representing gel electrophoresis experiments performed in proteomics investigations. The format closely follows the reporting guidelines for gel electrophoresis, which are part of the Minimum Information About a Proteomics Experiment (MIAPE) set of modules. GelML supports the capture of metadata (such as experimental protocols) and data (such as gel images) resulting from gel electrophoresis so that laboratories can be compliant with the MIAPE Gel Electrophoresis guidelines, while allowing such data sets to be exchanged or downloaded from public repositories. The format is sufficiently flexible to capture data from a broad range of experimental processes, and complements other PSI formats for MS data and the results of protein and peptide identifications to capture entire gel-based proteome workflows. GelML has resulted from the open standardisation process of PSI consisting of both public consultation and anonymous review of the specifications.


Outcome results for the Ma'alahi Youth Project, a Tongan community-based obesity prevention programme for adolescents.
Tonga has a very high prevalence of obesity with steep increases during youth, making adolescence a critical time for obesity prevention. The Ma'alahi Youth Project, the Tongan arm of the Pacific Obesity Prevention in Communities project, was a 3-year, quasi-experimental study of community-based interventions among adolescents in three districts on Tonga's main island (Tongatapu) compared to the island of Vava'u. Interventions focused mainly on capacity building, social marketing, education and activities promoting physical activity and local fruit and vegetables. The evaluation used a longitudinal design (mean follow-up duration 2.4 years). Both intervention and comparison groups showed similar large increases in overweight and obesity prevalence (10.1% points, n = 815; 12.6% points, n = 897 respectively). Apart from a small relative decrease in percentage body fat in the intervention group (-1.5%, P < 0.0001), there were no differences in outcomes for any anthropometric variables between groups and behavioural changes did not follow a clear positive pattern. In conclusion, the Ma'alahi Youth Project had no impact on the large increase in prevalence of overweight and obesity among Tongan adolescents. Community-based interventions in such populations with high obesity prevalence may require more intensive or longer interventions, as well as specific strategies targeting the substantial socio-cultural barriers to achieving a healthy weight.


Magnetic resonance imaging of flow-distributed oscillations.
The formation of stationary concentration patterns in a packed-bed reactor (PBR), using a manganese-catalyzed Belousov-Zhabotinsky (BZ) reaction in a mixed sulfuric-phosphoric acid medium, was studied using magnetic resonance imaging (MRI). The PBR was composed of a column filled with glass beads, which was fed by a continuous stirred tank reactor (CSTR). As the reactor is optically opaque, investigation of the three-dimensional (3D) structure of these reaction-diffusion-advection waves is not possible using conventional image capture techniques. MRI has been used to probe this system and the formation, 3D structure, and development of these waves has been studied. At reactor startup, traveling waves were observed. After this initial period the waves stabilized and became stationary. Once fixed, they were found to be remarkably stable. There was significant heterogeneity of the reaction fronts, which were not flat, as would be expected from a plug-flow reactor. Instead, the reaction wave fronts were observed to be conical in shape due to the local hydrodynamics of the bed and specifically the higher velocities and therefore lower residence times close to the wall of the reactor.


Evaluation of an evidence-based tobacco treatment curriculum for psychiatry residency training programs.
Smokers with mental illness and addictive disorders account for nearly one in two cigarettes sold in the United States and are at high risk for smoking-related deaths and disability. Psychiatry residency programs provide a unique arena for disseminating tobacco treatment guidelines, influencing professional norms, and increasing access to tobacco cessation services among smokers with mental illness. The current study evaluated the Rx for Change in Psychiatry curriculum, developed for psychiatry residency programs and focused on identifying and treating tobacco dependence among individuals with mental illness. The 4-hour curriculum emphasized evidence-based, patient-oriented cessation treatments relevant for all tobacco users, including those not yet ready to quit. The curriculum was informed by comprehensive literature review, consultation with an expert advisory group, faculty interviews, and a focus group with psychiatry residents. This study reports on evaluation of the curriculum in 2005-2006, using a quasi-experimental design, with 55 residents in three psychiatry residency training programs in Northern California. The curriculum was associated with improvements in psychiatry residents' knowledge, attitudes, confidence, and counseling behaviors for treating tobacco use among their patients, with initial changes from pre- to posttraining sustained at 3-months' follow-up. Residents' self-reported changes in treating patients' tobacco use were substantiated through systematic chart review. The evidence-based Rx for Change in Psychiatry curriculum is offered as a model tobacco treatment curriculum that can be implemented in psychiatry residency training programs and disseminated widely, thereby effectively reaching a vulnerable and costly population of smokers.


Synergistic and antagonistic effects of salinity and pH on germination in switchgrass (Panicum virgatum L.).
The effects of salt-alkaline mixed stress on switchgrass were investigated by evaluating seed germination and the proline, malondialdehyde (MDA) and soluble sugar contents in three switchgrass (Panicum virgatum L.) cultivars in order to identify which can be successfully produced on marginal lands affected by salt-alkaline mixed stress. The experimental conditions consisted of four levels of salinity (10, 60, 110 and 160 mM) and four pH levels (7.1, 8.3, 9.5 and 10.7). The effects of salt-alkaline mixed stress with equivalent coupling of the salinity and pH level on the switchgrass were explored via model analyses. Switchgrass was capable of germinating and surviving well in all treatments under low-alkaline pH (pH≤8.3), regardless of the salinity. However, seed germination and seedling growth were sharply reduced at higher pH values in conjunction with salinity. The salinity and pH had synergetic effects on the germination percentage, germination index, plumular length and the soluble sugar and proline contents in switchgrass. However, these two factors exhibited antagonistic effects on the radicular length of switchgrass. The combined effects of salinity and pH and the interactions between them should be considered when evaluating the strength of salt-alkaline mixed stress.


The alpha- and beta-subunits are required for expression of catalytic activity in the hetero-dimeric glucosidase II complex from human liver.
The alpha- and beta-subunits of the hetero-dimeric glucosidase II complex from human liver were cloned and expressed in COS-1 cells. The 4106 bp full-length cDNA for the alpha-subunit contained a 2835 bp ORF encoding a 107 kDa polypeptide. The 2095 bp cDNA for the beta-subunit encodes a approximately 60 kDa protein in a continuous 1605 bp ORF. The alpha- and beta-subunits each contain two potential Asn-Xaa-Thr/Ser acceptor sites, with only one site in the alpha-subunit (Asn97) being glycosylated. Additional lambda-clones were isolated for each subunit containing in-frame insertions/deletions within the coding region, indicating alternative splicing. Analysis of different human tissues revealed approximately 4.4 kb and approximately 2.4 kb transcripts for alpha- and beta-subunit, respectively, consistent with their full-length cDNA. Coexpression of the alpha- and beta-subunits in COS-1 cells resulted in >4-fold increase of glucosidase II activity. An inactive protein was obtained, however, after transfection with the alpha-subunit alone, showing that both subunits are essential for expression of active glucosidase II. The observation that the enzyme, previously purified from pig liver and lacking the beta-subunit, was catalytically active indicates that the beta-subunit is involved in alpha-subunit maturation rather than being required for enzymatic activity once the alpha-subunit has acquired its mature form. The alpha-subunit is expressed in COS-1 cells as an ER-located protein, whether inactive or part of a catalytically active complex. This suggests that ER-localization of the alpha-subunit, when associated with the dimeric enzyme complex, is mediated by the C-terminal HDEL-signal in the beta-subunit, whereas the apparently incompletely folded form of the inactive alpha-subunit could be retained in the ER by the putative "glycoprotein-specific quality control machinery. "


5-Fluorouracil treatment of a human colon adenocarcinoma implanted in the subrenal capsule site of athymic mice.
5-Fluorouracil was used in the treatment of a recently established. human adenocolon carcinoma implanted in the subrenal capsule site of athymic mice as part of a program to determine the relative effectiveness of antitumor agents in the treatment of human tumors grown as xenografts and in the clinic. 5-Fluorouracil treatments resulted in a dose-dependent inhibition of tumor growth under the two examined schedules (five daily doses and three doses every 4 days) when administered either i.v. or i.p. The various schedules and routes resulted in similar patterns of antitumor effects when the data were based on the endpoints of either final tumor weight, change in tumor weight, or relative tumor weight. Assessments of cell viability based on histological examination of tumored kidneys resulted in a downward displacement of the dose-response curves but did not alter their shape or the interpretation of the data. Although approximately 1-mm3 tumor fragments were implanted, variability of size was allowed. Tumor growth was not dependent on the initial size of the implant, as shown by a comparison between the initial individual or average tumor sizes and the final individual or average tumor sizes, respectively. The antitumor effects of the 5-fluorouracil could have been determined on the basis of final tumor weight alone. The inhibition of tumor growth was accompanied by weight loss in treated mice as compared with controls. At lower doses, however, the weight loss of the mice was not extensive, which indicated that the inhibition of tumor growth was, at least in part, attributable to some selective antitumor action of the 5-FUra. The greatest inhibition of tumor growth, however, was accompanied by the most extensive animal weight loss. This correlation raised the question of the degree to which the tumor growth inhibition observed was a consequence of nonspecific drug toxicity to the host. The current results stress the importance of detailed investigations into the nature of host-tumor parameters in order to assess the antitumor activity of candidate drugs in the treatment of human tumor xenografts.


A cohort study of the effectiveness of insecticide-treated bed nets to prevent malaria in an area of moderate pyrethroid resistance, Malawi.
Insecticide-treated bed nets (ITNs) are the cornerstone of malaria control in sub-Saharan Africa but their effectiveness may be compromised by the spread of pyrethroid resistance among malaria vectors. The objective of this investigation was to assess the effectiveness of ITNs to prevent malaria in an area of Malawi with moderate pyrethroid resistance. One deltamethrin ITN was distributed in the study area for every two individuals in each household plus one extra ITN for households with an odd number of residents. A fixed cohort of 1,199 children aged six to 59 months was seen monthly for one year and at sick visits to measure malaria infection and use of ITNs. Insecticide resistance among malaria vectors was measured. The effect of ITN use on malaria incidence was assessed, adjusting for potential confounders using generalized estimating equations accounting for repeated measures. There were 1,909 infections with Plasmodium falciparum over 905 person-years at risk (PYAR), resulting in an observed incidence of 2.1 infections per person-year (iPPY). ITNs were used during 97% of the PYAR. The main vector was Anopheles funestus: mortality in WHO tube assays after exposure to 0.05% deltamethrin was 38% (95% confidence interval (CI) 29-47), and resistance was due to elevated oxidase enzymes. After adjusting for potential confounders, the incidence of malaria infection among ITN users was 1.7 iPPY (95% CI 1.5-2.1) and among non-bed net users was 2.6 iPPY (95% CI 2.0-3.3). Use of ITNs reduced the incidence of malaria infection by 30% (rate ratio 0.7; 95% CI, 0.5-0.8) compared to no bed nets. ITNs significantly reduced the incidence of malaria infection in children in an area with moderate levels of pyrethroid resistance and considerable malaria transmission. This is the first study to show that ITNs provide protection in areas where pyrethroid-resistant An. funestus is the major malaria vector. Malaria control programmes should continue to distribute and promote ITNs in areas with low to moderate pyrethroid resistance; however, insecticide resistance may intensify further and it is not known whether ITNs will remain effective at higher levels of resistance. There is an urgent need to identify or develop new insecticides and technologies to limit the vulnerability of ITNs to insecticide resistance.


Non-linear Raman scattering intensities in graphene.
We show that the Raman scattering signals of the two dominant Raman bands G and 2D of graphene sensitively depend on the laser intensity in opposite ways. High electronic temperatures reached for pulsed laser excitation lead to an asymmetric Fermi-Dirac distribution at the different optically resonant states contributing to Raman scattering. This results in a partial Pauli blocking of destructively interfering quantum pathways for G band scattering, which is observed as a super-linear increase of the G band intensity with laser power. The 2D band, on the other hand, exhibits sub-linear intensity scaling due to the blocking of constructively interfering contributions. The opposite intensity dependencies of the two bands are found to reduce the observed 2D/G ratio, a key quantity used for characterizing graphene samples, by more than factor two for electronic temperatures around 3000 K.


Genetic inactivation of β-catenin is salubrious whereas its activation is deleterious in desmoplakin cardiomyopathy.
Mutations in the DSP gene encoding desmoplakin, a constituent of the desmosomes at the intercalated discs (IDs), cause a phenotype that spans arrhythmogenic cardiomyopathy (ACM) and dilated cardiomyopathy (DCM). It is typically characterized by biventricular enlargement and dysfunction, myocardial fibrosis, cell death, and arrhythmias.The canonical WNT (cWNT)/β-catenin pathway is implicated in the pathogenesis of ACM. The β-catenin is an indispensable co-transcriptional regulator of the cWNT pathway and a member of the IDs. We genetically inactivated or activated β-catenin to determine its role in the pathogenesis of desmoplakin cardiomyopathy. The Dsp gene was conditionally deleted in the 2-week-old post-natal cardiac myocytes using tamoxifen-inducible MerCreMer mice (Myh6-McmTam:DspF/F). The cWNT/β-catenin pathway was markedly dysregulated in the Myh6-McmTam:DspF/F cardiac myocytes, as indicated by a concomitant increase in the expression of cWNT/β-catenin target genes, isoforms of its key co-effectors, and the inhibitors of the pathway. The β-catenin was inactivated or activated upon inducible deletion of its transcriptional or degron domain, respectively, in the Myh6-McmTam:DspF/F cardiac myocytes. Genetic inactivation of β-catenin in the Myh6-McmTam:DspF/F mice prolonged survival, improved cardiac function, reduced cardiac arrhythmias, and attenuated myocardial fibrosis, and cell death caused by apoptosis, necroptosis, pyroptosis, i.e., PANoptosis. In contrast, activation of β-catenin had the opposite effects. The deleterious and the salubrious effects were independent of changes in the expression levels of the cWNT target genes and were associated with changes in several molecular and biological pathways, including cell death programs. The cWNT/β-catenin was markedly dysregulated in the cardiac myocytes in a mouse model of desmoplakin cardiomyopathy. Inactivation of β-catenin attenuated whereas its activation aggravated the phenotype, through multiple molecular pathways, independent of the cWNT transcriptional activity. Thus, suppression but not activation of β-catenin might be beneficial in desmoplakin cardiomyopathy.


Immunity and skin cancer.
Observations in humans and animal studies support the theory that immunologic surveillance plays an important role in limiting the development of skin malignancies. These immune responses undergo progressive diminution with age. In addition, other factors, such as bereavement, poor nutrition, and acute and chronic exposure to ultraviolet light, can further diminish immune mechanisms. We believe these decreases in immune function in combination with the carcinogenic effect of ultraviolet light are important in the production of skin cancers.


Early glycemic control in critically ill emergency department patients: pilot trial.
Glycemic control in the critically ill intensive care unit (ICU) patient has been shown to improve morbidity and mortality. We sought to investigate the effect of early glycemic control in critically ill emergency department (ED) patients in a small pilot trial. Adult non-trauma, non-pregnant ED patients presenting to a university tertiary referral center and identified as critically ill were eligible for enrollment on a convenience basis. Critical illness was determined upon assignment for ICU admission. Patients were randomized to either ED standard care or glycemic control. Glycemic control involved use of an insulin drip to maintain blood glucose levels between 80-140 mg/dL. Glycemic control continued until ED discharge. Standard patients were managed at ED attending physician discretion. We assessed severity of illness by calculation of APACHE II score. The primary endpoint was in-hospital mortality. Secondary endpoints included vasopressor requirement, hospital length of stay, and mechanical ventilation requirement. Fifty patients were randomized, 24 to the glycemic group and 26 to the standard care cohort. Four of the 24 patients (17%) in the treatment arm did not receive insulin despite protocol requirements. While receiving insulin, three of 24 patients (13%) had an episode of hypoglycemia. By chance, the patients in the treatment group had a trend toward higher acuity by APACHE II scores. Patient mortality and morbidity were similar despite the acuity difference. There was no difference in morbidity and mortality between the two groups. The benefit of glycemic control may be subject to source of illness and to degree of glycemic control, or have no effect. Such questions bear future investigation.


Implementing Evidence-Based HIV Prevention for Female Sex Workers in Mexico: Provider Assessments of Feasibility and Acceptability.
This study examined service provider perceptions of feasibility and acceptability of implementing evidence-based practices for preventing HIV/AIDS and STIs in female sex workers (FSWs) in Mexico. Semi-structured interviews were conducted with 124 directors, supervisors and counselors from 12 reproductive health clinics located throughout Mexico participating in a large randomized controlled trial to scale-up the use of a psychoeducational intervention designed to promote FSW condom use and enhance safer sex negotiation skills. Feasibility was based on assessment of personal, organizational and social costs, benefits, and capacity. Costs included anxiety over intervention competency, purchase of condoms and other supplies, expenses of laboratory tests for HIV/STIs, and stigma associated with FSWs. Benefits included increase in personal knowledge and experience, enhanced clinic reputation and service delivery capacity, and increased public health. Capacity was expressed in terms of provider skills to deliver the intervention and additional workload. Acceptability was expressed in terms of provider enthusiasm in delivering these services and FSWs willingness to receive the services. Service provider evaluations of feasibility and acceptability of implementing evidence-based prevention interventions are grounded in local contexts that define costs and benefits of and capacity for implementation and determine features of the intervention and its implementation that are acceptable.


Limb length change after total knee arthroplasty for valgus deformity does not affect the clinical score and is not affected by fixed flexion deformity.
The limb length change (LLC) after total knee arthroplasty (TKA) is especially significant in valgus deformity. The higher LLC could cause higher incidences of lower limb length discrepancy (LLD) and low clinical score. However, studies about LLC after TKA for valgus deformity are limited, and there are none on the relationship between LLC and fixed flexion deformity (FFD) in valgus deformity. (1) The amount of LLC would affect the postoperative LLD, (2) the improvement of fixed flexion deformity (FFD) would affect the amount of LLC, (3) The amount of LLC would affect the improvement in the clinical score after TKA for valgus deformity. Fifty knees of 50 patients who underwent primary unilateral TKA for valgus-type osteoarthritis between January 2000 and October 2021 were included. A radiological and clinical assessment were performed the day before the operation and at 12 months post-operatively. Full-length standing anteroposterior radiographs were used to measure HKA and LLC. FFD and Hospital for Special Surgery (HSS) score were measured in the outpatient department. The incidence of lengthening was 92.0% and the mean LLC was 18.85mm (SD, 19.60mm). Postoperative LLD over 10 mm occurred in 26% and the mean of postoperative LLD was 4.21mm (SD, 7.96mm). The LLC was correlated with postoperative LLD (rs=0.357, p=0.011) and the HKA change (rs=0.375, p=0.007), but not with the FFD improvement (rs=0.164, p=0.255) and HSS improvement (rs=0.076, p=0.613) or postoperative HSS (rs=0.094, p=0.528). LLC was affected by HKA improvement but not by FFD improvement after TKA for patients with valgus deformity. Additionally, LLC did not affect the clinical score. III; retrospective cohort study.


Leptin Signaling in Liver Tissue of a Transgenic Breast Cancer Mouse Model.
Leptin, an adipocytokine, is secreted from various tissues including the liver. The roles of both leptin and leptin receptor (ObR) in numerous pathophysiological conditions including mammary tumor (MT) development have been reported. However, the roles of leptin signaling-related proteins in the liver have not been reported previously in MT development. The objective of this study was to examine the expression levels of leptin and ObR in liver tissue of a transgenic breast cancer mouse model to investigate whether the roles of leptin in MT development are systemic or local. MMTV-TGF-α transgenic female mice were fed ad-libitum from week 10 up to week 74. Protein expression levels of leptin and ObR were measured in liver tissues of 74-week-old MMTV-TGF-α mice with and without MT by western blot. Serum leptin and insulin levels were measured using a enzyme-linked immunosorbent assay. Protein expression levels of leptin and ObR were similar in mice with MT compared to the ones without MT. Serum leptin and insulin levels were also not significantly different between the two groups. These results indicate that the effects of leptin signaling in MT development might be important at a local tissue level, such as mammary fat pad, and not as important at a systemic level.


The Drosophila tumor necrosis factor receptor-associated factor-1 (DTRAF1) interacts with Pelle and regulates NFkappaB activity.
A member of the tumor necrosis factor (TNF) receptor-associated factor (TRAF) family was identified in Drosophila. DTRAF1 contains 7 zinc finger domains followed by a TRAF domain, similar to mammalian TRAFs and other members of the family identified in data bases from Caenorhabditis elegans, Arabidopsis, and Dictyostelium. Analysis of DTRAF1 binding to different members of the human TNF receptor family showed that this protein can interact through its TRAF domain with the p75 neurotrophin receptor and weakly with the lymphotoxin-beta receptor. DTRAF1 can also self-associate and binds to human TRAF1, TRAF2, and TRAF4. Interestingly, DTRAF1 interacts with human cIAP-1 and cIAP-2 but not with Drosophila DIAP-1 and -2. By itself, DTRAF1 did not induce significant NFkappaB activation when overexpressed in mammalian cells, although it specifically increased NFkappaB induction by TRAF6. In contrast, TRAF2-mediated NFkappaB induction was partially inhibited by DTRAF1. Mutants of DTRAF1 lacking the N-terminal region inhibited NFkappaB induction by either TRAF2 or TRAF6. DTRAF1 specifically associated with the regulatory N-terminal domain of Pelle, a Drosophila homolog of the human kinase interleukin-1 receptor-associated kinase (IRAK). Interestingly, though Pelle and DTRAF1 individually were unable to induce NFkappaB in a human cell line, co-expression of Pelle and DTRAF1 resulted in significant NFkappaB activity. Interactions of DTRAF1 with human TRAF-, TNF receptor-, and IAP-family proteins imply strong evolutionary conservation of TRAF protein structure and function throughout Metazoan evolution.


Acute feasibility study of a novel device for the treatment of mitral regurgitation in a normal canine model.
: The purpose of this study was to evaluate the implantability of a novel epicardial mitral annuloplasty device and its ability to reduce the septal-lateral (S-L) dimension of the mitral annulus. : The devices were implanted on the beating heart in 2 healthy dogs (the 24-mm long device in dog A and the 27-mm and 24-mm standard devices in dog B) by sliding the anterior arm onto the floor of the transverse sinus and positioning the posterior arm just apical to the atrioventricular groove on the left ventricular posterolateral wall. The devices were secured with titanium helical tacks driven through the device into the ventricular wall. Two-dimensional epicardial echocardiograms were performed before and after device implantation to evaluate the degree of mitral regurgitation (MR) and the S-L dimension. : Device implantation was uneventful, taking only ∼30 seconds to deploy. MR (1+) in both dogs at baseline was reduced to zero after implant. The reductions in S-L dimension in systole for the 24-mm device were 7.5% in dog A and 30.5% in dog B. For the 27-mm device in dog B, S-L reduction in systole was 29.9%. The leaflet coaptation length was increased in both cases. : The new device was effective in reducing S-L dimension and 1+ MR without requiring the use of cardiopulmonary bypass. We are currently evaluating this device for the treatment of MR in a rapid-pacing canine heart failure model.


Water Transport and Thermomechanical Properties of Ti3C2Tz MXene Epoxy Nanocomposites.
Herein, we present the fabrication of dispersed, 5.0 wt % (1.74 vol %) Ti3C2Tz MXene epoxy nanocomposites (NCs), and report on their water transport and mechanical properties. To make the composites, Li+ ions between Ti3C2Tz MXene multilayers, MLs, present after the etching step were exchanged with either 12-aminolauric acid, ALA, or di(hydrogenated tallow)benzyl methyl ammonium chloride, DHT. After drying, the resulting ML powders were added at room temperature to the epoxy resin (diglycidyl ether of bisphenol A), followed by the curing agent, triethylenetetramine. The NCs were characterized by X-ray diffraction, thermogravimetric analysis, dynamic vapor sorption, dynamic mechanical analysis, scanning and transmission electron microscopies, and infrared spectroscopy. From XRD, the lack of signature MXene basal peaks, as well as evidence of exfoliation supported by TEM micrographs, we conclude that the MXene ML had indeed been intercalated by the epoxy. The distribution of the exfoliated multilayers, MLs, however, was not uniform. Nevertheless, our relative permeabilities, with a 1.74 vol % loading, are 5 times lower than results obtained in the carbon- or clay-reinforced epoxy NC literature. The lower permeabilities are due to reductions in both solubilities and diffusivities relative to the neat polymer. In the case of DHT, the water solubility at all temperatures was almost halved. The mechanical properties and thermal stability are found to be slightly improved with the addition of DHT-MXene. As far as we are aware, this is the first report of exfoliation of MXene in an epoxy matrix. Additionally, this study is the first to measure the diffusion of water in MXene epoxy NCs. More work on better dispersion of the MLs is indicated and ongoing.


Histone deacetylase 3 inhibition alleviates 2,4-dinitrochlorobenzene-induced atopic dermatitis via epigenetically upregulating Nrf2/HO-1 signaling pathway.
Atopic dermatitis (AD) is a frequent skin disorder that is associated with immune dysfunction and skin inflammation. Histone deacetylase 3 (HDAC3) possesses strong immune and inflammatory modulatory properties in multiple diseases. However, the role and mechanism of HDAC3 in AD remain unknown. Here, we reported that HDAC3 expression was aberrantly upregulated in 2,4-dinitrochlorobenzene (DNCB)-induced lesional AD skin in mice. Inhibition of HDAC3 by RGFP966 protected against DNCB-induced AD, indicated by improved histological damages, relieved inflammatory and immune dysfunction. Nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) signaling pathway activity in lesional AD skin was significantly decreased and RGFP966 attenuated the decrease. Inhibition of Nrf2/HO-1 signaling pathway via Nrf2 inhibitor ML385 blunted anti-AD effect of RGFP966 in DNCB-treated mice. Mechanistically, RGFP966 promoted Nrf2 expression and upregulated H3K27ac deposition on the promoter region of Nrf2. Collectively, HDAC3 inhibition protects against AD via epigenetically activating Nrf2 transcription to upregulate Nrf2/HO-1 signaling pathway activity. HDAC3 may act as a promising therapeutic target for the treatment of AD.


Efficacy of fibrin sealant (human) (Evicel) in rhinoplasty: a prospective, randomized, single-blind trial of the use of fibrin sealant in lateral osteotomy.
To determine whether patients receiving fibrin sealant placed in a single lateral osteotomy site during rhinoplasty will note substantial improvement in pain, bruising, swelling, and overall healing compared with the untreated side and to determine whether blinded observers detect a substantial difference in bruising and swelling on the basis of review of standard postoperative photographs. We conducted a prospective, randomized, single-blind, controlled trial of the use of fibrin sealant (human) (Evicel; Johnson & Johnson-Wound Management, Somerville, New Jersey) in 10 consecutive patients undergoing lateral osteotomy in rhinoplasty. Written consent was obtained from all participants. Each patient was randomized for the use of fibrin sealant on either the right or the left side with the contralateral side acting as the control. Patients were evaluated on postoperative days 1, 7, and 21 with standard photographic views and a patient questionnaire. The blinded observers consisted of 5 raters familiar with the outcomes and results of rhinoplastic surgery. The observers evaluated all photographs and completed a grading scale to define bruising and swelling on each side. The mean patient age was 41 years (age range, 21-66 years). Half of the patients were women. The blinded observer Wilcoxon rank sum test revealed a statistically significant difference on postoperative day 1 for bruising (P< .03; Wilcoxon critical z value, 1.99) and swelling (P< .01; 2.41). Similar findings were discovered on postoperative day 7 for both bruising and swelling (P< .03). On postoperative day 21, bruising retained statistical significance (P< .05); however, swelling did not achieve statistical significance. Patient questionnaires were evaluated and significance was determined for the treated compared with the untreated side of the nose on postoperative days 1, 7, and 21. Categories included pain, bruising and swelling, and overall rate of healing. The Wilcoxon rank sum test revealed no significance for pain or overall rate of recovery (P> .06) on postoperative days 1, 7, or 21. However, bruising and swelling both achieved statistical significance. On postoperative day 1, both pain and swelling scales achieved a significance of P< .01 (Wilcoxon critical z value, 2.34). On postoperative day 7, bruising achieved significance at P< .005 (Wilcoxon critical z value, 2.63) and swelling achieved significance at P< .01 (2.45). Both bruising and swelling achieved equal significance on postoperative day 21 (P< .01; Wilcoxon critical z value, 2.57 and 2.45, respectively). Fibrin sealant applied to a lateral osteotomy site significantly reduced bruising and swelling per patient report on postoperative days 1, 7, and 21. Physician observation reported significant reduction in bruising on postoperative days 1, 7, and 21 and reduction in swelling on postoperative days 1 and 7. The ease of application and versatility of fibrin sealant enable rapid healing after rhinoplasty and produce increased patient satisfaction.


The prevalence of familial testicular cancer: an analysis of two patient populations and a review of the literature.
Undescended testes and antecedent testicular tumor are recognized risk factors for testicular germ cell cancer. It has been suggested that a family history of testicular cancer constitutes another major risk indicator. This postulation is mainly based on clinical observations and on very few systematic investigations. In the current study, the authors analyzed the proportion of familial testicular cancer in their study population and estimated the relative risk created by a family history of the disease. The proportion of familial testicular cancer was analyzed in a prospective multicentric study involving 1692 patients. The median ages of patients with and without a family history of the disease were compared. In a different patient population consisting of 518 patients and 531 age-matched controls, the frequency of family history was investigated and the relative risk calculated. In addition, a literature survey was performed to look for previous systematic reports on familial testicular cancer. In the prospective study, 18 patients (1.1%; 95% confidence interval, 0.63-1.68%) had a first-degree relative afflicted with testicular cancer. Age at presentation was not significantly different between patients with a family history and those without. In the retrospective series, the proportion of those with a family history was 1.7% (95% confidence interval [CI], 0.80-3.27%). There was a 3.1-fold increased relative risk (95% CI, 0.77-17.95) for first-degree relatives of patients with testicular cancer. Ten previous reports on familial testicular cancer were identified in the literature. Combining the results of those previous reports and the current study led to a weighted mean prevalence of familial testicular cancer of 1.35% (95% CI, 1.12-1.58%). The current study underscores that susceptibility to testicular germ cell cancer is influenced by genetic factors. A family history of testicular cancer is encountered in about 1.35% of patients. The relative risk for first-degree relatives of patients with the disease is increased by a factor of 3-10.


Crosstalk between Iron Metabolism and Erythropoiesis.
Iron metabolism and erythropoiesis are inextricably linked. The majority of iron extracted from circulation daily is used for hemoglobin synthesis. In the last 15 years, major advances have been made in understanding the pathways regulating iron metabolism. Hepcidin is a key regulator of iron absorption and recycling and is itself regulated by erythropoiesis. While several viable candidates have been proposed, elucidating the "erythroid regulator" of hepcidin continues to generate significant experimental activity in the field. Although the mechanism responsible for sensing iron demand for erythropoiesis is still incompletely understood, evaluating diseases in which disordered erythropoiesis and/or iron metabolism are showcased has resulted in a more robust appreciation of potential candidates coordinated erythroid iron demand with regulators of iron supply. We present data drawn from four different conditions-iron deficiency, congenital hypotransferrinemia, beta-thalassemia, and hereditary hemochromatosis-both in human and non-human models of disease, together suggesting that erythroid iron demand exerts a stronger influence on circulating iron supply than systemic iron stores. Greater understanding of the interplay between the key factors involved in the regulation of iron metabolism and erythropoiesis will help develop more effective therapies for disorders of iron overload, iron deficiency, and hemoglobin synthesis.


Uptake of core outcome sets by clinical trialists publishing in major medical journals: Protocol.
Background: Outcome heterogeneity, selective reporting, and choosing outcomes that do not reflect needs and priorities of stakeholders, limit the examination of health intervention effects, particularly in late phase trials. Core outcome sets (COS) are a proposed solution to these issues. A COS is an agreed-upon, standardised set of outcomes that should be measured and reported as a minimum in all trials in a specific area of health or healthcare. COS are intended to increase standardisation of outcome measurement and reporting to better enable comparisons between, and synthesis of findings of trials in a particular health area. Methods: This study will examine late phase trials, published between October 2019 and March 2020 (inclusive), in the following five medical journals: New England Journal of Medicine, Journal of the American Medical Association, Lancet, BMJ, and Annals of Internal Medicine. Trials will be examined to determine if they refer to a COS, and whether they use a COS. Trialists for each identified trial will subsequently be contacted to complete an online survey examining trialists' awareness of, and decisions to search for and use a COS. Discussion: This study will provide important information on uptake of COS by later phase trialists in major medical journals, and the views of these trialists on COS use in trials. These findings will inform approaches to increasing awareness and uptake of COS in future health trials.


[Restoring treatment with continuous insulin infusion pump (BICI)].
Significant advances in the treatment of diabetes have been made over the past few decades, although insulin treatment is the only option available until now for Type I diabetes patients. Type I diabetes is generally diagnosed in youths; therefore, micro insulin injectors enable doctors to offer them a better life style, preventing future complications. Medical professionals contemplate the possibility to use this treatment on patients over age 60 with satisfactory results. Continuous insulin infusion pumps (BICI) provide physiologically programmable subcutaneous administration of rapid acting insulin. Due to technological advances, these pumps have become safer more efficient and easier to use which improves the glucose level in blood and enables diabetes patients to carry out many activities without losing metabolic control, reduces the incidences of hypoglycemia and improves diabetes patients' life styles.


Linc01105 acts as an oncogene in the development of neuroblastoma.
Previous research from our group revealed that the long coding RNA (lncRNA) linc01105 is associated with neuroblastoma proliferation and apoptosis, and that its expression is correlated with the International Neuroblastoma Staging System stage. The purpose of the present study was to investigate the functions of Linc01105 in neuroblastoma. Lentivirus‑mediated linc01105 knockdown was performed in the neuroblastoma cell line SH‑SY5Y. The expression levels of linc01105 and of other associated genes were measured by reverse transcription‑quantitative PCR. Cell Counting Kit‑8 assay and flow cytometry were used to determine cell viability and apoptosis. The levels of proteins were detected using western blot analysis. Bioinformatics analysis and luciferase reporter assays were used to examine the relationship between linc01105, miR‑6769b‑5p and vascular endothelial growth factor A (VEGFA). Angiogenesis ability was measured using a tube formation assay. The results demonstrated that HIF‑1α overexpression promoted the transcription of linc01105 by acting as a transcription factor. Knockdown of linc01105 inhibited neuroblastoma cell proliferation, migration and invasion, and it induced apoptosis. In addition, linc01105 affected the expression of p53 and Bcl‑2 family proteins and activated the caspase signaling pathway. Further functional experiments revealed that linc01105 promoted the expression of the miR‑6769b‑5p target gene VEGFA by acting as a sponge of miR‑6769b‑5p. In conclusion, linc01105 may contribute to neuroblastoma tumorigenesis and development. The present findings indicated that the interplay between the p53/caspase pathway and the linc01105/miR‑6769b‑5p/VEGFA axis may have important roles in the development of neuroblastoma.


A new subfamily of the Nudix hydrolase superfamily active on 5-methyl-UTP (ribo-TTP) and UTP.
A new subfamily of the Nudix hydrolases, identified by conserved amino acids upstream and downstream of the Nudix box, has been characterized. The cloned, expressed, and purified orthologous enzymes have major activities on the non-canonical nucleoside triphosphate 5-methyl-UTP (ribo-TTP) and the canonical nucleotide UTP. In addition to their homologous signature sequences and their similar substrate specificities, the members of the subfamily are inhabitants of or are related to the bacterial rhizosphere. We propose the acronym and mnemonic, utp, for the gene designating this unique UTPase.


Percutaneous ultrasonic lithotripsy (PUL) after shock wave lithotripsy (SWL) failure.
Shock wave lithotripsy (SWL) is an attractive initial treatment for nephrolithiasis. Unfortunately, a significant number of stones are resistant to SWL therapy and require subsequent percutaneous ultrasonic lithotripsy (PUL) for definitive treatment. Our objective was to determine if previous SWL had adverse effects on PUL success and if there were differences between the patients undergoing primary PUL and those undergoing PUL after SWL failure. In 2001, 108 PULs were performed at our institution, of which 40 (37%) were performed after SWL failure. Stone location, anesthesia time, stone composition and size, and complication rates were compared between patients who had PUL alone and those who underwent PUL after SWL failure. Anesthesia time was considered a reflection of technical difficulty of the case. Stone composition differed between the SWL failure and primary PUL groups. Cystine stones were found to be more common in the ESWL failure group and calcium oxalate monohydrate stones more common in the primary PUL group. There was not a statistically significant difference in stone size, anesthesia time or complication rates, between the PUL alone and PUL after SWL failure groups. PUL remains a reliable and safe treatment of nephrolithiasis. Prior SWL does not affect efficacy, technical difficulty, or postoperative complications of subsequent PUL. Based on our data, prior SWL should not be considered a negative factor in PUL outcomes. Stone composition should be considered a prognostic indicator of SWL failure and alert the surgeon that primary PUL may be indicated.


An ethnographic exploration of influences on prescribing in general practice: why is there variation in prescribing practices?
Prescribing is a core activity for general practitioners, yet significant variation in the quality of prescribing has been reported. This suggests there may be room for improvement in the application of the current best research evidence. There has been substantial investment in technologies and interventions to address this issue, but effect sizes so far have been small to moderate. This suggests that prescribing is a decision-making process that is not sufficiently understood. By understanding more about prescribing processes and the implementation of research evidence, variation may more easily be understood and more effective interventions proposed. An ethnographic study in three Scottish general practices with diverse organizational characteristics. Practices were ranked by their performance against Audit Scotland prescribing quality indicators, incorporating established best research evidence. Two practices of high prescribing quality and one practice of low prescribing quality were recruited. Participant observation, formal and informal interviews, and a review of practice documentation were employed. Practices ranked as high prescribing quality consistently made and applied macro and micro prescribing decisions, whereas the low-ranking practice only made micro prescribing decisions. Macro prescribing decisions were collective, policy decisions made considering research evidence in light of the average patient, one disease, condition, or drug. Micro prescribing decisions were made in consultation with the patient considering their views, preferences, circumstances and other conditions (if necessary).Although micro prescribing can operate independently, the implementation of evidence-based, quality prescribing was attributable to an interdependent relationship. Macro prescribing policy enabled prescribing decisions to be based on scientific evidence and applied consistently where possible. Ultimately, this influenced prescribing decisions that occur at the micro level in consultation with patients. General practitioners in the higher prescribing quality practices made two different 'types' of prescribing decision; macro and micro. Macro prescribing informs micro prescribing and without a macro basis to draw upon the low-ranked practice had no effective mechanism to engage with, reflect on and implement relevant evidence. Practices that recognize these two levels of decision making about prescribing are more likely to be able to implement higher quality evidence.


[Lysosomal component in the mechanism of the toxic effect of sporofusarin].
The effect of Fusarium sporotrichiella v. sporotrichioides mycotoxin (sporofusarin) on the total and non-sedimentary supernatant activity of 13 marker-enzymes of subcellular particles (2 mitochondrial enzymes-cytochrome oxidase and malate dehydrogenase; 8 lysosomal enzymes -- acid phosphatase, acid RNAase, acid DNAase, arylsulphatases A and B, beta-N-acetylglucosaminidase, beta-glucuronidase, beta-galactosidase and beta-glucosidase; 2 microsomal enzymes -- glucose-6-phosphatase and acetylesterase; plasma membrane enzyme -- alkaline phosphatase) of the rat liver, kidney, spleen and bone-marrow was studied in in vivo experiments. The latter demonstrated that sporofusarin effects were characterized by a significant organ and organella specificity, viz. the toxin caused a sharply increased activity, mainly of lysosomes enzymes and labilization of the lysosomal membranes, primarily in the spleen and the bone-marrow. A conclusion is drawn that the discovered selective destructive action of sporofusarin on the lysosomes may be regarded as a new phenomenon that, possibly is directly related to the characterization of the mechanism responsible for a specific effect produced by sporofusarin.


Analysis of the dominant mutation N188T of human connexin46 (hCx46) using concatenation and molecular dynamics simulation.
Connexins (Cx) are proteins that form cell-to-cell gap junction channels. A mutation at position 188 in the second extracellular loop (E2) domain of hCx46 has been linked to an autosomal dominant zonular pulverulent cataract. As it is dominantly inherited, it is possible that the mutant variant affects the co-expressed wild-type Cx and/or its interaction with other cellular components. Here, we proposed to use concatenated hCx46wt-hCx46N188T and hCx46N188T-hCx46wt to analyze how hCx46N188T affected co-expressed hCx46wt to achieve a dominant inheritance. Heterodimer hCx46wt-hCx46N188T formed fewer gap junction plaques compared to homodimer hCx46wt-hCx46wt, while the hCx46N188T-hCx46N188T homodimer formed almost no gap junction plaques. Dye uptake experiments showed that hemichannels of concatenated variants were similar to hemichannels of monomers. Molecular dynamics simulations revealed that for docking, the N188 of a protomer was engaged in hydrogen bonds (HBs) with R180, N189, and D191 of the counterpart protomer of the adjacent hemichannel. T188 suppressed the formation of HBs between protomers. Molecular dynamics simulations of an equimolar hCx46wt/hCx46N188T gap junction channel revealed a reduced number of HBs between protomers, suggesting reduction of gap junction channels between lens fibers co-expressing the variants.


Parental drinking in Australia: Does the age of children in the home matter?
Parental role modelling of alcohol use is known to influence alcohol consumption in adolescence and in later life. This study aimed to assess relationships between parental status, child age and alcohol consumption, which have not been well documented. Data were sourced from the 2013 Australian National Drug Strategy Household Survey. Analyses were conducted for 25-55 year olds (n = 11 591) by parental status, gender and age of youngest child in the household, controlling for socio-demographic factors. Parents were less likely than non-parents to exceed the alcohol guideline for increased lifetime risk (18.2% vs. 24.2%) and short-term risk: at least weekly (14.2% vs. 21.2%); and at least monthly (27.5% vs. 35.9%). Fathers were just as likely to exceed the guidelines for lifetime risk as other men, but those with children aged 0-2, were less likely to exceed the guideline for short-term risk. Women were least likely to exceed the guideline for lifetime risk if they had children aged 0-2, 6-11 or 15 years and over, or the guideline for short-term risk, if they had children aged 0-2, or 15 years and over in the household. Parents were more likely to report drinking in the home. Parents were less likely to exceed alcohol guidelines than non-parents, especially mothers whose youngest child was an infant or in high school or older. Consistent with population rates in men, fathers were more likely to exceed alcohol guidelines than mothers, and this excess consumption warrants public health attention.


Evaluation of changes in the vertical facial dimension with different anchorage systems in extraction and non-extraction subjects treated by Begg fixed appliances: a retrospective study.
The aim of the study was to evaluate and compare the effects of mesial movement of the maxillary and mandibular molars on the facial vertical dimensions following orthodontic treatment. Patients with an Angle Class I malocclusion were treated by four first premolar extractions and Begg fixed appliances (moderate and maximum anchorage groups), while those with an Angle Class II malocclusion were treated by Begg intraoral distalization mechanics without extractions (distalization group). Following treatment, the patients were grouped according to the mesial movement of the mandibular first molars and compared with an Angle Class I control group. All groups comprised 15 patients, their mean pre-treatment ages were 14.95 years for the moderate (13 females, 2 males), 14.88 years for the maximum (13 females, 2 males), 14.41 years for the distalization (10 females, 5 males), and 14.38 years for the control (13 females, 2 males) groups. Lateral cephalometric measurements were performed at two time points (T(1): pre-treatment/control, T(2): post-treatment/control). A paired t-test was used for within-group comparisons and non-parametric Kruskal-Wallis and Dunn's multiple-comparison tests to determine any differences. The increases in anterior and posterior face heights were similar between groups. The mandibular plane angle (SN/GoGn) was increased in all treatment groups, while it decreased in the control group. Although SN/GoGn increased more in the distalization and less in the moderate anchorage groups, these differences were not statistically significant. The change in SN/GoGn was significantly different only between the distalization and control groups. Facial vertical dimensions were not significantly affected by the amount of mesial movement of the molar teeth.