Phospholipases modulate the rat testicular androgen biosynthetic pathway in vitro.
The role of membrane phospholipids in testicular androgen biosynthesis was investigated by monitoring the effects of phospholipase treatments on the activities of the steroid transforming enzymes. Androgen biosynthesis in untreated rat testicular microsomes was examined by monitoring the temporal appearance of pregnenolone metabolites and was found to proceed through the 4-ene route. When phospholipase A2 was included, the 5-ene steroids 17-hydroxypregnenolone and dehydroepiandrosterone (DHEA) were formed in greater quantities, and the production of 4-ene steroids was reduced indicating that the conversion of 5-ene steroids to the 4-ene configuration was inhibited by phospholipase A2 treatment. Phospholipase C, in addition to inhibiting this step, also inhibited the conversion of C21 steroids to C19 steroids. When the enzymatic steps were measured individually, phospholipase A2 inhibited 3 beta-hydroxysteroid dehydrogenase-isomerase (3 beta-HSD-Isomerase) with an ED50 of 73 mU/ml but had no effect on the activities of 17-hydroxylase, C-17, 20 lyase, or 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD). However, though phospholipase C treatment inhibited 3 beta-HSD-Isomerase, it caused less inhibition (the ED50 value was 149 mU/ml). Furthermore, 17-hydroxylase and C-17, 20 lyase activities were also inhibited by phospholipase C treatment (ED50 values were 410 and 343 mU/ml, respectively), but no effect on 17 beta-HSD was observed. The differences in the apparent phospholipid requirements of the steroidogenic enzymes provides the possibility that the metabolic fate of pregnenolone may be regulated by changes in the phospholipid composition of the microenvironment.


[Study on the prevalence rate of hypertension in Guangxi Hei Yi Zhuang population].
To study the prevalence rate of hypertension and the risk factors in Guangxi Hei Yi Zhuang population. A total of 1068 people of Hei Yi Zhuang nationality aged 20 and over were surveyed. Blood pressure, height, weight, serum lipid and apolipoprotein levels were measured ,and both body surface areas and body mass index were calculated. Results were compared with those in 933 people of Han nationality living in the same district. The prevalence rates of hypertension in Hei Yi Zhuang population were significantly higher than that in Han population (25.2% vs. 17.3%, P < 0.001) ,as well as the isolated systolic hypertension (12.5% vs. 3.9%, P < 0.001). The mean levels of systolic pressure and pulse pressure in Hei Yi Zhuang population were significantly higher than those in Han population (125.20 +/- 18.62 vs. 121.88 +/- 15.99 mm Hg, P < 0.001 and 48.64 +/- 14.75 vs. 44.98 +/- 11.12 mm Hg, P <.001; respectively). The prevalence rate of hypertension in Hei Yi Zhuang population was positively correlated with triglyceride, sex and age whereas the prevalence rate of hypertension in Han population was positively correlated with total cholesterol, sex, age, and alcohol consumption. Rates of awareness on hypertension in Hei Yi Zhuang and Han population were 8.6% vs. 21.1% (P <.001), patients with treatment rate of established hypertension were 4.5 vs. 15.5 (P < 0.001) with the control rates as 1.9 vs. 10.6 (P < 0.001). The prevalence rate of hypertension in Guangxi Hei Yi Zhuang population was significantly higher than that in Han ethnic group. The causes of high prevalence rate of hypertension might be ascribed to special geographical surroundings, unhealthy life style, high sodium intake, low education, and possibly genetic factors. The rates on awareness, treatment and control of hypertension were still under satisfaction.


Real-time noise cancellation for speech acquired in interactive functional magnetic resonance imaging studies.
To present online scanner noise cancellation for speech acquired in functional magnetic resonance imaging (fMRI) studies. An online active noise cancellation method for speech acquired in fMRI studies was developed. The approach consists of two automated steps: 1) creation of an MR noise template in a short "test" fMRI scan; 2) application of the template for automatic recognition and subtraction of the MR noise from the acquired microphone signal during an fMRI study. The method was applied in an experimental paradigm where a subject and an investigator communicated in an interactive verbal generation task during fMRI. By applying online active noise cancellation, the quality of the subject's speech was substantially improved. The present approach was found to be flexible, reliable, and easy to implement, providing a method for fMRI studies that investigate the neural correlates of interactive speech communication. Using online noise cancellation it is possible to improve the quality of acquired speech in fMRI. This approach may be recommended for interactive fMRI studies.


Attitudes amongst Australian hospital healthcare workers towards seasonal influenza and vaccination.
Amongst healthcare workers (HCWs), compliance rates with influenza vaccination are traditionally low. Although a safe and effective vaccine is available, there is little Australian data on reasons for poor compliance, especially amongst allied health and ancillary support staff. Cross-sectional investigation of a sample of clinical and non-clinical HCWs from two tertiary-referral teaching hospitals in Sydney, Australia was conducted between June 4 and October 19, 2007. The self-administered questionnaire was distributed to hospital personal from 40 different wards and departments. The main outcome measures were personal beliefs about influenza vaccination and self-reported vaccination status. Respondents (n = 1079) were categorized into four main groups by occupation: nurses (47.5%, 512/1079), physicians (26.0%, 281/1079), allied health (15.3%, 165/1079) and ancillary (11.2%, 121/1079). When asked whether they felt the influenza vaccine was safe or effective, 81% (879/1079) and 68% (733/1079), respectively, replied in the affirmative. Participants felt that it was more important to get vaccinated to protect patients (74%, 796/1079) than family (68%, 730/1079) or self-protection (66%, 712/1079). However, only 22% (241/1079) of the HCWs who replied reported receiving the vaccine the year the survey was conducted. Although HCWs had an adequate level of knowledge towards influenza vaccination, only 22% of them were vaccinated. The approach to improving influenza vaccination rates amongst HCWs and to tackling misconceptions must be multifaceted, adaptable and must evolve regularly to increase coverage.


Concanavalin (ConA)-reactive human epidermal glycoproteins.
The possibility that glycoconjugates play a role in epidermal differentiation has stimulated attempts at their biochemical identification. We have examined the Concanavalin (ConA)-binding glycoproteins present: in whole human epidermis, epidermal cells obtained by trypsinization of human skin, non dissociated layers (the most differentiated ones) remaining after preparation of these cells and human epidermal cell cultures. 125I-ConA was applied to NP-40 soluble molecules separated by SDS-PAGE. ConA labelling of the whole epidermal extract visualized numerous bands. That of trypsinized cells differed in the intensity of some bands and the absence of others. However most of the latter bands could be identified in the most differentiated layers. These glycoproteins were absent or poorly labelled in the cultures of keratinocytes. The restriction of some ConA-binding glycoproteins to the most differentiated epidermal layers, suggests that these molecules are involved in, or can be considered as markers of the human epidermal differentiative process.


The diagnostic role of 99mTc-dual receptor targeted probe and targeted peptide bombesin (RGD-BBN) SPET/CT in the detection of malignant and benign breast tumors and axillary lymph nodes compared to ultrasound.
This study aimed to explore the diagnostic role of a new dual receptor-targeted probe, integrin ανβ3 and gastrin releasing peptide receptor (GRPR) targeted peptide Glu-c(RGDyK)-bombesin (RGD-BBN) labeled with technetium-99m ((99m)Tc-RGD-BBN), using single photon emission tomography/computed tomography (SPET/CT) in the detection of breast tumor in comparison to ultrasound (US). One hundred and twenty six female patients with suspicious breast lesions who had already been scheduled for biopsy or surgery were enrolled in this study. All patients had previously underwent breast US and (99m)Tc-RGD-BBN SPET/CT. The US findings were evaluated according to the breast imaging report and the data system (BI-RADS). Technetium-99m-RGD-BBN SPET/CT images were interpreted independently by two experienced nuclear medicine physicians. A final diagnosis was made by histopathology of the specimens. A total of 130 lesions, 77 malignant and 53 benign lesions were ascertained. One hundred and twelve breast lesions, 69 malignant and 43 benign lesions were above 10mm in diameter and 18 breast lesions (8 malignant lesions and 10 benign lesions) were below 10mm. The overall sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of (99m)Tc-RGD-BBN SPET/CT and US for breast lesions were 93.5% vs. 81.8% (P<0.05), 79.2% vs. 75.5% (P>0.05), 86.7% vs. 82.9% (P>0.05), 89.4% vs. 74.1% (P<0.05) and 87.7% vs. 79.2% (P>0.05). Technetium-99m-RGD-BBN SPET/CT detected all lesions ≥10mm and US only detected 57 (P<0.05). In malignant lesions <10mm, US was superior than (99m)Tc-RGD-BBN SPET/CT (75.0% vs. 37.5%, P<0.05). There was no significant difference between the two methods no matter the size of the benign lesions. The overall sensitivity and specificity of (99m)Tc-RGD-BBN SPET/CT and US for axillae lymph nodes were 87.5% vs. 71.9% (P<0.05) and 77.6% vs. 68.9% (P>0.05), respectively. For the metastatic lymph nodes of <10mm, the sensitivity of (99m)Tc-RGD-BBN SPET/CT and of US was 88.5% and 72.1% respectively (P<0.05). Statistical analysis was not performed due to the small number of metastatic lesions of <10mm. The specificity of (99m)Tc-RGD-BBN SPET/CT and of US was not different, no matter the size of the axilla lymph nodes that had no metastases (P>0.05). Technetium-99m-RGD-BBN SPET/CT had higher sensitivity and NPV than US in detecting primary breast tumors and axilla lymph nodes and it also showed an advantage in distance metastatic lesions detection. On the contrary, specificity and PPV of the two methods were not different. Technetium-99m-RGD-BBN SPET/CT cannot totally replace US in the detection of primary breast cancer and axillary lymph nodes metastases. It can be used as an additional imaging tool of eliminating the necessity of surgical biopsy and histopathologic examination because of its high NPV.


Postoperative Patient Reported Outcomes After Cholesteatoma Surgery.
Results and success measures of cholesteatoma surgery are generally described using objective data whereas subjective data are mostly lacking. Patients experiences and complaints are becoming more important alongside clinical and audiometric outcome measures in cholesteatoma care. To investigate the course of patient-reported complaints, the impact of complaints, audiometric measures and the stability of audiometric measures, and complaints over time after primary and recurrent/residual cholesteatoma surgery. Postoperative patients were prospectively included and divided into primary acquired and recurrent/residual cholesteatoma. The EuroQol 5D (EQ-5D-3L), Otology Questionnaire Amsterdam (OQUA), and the Speech Spatial Questionnaire (SSQ) were completed by 144 patients up to 2 years postoperative. Patient-reported complaints divided in eight separate domains, postoperative hearing and impact on daily life were longitudinally assessed by means of linear mixed models. Hearing loss and tinnitus are the most reported postoperative complaints over time. Patient-reported loss of taste and the impact of all complaints decline over time. All other patient-reported complaints remain stable over time, only itch complaints fluctuate. Primary cholesteatoma patients score significantly higher on hearing loss complaints compared with recurrent/residual patients although they have comparable mean audiometric hearing loss. Furthermore, pure-tone hearing threshold, instead of asymmetric hearing loss, is correlated with the localization domain of the SSQ. This study provides important insights in the course of complaints and its impact on daily life after cholesteatoma surgery. Overall, the postoperative patient-reported complaints after cholesteatoma surgery are generally low in the studied population.


Reversible posterior leukoencephalopathy caused by azathioprine in systemic lupus erythematosus.
The authors report a case of systemic lupus erythematosus with posterior leukoencephalopathy who presented with headache, tonic-clonic seizure, loss of consciousness and bilateral loss of vision, after taking azathioprine for three weeks. The patient had hypertension with normal eye grounds. The brain CT showed a hypodensity lesion at both bilateral occipital lobes, mainly in the white matter The symptoms and follow-up MRI were improved after the control of hypertension and discontinuation of azathioprine.


[Peripheral intravenous catheter-related phlebitis].
Phlebitis is a very common complication of the use of intravenous catheters. Two patients with an i.v. catheter complicated by thrombophlebitis are described. Patient A was immunocompromised due to chronic lymphatic leukaemia and developed septic thrombophlebitis with positive blood cultures for S. Aureus. Patient B was being treated with flucloxacillin because of an S. Aureus infection and developed chemical phlebitis. Septic phlebitis is rare, but potentially serious. Chemical or mechanical types of thrombophlebitis are usually less severe, but happen very frequently. Risk factors include: female sex, previous episode of phlebitis, insertion at (ventral) forearm, emergency placement and administration of antibiotics. Until recently, routine replacement of peripheral intravenous catheters after 72-96 h was recommended, but randomised controlled trials have not shown any benefit of this routine. A recent Cochrane Review recommends replacement of peripheral intravenous catheters when clinically indicated only.


Embolic Showering from Non-Bacterial Thrombotic Endocarditis and Adenocarcinoma of the Lung.
Non-bacterial thrombotic endocarditis (NBTE) is a paraneoplastic phenomenon with sterile vegetations. It is associated with adenocarcinoma and can shower emboli, which can be the presenting symptom. A 44-year-old woman with adenocarcinoma of the lung presented with chest pain, left hand weakness, and ataxia due to repeated embolic showering from NBTE to the central nervous system (CNS) and spleen. NBTE is a rare condition that should be on the differential diagnosis in patients with culture-negative endocarditis and a history of adenocarcinoma. Differentiating non-bacterial thrombotic endocarditis (NBTE) from infective endocarditis can be a diagnostic challenge due to slow growing organisms; laboratory findings that suggest NBTE include the lack of leucocytosis, normal C-reactive protein, negative blood culture sets, and positive antiphospholipid antibodies.Serial transesophageal echocardiogram (TEE) should be performed if suspicion of valvular vegetations is high despite the initial TEE showing no vegetations.The mainstay treatment of NBTE is anticoagulation and addressing the underlying condition.


Children's conceptions of AIDS: a developmental analysis.
Examined causal reasoning about AIDS in children representing three major phases of cognitive development: prelogical, concrete logical, and formal logical thinking. 60 Ss (age groups: 5-7 years, 8-10 years, and 11-13 years) were administered the Concepts of AIDS Protocol. Responses were scored using the developmentally ordered Concepts of Illness Category System. The data confirm that, as a group, children's causal thinking about AIDS parallels the ways in which children think about illness in general. More specifically, the data could be organized in terms of 6 major categories or ways in which children conceptualize AIDS and its causes. The findings provide an initial empirical foundation for AIDS education curricula.


Psychotic spectrum disorders and alcohol abuse: a review of pharmacotherapeutic strategies and a report on the effectiveness of naltrexone and disulfiram.
The rate of substance-use disorders in patients with mental illnesses within the psychotic spectrum, such as schizophrenia, schizoaffective disorder, and bipolar disorder, is higher than the rate observed in the general population and is associated with significant morbidity and mortality. Although there are currently 3 medications approved by the Food and Drug Administration for the treatment of alcohol dependence, no medications have been approved for the specific treatment of dually diagnosed patients. A small but growing body of literature supports the use of 2 of these medications, disulfiram and naltrexone, in dually diagnosed individuals. This article outlines a review of the literature about the use of disulfiram and naltrexone for alcoholism and in patients with comorbid mental illness. In addition, results are presented of a 12-week randomized clinical trial of disulfiram and naltrexone alone and in combination for individuals with Axis I disorders and alcohol dependence who were also receiving intensive psychosocial treatment. Individuals with a psychotic spectrum disorder, including schizophrenia, schizoaffective disorder, and bipolar disorder, had worse alcohol outcomes than those without a psychotic spectrum disorder. Individuals with a psychotic spectrum disorder had better alcohol-use outcomes on an active medication compared with placebo, but there was no clear advantage of disulfiram or naltrexone or of the combination. Retention rates and medication compliance in the study were high and exceeded 80%. Pharmacotherapeutic strategies should take into account the advantages and disadvantages of each medication. Future directions of pharmacotherapeutic options are also discussed.


Platelet GP IIb-IIIa Receptor Antagonists in Primary Angioplasty: Back to the Future.
Coronary artery disease and acute myocardial infarction still represent the leading cause of mortality in developed countries. Therefore, great efforts have been made in the last decades to improve reperfusion strategies and adjunctive antithrombotic therapies. In fact, despite optimal epicardial recanalisation, a large proportion of patients still experience impaired reperfusion and in-stent thrombosis. The adjunctive use of glycoprotein (GP) IIb-IIIa inhibitors may certainly contribute in the reduction of such complications, especially when administered in the early phase of infarction. In fact, in this phase a larger platelet composition of the thrombus and the presence of a larger amount of viable myocardium, as compared to a delayed phase, may increase the benefits from this therapy and counterbalance the potential higher risk of bleeding. A large body of evidence has been accumulated on the benefits from GP IIb-IIIa inhibitors in terms of prevention of stent thrombosis, and benefits in mortality, especially among high-risk patients and as upstream strategy. Therefore, based on current available data, GP IIb-IIIa inhibitors can be recommended as early as possible (upstream strategy) among high-risk patients, such as those with advanced Killip class or anterior myocardial infarction (MI), and those presenting within the first three hours. Even though it is not universally accepted, in our opinion this strategy should be implemented in a pre-hospital setting (in ambulance) or at first hospital admission (Emergency Room or Coronary Care Unit, irrespective of whether they are in the spoke or hub hospitals). Peri-procedural intracoronary administration of GP IIb-IIIa inhibitors has not provided additional benefits as compared to intravenous administration and therefore cannot be recommended. Even though the vast majority of trials have been conducted with abciximab, several meta-analyses comparing small molecules (mainly high-dose tirofiban rather than eptifibatide) versus abciximab showed similar angiographic and clinical results between the molecules. Several recent investigations and meta-analyses have documented the higher risk of stent thrombosis associated with bivalirudin as compared to unfractionated heparin (UFH). Being that these results are independent from the use of GP IIb-IIIa inhibitors, UFH should still remain the anticoagulation therapy of choice in ST-segment elevation myocardial infarction (STEMI) patients. Minimisation of bleeding complications by extensive use of the radial approach, in the setting of STEMI, may further contribute to the adoption of a more aggressive antithrombotic and antiplatelet therapy incorporating the use of GP IIb-IIIa inhibitors. The establishment of dedicated networks for STEMI, and the large STEMI campaign, will certainly contribute to increase the proportion of patients presenting at first medical contact within the early phase (3 h) of infarction and therefore highly suitable for a more aggressive pharmacoinvasive approach with upstream administration of GP IIb-IIIa inhibitors. In fact, although the current therapeutic targets of increased rates of timely reperfusion, mainly by primary percutaneous coronary intervention (PCI), has been achieved, a deep look into the future in the fight against MI will certainly put aborting infarction as the major desirable target to be achieved.


Breaking and entering: predators invade the shelter of their prey and gain protection.
Many herbivorous arthropods construct shelters on their host plant that offer protection against natural enemies. This has resulted in selection on natural enemies to enter these shelters, where they can feed on prey that are inaccessible for competing predators and parasitoids. The spider mite Tetranychus evansi produces a shelter consisting of a dense web that is impenetrable for most predators; the only known natural enemy that can penetrate the web and can forage efficiently on this pest is Phytoseiulus longipes. We show that this predator preferentially foraged and oviposited in the web of its prey. Moreover, intraguild predation on juveniles of these predators was significantly higher outside this web and in the less dense web of a closely related prey species (T. urticae) than inside the web of T. evansi. Although the production of shelters by herbivores may be profitable at first, their adapted natural enemies may reap the benefit in the end.


QTL associated with Gummy Stem Blight (GSB) resistance in watermelon.
Gummy stem blight (GSB), caused by Didymella bryoniae (syn. Stagonosporopsis cucurbitacearum), produces devastating symptoms on whole plants of watermelon (Citrullus lanatus) and other cucurbits, significantly reducing yield and quality. Identification of genetic determinants and sources of resistance to this devastating GSB disease in watermelon is essential for developing resistant varieties. In this study, we aimed at identifying quantitative trait loci (QTLs) linked to GSB resistance in melon. We identified the genome-wide single nucleotide polymorphisms (SNPs) by genotyping by sequencing (GBS) of an F2 population developed from C. lanatus lines, 'PI 279461' (resistant) ✕ 'PI 223764' (susceptible). Inheritance analysis indicated that resistance to GSB is a multi-genic trait in this population. Three QTLs namely, ClGSB1.1, ClGSB10.1, and ClGSB11.1 associated with GSB resistance, explaining approximately 10% of the phenotypic variation, were identified. Among these, the QTL ClGSB1.1 on chromosome 1 is identified as a major QTL harboring five candidate genes associated with GSB resistance including two RLKs (ClC01G014900 and ClC01G015010), two WRKY transcription factors (ClC01G014910 and ClC01G014990), and one AvrRpt-cleavage domain protein (ClC01G015130). Two high resolution melting (HRM) markers, WmGSB1.1-2 and WmGSB1.1-7 having a high positive correlation with the phenotypic variations, were developed. Five potential candidate genes were predicted to be associated with GSB resistance. These findings will help breeders to develop watermelon cultivars resistant to GSB.


Nonrestrictive developmental regulation of microRNA gene expression.
During different periods of mammalian development, global changes in gene expression occur. Developmental changes in global gene expression have been modeled as a restrictive process. To test the restriction model of global changes in gene expression, we have used embryonic stem (ES) cells as a model system for the early mammalian embryo. ES cells are pluripotent cells that can contribute to all cellular lineages of the developing mammalian fetus and are derived from early embryonic cells. Using this model system, we have studied a new class of RNAs called microRNAs that have been identified and shown to play a role in the direct regulation of messenger RNAs. Here we report the expression signature for 248 microRNAs in 13 independent murine ES cells, embryoid bodies, and somatic tissues. The expression profile for 248 mouse microRNAs was determined for embryonic stem cells, embryoid bodies, mouse embryos, mature heart, lung, liver, kidney, and brain. Characteristic microRNA expression signatures were observed for each evaluated sample. When the characteristic microRNA signatures for developmentally ordered samples were compared, immature samples exhibited a less complex microRNA transcript profile than did mature samples. Our data support a progressive model of microRNA gene expression. Based on the progressive increase in complexity of micro- RNA expression, we hypothesize that the mammalian developmental program requires a temporal coupling of expression between microRNAs and messenger RNAs to enable the developmental potential observed in mammalian ontogeny.


Development of radiation resistance in Salmonella cultures.
Cultures of Salmonella subjected to repeated cycles of gamma-irradiation and subculture developed radiation resistance sooner at a low cycling dose ( approximately 1% survival) compared to a high cycling dose ( approximately 0.001% survival). Radioresistant cells in a population of radiosensitive cells of Salmonella newport or S. typhimurium could be selected by the double-irradiation plate method. The frequency of radioresistant cells in a population of S. newport was found to be about 1 per 8.9 million. Radio-resistant cells obtained by cyclic irradiation and subculture were larger (plumper) than the parent strain and showed a marked pleomorphism.


Different promoter regions control level and tissue specificity of expression of Agrobacterium rhizogenes rolB gene in plants.
Expression of the rolB gene of A. rhizogenes T-DNA triggers root differentiation in transformed plant cells. In order to study the regulation of this morphogenetic gene, the GUS reporter gene was placed under the control of several deleted fragments of the rolB 5' non-coding region: carrot disc transformations and the analysis of transgenic tobacco plants containing these constructions identified the presence of distinct regulatory domains in the rolB promoter. Two regions (located from positions -623 to -471 and from -471 to -341, from the translation start codon) control the level but not the tissue specificity of rolB expression: progressive deletions of the rolB promoter starting from position -1185 to -341, although at different levels, maintained the same pattern of GUS expression-maximal in root meristems and less pronounced in the vascular tissue of aerial organs. Further deletion of 35 bp, from -341 to -306, drastically affected tissue specificity: GUS activity was still clearly detectable in the vascular tissue of the aerial organs while expression in the root meristem was totally suppressed. Analysis of transgenic embryos and seedlings confirmed that distinct promoter domains are responsible for meristematic (root) and differentiated (vascular) expression of rolB. Finally, we present data concerning the effects of plant hormones on the expression of rolB-GUS constructions.


Comparative anatomical and developmental analysis of dry and fleshy fruits of Solanaceae.
An anatomical examination of dry and fleshy fruits within the Solanaceae was carried out to identify comparable stages throughout development as well as features exclusive to each type of fruit. We studied fruit development of Nicotiana and Petunia, which have the plesiomorphic capsular fruit; Solanum and Iochroma, characterized by a derived fleshy berry; Cestrum, an independent origin of a fleshy fruit; and Datura, a reversion to a dry fruit. • Pre- and postanthesis carpels and fruits of all species were collected, sectioned, stained, and examined using light microscopy. • Comparable stages of carpel and fruit development were identified in all species. Furthermore, anatomical and developmental features were identified that characterize capsules in Solanaceae, including lack of increase in the number of pericarp cell layers, formation of a sclerified endocarp, and elongation of the epidermal cells of the placenta. Pericarps of fleshy fruits of the Solanoideae are characterized by abundant collenchyma, an increase in the number of cell layers, and a parenchymatous endocarp often expanding into the locules. Anatomical data show that early developmental stages of the fruit of Cestrum, a berry, are similar to the capsular fruits of Petunia and Nicotiana; similarly, Datura, one of the few capsular members of the Solanoideae, shares several anatomical features with closely related berried taxa. • Ontogenetically, all fleshy or all dry fruits do not necessarily share a common developmental ground plan. Independent evolution of fleshiness, sclerification, dryness, and dehiscence are discussed in a phylogenetic context.


Persistent binocular diplopia in childhood following successful treatment for esotropia: a case series.
Diplopia is rare in children with nonpathological esotropia because facultative suppression develops prior to visual maturation. Our aim is to present five cases of children with binocular diplopia following treatment of childhood esotropia, exploring possible mechanisms for the development of diplopia based upon the patient's history and findings, and discussing treatment for diplopia in this setting. A retrospective chart review of five sequential patients presenting with binocular diplopia following treatment of esotropia was performed. Treatment of esotropia included correction of hyperopic error, correction of anisometropic amblyopia to 20/30 or better, and strabismus surgery as indicated. Diplopia developed in two children with accommodative esotropia, one child with infantile esotropia and partially accommodative esotropia, and two children with a history of infantile esotropia and consecutive exotropia following surgical correction. Diplopia onset occurred between ages 7 and 11. Four of the five children experienced resolution of diplopia with prism glasses. Children with esotropia before visual maturation may develop persistent diplopia. Clinical features of the children with diplopia in our case series include poor fusion, poor stereopsis, consecutive exotropia, refractive changes, onset of diplopia after 7 years of age, and occlusion therapy for amblyopia. Prisms may be beneficial in controlling the diplopia.


[Psychopathologic and psychophysiologic follow-up of inpatient depressed patients with standardized treatment with clomipramine and oxaprotiline].
The aim of this study was to show psychopathological and especially psychophysiological data of depressed inpatients which were collected during a 28-day double-blind study with 68 depressed inpatients given clomipramine 150 mgr/die) or maprotiline/oxaprotiline 150 mgr/die). Using Hamilton-Depression-Scale, Self-depression-scale (SDS by Zung), Beschwerdeliste (BL) and Befindlichkeitsskala (Bf-S) by v. Zerssen an impressive reduction of depression scores was observed in all rating scales with some superiority of clomipramine. But at the end of the study at treatment day 28 both groups - clomipramine and maprotiline/oxaprotilin - had similar results. During treatment each patient was part of a weekly habituation experiment using electrodermal activity in an orientation reaction as measure. Contrary to other studies (Lader 1975 or Heimann 1976, 1980) there was no difference in the course of psychophysiological data (EDA) between both treatment groups or between depressive agitation and psychomotor retardation. There was also no relationship between reactivity in EDA at study begin and treatment results.


Sugar-Free Dark Chocolate Consumption Results in Lower Blood Glucose in Adults With Diabetes.
Diabetes is characterized by an impaired ability to appropriately control blood glucose. Postprandial hyperglycemia, in particular, is associated with complications in people with type 1 diabetes (T1D) and type 2 diabetes (T2D). The objective of this study was to determine how sugar-free dark chocolate sweetened with stevia, erythritol, and inulin impacts postprandial blood glucose levels in individuals with diabetes compared to conventional dark chocolate. In a randomized crossover design, 13 participants consumed 1 bar (34 g) of sugar-free dark chocolate or 1 bar (34 g) of conventional dark chocolate with glucose levels measured before and throughout a 120-min postprandial period. The incremental area under the curve (iAUC) was lower after the consumption of sugar-free dark chocolate (-65%, P = .04) compared to conventional dark chocolate. No significant differences between chocolates were found for peak glucose value above baseline, the total area under the curve, or peak glucose values. Our results suggest that a sugar-free dark chocolate bar sweetened with stevia, erythritol and inulin led to a lower blood glucose iAUC compared to the conventional dark chocolate bar in people with diabetes, whilst longer-term effects on glucose control remain to be determined.


Antibody production using genetic immunization.
Extract: Antibodies are key tools in proteomics. Their high specificity and affinity allow them to be used to measure individual proteins in complex mixtures, and most importantly to provide quantitative information. There is now great interest in generating proteome-wide sets of antibodies in order to discover biomarkers of disease and to enhance drug development. Antibodies are traditionally produced by injecting a pure form of the protein mixed with an adjuvant into an animal. The pure protein is usually produced either by overexpression in a host such as E. coli and purifying it using a fused tag (e.g., GST, 6-histidines), or by synthesizing small peptide sequences from the whole protein. The breakthrough effort in the last decade of determining the sequences of entire genomes has discovered tens of thousands of new proteins. Antibodies are now the major limiting factor in quantitative proteomics. The protein-based technology has proven too slow and expensive to make it feasible to produce antibodies on the scale now needed by the pharmaceutical and biotechnology industries.


Histamine-releasing factors and inhibitory factors.
Peripheral-blood mononuclear cells (MNC) synthesize several histamine-releasing factors (HRF) spontaneously and when stimulated. Some of the characterized cytokines have histamine-releasing activity, especially connective tissue activating peptide III, neutrophil-activating peptide 2 and interleukin-3 (IL-3). At least two species of HRF remain to be characterized. MNC also secrete a histamine release-inhibitory factor (HRIF), which is a highly specific inhibitor, because it antagonizes only HRF. IL-8 resembles the low-molecular-weight species of HRIF in terms of size and specificity.


Treatment Adherence of Tuberculosis Patients Attending Two Reference Units in Equatorial Guinea.
Equatorial Guinea has one of the highest burden of tuberculosis (TB) in Africa. Incomplete adherence to TB treatment has been identified as one of the most serious remaining problem in tuberculosis control. The following study is aimed at determining the adherence to anti-tuberculosis treatment in Equatorial Guinea and its determinants, as well as at assessing the knowledge of the people about the disease. In this cross-sectional study, participants were recruited by non-probabilistic consecutive sampling amongst patients who attended the reference units for TB in Bata and Malabo between March and July 2015. Socio-demographic and clinical data were collected. Adherence to treatment and knowledge about TB were assessed by Morisky-Green-Levine and Batalla tests and a questionnaire on adherence related factors specifically prepared for this research. Descriptive statistics were computed to summarize the data and bivariate analyses by adherence profile were performed with χ2 test for categorical data. A total of 98 patients with TB were interviewed. 63.27% of interviewees had good knowledge about TB (Batalla test) while 78.57% of respondents were adherent according to the Morisky-Green-Levine test. A low educational level, lack of family support and lack of medical advice about the disease were significantly associated to lower adherence level. Patients with re-infection (due to relapse or treatment failure) and those who have suffered from drug shortages were also less adherents. The National Programme for TB Control should consider improving the early diagnosis and follow-up of TB cases, as well as the implementation of all components of DOTS (Directly observed Treatment, short-course) strategy all over the country.


Squamous cell carcinoma of the external auditory canal: long-term clinical outcomes using surgery and external-beam radiotherapy.
Squamous cell carcinoma (SCCA) of the external auditory canal (EAC) is often treated with a combination of surgery and radiotherapy (RT) to optimize the chance of achieving locoregional control. This retrospective review describes a 27-year experience of treating these tumors at the University of Florida. Thirty patients with histologically confirmed SCCA of the EAC received external-beam radiation (RT) alone or combined with surgical resection between 1976 and 2003. Seven patients were treated with RT alone, 22 with postoperative RT, and 1with preoperative RT. Patients were grouped according to nodal status (N0/N1) and the Stell staging system for tumors of the EAC and middle ear. Early stage was defined as T1/T2 (n = 12) and advanced stage as T3 (n = 18). Median follow-up was 2 years (range, 0.1-19.4 years) with no patients lost to follow-up. The 5-year actuarial probabilities for local control, locoregional control, and cause-specific survival for patients with early stage (T1/T2) versus advanced-stage (T3) tumors were 74% and 55% (P = 0.27), 63% and 38% (P = 0.16), and 70% and 41% (P = 0.04), respectively. The regional control rate was 83% (P = 0.6). There were 12 local recurrences and 4 neck recurrences as the first site of failure. One failure was successfully salvaged with surgery. Five of 23 (21%) patients undergoing surgery had significant complications (grade 3 or 4), whereas 2 of 30 (7%) patients receiving RT experienced grade 3 complications. Patients with early stage disease achieved better local control, locoregional control, and cause-specific survival than those with advanced tumors. Less than half of the patients (13 of 30; 43%) were cured without significant complications, suggesting a suboptimal therapeutic ratio, using current treatment methods.


Platelet cyclooxygenase expression in normal dogs.
Human platelets express both cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). Variation in COX-2 expression could be a mechanism for variable response to aspirin. The hypotheses were that circulating canine platelets express COX-1 and COX-2, and that aspirin alters COX expression. The objective was to identify changes in platelet COX expression and in platelet function caused by aspirin administration to dogs. Eight female, intact hounds. A single population, repeated measures design was used to evaluate platelet COX-1 and COX-2 expression by flow cytometry before and after aspirin (10 mg/kg Q12h for 10 days). Platelet function was analyzed via PFA-100(®) (collagen/epinephrine), and urine 11-dehydro-thromboxane B(2) (11-dTXB(2)) was measured and normalized to urinary creatinine. Differences in COX expression, PFA-100(®) closure times, and urine 11-dTXB(2 ): creatinine ratio were analyzed before and after aspirin administration. Both COX-1 and COX-2 were expressed in canine platelets. COX-1 mean fluorescent intensity (MFI) increased in all dogs, by 250% (range 63-476%), while COX-2 expression did not change significantly (P = 0.124) after aspirin exposure, with large interindividual variation. PFA-100(®) closure times were prolonged and urine 11-dTXB(2) concentration decreased in all dogs after aspirin administration. Canine platelets express both COX isoforms. After aspirin exposure, COX-1 expression increased despite impairment of platelet function, while COX-2 expression varied markedly among dogs. Variability in platelet COX-2 expression should be explored as a potential mechanism for, or marker of, variable aspirin responsiveness.


Synchronous infiltrating ductal carcinoma and primary extramedullary plasmacytoma of the breast.
Extramedullary plasmacytomas are seldom solitary and usually progress to diffuse myelomatosis. Plasmacytomas of the breast are rare, especially when not associated multiple myeloma. Synchronous infiltrating ductal carcinoma and primary extramedullary plasmacytoma of the breast have not previously reported. A 27-years-old woman with an untreated upper outer quadrant breast mass for 1-year was referred to our cancer hospital for surgical evaluation of increasing breast pain. Postoperatively, microscopic examination revealed an infiltrating ductal carcinoma complicated by an extramedullary plasmacytoma divided by fibrous tissue in one section. Following surgery, the patient received chemotherapy for the carcinoma and radiotherapy for the plasmacytoma. In this case, careful histopathology examination was essential to make the correct diagnosis and therapy for these synchronous lesions. The patient finished chemotherapy and radiotherapy without significant adverse effects.


Bluetongue virus viral protein 7 stability in the presence of glycerol and sodium chloride.
The Orbivirus Bluetongue virus (BTV) is an economically significant disease that affects mainly wild and domestic ruminants. BTV is most often seen symptomatically in sheep, but is easily carried by goats, cattle, and wild ruminants. To date there are several problems with the vaccines currently available for BTV, and one of the most promising candidates to increase vaccine efficacy is a protein-based vaccine, for which viral protein 7 (VP7) is a great candidate to be included in it. In order to further these studies, the stability of BTV VP7 in common vaccine additives needs to be investigated. Recombinant BTV VP7 was expressed in a bacterial cell system and purified before being analysed using spectroscopic techniques including far-ultraviolet (UV) circular dichroism and intrinsic tryptophan fluorescence. BTV was analysed in a number of different buffer conditions. We report here that BTV VP7 maintains its native secondary structure until at least 52℃ and native-like tertiary structure to at least 80℃. Far-UV circular dichroism and intrinsic tryptophan fluorescence emission spectra indicate significant secondary and tertiary structure remaining even at 90℃, respectively. Six M guanidinium chloride is able to unfold BTV VP7 while 8 M urea could not. Twenty percent glycerol and 300 mM sodium chloride appear to have a protective effect on BTV VP7's structure, as significantly more structure is seen at 90℃ when compared to BTV VP7 without the addition of these chemicals. Both glycerol and sodium chloride are common vaccine additives.


Cloning of a recombinant complementary DNA to a baboon (Papio anubis) estradiol-dependent oviduct-specific glycoprotein.
The estradiol-dominated baboon oviduct synthesizes and secretes a glycoprotein (mol wt, 120,000) that binds to oocyte zona pellucida in vivo. This glycoprotein separates into two major isoelectric variants (pl 8.0 and 4.5) on two-dimensional electrophoretic gels. This study was undertaken to further characterize the steroid-controlled gene expression of this glycoprotein. A recombinant cDNA library was prepared to poly(A)+ RNA isolated from oviducts obtained from estradiol-treated baboons. The library was screened with a polyclonal antibody prepared against the acidic component (pl 4.5) of the glycoprotein. A single positive plaque was purified. Digestion of the recombinant plasmid with EcoRI revealed fragments of 230 and 1000 basepairs. The antibody used to screen the library was epitope selected against the fusion protein produced by the purified recombinant phage. The epitope-selected antibody, when incubated with Western blots of oviductal explant culture medium obtained from estradiol-treated baboons, recognized both the acidic and the basic variants of the glycoprotein. Northern blot hybridization with a 32P-labeled insert indicated that a single message of 2.8 kilobases was present in oviducts obtained from estradiol-treated baboons; no hybridization was detectable to RNA from oviducts obtained from progesterone-treated baboons. A mRNA of comparable size was also found in human, hamster, rabbit, and mouse oviduct tissue. In vitro translation of oviductal poly(A)+ RNA from an estradiol-treated baboon followed by immunoprecipitation with the polyclonal antibody against the acidic component indicated that the protein component of the oviductal glycoprotein had a mol wt of 66,000.(ABSTRACT TRUNCATED AT 250 WORDS)


On the Relation Between G-formula and Inverse Probability Weighting Estimators for Generalizing Trial Results.
When generalizing inferences from a randomized trial to a target population, two classes of estimators are used: g-formula estimators that depend on modeling the conditional outcome mean among trial participants and inverse probability (IP) weighting estimators that depend on modeling the probability of participation in the trial. In this article, we take a closer look at the relation between these two classes of estimators. We propose IP weighting estimators that combine models for the probability of trial participation and the probability of treatment among trial participants. We show that, when all models are estimated using nonparametric frequency methods, these estimators are finite-sample equivalent to the g-formula estimator. We argue for the use of augmented IP weighting (doubly robust) generalizability estimators when nonparametric estimation is infeasible due to the curse of dimensionality, and examine the finite-sample behavior of different estimators using parametric models in a simulation study.


Bioactive compounds from orange epicarp to enrich fish burgers.
The orange industry produces considerable amounts of by-products, traditionally used for animal feed or fuel production. Most of these by-products could be used as functional ingredients. To assess the potential food application of orange epicarp, different percentages of micro-encapsulated orange extract were added to fresh fish burgers. Then, an in vitro digestion was also carried out, before and after micro-encapsulation, to measure the bio-accessibility of the active compounds. A significant increase of bio-accessibility of bioactive compounds has been observed in the orange epicarp extract after micro-encapsulation by spray-drying. From the sensory point of view, the fish sample enriched with 50 g kg-1 micro-encapsulated extract was the most comparable to the control burger, even if it showed a higher phenolic, flavonoid and carotenoid bio-accessibility. Orange epicarp may be used as a food additive to enhance the health content of food products. The micro-encapsulation is a valid technique to protect the bioactive compounds and increase their bio-accessibility. © 2017 Society of Chemical Industry.


General organization of the perinatal and adult accessory olfactory bulb in mice.
The vomeronasal system is currently a topical issue since the dual functional specificity, vomeronasal system-pheromones, has recently been questioned. Irrespective of the tools used to put such specificity in doubt, the diversity of the anatomy of the system itself in the animal kingdom is probably of more importance than has previously been considered. It has to be pointed out that a true vomeronasal system is integrated by the vomeronasal organ, the accessory olfactory bulb, and the so-called vomeronasal amygdala. Therefore, it seems reasonable to establish the corresponding differences between a well-developed vomeronasal system and other areas of the nasal cavity in which putative olfactory receptors, perhaps present in other kinds of mammals, may be able to detect pheromones and to process them. In consequence, a solid pattern for one such system in one particular species needs to be chosen. Here we report on an analysis of the general morphological characteristics of the accessory olfactory bulb in mice, a species commonly used in the study of the vomeronasal system, during growth and in adults. Our results indicate that the critical period for the formation of this structure comprises the stages between the first and the fifth day after birth, when the stratification of the bulb, the peculiarities of each type of cell, and the final building of glomeruli are completed. In addition, our data suggest that the conventional plexiform layers of the main olfactory bulb are not present in the accessory bulb.


Diagnostic methods for the etiological assessment of infectious corneal pathology (Review).
Infectious keratitis is a leading cause of visual morbidity, including blindness, all across the globe, especially in developing countries. Prompt and adequate treatment is mandatory to maintain corneal integrity and to recover the best possible final visual acuity. Although in most of the cases practitioners chose to employ empirical broad-spectrum antimicrobial medication that is usually effective, in some instances, they face the need to identify the causative agent to establish the appropriate therapy. An extensive search was conducted on published literature before December 2020 concerning the main laboratory investigations used to identify the microbial agents found in infectious keratitis, their indications, advantages, and disadvantages, as well as the results reported by other studies concerning different diagnostic tools. At present, the gold standard for diagnosis is still considered to be the isolation of microorganisms in cultures, along with the examination of smears, but other newer techniques, such as polymerase chain reaction (PCR), next-generation sequencing (NGS), and in vivo confocal microscopy (IVCM) have gained popularity in the last decades. Currently, these newer methods have proved to be valuable adjuvants in making the diagnosis, but technological advances hold promise that, in the future, these methods will have increased performance and availability, and may become the new gold standard, replacing the classic cultures and smears.


Plasma norepinephrine in normal young and aged men: relationship with sleep.
This study explored the relationship between age-related elevations of plasma norepinephrine (NE) levels (thought to reflect heightened sympathetic nervous system activity) and sleep. Plasma NE levels were consistently and significantly greater in 8 aged than 10 young men across a 24-hr study period. For both groups, plasma catecholamine levels remained stable near bedtimes and rise times and during spontaneous and experimentally induced nighttime awakenings; reversal of the sleep/wake pattern (nighttime wakefulness, daytime sleep) had little effect on the 24-hr plasma NE or sleep patterns, indicating that elevated NE cannot be attributed to altered sleep/wakefulness per se. The aged group had significantly more wakefulness and significantly higher plasma NE levels while in bed in all study conditions. These two variables were significantly correlated, suggesting that heightened sympathetic activity may fragment sleep with wakefulness in the aged.


In vivo effect of industrial titanium dioxide nanoparticles experimentally exposed to hairless rat skin.
We morphologically investigated animal skin exposed to W/O emulsion containing 10 wt % ultrafine TiO(2) particles that had been characterized. After 4 h, exposed skin was investigated by light microscopy, confocal laser scanning microscopy (CLSM) and electron microscopy with energy-dispersive X-ray spectrometry (EDX). Light microscopic evaluation was also performed on the exposed skin after 24, 72 and 168 h. Light microscopy did not show any morphological and immunohistochemical changes in the skin. Electron microscopy revealed that the most TiO(2) particles were localized in the interfollicular stratum disjunctum and the keratinized layer of follicular infundibulum. No TiO(2) particles were detected in the viable skin, which was confirmed by EDX. Furthermore, we demonstrated a specific TiO(2) affinity to the follicular opening area by light microscopy and low-vacuum scanning electron microscopy with EDX. Our study suggests that TiO(2) particles neither penetrate into viable cell layers nor biologically cause any cellular changes.


Total syntheses of (+/-)-beta-erythroidine and (+/-)-8-oxo-beta-erythroidine by an intramolecular Diels-Alder cycloaddition of a 2-amidoacrolein.
[reaction: see text] The total syntheses of (+/-)-beta-erythroidine and (+/-)-8-oxo-beta-erythroidine are described. The tetracyclic ring system of the natural products was quickly assembled by a strategy that features a retrocycloaddition/cycloaddition reaction of an amidodioxin, an intramolecular Heck reaction, and a 6pi-electrocyclic ring closure of a dienoic acid.


Does Cue Focality Modulate Age-related Performance in Prospective Memory? An fMRI Investigation.
How prospective memory (PM) weakens with increasing age has been largely debated. We hypothesized that automatic and strategic PM processes, respectively mediated by focal and non-focal cues, are differently affected by aging, even starting from 50-60 years of age. We investigated this issue using a 2 × 2 design in which focal and non-focal experimental conditions were created by varying the conjoint nature of the ongoing task (lexical decision vs. syllable matching tasks) and the PM cue (words vs. syllables). In the whole-brain analysis we found that the left inferior frontal gyrus and the middle cingulate cortex were more activated when young compared to older individuals performed a PM task; moreover, the anterior cingulate cortex was selectively activated during non-focal PM when the cues were words. In a region-of-interest analysis we observed that the medial and the lateral portions of the rostral prefrontal cortex were associated with the focal and non-focal conditions respectively, more in young than in older adults. Our findings provide evidence in support of early age-related differences in automatic/strategic PM functioning.


Proposal of a Geometric Calibration Method Using Sparse Recovery to Remove Linear Array Push-Broom Sensor Bias.
The rational function model (RFM) is widely used in the most advanced Earth observation satellites, replacing the rigorous imaging model. The RFM method achieves the desired calibration performance when image distortion is caused by long-period errors. However, the calibration performance of the RFM method deteriorates when short-period errors-such as attitude jitter error-are present, and the insufficient and uneven ground control points (GCPs) can also lower the calibration precision of the RFM method. Hence, this paper proposes a geometric calibration method using sparse recovery to remove the linear array push-broom sensor bias. The most important issue regarding this method is that the errors related to the imaging process are approximated to the equivalent bias angles. By using the sparse recovery method, the number and distribution of GCPs needed are greatly reduced. Meanwhile, the proposed method effectively removes short-period errors by recognizing periodic wavy patterns in the first step of the process. The image data from Earth Observing 1 (EO-1) and the Advanced Land Observing Satellite (ALOS) are used as experimental data for the verification of the calibration performance of the proposed method. The experimental results indicate that the proposed method is effective for the sensor calibration of both satellites.


(99m)Tc-MDP scintigraphic findings in children with leukemia: value of early and delayed whole-body imaging.
The purpose of this study was to reveal the bone scan abnormalities in children with leukemia and to show the value of whole-body scanning in early and delayed phases. From a database of all patients with a diagnosis of leukemia from January 1990 to April 2000, 12 children (9 male, 3 female; mean age, 8.0 y; age range, 4.7--13.2 y) were identified for whom the diagnosis of leukemia was suggested on the basis of bone scans obtained as part of the initial work-up for unexplained skeletal pain. Early and delayed whole-body bone scans and radiographs were reviewed retrospectively. Areas of abnormal uptake on early and delayed phases were categorized into locations: metaphysis--diaphysis--epiphysis (MDE), pelvis, ribs, spine, and others. MDE lesions included abnormalities in the metaphysis extending into the diaphysis for some length: metaphysis/diaphysis, metaphysis only, diaphysis only, epiphysis only, and the entire bone. Pelvic and spine lesions were further characterized as focal or diffuse. Ten patients had lesions in 2 or more locations on both phases. Two patients had multiple lesions on the early scans but only rib lesions on the delayed scans. Lesions correlated with symptomatic sites in 8 patients on the delayed scans and in 11 patients on the early scans. The most common sites of abnormalities on the delayed scans were metaphyseal/diaphyseal, pelvis (focal), and ribs. The most common locations of lesions on the early scans were metaphyseal/diaphyseal, pelvis (diffuse or focal), and spine. More metaphyseal/diaphyseal lesions were seen on the early scans than on the delayed scans. Diffuse involvement of the pelvis and spine was seen only on the early phase. However, rib lesions were seen more frequently on the delayed scan. Early whole-body imaging in conjunction with delayed whole-body scanning may enhance the diagnostic accuracy of bone scanning in the evaluation of children with skeletal pain of obscure etiology, such as that associated with leukemia.


Tissue culture-induced DNA methylation polymorphisms in repetitive DNA of tomato calli and regenerated plants.
The propagation of plants through tissue culture can induce a variety of genetic and epigenetic changes. Variation in DNA methylation has been proposed as a mechanism that may explain at least a part of these changes. In the present study, the methylation of tomato callus DNA was compared with that of leaf DNA, from control or regenerated plants, at MspI/HpaII sites around five middle-repetitive sequences. Although the methylation of the internal cytosine in the recognition sequence CCGG varied from zero to nearly full methylation, depending on the probe used, no differences were found between callus and leaf DNA. For the external cytosine, small differences were revealed between leaf and callus DNA with two probes, but no polymorphisms were detected among DNA samples of calli or DNA samples of leaves of regenerated plants. When callus DNA cut with HindIII was studied with one of the probes, H9D9, most of the signal was found in high-molecular-weight DNA, as opposed to control leaf DNA where almost all the signal was in a fragment of 530 bp. Also, an extra fragment of 630 bp was found in the callus DNA that was not present in control leaf DNA. Among leaves of plants regenerated from tissue culture, the 630-bp fragment was found in 10 of 68 regenerated plants. This 630-bp fragment was present among progeny of only 4 of these 10 plants after selfing, i.e. it was partly inherited. In these cases, the fragment was not found in all progeny plants, indicating heterozygosity of the regenerated plants. The data are interpreted as indicating that a HindIII site becomes methylated in callus tissue, and that some of this methylation persists in regenerated plants and is partly transmitted to their progeny.


Molecular Cloning, Characterization, and Expression Analysis of Cathepsin A in the Chinese Giant Salamander Andrias davidianus.
Cathepsin A (CTSA) is serine carboxypeptidase, an important protease in the lysosome. In this study, the full complementary DNA (cDNA) sequence of CTSA in Chinese giant salamanders Andrias davidianus was cloned, and its sequence features were analyzed. Tissue expression patterns of CTSA in healthy and Aeromonas hydrophila-infected salamanders were also investigated. The full cDNA sequence of salamander CTSA was 1,620 base pairs in length, encoding 472 amino acids. Salamander CTSA shared high sequence identities with other vertebrates' CTSAs, ranging from 62.7% to 68.9%. In healthy salamanders, CTSA was highly expressed in spleen, followed by brain, intestine, and stomach. After A. hydrophila infection, salamander CTSA was significantly upregulated in lung, heart, muscle, and kidney; was downregulated in liver, spleen, and intestine; and exhibited no significant changes in stomach and skin, indicating that salamander CTSA might play defense roles in multiple tissues during bacterial infection. These results provide a solid basis for further study of the immune function of amphibian CTSA. Received September 18, 2016; accepted June 18, 2017.


Control of the functional activity of an antisense RNA by a tetracycline-responsive derivative of the human U6 snRNA promoter.
In an effort to develop a regulatable derivative of the promoter of the human gene for U6 snRNA, we generated several constructs composed of the human U6 snRNA promoter and sequences derived from the gene for the tetracycline operator of a prokaryotic tetracycline resistance transposon. One of the constructs had strong transcriptional activity in the presence of tetracycline that was equivalent to 80% of the activity of the wild-type promoter. Furthermore, the transcriptional activity was almost completely repressed in the absence of tetracycline. Transcriptional activity became detectable within 4 hr after the addition of tetracycline to the culture medium. We used this system to control the functional activity of an antisense RNA for a chimeric gene derived from genes for the epidermal growth factor receptor (EGFR) and green fluorescent protein (GFP). A plasmid that expressed the chimeric gene and a plasmid that expressed the antisense RNA under the control of the inducible U6 promoter were used to cotransfect HeLa cells that were producing the tetracycline repressor protein (Tet R). Addition of tetracycline to the culture medium 12 hr after transfection resulted in the almost complete disappearance of the fluorescent signal due to the chimeric protein within 24 hr. Our results suggest that this expression system might be a useful tool for controlling the expression of functional RNAs, such as aptamers and antisense RNAs, both in basic research and in gene therapy.


Antral choanal polyp presenting as obstructive sleep apnea syndrome.
Obstructive sleep apnea syndrome (OSAS) in children is commonly caused by adenotonsillar hypertrophy. The diagnostic criteria of OSAS in children are not so well delineated as in adults. We report the first case of antral choanal polyp presenting as OSAS in a 10-year-old boy that initially presented to the child psychiatry service for behavior disturbance, enuresis, and daytime somnolence. Overnight electroencephalogram sleep study revealed events consistent with OSAS. Multiple inhalant allergies, chronic maxillary sinusitis, and obstructive adenoid hypertrophy were diagnosed by the allergy and otolaryngology services. The child was scheduled for adenoidectomy when his sleep apnea symptoms persisted following antimicrobial therapy. Examination under anesthesia revealed a normal adenoid bed and a large left antral choanal polyp. Polypectomy was performed as dictated by parental consent. Postoperatively treatment with an intranasal steroid was begun. However, polypoid nasal mucosa recurred in 2 months and a Caldwell-Luc procedure was performed. Subjective reports following surgery indicated improvement in daytime irritability, attention, and mood. A follow-up overnight electroencephalogram sleep study confirmed resolution of OSAS.


Identification of bacterial protein O-oligosaccharyltransferases and their glycoprotein substrates.
O-glycosylation of proteins in Neisseria meningitidis is catalyzed by PglL, which belongs to a protein family including WaaL O-antigen ligases. We developed two hidden Markov models that identify 31 novel candidate PglL homologs in diverse bacterial species, and describe several conserved sequence and structural features. Most of these genes are adjacent to possible novel target proteins for glycosylation. We show that in the general glycosylation system of N. meningitidis, efficient glycosylation of additional protein substrates requires local structural similarity to the pilin acceptor site. For some Neisserial PglL substrates identified by sensitive analytical approaches, only a small fraction of the total protein pool is modified in the native organism, whereas others are completely glycosylated. Our results show that bacterial protein O-glycosylation is common, and that substrate selection in the general Neisserial system is dominated by recognition of structural homology.


Ascorbic acid augments the adenylyl cyclase-cAMP system mediated POMC mRNA expression and beta-endorphin secretion from hypothalamic neurons in culture.
Besides acting as an important cofactor in the biosynthesis of catecholamine, ascorbic acid (AA) also modulates the activity of peptidylglycine-alpha-amidating monooxygenase for the post-translational modification of neuropeptides such as alpha-MSH and TRH. We report here a novel action of AA in modulating the secretion of immunoreactive beta-endorphin (ir-beta EP) and mRNA expression of proopiomelanocortin (POMC) following the activation of cAMP-dependent protein kinase A pathway in rat hypothalamic neurons. Primary cultures of hypothalamic neurons from neonatal rats as previously described were employed in the present studies. Six days after plating, cultures were replenished with serum-free media and incubated with vehicle or various doses of AA in the presence or absence of forskolin, 3-isobutyl-1-methylxanthine (IBMX), N6,2'-O-dibutyryladenosine 3'5'-(cyclic)monophosphate [(Bu)2cAMP]. Whereas the basal ir-beta EP release was 22.0 +/- 0.4 pg/well (mean +/- S.E.; n = 3), 10 microM of forskolin treatment increased ir-beta EP release approximately 4.2-fold. Co-incubation with AA enhanced forskolin induced ir-beta EP release and that this enhancing effect of AA was both time related and dose-dependent, with an ED50 of approximately 10 microM and an Emax of 100 microM. At the concentration of 10 microM, AA augmented ir-beta EP release approximately 6.1-fold that of cultures treated with forskolin alone. A similar potentiating effect of AA was also seen in cultures co-treated with IBMX or with (Bu)2cAMP. These enhancing effects of AA were similarly found in the abundance of total cAMP and of POMC mRNA of cultures which received identical treatments. However, it is important to point out that AA alone did not modulate ir-beta EP release or the abundance of POMC mRNA or total cAMP levels of the hypothalamic cultures when protein kinase A pathway was not activated. We thus conclude that AA augments cAMP-dependent protein kinase A pathway-induced production and release of beta EP from rat hypothalamic neurons in culture. Furthermore, this biological effect of AA is, at least in part, mediated through enhancing the responsiveness of the adenylyl cyclase-cAMP system.


Language-based measures of mindfulness: initial validity and clinical utility.
This study examined relationships among language use, mindfulness, and substance-use treatment outcomes in the context of an efficacy trial of mindfulness-based relapse prevention (MBRP) for adults with alcohol and other drug use (AOD) disorders. An expert panel generated two categories of mindfulness language (ML) describing the mindfulness state and the more encompassing "mindfulness journey," which included words describing challenges of developing a mindfulness practice. MBRP participants (n = 48) completed baseline sociodemographic and AOD measures, and participated in the 8-week MBRP program. AOD data were collected during the 4-month follow-up. A word count program assessed the frequency of ML and other linguistic markers in participants' responses to open-ended questions about their postintervention impressions of mindfulness practice and MBRP. Findings supported concurrent validity of ML categories: ML words appeared more frequently in the MBRP manual compared to the 12-step Big Book. Further, ML categories correlated with other linguistic variables related to the mindfulness construct. Finally, predictive validity was supported: greater use of ML predicted fewer AOD use days during the 4-month follow-up. This study provided initial support for ML as a valid, clinically useful mindfulness measure. If future studies replicate these findings, ML could be used in conjunction with self-report to provide a more complete picture of the mindfulness experience.


Identification, characterization, and expression pattern of the chicken EKLF gene.
EKLF/KLF1 was the first of the Krüppel-like factors (KLFs) to be identified in mammals and plays an important role in primitive and definitive erythropoiesis. Here, we identify and characterize EKLF in the chicken (cEKLF). The predicted amino acid sequence of the zinc finger region of cEKLF is at least 87.7% similar to mammalian EKLF proteins and is 98.8% and 95% similar to the EKLF orthologues in Xenopus and zebrafish, respectively. During early embryonic development, cEKLF expression is seen in the posterior primitive streak, which gives rise to hematopoietic cells, and then in the blood islands and in circulating blood cells. cEKLF mRNA is expressed in blood cells but not in brain later in chicken embryonic development. cEKLF mRNA is increased in definitive compared with primitive erythropoiesis. The conserved sequence and expression pattern of cEKLF suggests that its function is similar to its orthologues in mammals, Xenopus, and zebrafish.


A Kalman Filtering and Nonlinear Penalty Regression Approach for Noninvasive Anemia Detection with Palpebral Conjunctiva Images.
Noninvasive medical procedures are usually preferable to their invasive counterparts in the medical community. Anemia examining through the palpebral conjunctiva is a convenient noninvasive procedure. The procedure can be automated to reduce the medical cost. We propose an anemia examining approach by using a Kalman filter (KF) and a regression method. The traditional KF is often used in time-dependent applications. Here, we modified the traditional KF for the time-independent data in medical applications. We simply compute the mean value of the red component of the palpebral conjunctiva image as our recognition feature and use a penalty regression algorithm to find a nonlinear curve that best fits the data of feature values and the corresponding levels of hemoglobin (Hb) concentration. To evaluate the proposed approach and several relevant approaches, we propose a risk evaluation scheme, where the entire Hb spectrum is divided into high-risk, low-risk, and doubtful intervals for anemia. The doubtful interval contains the Hb threshold, say 11 g/dL, separating anemia and nonanemia. A suspect sample is the sample falling in the doubtful interval. For the anemia screening purpose, we would like to have as less suspect samples as possible. The experimental results show that the modified KF reduces the number of suspect samples significantly for all the approaches considered here.


Epidemiology, patient profile, and health care resource use for hepatitis C in Italy.
The objectives of this study were to estimate the prevalence of Hepatitis C among six Italian Local Health Units (LHUs), to describe patient and antiviral drug characteristics, and to estimate the health care consumption rates and related costs for the management of patients affected by hepatitis C virus (HCV) infection by using data from routine clinical practice. We conducted a retrospective study using administrative databases of six Italian LHUs. All patients who had a record related to HCV during the enrollment period (July 1, 2009, to December 31, 2014) and who had at least 6 months of data available prior to the first HCV record were included. The date of the first record related to HCV during the enrollment period was considered as a proxy of diagnosis and used as the index date. Patients were followed from the index date up to 1 year, death, or exiting the database. Using the overall cohort of HCV adult patients as the numerator, we estimated the prevalence of HCV among six LHUs. The denominators were obtained from the National Institute of Statistics (N=1,665,682). We also evaluated descriptive patient's characteristics and treatment patterns, and estimated health care consumption rates and related costs for the management of the HCV patients. A total of 7,550 patients were analyzed, of whom 57% were male with a mean age of 57.6±16.4 years. The prevalence of HCV was estimated to be 0.45% (95% confidence interval 0.44-0.46). During the follow-up period, 78.6% of HCV patients had received no antiviral treatment. The annual health care cost associated with HCV infection was €6,022.7 (±7,922.6) while the cost specific to HCV care was €3,154.6 (±4,972.0). Our findings showed that, in the Italian real-world setting, only a small proportion of HCV-infected patients received an antiviral treatment. Despite the current low prevalence of HCV, the economic impact of such disease remains high.


The breast screening programme and misinforming the public.
The information provided to the public by the NHS Breast Screening Programme has been criticized for lack of balance, omission of information on harms and substantially exaggerated estimates of benefit. These shortcomings have been particularly evident in the various invitation leaflets for breast screening and in the Programme's own 2008 Annual Review, which celebrated 20 years of screening. The debate on screening has been heated after new data published in the last two years questioned the benefit and documented substantial harm. We therefore analysed whether the recent debate and new pivotal data about breast screening has had any impact on the contents of the new 2010 leaflet and on the 2010 Annual Review. We conclude that spokespeople for the Programme have stuck to the beliefs about benefit that prevailed 25 years ago. Concerns about over-diagnosis have not been addressed either and official documents still downplay this most important harm of breast cancer screening.


The delta F508 mutation in cystic fibrosis patients of southern Italy.
Fifty one independent cystic fibrosis (CF) families originating from a restricted area of Southern Italy (Campania) have been analyzed for KM19 and XV2c haplotypes and the delta F508 mutation: 54% of the total CF chromosomes show the delta F508 mutation. No significative correlations were obtained when clinical score, radiological score, Pseudomonas colonization, or clinical symptoms at presentation were matched with the presence or absence of the delta F508 mutation.


Accessory Ossicles in the Region of the Foot and Ankle: An Epidemiologic Survey in a Jordanian Population.
Background and Objectives: The incidence of accessory bones in the region of foot and ankle is quite variable between studies and are often confused with avulsion fractures in trauma patients with musculoskeletal injuries. The aim of this study was to assess the incidence of accessory ossicles of the foot and ankle according to gender, side and coexistence, and to determine how frequently accessory ossicles were misdiagnosed as avulsion fractures. Materials and Methods: Oblique and/or lateral foot radiographs of 1000 adult patients referred from emergency departments to foot and ankle clinic were retrospectively reviewed for the presence of accessory ossicles. The Kappa statistic was used in order to assess the validity of radiographic interpretation for the presence of these bones. Results: Accessory ossicles were detected in 40.2% of the radiographs. The incidence rates for the accessory ossicles in order of frequency were: Os trigonum (15.4%), accessory navicular (13.7%), os peroneum (11.5%), os vesalianum (1.1%), os supranaviculare (0.7%), os subfibulare (0.6%), os talotibiale (0.4%), os calcaneus secundarius (0.3%), os supratalare (0.3%), os infranaviculare (0.3%), os intermetatarseum (0.2%), and os subtibiale (0.1%). Coexistence of two or three ossicles in the same foot was observed in 4.4% of the cases, mostly coexistence with os peroneum (2.9%), followed by accessory navicular (1.6%). 2.7% of accessory ossicles were initially misdiagnosed as avulsion fractures at emergency departments. Interrater agreement over identification of different accessory ossicles was found to be reasonably reliable, with a Kappa greater than 0.80 for all assessed bones. Conclusions: In clinical practice, a thorough knowledge of normal anatomical variants is essential to facilitate appropriate diagnosis and treatment and can help to prevent diagnostic errors.


Ductal carcinoma in situ of the breast: correlation between histologic classifications and biologic markers.
The expression of hormone receptors (estrogen and progesterone), c-erb-B2 (neu), p53, and Ki-67) was studied by immunohistochemical analysis in a series of 94 carcinomas in situ of the breast. The tumors were classified in two (high grade vs. low grade) and three (high grade vs. intermediate grade vs. low grade) categories. High-grade carcinomas were well defined by biologic markers showing statistically significant differences for all of the parameters tested. No clear biologic distinction, however, between low-grade and intermediate-grade cancers could be obtained. Negative progesterone receptors and a high proliferative index are the best discriminant parameters for the final assignment of doubtful cases.


Cell surface expression of GRP78 and CXCR4 is associated with childhood high-risk acute lymphoblastic leukemia at diagnostics.
Acute lymphocytic leukemia is the most common type of cancer in pediatric individuals. Glucose regulated protein (GRP78) is an endoplasmic reticulum chaperone that facilitates the folding and assembly of proteins and regulates the unfolded protein response pathway. GRP78 has a role in survival of cancer and metastasis and cell-surface associated GRP78 (sGRP78) is expressed on cancer cells but not in normal cells. Here, we explored the presence of sGRP78 in pediatric B-ALL at diagnosis and investigated the correlation with bona fide markers of leukemia. By using a combination of flow cytometry and high multidimensional analysis, we found a distinctive cluster containing high levels of sGRP78, CD10, CD19, and CXCR4 in bone marrow samples obtained from High-risk leukemia patients, which was absent in the compartment of Standard-risk leukemia. We confirmed that sGRP78+CXCR4+ blood-derived cells were more frequent in High-risk leukemia patients. Finally, we analyzed the dissemination capacity of sGRP78 leukemia cells in a model of xenotransplantation. sGRP78+ cells emigrated to the bone marrow and lymph nodes, maintaining the expression of CXCR4. Testing the presence of sGRP78 and CXCR4 together with conventional markers may help to achieve a better categorization of High and Standard-risk pediatric leukemia at diagnosis.


TEMPO-catalyzed aerobic oxygenation and nitrogenation of olefins via C═C double-bond cleavage.
A novel TEMPO-catalyzed aerobic oxygenation and nitrogenation of hydrocarbons via C═C double-bond cleavage has been disclosed. The reaction employs molecular oxygen as the terminal oxidant and oxygen-atom source by metal-free catalysis under mild conditions. This method can be used for the preparation of industrially and pharmaceutically important N- and O-containing motifs, directly from simple and readily available hydrocarbons.


Taxonomic and Functional Diversity of Rhizosphere Microbiome Recruited From Compost Synergistically Determined by Plant Species and Compost.
Compost is frequently served as the first reservoir for plants to recruit rhizosphere microbiome when used as growing substrate in the seedling nursery. In the present study, recruitment of rhizosphere microbiome from two composts by tomato, pepper, or maize was addressed by shotgun metagenomics and 16S rRNA amplicon sequencing. The 16S rRNA amplicon sequencing analysis showed that 41% of variation in the rhizosphere bacterial community was explained by compost, in contrast to 23% by plant species. Proteobacterial genera were commonly recruited by all three plant species with specific selections for Ralstonia by tomato and Enterobacteria by maize. These findings were confirmed by analysis of 16S rRNA retrieved from the shotgun metagenomics library. Approximately 70% of functional gene clusters differed more than sevenfold in abundance between rhizosphere and compost. Functional groups associated with the sensing and up-taking of C3 and C4 carboxylic acids, amino acids, monosaccharide, production of antimicrobial substances, and antibiotic resistance were over-represented in the rhizosphere. In summary, compost and plant species synergistically shaped the composition of the rhizosphere microbiome and selected for functional traits associated with the competition on root exudates.


Spatial resolution and contrast sensitivity of single neurons in area 19 of split-chiasm cats: a comparison with primary visual cortex.
Electrophysiological recordings were carried out in the callosal recipient zone of area 19 in normal and split-chiasm cats and, for comparison purposes, at the border of areas 17 and 18 of split-chiasm cats. The influences of retinothalamic and callosal inputs on a single cortical neurons were thereby evaluated. Extracellular recordings of single cells were made in anaesthetized and paralysed cats in the zone representing the central visual field. Receptive field properties were assessed using sine wave gratings drifting in optimal directions. Results showed that in area 19 and areas 17/18 one-third of the cells were binocularly driven after section of the optic chiasm. In area 19, the spatial resolution and contrast sensitivity of cells driven via the dominant eye were similar in the normal and split-chiasm groups. In areas 17/18 and area 19 of split-chiasm cats, binocular cells showed significant interocular matching of their receptive field properties (spatial resolution and contrast threshold), although small differences were observed. These small interocular differences were related to the cell's ocular dominance rather than to the signal transmission route (thalamic or callosal).


The financial implications of HMOs' partial county carve-out option.
This study examined the Health Care Financing Administration's policy allowing Medicare HMO risk contracts to carve out certain portions of counties from their service areas without adjusting the HMOs' capitation rate. In 1999, the policy resulted in 2.2 million Medicare enrollees losing access to HMOs and 2.3 million Medicare enrollees left with fewer HMO options. Although the majority of Medicare HMOs did not appear to be adopting the policy, there did seem to be a general tendency to exclude higher cost areas, which resulted in an estimated loss to the Medicare Trust Fund of $769 million in 1999. Of particular concern is the magnitude of profits and losses this policy generated for some individual HMOs.


Cholesteric liquid crystalline DNA; a comparative analysis of cryofixation methods.
The ultrastructure of liquid crystalline phases of DNA raises numerous problems because of the structure itself which is fluid and which nature depends on the relative amount of DNA, water and ions. Different cryofixation methods were tested and compared after freeze-fracture of the specimen. A good ultrastructural preservation of the samples can be achieved without addition of any cryoprotectant by quick-freezing against a copper block cooled down to liquid helium temperature. Then, molecular orientations can be followed very accurately and the local disorder around a mean direction which exists in the liquid state is kept in the frozen structure.


Functional Traits Are Good Predictors of Tree Species Abundance Across 101 Subtropical Forest Species in China.
What causes variation in species abundance for a given site remains a central question in community ecology. Foundational to trait-based ecology is the expectation that functional traits determine species abundance. However, the relative success of using functional traits to predict relative abundance is questionable. One reason is that the diversity in plant function is greater than that characterized by the few most commonly and easily measurable traits. Here, we measured 10 functional traits and the stem density of 101 woody plant species in a 200,000 m2 permanent, mature, subtropical forest plot (high precipitation and high nitrogen, but generally light- and phosphorus-limited) in southern China to determine how well relative species abundance could be predicted by functional traits. We found that: (1) leaf phosphorus content, specific leaf area, maximum CO2 assimilation rate, maximum stomata conductance, and stem hydraulic conductivity were significantly and negatively associated with species abundance, (2) the ratio of leaf nitrogen content to leaf phosphorus content (N:P) and wood density were significantly positively correlated with species abundance; (3) neither leaf nitrogen content nor leaf turgor loss point were related to species abundance; (4) a combination of N:P and maximum stomata conductance accounted for 44% of the variation in species' abundances. Taken together, our findings suggested that the combination of these functional traits are powerful predictors of species abundance. Species with a resource-conservative strategy that invest more in their tissues are dominant in the mature, subtropical, evergreen forest.


Raman Linewidth Contributions from Four-Phonon and Electron-Phonon Interactions in Graphene.
The Raman peak position and linewidth provide insight into phonon anharmonicity and electron-phonon interactions in materials. For monolayer graphene, prior first-principles calculations have yielded decreasing linewidth with increasing temperature, which is opposite to measurement results. Here, we explicitly consider four-phonon anharmonicity, phonon renormalization, and electron-phonon coupling, and find all to be important to successfully explain both the G peak frequency shift and linewidths in our suspended graphene sample over a wide temperature range. Four-phonon scattering contributes a prominent linewidth that increases with temperature, while temperature dependence from electron-phonon interactions is found to be reversed above a doping threshold (ℏω_{G}/2, with ω_{G} being the frequency of the G phonon).


A Novel, Validated Stability-Indicating UPLC Method for the Estimation of Lansoprazole and its Impurities in Bulk Drug and Pharmaceutical Dosage Forms.
A novel, reversed-phase ultra-performance liquid chromatographic method was developed and validated for the determination of the assay and related substances of Lansoprazole (LAN) in bulk drug and capsule dosage forms. The related substances include degradation and process-related impurities. The method was developed using the Waters Acquity BEH C18 column and gradient program with mobile phase A as a pH 7.0 phosphate buffer and methanol in the ratio of 90: 10 (v/v), and mobile phase B as methanol and acetonitrile in the ratio of 50:50 (v/v). Lansoprazole and its impurities were monitored at 285 nm. Lansoprazole was subjected to the stress conditions of oxidative, acid, base, hydrolytic, thermal, humidity, and photolytic degradation and found to degrade significantly under acid and oxidative stress conditions. The degradation products were well-resolved from the main peak and its impurities, proving the stability-indicating power of the method. The performance of the method was validated according to the present ICH guidelines for specificity, limit of detection, limit of quantification, linearity, accuracy, precision, ruggedness, and robustness.


Performance of a novel ABR-bioelectricity-Fenton coupling reactor for treating traditional Chinese medicine wastewater containing catechol.
In this study, a novel anaerobic baffled reactor-bioelectricity-Fenton (ABR-BEF) coupling reactor, combining an ABR, microbial fuel cell (MFC), and Fenton system, was used to treat traditional Chinese medicine (TCM) wastewater containing catechol. The bio-electrochemical degradation of the catechol reached 99.7% after 8 h at dissolved oxygen (DO) concentration of 4 mg/L in the cathodic chamber. The removal rates of chemical oxygen demand (COD) reached 91.7%, when the ratio rate was 1 and the DO concentration was 4 mg/L. Moreover, the maximum open-circuit voltage and power density of the coupling reactor reached 424.9 mV and 77.1 mW/m3, respectively. According to the PICRUSt analysis, carbohydrate metabolism took up the most abundant function of metabolism and the enrichment of membrane transporters may relieve TCM wastewater toxicity. These results suggest that the ABR-BEF coupling reactor could be applied as an efficient approach to treat TCM wastewater.


Treatment outcomes of 156 patients with cervical esophageal cancers treated with definitive radiation therapy- A single-institution experience of a rare cancer.
Esophageal cancer is the most prevalent malignant tumor. The incidence of cervical esophageal cancer is low and there are insufficient data on the efficacy of radical radiotherapy. The purpose of this study was to clarify the efficacy with radical IFI radiotherapy, to analyze the pattern of initial lymph node metastasis and recurrence under the new lymph node zoning of esophageal cancer. We reviewed cervical esophageal cancer treated with radical radiotherapy. The inclusion criteria were diagnosis of esophageal cancer by pathology; receiving radical radiotherapy or chemoradiotherapy; tumor location in accordance with definition of cervical esophageal cancer. Three dimensional radiotherapy was used. The target area was IFI. 156 patients entered the final analysis. The proportion of no failure was 42.31%, local esophageal failure was 30.13%, in-field lymph node metastasis was 10.26%, out-field lymph node metastasis was 1.28% and distant organ metastasis was 23.72%, second primary tumor was 2.56%. The median OS and DFS was 49.0 months (35.27-62.73) and 31.0 months (14.47-47.53). The results of initial LN metastasis pattern analysis showed the supraclavicular and upper mediastinum were the main sites of cervical esophageal cancer metastasis. In patients with recurrent LN, the results showed that the cervical, supraclavicle, upper mediastinum and abdomen were the main sites of recurrence. Our study is a retrospective study of a large sample of radical radiotherapy for cervical esophageal cancer. Failure in irradiation field is the main failure pattern. Concurrent radiotherapy and chemotherapy under IFI radiation is a considerable treatment option for cervical esophageal cancer.


Study on pharmacokinetics of scutellarin in rabbits.
To establish a solid phase extraction-high performance liquid chromatographic (SPE-HPLC) method for determining plasma scutellarin concentration, and study its pharmacokinetics after i.v. breviscapine in rabbits. Methanol-water-phosphoric acid (50:50:0.5) mixture was used as mobile phase, Nucleosil C18 column (250 mm x 4.6 mm ID) was selected. The wavelength of UV detection was 335 nm. Fifteen rabbits, randomized into 3 groups, were given breviscapine i.v. at the dose of 10, 20 and 40 mg.kg-1. Scutellarin in plasma was determined by SPE-HPLC method. Linearity was obtained over the range of 0.02-10.0 mg.L-1 of scutellarin. The method recovery was 96.15%-99.31%; the within-day and between-day RSDs were all below 10%. After i.v. 10, 20 and 40 mg.kg-1 of breviscapine to rabbits, the concentration-time curve of scutellarin fitted to a three compartment model. The main pharmacokinetic parameters showed no significant difference between low and medium doses, but the difference was significant between high dose and other doses. This assay method was accurate, sensitive, simple and suitable for the measurement of plasma scutellarin concentration. The pharmacokinetic characteristics were found to fit a three-compartment model following i.v. injection of breviscapine to rabbits. The changes of drug concentration in vivo exhibited linear kinetics ove the dosage range of 10-20 mg.kg-1, but when the dosage was 40 mg.kg-1, the linear kinetic properties disappeared.


[Study on pathogenesis of CCl4 induced cirrhosis formation in rats based on the recipe used].
To investigate the recipe-based pathogenesis and effects of Xiayuxue Decoction (XD), Yinchenhao Decoction (YcD), Yiguanjian Decoction (YgD) and Huangqi Decoction (HD) on carbon tetrachloride (CCl4) induced liver cirrhosis formation in rats on the basis of the recognition of basic pathogenesis of liver cirrhosis in TCM and train of thoughts of detecting the TCM syndrome by recipe. Model rats of liver cirrhosis were established by subcutaneous injecting of 100% CCl4 3ml/kg followed by 50% CCl4 olive solution 2ml/kg, twice a week for 12 weeks. They were randomly divided into the model group, the XD treated group, the YcD treated group, the YgD treated group and the HD treated group. Rats in the three treated group received the treatment starting from the 9th week of modeling with the corresponding decoctions. All animals were sacrificed by the end of the 12th week, and their hepatic function, liver pathological changes and hydroxyproline (Hyp) content of hepatic tissue were detected. (1) Typical chronic liver injury and fibrosis became evident in the model rat at the 8th week and cirrhosis came into being at the 12th week. (2) Compared with the rats in the model group, hepatic pathological changes were alleviated significantly, content of Hyp in hepatic tissue was decreased markedly and hepatic function improved remarkably in the XD group and YgD group. The improvement in the XD group was superior to that in the YgD group, while the serum albumin level elevated more significant in the YgD group. The main pathological changes during CCl4 induced liver cirrhosis formation in rats is the rapid hyperplasia of hepatic fibrous connective tissue and obstruction of collaterals by blood stasis, thus induced reconstruction of the tissue structure, which could be treated with XD effectively, while the severe injury of liver parenchyma in this phase is another pathological change of Gan-yin deficiency syndrome, which could be effectively treated with YgD by its Yin-nourishing action.


Ornithine decarboxylase encoded by chlorella virus PBCV-1.
Sequence analysis of the 330-kb genome of chlorella virus PBCV-1 revealed an open reading frame, A207R, which encodes a protein with 37-41% amino acid identity to ornithine decarboxylase (ODC) from many eukaryotic organisms. The a207r gene was cloned and the protein was expressed as a His-A207R fusion protein in Escherichia coli. The recombinant protein catalyzes pyridoxal 5'-phosphate-dependent decarboxylation of ornithine to putrescine, the first step in the polyamine biosynthetic pathway. The enzyme has a pH optimum of 9.0 and a temperature optimum of 42 degrees C, and it requires dithiothreitol for maximal activity. The enzyme has a K(m) for ornithine of 0.78 mM and a specific activity of 100 micromol/min/mg protein. PBCV-1 ODC is quite sensitive to the competitive inhibitor L-arginine and the irreversible inhibitor difluoromethylarginine but it is less sensitive to the irreversible inhibitor difluoromethylornithine. The a207r gene is expressed both early and late in PBCV-1 infection and is highly conserved among the chlorella viruses. The 42-kDa PBCV-1 ODC (372 amino acids) is the smallest ODC in the databases and, to our knowledge, is the first virus-encoded ODC.


Inhaled corticosteroids in the management of chronic bronchial asthma.
Inhaled corticosteroids have become a standard treatment for chronic severe asthma in adults and in children. In patients whose symptoms are not controlled by beta 2 agonist, theophylline preparations and disodium cromoglycate, inhaled corticosteroids can either reduce the need of or substitute for oral corticosteroids. Inhaled corticosteroids are not effective in an acute asthmatic attack when oral corticosteroids are indicated. Patients who are not adequately treated with conventional doses of inhaled corticosteroids may benefit from high dose inhaled corticosteroids.


Acanthamoeba keratitis with two species of Acanthamoeba.
We describe a case of Acanthamoeba keratitis related to soft contact lens wear. The patient presented with a 3-week history of severe uniocular pain, radial stromal infiltrates and subepithelial infiltrates with no epithelial defect. Acanthamoeba was cultured from the corneal biopsy specimen, contact lens and lens case. The corneal biopsy culture grew both A. castellani and A. polyphaga as well as Escherichia coli. The patient was treated with topical dibromopropamidine isethionate (Brolene) drops, neomycin and polymyxin B drops and fortified gentamicin drops. Gradual clinical improvement ensued.


Brief Report: impaired Flexible Item Selection Task (FIST) in school-age children with autism spectrum disorders.
Cognitive flexibility has been measured with inductive reasoning or explicit rule tasks in individuals with autism spectrum disorders (ASD). The Flexible Item Selection Task (FIST) differs from previous cognitive flexibility tasks in ASD research by giving children an abstract, ambiguous rule to switch. The ASD group (N = 22; Mean age = 8.28 years, SD = 1.52) achieved a lower shift percentage than the typically developing verbal mental-age control group (N = 22; Mean age = 6.26 years, SD = 0.82). There was a significant positive correlation between verbal mental age and shift percentage for children with ASD. Group differences on the FIST converge and extend prior evidence documenting an impaired ability to adapt rapidly to changes in task demands for individuals with ASD.