Alpha-adrenergic stimulation of glutamine metabolism in isolated rat hepatocytes.
The mechanisms by means of which phenylephrine stimulates glutamine metabolism were studied in isolated rat hepatocytes. In the first 2 min after phenylephrine addition there was a rapid fall in the concentrations of intracellular 2-oxoglutarate and glutamate, presumably owing to activation of 2-oxoglutarate dehydrogenase. This was followed 2-3 min later by activation of glutaminase and by increases in glutamate and 2-oxoglutarate. Activation of glutaminase by phenylephrine was due to direct stimulation of the enzyme rather than to reversal of inhibition by the decrease in 2-oxoglutarate and glutamate. The stimulation of glutaminase by phenylephrine is partly due to an increase in the affinity of the enzyme for ammonia, its essential activator. It is concluded that stimulation of steady-state flux through the pathway from glutamine to glucose and urea can only be achieved by stimulation of glutaminase, the first enzyme in the pathway.


ATXN3 promotes proliferation, stemness and motility of clear cell renal cell carcinoma cells by regulating S100A8 ubiquitination.
Clear cell renal cell carcinoma (ccRCC) is a dominant subtype of kidney cancer with a dismal outcome at advanced stages. Ataxin 3 (ATXN3) has been proven to play a cancer-promoting role in several tumors and is upregulated in the patients with renal cell carcinoma. Thus, the objective of this research is to examine the biological roles and underlying mechanisms of ATXN3 in ccRCC. Bioinformatics analysis was carried out to analyze ATXN3 expression in ccRCC tissues and patient survival. Gain- and loss-of-function assays were applied to explore the effect of ATXN3 on ccRCC cell malignant behavior in vitro. The effect of ATXN3 on the NF-κB pathway was assessed by Western blot and immunofluorescence staining. The binding between ATXN3 and S100A8 and the effect of ATXN3 on S100A8 ubiquitination were verified using coimmunoprecipitation. ATXN3 was upregulated in ccRCC tissues and correlated with adverse patient outcome. ATXN3 overexpression facilitated the proliferation, stemness, invasion and migratory capacity of ccRCC cells, whereas silencing had the opposite effect. ATXN3 enhanced the activity of the NF-κB pathway. Silencing ATXN3 facilitated S100A8 ubiquitination. Rescue experiments demonstrated that S100A8 downregulation reversed the promoting effect of ATXN3 on malignant behavior and NF-κB pathway activation in ccRCC cells. ATXN3 exerts a cancer-promoting effect in ccRCC by regulating S100A8 ubiquitination. Therefore, targeting the ATXN3/S100A8/NF-κB axis may provide a novel underlying therapeutic strategy for ccRCC.


Methods in molecular cardiology: proteome analysis.
Proteins are involved in virtually every cellular function, they control regulatory mechanisms and are modified in diseases (either cause or effect). To understand the function and adaptation of a cell, the researcher has to be able to identify proteins and visualise the concentrations and form in which the proteins are expressed. The technique is called 'proteomics' or 'proteome analysis'. In this article proteomics will be explained from starting material to detection and analysis of the individual proteins. It will give an indication of the work involved and how it can be implemented in cardiovascular research.


Prevalence of HIV and HCV infections in two populations of Malian women and serological assays performances.
To estimate the prevalence of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) infections in women in Mali and to evaluate the performance of serological assays. Two prospective studies were conducted in 2009 and 2010 in Mali. They concerned first, 1000 pregnant women attending six reference health centers in Bamako (Malian capital) between May 26 and June 16, 2009; and secondly, 231 women over 50 years who consulted general practitioners of two hospitals in Bamako between October 25 and December 24, 2010. Blood samples were collected and kept frozen in good condition before analysis. All samples depicted as positive using HIV/HCV enzyme immuno-assay screening assays were submitted to confirmation analysis. Molecular markers of HCV were characterized. The seroprevalence of HIV and HCV in the population of pregnant women was 4.1% and 0.2% respectively. Among older women the seroprevalence was higher and similar for HIV and HCV (6.1% vs 6.5%). The anti-HIV prevalence was not different in young and older women (4.1% vs 6.1%). In contrast, the anti-HCV prevalence was higher in older compared to younger women (6.5% vs 0.2%, P < 0.01). Of 2 pregnant women who were HCV seropositive, only one was polymerase chain reaction (PCR) reactive and infected by genotype 2, with a viral load of 1600 IU/mL. Regarding older women who were HCV seropositive, 13 out of 15 were PCR reactive, infected by genotype 1 or 2. Globally HCV genotype 2 was predominant. The positive predictive value (PPV) measured with VIKIA HIV test in young women was 100% therefore significantly higher than the 87.5% measured in older women (P < 0.05). Conversely, the PPV measured with Monolisa HCV assay in older women was 88.2% and higher than the 14.3% measured in younger women (P < 0.01). Whereas HIV prevalence was similar in both subpopulations HCV was more frequent among older women (P < 0.01). The PPV of screening assays varied with the age of the subjects.


Association of Stunting with Socio-Demographic Factors and Feeding Practices among Children under Two Years in Informal Settlements in Gauteng, South Africa.
Despite improvements in childhood undernutrition through integrated nutritional programs in South Africa, stunting among children remains persistent, and is attributed to poor socio-demographic status. This context has been implicated in disrupting mothers' decisions regarding effective infant feeding, ultimately meaning that children's nutritional demands remain unmet. In view of this, we conducted a cross-sectional study to determine the association between socio-demographic factors and infant and young child feeding (IYCF) practices and stunting among children under two years receiving primary health care in informal settlements in Gauteng, South Africa. A validated questionnaire was used to assess mothers' socio-demographic status and feeding practices using WHO core indicators. Stunting was defined as length-for age z-scores (LAZ) below -2 standard deviation, computed using WHO Anthro software version 188.8.131.52 using age, sex, and anthropometric measurements of children. Univariate and multivariate analyses were stratified by stunting to determine the relationship with socio-demographic, infant, and IYCF factors using STATA 17. The prevalence of stunting was 16% among surveyed children under two years (with a mean age of 8 ± 5 months) living in poor socio-demographic households. Poor feeding practices were characterized by delayed initiation of breastfeeding (58%), sub-optimal exclusive breastfeeding (29%), discontinued breastfeeding (44%), early introduction of solid foods (41%), and low dietary diversity (97%). Significant differences in terms of child's age, monthly household income, and ever being breastfed were observed (Chi square test and univariate analysis). After controlling for potential confounders, stunting was significantly associated with child's age [12-23 months: AOR = 0.35, 95% CI: 0.16-0.76], and monthly household income [ZAR 3000-ZAR 5000: AOR = 0.47, 95% CI: 0.26-0.86]. Despite the few aforementioned socio-demographic and IYCF factors associated with stunting, this study reiterates stunting as the commonest poor nutritional status indicator among children under two years, suggesting the presence of chronic undernutrition in these poverty-stricken informal settlements. A multisectoral approach to address stunting should be context-specific and incorporate tailor-made interventions to promote optimal infant-feeding practices. Conducting future nutrient assessments focusing on children is imperative.


Therapeutic potential of antisense oligonucleotides as modulators of alternative splicing.
An estimated 60% of all human genes undergo alternative splicing, a highly regulated process that produces splice variants with different functions. Such variants have been linked to a variety of cancers, and genetic diseases such as thalassemia and cystic fibrosis. This Perspective describes a promising approach to RNA repair based on the use of antisense oligonucleotides to modulate alternative splicing and engender the production of therapeutic gene products.


Clinical presentation and outcomes of acute coronary syndromes in the gulf registry of acute coronary events (Gulf RACE).
To identify the characteristics, treatments and hospital outcomes for patients diagnosed with acute coronary syndromes (ACS) in the Gulf area. Prospective, multinational, multicentre, observational survey of consecutive ACS patients who were admitted to 65 hospitals during May 2006. A total of 1484 ACS patients were recruited. The mean age was 55 years, and 76% were men. The final discharge diagnosis was ST-segment elevation myocardial infarction (STEMI) in 37%, non-ST-segment elevation myocardial infarction (NSTEMI) in 32%, left bundle branch block myocardial infarction (LBBB MI) in 2%, and unstable angina in 29%. Among patients with STEMI and LBBB MI, the reperfusion rate was 65%, with use of primary percutaneous coronary intervention in 7% and thrombolytic therapy in 93%. When thrombolytic therapy was used, the median door to needle time was 45 minutes, with 37% receiving it within 30 minutes of hospital presentation. During the first day of hospitalization, aspirin was administered to 94%, clopidogrel to 51%, and beta blockers to 65%. Angiotensin converting enzyme inhibitors/Angiotensin receptor blockers and statins were used in 62% and 82%, respectively. Coronary angiography during hospitalization was performed in 21%. In-hospital mortality was 3%. We were able to determine the characteristics, treatments and in-hospital outcomes of patients hospitalized with ACS in our region. There is room for improvement in using medications, reducing needle to door time and utilizing more cardiac catheterization services.


Interdomain orientation of cardiac troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state.
Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations.


Quality of Sleep among Portuguese Anaesthesiologists: A Cross-Sectional Study.
Sleeping is essential to maintain proper relationships with others, keep alertness, and execute responsibilities, among many other functions. In the medical profession, there are several studies linking sleep deprivation with a decrease in responsiveness, cognition and attention. With this study we intended to characterize the sleep pattern of Portuguese anaesthesiologists and identify independent factors associated with sleep quality in this population. An observational, cross-sectional study of senior and resident anesthesiologists working in Portugal was carried out through an online questionnaire. Individuals working exclusively in intensive care units, emergency departments or with previously diagnosed sleep disorders were excluded. Socio-demographic data, Pittsburgh Sleep Quality Index, Epworth Sleepiness Scale and Perceived Stress Scale were applied. Statistical significance was assessed using the Mann-Whitney test and the chi-square test. A multivariable analysis was performed to examine the association between the Pittsburgh Sleep Quality Index and certain variables. Among 256 respondents, 46.1% reported "poor" quality of sleep (Pittsburgh Sleep Quality Index > 5). Within these individuals, 77.1% slept less than 7 hours per night (p < 0.001). Excessive daytime sleepiness (Epworth Sleepiness Scale > 10) was present in 41.0% of the sample, and the median Perceived Stress Scale score was 17.0. The independent factors associated with worse quality ofsleep were the number of working hours/week (OR 1.03, 95% CI 1,01 to 1,06), perceived stress (OR 1.18, 95% CI 1.11 to 1.26), taking sleep medication (OR 14.72, 95% CI 5.55 to 39.08), and sleep hours/night (OR 0.25, 95% CI 0.15 to 0.42). This fraction of Portuguese anaesthesiologists presented a poorer quality of sleep, with excessive daytime somnolence, perceived stress and higher sedative use compared to previously studied populations. Our study characterizes sleep patterns and identifies potential risk factors linked to sleep disturbances in a sample of Portuguese anaesthesiologists. Government and institutional policies can endorse sleep hygiene practices and habits, promoting healthier working environments.


Efficacy of endoscopic ultrasonography-guided fine needle aspiration for pancreatic neuroendocrine tumor grading.
To evaluate the efficacy of endoscopic ultrasonography-guided fine needle aspiration (EUS-FNA) for grading pancreatic neuroendocrine tumors (PNETs). A total of 22 patients were diagnosed with PNET by EUS-FNA between October 2001 and December 2013 at Fukushima Medical University Hospital. Among these cases, we targeted 10 PNET patients who were evaluated according to the World Health Organization (WHO) 2010 classification. Surgery was performed in eight patients, and chemotherapy was performed in two patients due to multiple liver metastases. Specimens obtained by EUS-FNA were first stained with hematoxylin and eosin and then stained with chromogranin, synaptophysin, CD56, and Ki-67. The specimens were graded by the Ki-67 index according to the WHO 2010 classification. Specimens obtained by surgery were graded by the Ki-67 index and mitotic count (WHO 2010 classification). For the eight specimens obtained by EUS-FNA, the Ki-67 index results were compared with those obtained by surgery. In the two cases treated with chemotherapy, the effects and prognoses were evaluated. The sampling rate for histological diagnosis by EUS-FNA was 100%. No adverse effects were observed. The concordance rate between specimens obtained by EUS-FNA and surgery was 87.5% (7/8). For the two cases treated with chemotherapy, case 1 received somatostatin analog therapy and transcatheter arterial infusion (TAI) targeting multiple liver metastases. Subsequent treatment consisted of everolimus. During chemotherapy, the primary tumor remained unconfirmed, although the multiple liver metastases diminished dramatically. Case 2 was classified as neuroendocrine carcinoma (NEC) according to the Ki-67 index of a specimen obtained by EUS-FNA; therefore, cisplatin and irinotecan therapy was started. However, severe adverse effects, including renal failure and diarrhea, were observed, and the therapy regimen was changed to cisplatin and etoposide. TAI targeting multiple liver metastases was performed. Although the liver metastases diminished, the primary tumor remained unconfirmed. These chemotherapy regimens had immediate effects for both unresectable neuroendocrine tumor (NET) and NEC cases. These two subjects are still alive. EUS-FNA was effective for PNET diagnosis and Ki-67 index grading for WHO 2010 classification, enabling informed decisions on unresectable PNET treatment by identifying NET or NEC.


Clinics in diagnostic imaging (70). Bilateral sacroiliitis due to ankylosing spondylitis.
A 15-year-old boy was incidentally found to have bilateral sacroiliitis. Radiographs showed erosions and subchondral sclerosis of both sacroiliac joints. His ESR was raised and HLA-B27 was positive. Presumptive diagnosis was ankylosing spondylitis. The clinical and imaging features, including the radiographical classification, of sacroiliitis are discussed. Knowledge of the patterns of distribution and correlation with clinical manifestations aid in differentiating among the various causes of sacroiliitis.


Androgen Receptor and PI3K Pathway Activity in Ovarian Cancer.
We sought to evaluate androgen receptor (AR) and PI3K pathway activity in ovarian cancer cell lines and tissue and determine if either pathway was correlated with growth of ovarian cancers. AR expression and activity were quantified using immunohistochemistry (IHC) and RT-qPCR in six ovarian cancer cell lines and 51 tissue samples. Phospho-mTOR and AKT expression were quantified by IHC as well. Cell growth was assessed in the presence of AR modulating drugs and metformin. We found that despite robust AR expression and activity, no cell line was dependent on androgen for growth. However, metformin inhibited activity in five of the six cell lines. Patient tissues had large variation in AR expression and activity, as well as in expression of phospho-mTOR and AKT, but none of these variables correlated with progression-free survival (PFS). AR expression and activity did not predict the dependence of ovarian cancer cell lines on androgens for growth, and AR expression and activity did not correlate with PFS. This result suggests that AR expression as a criterion for patient selection for clinical trials evaluating molecules targeting AR may not predict response for ovarian cancer patients.


Folic acid administration reduces neointimal thickening, augments neo-vasa vasorum formation and reduces oxidative stress in saphenous vein grafts from pigs used as a model of diabetes.
There is evidence that plasma homocysteine augments vein graft failure and that it augments both micro- and macro-angiopathy in patients with diabetes mellitus. It is therefore suggested that homocysteine may augment vein graft thickening, a major cause of vein graft failure, in diabetic patients, as well as impairing adaptive growth of a new vasa vasorum, possibly through overproduction of superoxide. In order to test these proposals, the effect of folic acid administration, which lowers plasma homocysteine, on vein graft thickening and microvessel density was studied in pigs used as a model of diabetes. Non-ketotic hyperglycaemia was induced in Landrace pigs by intravenous injection of streptozotocin, and folic acid was fed daily for 1 month. Vein grafts were excised and the thickness of the neointima and media and microvessel density were assessed by planimetry and superoxide formation. Plasma total homocysteine was significantly reduced by folic acid in both control and diabetic pigs, whereas glucose was unchanged. Compared with controls, diabetic pigs showed increased neointimal thickness and superoxide formation and decreased adventitial microvessel density. Folic acid reduced neointimal thickness and superoxide formation and augmented microvessel density in diabetic but not in control pigs. Folic acid administration reduces neointimal thickening, augments vasa vasorum neoformation and reduces oxidative stress in saphenous vein grafts from diabetic pigs. Folic acid may therefore be particularly effective in reducing vein graft failure in diabetic patients.


Association between duration of humid season and geographical distribution patterns of different species of chewing lice (Mallophaga:Insecta) infesting domestic chickens in Nigeria.
In a countrywide study of the geographical distribution patterns of chicken lice in Nigeria, 3 species, Menacanthus pallidulus (Neumann 1912), Goniodes dissimilis Denny, 1842, and Menopon gallinae (Linnaeus, 1758), were completely restricted to within areas with a long humid season (6-12 humid mo per yr). In constrast, 4 species, Cuclotogaster occidentalis Tendeiro, 1954, Menacanthus cornutus (Schommer, 1913), Amyrsidea powelli (Bedford, 1920), and Goniodes gigas (Taschenberg, 1879), thrived only in areas where the humid season was of a short duration (< or = 5 mo). The last 3 named species extended also into the fringes of areas with a long humid season (6-8 humid mo) at significantly reduced numbers. They were entirely absent in areas with a particularly protracted humid season (9-12 humid mo). Three other species. Numidilipeurus tropicalis (Peters, 1931), Goniocotes gallinae (DeGeer, 1778), and Lipeurus caponis (Linnaeus, 1758), were just as abundant in areas of a long humid season as they were in areas with a short humid season. It was concluded that there was an association between duration of the humid season and the geographical distribution patterns of certain species of chicken lice in Nigeria. The duration of the humid season might thus provide a means of predicting the potential occurrence of some species of chicken lice in a particular area.


What is eHealth (6): perspectives on the evolution of eHealth research.
The field of eHealth holds promise for supporting and enabling health behavior change and the prevention and management of chronic disease. In order to establish areas of congruence and controversy among contributors to the early development, evaluation, and dissemination of eHealth applications, as well as the desire to inform an evaluation research funding agenda, 38 semistructured, qualitative interviews were conducted among stakeholders in eHealth between May 2002 and September 2003. Participants were asked about their perspectives on the credibility, value, and future potential of information technology for health behavior change and chronic disease management. Interviews were coded and analyzed for emergent themes using qualitative methods. Consistent themes were identified across stakeholder groups, with slight differences in emphasis. These topics included the following: (1) consensus and standardization-most stakeholders expressed a strong desire for a more coordinated, rigorous effort to define and integrate the field; (2) evaluation methods and challenges-demonstrating outcomes is required to establish eHealth quality and efficacy, but stakeholders were not satisfied with the sensitivity, validity, and reliability of existing outcome measures; (3) quality, value, and future potential-the intersection between eHealth's potential cost-effectiveness, efficiency, and improved clinical status among users generated a high degree of interest; and (4) health disparities-many stakeholders contended that traditionally underserved populations will particularly benefit from eHealth applications, although others argued that the underserved are also disadvantaged in terms of access to technology. Recommendations included the need for improvement and formalization of development and evaluation standards across private and public sectors, additional research on the technology needs and preferences of traditionally underserved populations, and long-term epidemiologic studies of the impact of eHealth on outcomes and cost-effectiveness.


Structure and regulation of gene expression of a Clo DF13 plasmid DNA region involved in plasmid segregation and incompatibility.
The bacteriocinogenic plasmid Clo DF13 contains genetic information involved in the accurate partitioning of the plasmid (parA and parB) as well as in incompatibility phenomena (incA, B, C and D). In this paper we report on the primary structure and regulation of gene expression of the 29% - 50% part of Clo DF13, containing the DNA regions incA, incB and parB as well as genes K and L. According to the results of our DNA sequence analysis, mapping of transposon insertions, RNA blotting and S1 mapping experiments, we conclude that: a) genes K and L are transcribed as one operon; transcription of this operon is initiated at a promoter (P2) located at 32.5% and proceeds in a clockwise direction. b) treatment of cells with mitomycin-C, significantly enhances transcription from P2, although this promoter is probably not directly repressed by lexA protein. c) Termination of transcription of this operon occurs between genes K and L, as well as distal to gene L. The possible role of gene products and/or sites, located within the 29-50% DNA region, in plasmid incompatibility and segregation is discussed.


Estimating the Number of Pregnant Women Infected With Zika Virus and Expected Infants With Microcephaly Following the Zika Virus Outbreak in Puerto Rico, 2016.
Zika virus (ZIKV) infection during pregnancy is a cause of congenital microcephaly and severe fetal brain defects, and it has been associated with other adverse pregnancy and birth outcomes. To estimate the number of pregnant women infected with ZIKV in Puerto Rico and the number of associated congenital microcephaly cases. We conducted a modeling study from April to July 2016. Using parameters derived from published reports, outcomes were modeled probabilistically using Monte Carlo simulation. We used uncertainty distributions to reflect the limited information available for parameter values. Given the high level of uncertainty in model parameters, interquartile ranges (IQRs) are presented as primary results. Outcomes were modeled for pregnant women in Puerto Rico, which currently has more confirmed ZIKV cases than any other US location. Zika virus infection in pregnant women. Number of pregnant women infected with ZIKV and number of congenital microcephaly cases. We estimated an IQR of 5900 to 10 300 pregnant women (median, 7800) might be infected during the initial ZIKV outbreak in Puerto Rico. Of these, an IQR of 100 to 270 infants (median, 180) may be born with microcephaly due to congenital ZIKV infection from mid-2016 to mid-2017. In the absence of a ZIKV outbreak, an IQR of 9 to 16 cases (median, 12) of congenital microcephaly are expected in Puerto Rico per year. The estimate of 5900 to 10 300 pregnant women that might be infected with ZIKV provides an estimate for the number of infants that could potentially have ZIKV-associated adverse outcomes. Including baseline cases of microcephaly, we estimated that an IQR of 110 to 290 total cases of congenital microcephaly, mostly attributable to ZIKV infection, could occur from mid-2016 to mid-2017 in the absence of effective interventions. The primary limitation in this analysis is uncertainty in model parameters. Multivariate sensitivity analyses indicated that the cumulative incidence of ZIKV infection and risk of microcephaly given maternal infection in the first trimester were the primary drivers of both magnitude and uncertainty in the estimated number of microcephaly cases. Increased information on these parameters would lead to more precise estimates. Nonetheless, the results underscore the need for urgent actions being undertaken in Puerto Rico to prevent congenital ZIKV infection and prepare for affected infants.


[Cholestatic hepatitis in situs inversus abdominalis].
A 75 year old patient was admitted to hospital due to painless jaundice and fatigue. Patient's history was not remarkable for chronic viral hepatitis, autoimmune hepatitis, hereditary metabolic disorders or a hepatotoxic damage. Laboratory tests revealed significant elevation of the parameters of cholestasis and aminotransferase activity, a significantly elevated titer of both antinuclear (ANA) and antimitochondrial (AMA) antibodies. Abdominal ultrasound and computertomography showed no mechanic cholestasis nor tumorous process but a situs inversus abdominalis of the abdominal parenchymatous organs. Portal and periportal inflammation, consistent with autoimmune hepatitis, was diagnosed histologically after performing laparoscopic liver biopsy. Regarding the significantly elevated cholestatic parameters and the positive AMA-autoantibodies AIH-PBC overlap syndrome was highly probable so medical treatment including both ursodeoxycholic acid and immunosuppressive therapy with corticosteroids and azathioprine was started resulting in a continous downward tendency of liver enzymes and an improvement of the patient's clinical condition. Finding the correct diagnosis and therapy of autoimmune liver disease is not always easy and unproblematic. Regarding its sometimes fatal progression and good response towards medical treatment prompt diagnosis and institution of autoimmune hepatitis--alone or combined as PBC/AIH overlap syndrome--should not be deferred. Laparoscopic liver biopsy should be the method of choice in patients's with situs inversus abdominalis regading the variable anatomy.


Substrate curvature governs texture orientation in thin films of smectic block copolymers.
Self-assembly of ordered nanometer-scale patterns is interesting in itself, but its practical value depends on the ability to predict and control pattern formation. In this paper we demonstrate theoretically and numerically that engineering of extrinsic as well as intrinsic substrate geometry may provide such a controllable ordering mechanism for block copolymers films. We develop an effective two-dimensional model of thin films of striped-phase diblock copolymers on general curved substrates. The model is obtained as an expansion in the film thickness and thus takes the third dimension into account, which crucially allows us to predict the preferred orientations even in the absence of intrinsic curvature. We determine the minimum-energy textures on several curved surfaces and arrive at a general principle for using substrate curvature as an ordering field, namely that the stripes will tend to align along directions of maximal curvature.


Potential impact of antiretroviral therapy and screening on cervical cancer mortality in HIV-positive women in sub-Saharan Africa: a simulation.
Despite having high cervical cancer incidence and mortality rates, screening for cervical precancerous lesions remains infrequent in sub-Saharan Africa. The need to screen HIV-positive women because of the higher prevalence and faster progression of cervical precancerous lesions may be heightened by the increased access to highly-active antiretroviral therapy (HAART). Policymakers need quantitative data on the effect of HAART and screening to better allocate limited resources. Our aim was to quantify the potential effect of these interventions on cervical cancer mortality. We constructed a Markov state-transition model of a cohort of HIV-positive women in Cameroon. Published data on the prevalence, progression and regression of lesions as well as mortality rates from HIV, cervical cancer and other causes were incorporated into the model. We examined the potential impact, on cumulative cervical cancer mortality, of four possible scenarios: no HAART and no screening (NHNS), HAART and no screening (HNS), HAART and screening once on HAART initiation (HSHI), and HAART and screening once at age 35 (HS35). Our model projected that, compared to NHNS, lifetime cumulative cervical cancer mortality approximately doubled with HNS. It will require 262 women being screened at HAART initiation to prevent one cervical cancer death amongst women on HAART. The magnitudes of these effects were most sensitive to the rate of progression of precancerous lesions. Screening, even when done once, has the potential of reducing cervical cancer mortality among HIV-positive women in Africa. The most feasible and cost-effective screening strategy needs to be determined in each setting.


Delivery of interferon-beta to the monkey nervous system following intranasal administration.
We determined the nervous system targeting of interferon-beta1b (IFN-beta1b), a 20 kDa protein used to treat the relapsing-remitting form of multiple sclerosis, following intranasal administration in anesthetized, adult cynomolgus monkeys. Five animals received an intranasal bolus of [(125)I]-labeled IFN-beta1b, applied bilaterally to the upper nasal passages. Serial blood samples were collected for 45 min, after which the animals were euthanized by transcardial perfusion-fixation. High resolution phosphor imaging of tissue sections and gamma counting of microdissected tissue were used to obtain the distribution and concentration profiles of [(125)I]-IFN-beta1b in central and peripheral tissues. Intranasal administration resulted in rapid, widespread targeting of nervous tissue. The olfactory bulbs and trigeminal nerve exhibited [(125)I]-IFN-beta1b levels significantly greater than in peripheral organs and at least one order of magnitude higher than any other nervous tissue area sampled. The basal ganglia exhibited highest [(125)I]-IFN-beta1b levels among CNS regions other than the olfactory bulbs. Preferential IFN-beta1b distribution to the primate basal ganglia is a new finding of possible clinical importance. Our study suggests both IFN-beta and IFN-alpha, which share the same receptor, may be bound with relatively high affinity in these structures, possibly offering new insight into a neurovegetative syndrome induced by IFN-alpha therapy and suspected to involve altered dopamine neurotransmission in the basal ganglia. Most importantly, our results suggest intranasally applied macromolecules may bypass the blood-brain barrier and rapidly enter the primate CNS along olfactory- and trigeminal-associated extracellular pathways, as shown previously in the rat. This is the first study to finely detail the central distribution of a labeled protein after intranasal administration in non-human primates.


Distributions of amino acids suggest that certain residue types more effectively determine protein secondary structure.
Exponential growth in the number of available protein sequences is unmatched by the slower growth in the number of structures. As a result, the development of efficient and fast protein secondary structure prediction methods is essential for the broad comprehension of protein structures. Computational methods that can efficiently determine secondary structure can in turn facilitate protein tertiary structure prediction, since most methods rely initially on secondary structure predictions. Recently, we have developed a fast learning optimized prediction methodology (FLOPRED) for predicting protein secondary structure (Saraswathi et al. in JMM 18:4275, 2012). Data are generated by using knowledge-based potentials combined with structure information from the CATH database. A neural network-based extreme learning machine (ELM) and advanced particle swarm optimization (PSO) are used with this data to obtain better and faster convergence to more accurate secondary structure predicted results. A five-fold cross-validated testing accuracy of 83.8 % and a segment overlap (SOV) score of 78.3 % are obtained in this study. Secondary structure predictions and their accuracy are usually presented for three secondary structure elements: α-helix, β-strand and coil but rarely have the results been analyzed with respect to their constituent amino acids. In this paper, we use the results obtained with FLOPRED to provide detailed behaviors for different amino acid types in the secondary structure prediction. We investigate the influence of the composition, physico-chemical properties and position specific occurrence preferences of amino acids within secondary structure elements. In addition, we identify the correlation between these properties and prediction accuracy. The present detailed results suggest several important ways that secondary structure predictions can be improved in the future that might lead to improved protein design and engineering.


Precancerous lesions and conditions of the prostate: from morphological and biological characterization to chemoprevention.
Prostatic intraepithelial neoplasia (PIN) is composed of dysplastic cells with a luminal cell phenotype, expressing the androgen receptor as well as prostate-specific antigen. PIN is characterized by progressive abnormalities of phenotype that are intermediate between normal prostatic epithelium and cancer, indicating impairment of cell differentiation and regulatory control with advancing stages of carcinogenesis. High-grade PIN is considered the most likely precursor of prostatic carcinoma, according to virtually all available evidence. Androgen deprivation decreases the prevalence and extent of PIN and the degree of capillary vascularization (e.g., angiogenesis) in the surrounding stroma via suppression of vascular endothelial growth factor production. Prostatic carcinoma is also likely to arise from precursor lesions other than high-grade PIN such as low-grade PIN, atypical adenomatous hyperplasia, malignancy-associated foci, and atrophy.


Insulin-like growth factor II blocks apoptosis of N-myc2-expressing woodchuck liver epithelial cells.
N-myc2 and insulin-like growth factor II (IGF-II) are coordinately overexpressed in the great majority of altered hepatic foci, which are the earliest precancerous lesions observed in the liver of woodchuck hepatitis virus carrier woodchucks, and these genes continue to be overexpressed in hepatocellular carcinomas (HCCs). We have investigated the function of these genes in woodchuck hepatocarcinogenesis by using a woodchuck liver epithelial cell line (WC-3). WC-3 cells react positively with a monoclonal antibody (12.8.5) against woodchuck oval cells, suggesting a lineage relationship with oval cells. Overexpression of N-myc2 in three WC-3 cell lines caused their morphological transformation and increased their growth rate and saturation density in medium containing 10% serum. Removal of serum from the medium increased cell death of the N-myc2-expressing lines, whereas cell death in control lines was minimal. The death of N-myc2-expressing WC-3 cells was accompanied by nucleosomal fragmentation of cellular DNA, and DAPI (4',6-diamidino-2-phenylindole) staining revealed condensation and fragmentation of the nuclei, suggesting that N-myc2-expressing WC-3 cells undergo apoptosis in the absence of serum. In colony regression assays, conducted in the absence of serum, control colonies were stable, while N-myc2-expressing colonies regressed to various degrees. Addition of recombinant human IGF-II to the serum-free medium blocked both cell death and colony regression in all the N-myc2-expressing lines. Therefore, coordinate overexpression of N-myc2 and IGF-II in woodchuck altered hepatic foci may allow cells which otherwise might die to survive and progress to hepatocellular carcinoma.


Is ribonucleotide reductase the transforming function of herpes simplex virus 2?
Transformation of cells by herpes simplex virus 2 (HSV-2) can be induced by the BglII C (0.43-0.58 map units) or N (0.58-0.625) fragments of the viral genome. Sequences partially overlapping both fragments (0.566-0.602) encode two 3' coterminal mRNAs; these in turn direct the synthesis of two related polypeptides of molecular weight 140,000 (140K) and 35K (refs 4, 7), which may be involved in transformation. Recently, a temperature-sensitive (ts) mutation affecting HSV-induced ribonucleotide reductase has been mapped within this common region (B.M. Dutia, personal communication). We have partially purified the induced reductase and raised a rabbit antiserum to it which inhibits the enzyme activity and immunoprecipitates from infected cells a 144K polypeptide and minor species including a 38K polypeptide. Here we show that a monoclonal antibody to the putative transforming proteins competes with the rabbit serum for the 144K and 38K antigens and also immunoprecipitates specifically the induced reductase activity. These results suggest a possible role for ribonucleotide reductase in HSV-2-induced transformation.


Localization of metastasis within the sentinel lymph node biopsies: a predictor of additional axillary spread of breast cancer?
To explore the relationship between morphological characteristics and histologic localization of metastasis within sentinel lymph nodes (SLN) and axillary spread in women with breast cancer. We selected 119 patients with positive SLN submitted to complete axillary lymph node dissection from July 2002 to March 2007. We retrieved the age of patients and the primary tumor size. In the primary tumor, we evaluated histologic and nuclear grade, and peritumoral vascular invasion (PVI). In SLNs we evaluated the size of metastasis, their localization in the lymph node, number of foci, number of involved lymph nodes, and extranodal extension. Fifty-one (42.8%) patients had confirmed additional metastasis in non-sentinel lymph nodes (NLSN). High histologic grade, PVI, intraparenchymatous metastasis, extranodal neoplastic extension and size of metastasis were associated with positive NLSN. SLN metastasis affecting the capsule were associated to low risk incidence of additional metastasis. After multivariate analysis, PVI and metastasis size in the SLN remained as the most important risk factors for additional metastasis. The risk of additional involvement of NSLN is higher in patients with PVI and it increases progressively according the histologic localization in the lymph node, from capsule, where the afferent lymphatic channel arrives, to the opposite side of capsule promoting the extranodal extension. Size of metastasis greater than 6.0 mm presents higher risk of additional lymph node metastasis.


Have companies improved their health and safety approaches over the last decade? A longitudinal study.
Workplace level health and safety (H&S) policies, practices, and attitudes were compared longitudinally in 120 manufacturing firms. A mailed questionnaire for worker and management representatives in the sampled worksites was first completed in 1990 [Shannon et al. (1996) Am J Ind Med 29:258-268]. Workplaces that were still in business in 2001 were re-surveyed to assess change over time in key variables previously found to be related to lost-time injury (LTI) rates. Several variables differed between 1990 and 2001, e.g., increase in safety training, lower turnover rate, and more management involvement in H&S. Other variables previously associated with higher LTI rates also were more prevalent in 2001: more work stoppages for H&S issues, greater perception of risk from hazards, and an increase in workers' lobbying management for H&S improvements. There appears to be greater awareness of H&S issues today, and a movement to upper management becoming more involved in H&S and delegating less authority to individual workers.


Parathyroid hormone regulates the distribution and osteoclastogenic potential of hematopoietic progenitors in the bone marrow.
Parathyroid hormone (PTH) increases both the number of osteoclast in bone and the number of early hematopoietic stem cells (HSCs) in bone marrow. We previously characterized the phenotype of multiple populations of bone marrow cells with in vitro osteoclastogenic potential in mice. Here we examined whether intermittent administration of PTH influences these osteoclast progenitor (OCP) populations. C57BL/6 mice were treated with daily injections of bPTH(1-34) (80 µg/kg/day) for 7 or 14 days. We found that PTH caused a significant increase in the percentage of TN/CD115(+) CD117(high) and TN/CD115(+) CD117(int) cells (p < .05) in bone marrow on day 7. In contrast, PTH decreased the absolute number of TN/CD115(+) CD117(low) cells by 39% on day 7 (p < .05). On day 14, there was no effect of PTH on osteoclast progenitor distribution in vivo. However, PTH treatment for 7 and 14 days did increase receptor activator of NF-κB ligand (RANKL)- and macrophage colony-stimulating factor (M-CSF)-stimulated in vitro osteoclastogenesis and bone resorption in TN/CD115(+) cells. In the periphery, 14 days of treatment increased the percentage and absolute numbers of HSCs (Lin(-) CD117(+) Sca-1(+) ) in the spleen (p < .05). These data correlated with an increase in the percent and absolute numbers of HSCs in bone marrow on day 14 (p < .05). Interestingly, the effects on hematopoietic progenitors do not depend on osteoclast resorption activity. These results suggest that in vivo PTH treatment increased in vitro osteoclastogenesis and resorption without altering the number of osteoclast precursors. This implies that in vivo PTH induces sustained changes, possibly through an epigenetic mechanism, in the in vitro responsiveness of the cells to M-CSF and RANKL.


[Endarterectomy in arteriosclerotic stenosis of the internal carotid artery].
The efficacy of carotid endarterectomy (CE) was estimated in 50 patients with stenosis of the internal carotid artery due to atherosclerosis. In addition to neurological investigations, all the patients were subjected to USDG of the head vessels, cerebral angiography and CT of the brain. Patients with the signs of cerebral circulation disorders or those who suffered ischemic brain stroke with a reversible neurological deficit were the most prospective candidates for CE. According to the cerebral angiography data, the degree of stenosis ranged from 50 to 90%. Surgical intervention was performed within different times after derangement of brain circulation, most frequently from 3 months to 1 year. The overwhelming majority of the patients suffered operations without any complications. The lethality amounted to 4% (because of acute cardiovascular failure and myocardial infarction). The total mortality and morbidity due to brain stroke were 5.9%. Indications for carotid endarterectomy in stenosis of the ICA are under consideration.


Agents of newly recognized or infrequently encountered mycobacterial diseases.
This paper reviews recent information on the systematics and clinical significance of potentially pathogenic environmental mycobacteria. A short history of these mycobacteria is given. Information on species for which clinical and systematic aspects have already been well documented, i.e., Mycobacterium kansasii, M. marinum, M. scrofulaceum, M. simiae, M. szulgai, M. ulcerans, M. xenopi, and members of the M. fortuitum complex, is updated. Although the M. avium complex was extensively reviewed in earlier literature, major new systematic and clinical information is presented in some detail. Species that have received very limited prior coverage, i.e., M. asiaticum, M. haemophilum, M. malmoense, and M. shimoidei, are the main subjects of this review and are discussed in detail. The rare infections attributed to species that are normally considered nonpathogenic, i.e., M. gastri, M. gordonae, the M. terrae complex, and most of the rapidly growing mycobacteria outside of the M. fortuitum complex, are critically reviewed. Finally, suggestions are offered for practical measures that can minimize the risk of failing to isolate or misidentifying some of the more obscure potentially pathogenic environmental mycobacteria that are only infrequently recognized.


Molecular characterization of the Aedes aegypti odorant receptor gene family.
The olfactory-driven blood-feeding behaviour of female Aedes aegypti mosquitoes is the primary transmission mechanism by which the arboviruses causing dengue and yellow fevers affect over 40 million individuals worldwide. Bioinformatics analysis has been used to identify 131 putative odourant receptors from the A. aegypti genome that are likely to function in chemosensory perception in this mosquito. Comparison with the Anopheles gambiae olfactory subgenome demonstrates significant divergence of the odourant receptors that reflects a high degree of evolutionary activity potentially resulting from their critical roles during the mosquito life cycle. Expression analyses in the larval and adult olfactory chemosensory organs reveal that the ratio of odourant receptors to antennal glomeruli is not necessarily one to one in mosquitoes.


Human uterine lower segment myometrial cell and nuclear volume at term: influence of maternal age.
Little is known about the cytoarchitecture of human myometrial cells in pregnancy, and whether or not this may be influenced by maternal characteristics such as age, parity and body mass index (BMI). The aim of this study was primarily to evaluate human myometrial smooth muscle cell (SMC) and nuclear volume in the third trimester of human pregnancy, and secondarily to investigate if these parameters are altered in relation to the maternal characteristics outlined above. Myometrial biopsies were obtained from 30 women undergoing elective caesarean delivery at term. One-micrometer sections were prepared for light microscopy and 100-nm sections for electron microscopy. The nucleator technique was used to assess nuclear volume from the light microscopy images. Point-counting methodology was used on transmission electron micrographs to assess the percentage of the cell volume occupied by the nucleus. Cell volume was calculated from these measurements. The euchromatin to heterochromatin (Eu/Het) ratio was determined to ascertain whether differences in nuclear volume were due to an increased range of genes being transcribed. The mean (± SEM) nuclear volume was 175 ± 10 μm(3) , the nucleus occupied 1.5 ± 0.1% of the SMC and the mean cell size was 14 047 ± 1352 μm(3) . The Eu/Het ratio was 7.54 ± 0.4. The mean volume of heterochromatin and euchromatin in the nucleus was 21.5 ± 1.7 and 149 ± 9 μm(3) , respectively. A multivariate regression analysis revealed that advanced maternal age was associated with an increase in the percentage of the cell occupied by nucleus (R(2) = 0.32, P = 0.004). There were no other significant effects of maternal age, BMI or parity on the measured parameters. These findings provide reliable volumes for human myometrial cells and their nuclei at term gestation, and show that nuclear volume fraction may be influenced by maternal age.


The role of the excision-repair enzymes in mutation-induction by cis-Pt(NH3)2Cl2.
Mutation induction by cis-Pt(NH3)2Cl2 (cisplatin) has been shown to be absent in E.coli strains carrying a deletion of the uvrB gene (1). This suggested that excision-repair, which is normally thought to be error-free, is involved in mutation induction with cisplatin. Here, the role of the excision repair enzymes UvrA, UvrB and UvrC is investigated using E.coli strains with different repair capacities. It is shown that cisplatin induced mutagenesis is dependent both on UvrA and UvrB but not on UvrC. Of the UvrB enzyme the N-terminal 113 aminoacids are sufficient for mutation induction by cisplatin.


Assessment of the Association Between Cigarette Smoking and Cognitive Performance in Patients With Schizophrenia-Spectrum Disorders: A Case-Control Study.
The prevalence of cigarette smoking is significantly higher in patients with schizophrenia compared to the general population. Schizophrenia is also characterized by cognitive impairments that can be detected in the premorbid phase of illness. However, studies addressing the association between cigarette smoking and cognition in patients with psychosis have provided mixed findings. Therefore, the aim of this study was to assess the relationship between tobacco smoking and cognitive performance in patients with schizophrenia. In this case-control study, we recruited 67 inpatients with schizophrenia (34 cigarette smokers) and 62 healthy controls (30 cigarette smokers) at two clinical sites (Wroclaw and Szczecin, Poland). Cognitive performance was examined using the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS). Smoking dependence was determined using the Fagerström Test for Nicotine Dependence (FTND) and the pack-year index. Results show that, after adjustment for potential confounders, smokers with schizophrenia presented significantly lower scores on delayed memory tests compared to non-smokers with schizophrenia (F = 11.07, p = 0.002). In healthy controls, after adjustment for age, sex, and education level, smokers had significantly lower scores in immediate memory (47.1 ± 6.4 vs. 52.0 ± 4.0, F = 11.64, p = 0.001), visuospatial/constructional functions (34.8 ± 3.8 vs. 37.7 ± 1.8, F = 12.86, p = 0.001) and global cognition (177.0 ± 15.7 vs. 191.2 ± 14.0, F = 12.63, p = 0.001) compared to non-smokers. There were no significant correlations between FTND scores or pack-year index and cognitive performance neither in patient nor control group. Our results show that cigarette smoking is related to worse delayed memory performance in schizophrenia patients as well as deficits of immediate memory, visuospatial/constructional functions, and global cognition in controls. Longitudinal studies are required to establish causal interference between smoking and cognition in patients with schizophrenia.


[Calculation of the peak systolic wall stress at the equator of the left ventricle by coupled M mode echo and pressure recordings].
The peak systolic wall stress at the equator of the left ventricle (sigma max) is the maximum load that the myocardial fibres bear during contraction. It is an index of the adaptation of the left ventricle to cardiac disease, and, when elevated, it indicates cardiac decompensation. sigma max was calculated by coupled M mode echo-LV pressure recordings in 51 cases: 11 patients without LV disease, 14 patients with aortic stenosis (AS), 14 patients with aortic incompetence (AI), 7 patients with severe mitral incompetence (MI) and 5 patients with cardiomyopathy with dilatation (CMP). sigma max was calculated from Mirsky's formula, the length of the long axis being deduced from the short axis and the diastolic:systolic ratio of these two axes from ventriculography. The normal value of sigma max by this method is 220 dynes 10(3)/cm2 +/- 30 with an upper limit of normal of 280 dynes 10(3)/cm2. sigma max was normal in patients with AS and AI, and increased in the cases of MI and CMP, in positive correlation with LV volume (r = 0,47) and the shape of the LV (long:short axis ratio). No correlations were found between sigma max and maximum LV pressure. The relatively low values of sigma max compared to the results obtained from coupled echo-angio recordings are partly due to the thick walled LV model and, to a large extent, to the lower values of short axis when measured by echo compared to angiography.(ABSTRACT TRUNCATED AT 250 WORDS)


Sequence analysis and functional characterization of full-length hepatitis B virus genomes from Korean cirrhotic patients with or without liver cancer.
This study aimed to identify and characterize mutations in the hepatitis B virus (HBV) genome associated with advanced liver diseases. The 3.2-kb HBV genome of the C2 subgenotype was amplified from sera of 18 cirrhotic Korean patients with (10) or without (8) hepatocellular carcinoma (HCC), and two clones per patient were characterized by transient transfection experiments in human hepatoma cells. While A1762T/G1764A core promoter mutations were highly prevalent in both groups, the G1896A precore mutation to abolish hepatitis B e antigen (HBeAg) expression was more common in HCC clones (55% vs. 20%). High replication capacity was mostly found in HCC clones and associated with core promoter mutations, whereas more non-HCC clones harbored a nonfunctional core gene (34% vs. 8%). Large in-frame deletions in the preS region were found in 60% of HCC clones and 38% of non-HCC clones. They removed the first 11 residues of large envelope protein or impaired small envelope protein expression, or deleted a neutralizing epitope in the preS2 domain. Additional point mutations prevented middle envelope protein expression, or caused nonsense mutations in the preS or S region to truncate large and/or small envelope protein. Consequently, many clones were unable to express or secrete hepatitis B surface antigen (HBsAg). In conclusion, mutations associated with the advanced stage of chronic HBV infection are complex and diverse. Host immune pressure most likely selected for mutations in the HBV genome to abolish or reduce HBeAg or HBsAg production, to enhance genome replication, or to escape neutralizing antibodies. Some of these mutations may contribute to liver cirrhosis or HCC development.


Potential contribution of lifestyle and socioeconomic factors to healthy user bias in antihypertensives and lipid-lowering drugs.
Healthy user bias arises when users of preventive medications such as lipid-lowering drugs (LLDs), hormone replacement therapy and antihypertensive (AH) medications are healthier than non-users due to factors other than medication effects, making the medications appear more beneficial in observational studies of effectiveness and safety. The purpose of the study is to examine factors contributing to healthy user effect in patients taking AHs or LLDs. Among patients with hypertension or hyperlipidaemia in a population-based sample from the National Health and Nutrition Examination Survey (1999-2010), we assessed the association between socioeconomic and lifestyle factors and the use of AHs/LLDs by logistic regression with adjustment for demographics and comorbidities in a cross-sectional study. When 9715 AH/LLD users were compared with 3725 non-users, AH/LLD users were more likely to be: highly educated (OR 1.2, 95% CI 1.2 to 1.3), non-impoverished (OR 1.3, 95% CI 1.2 to 1.4), current non-smokers (OR 1.2, 95% CI 1.1 to 1.4), physically active (OR 1.1, 95% CI 1.0 to 1.2) and consume more calcium (OR 1.1, 95% CI 1.0 to 1.3) but less likely to have normal body mass index (OR 0.6, 95% CI 0.6 to 0.7) or to meet dietary sodium recommendations (OR 0.8, 95% CI 0.7 to 0.9). We identified several salutary lifestyle factors associated with AH/LLD use in a representative US population. Healthy user effect may be partly explained by better socioeconomic profiles and lifestyles in AH/LLD users compared with non-users.


Effect of simulated pulpal pressure on dentin permeability and adhesion of self-etch adhesives.
Dentin bonds made with one-bottle etch-and-rinse and self-etch adhesives are affected by the formation of interfacial blisters, porosities and deterioration. The first objective of this study was to evaluate the fluid flow through resin-dentin interfaces created by self-etching adhesives applied to deep dentin using a replica technique and by directly measuring dentin permeability (P). The second objective was to examine the effect of intrapulpal pressure on the microtensile bond strength of these adhesives. A fluid-transport model was used to measure the fluid permeability (%P) through different adhesives. Impressions of bonded dentin were taken with a polyvinylsiloxane impression material to monitor fluid transudation from the surface of the adhesive. Positive replicas were fabricated for SEM examination. Two groups of resin-bonded specimens (pulpal pressure versus no pulpal pressure) were created for microtensile bond strength evaluation. Adhesive application was performed under 0 cm H(2)O. Pulpal pressure group was submitted to 20 cm H(2)O of pulpal pressure during build-up procedures. Clearfil Protect Bond exhibited the lowest permeability and fewest numbers of fluid droplets over the surface of the bonded dentin. G-Bond and Clearfil-S3 Bond were more permeable than Clearfil Protect Bond. One Up Bond F was the most permeable adhesive. A highly significant correlation was observed between the relative permeability of these adhesives (%P) and the number of fluid droplets on the adhesive surfaces. The application of pulpal pressure significantly reduced bond strength. Resin-dentin bonds created by contemporary self-etch adhesives are susceptible to fluid permeation induced by pulpal pressure. HEMA-based adhesives showed the largest reductions in bond strengths after pulpal pressure application.


Uncovering the Repertoire of Endogenous Flaviviral Elements in Aedes Mosquito Genomes.
Endogenous viral elements derived from nonretroviral RNA viruses have been described in various animal genomes. Whether they have a biological function, such as host immune protection against related viruses, is a field of intense study. Here, we investigated the repertoire of endogenous flaviviral elements (EFVEs) in Aedes mosquitoes, the vectors of arboviruses such as dengue and chikungunya viruses. Previous studies identified three EFVEs from Aedes albopictus cell lines and one from Aedes aegypti cell lines. However, an in-depth characterization of EFVEs in wild-type mosquito populations and individual mosquitoes in vivo has not been performed. We detected the full-length DNA sequence of the previously described EFVEs and their respective transcripts in several A. albopictus and A. aegypti populations from geographically distinct areas. However, EFVE-derived proteins were not detected by mass spectrometry. Using deep sequencing, we detected the production of PIWI-interacting RNA-like small RNAs, in an antisense orientation, targeting the EFVEs and their flanking regions in vivo The EFVEs were integrated in repetitive regions of the mosquito genomes, and their flanking sequences varied among mosquito populations. We bioinformatically predicted several new EFVEs from a Vietnamese A. albopictus population and observed variation in the occurrence of those elements among mosquitoes. Phylogenetic analysis of an A. aegypti EFVE suggested that it integrated prior to the global expansion of the species and subsequently diverged among and within populations. The findings of this study together reveal the substantial structural and nucleotide diversity of flaviviral integrations in Aedes genomes. Unraveling this diversity will help to elucidate the potential biological function of these EFVEs.IMPORTANCE Endogenous viral elements (EVEs) are whole or partial viral sequences integrated in host genomes. Interestingly, some EVEs have important functions for host fitness and antiviral defense. Because mosquitoes also have EVEs in their genomes, characterizing these EVEs is a prerequisite for their potential use to manipulate the mosquito antiviral response. In the study described here, we focused on EVEs related to the Flavivirus genus, to which dengue and Zika viruses belong, in individual Aedes mosquitoes from geographically distinct areas. We show the existence in vivo of flaviviral EVEs previously identified in mosquito cell lines, and we detected new ones. We show that EVEs have evolved differently in each mosquito population. They produce transcripts and small RNAs but not proteins, suggesting a function at the RNA level. Our study uncovers the diverse repertoire of flaviviral EVEs in Aedes mosquito populations and contributes to an understanding of their role in the host antiviral system.


Women in agriculture: risks for occupational injury within the context of gendered role.
Women continue to make significant contributions to farming. Not only do women participate in the traditional roles of homemaker, caregiver, and wife, they also work side-by-side with their spouses in keeping the farm viable. More daughters are entering the farming business, either as partners with other family members or as independent operators. Each year since the United States Department of Agriculture began including gender in the Census of Agriculture, the percentage of women engaged in agriculture has increased, and women's participation in agriculture is increasing faster than in other business segments. This article examines the role of women in agriculture and how sociocultural, economic, and physical factors may affect women's exposure to injury-producing events and their knowledge and beliefs about injury prevention. To date, few studies have examined work-related unintentional injuries among farm women. Even less is known about the extent to which occupational risks are recognized when women seek medical care. Differences in size and stature, increased physical strain, and low maximal oxygen uptake may predispose women to ergonomic-related injuries. Limitations of current research and recommendations for future analyses are discussed.


Medical humanities in a rapidly changing world. Is there any worth in it?
This article was migrated. The article was marked as recommended. Humanism has shaped mankind during the last centuries, but technology and recent social changes have shifted the paradigm towards a data driven society where humans are no longer at the center of the world. Medicine is not alien to this new situation; clinical practice is rapidly changing, and medical curricula must adapt to it. Is there any place for humanities in medical training? In this paper we analyze the present situation of humanism in a technology flooded health environment, the roll of humanities in contemporary medical curricula and we propose a series of measures to be taken with the purpose of educating the next generation of doctors.


Crystal Structure of Tl2Ba2Ca2Cu3O10, a 125 K Superconductor.
There is now a new series of high-temperature superconductors that may be represented as (A(III)O)(2)A(2)(II)Can-1CunO2+2n where A(III) is Bi or Tl, A(II) is Ba or Sr, and n is the number of Cu-O sheets stacked consecutively. There is a general trend toward higher transition temperatures as n increases. The highest n value for a bulk phase is three and is found when A(III) is Tl. This compound, Tl(2)Ba(2)Ca(2)Cu(3)O(10), has the highest transition temperature( approximately 125 K) of any presently known bulk superconductor. The structure of Tl(2)Ba(2)Ca(2)Cu(3)O(10) has been determined from single-crystal x-ray diffraction data and is tetragonal, with a = 3.85 A and c = 35.9 A. No superstructure is observed, and the material is essentially twin-free. Electron microscopy in the Tl/Ba/Ca/Cu/O system has revealed intergrowths where n = 5; such regions may well be responsible for the superconducting onset behavior observed in this system at about 140 K.


Late-life depression and the association with multimorbidity and polypharmacy: a cross-sectional study.
Late-life depression often coincides with chronic somatic diseases and, consequently, with polypharmacy, which may complicate medical treatment. To determine the associations between patients diagnosed with late-life depression in primary care and multimorbidity and polypharmacy. This cross-sectional observational study was performed using 2012 primary care data. Depressed patients aged ≥60 years were compared to age and gender matched patients diagnosed with other psychological diagnoses and mentally healthy controls. Morbidity and prescription data were combined, and regression analyses were performed for the associations between depression and chronic disease and chronic drug use. We included 4477 patients; 1512 had a record of depression, 1457 of other mental health or psychological diagnoses and 1508 were controls. Depressed patients had a 16% [Prevalence Ratio (PR) 1.16; 95% confidence interval (95% CI) 10%-24%] higher rate of chronic somatic disease and higher odds for multimorbidity (OR 1.55; 95% CI 1.33-1.81) compared with controls. No differences existed between depressed patients and patients with other psychological diagnoses. Compared with controls, depressed patients had a 46% (95% CI 39-53%) higher rate of chronic drug use and higher odds for polypharmacy (OR 2.89; 95% CI 2.41-3.47). Depressed patients also had higher rates of chronic drug use and higher odds for polypharmacy compared with patients with other psychological diagnoses (PR 1.26; OR 1.75; both P < 0.001). Late-life depression in primary care patients is associated with more chronic drug use, even beyond the increased rates of comorbid somatic diseases. General practitioners should consider medication reviews to prevent unnecessary drug-related problems in these patients.


The effect of chemotherapy on the kinetics and proliferative capacity of normal and tumorous tissues in vivo.
The proliferative state of a given tissue is a major determinant of its sensitivity to both phase-specific and cycle-specific chemotherapeutic agents. To study the extent of injury induced by antitumor agents to normal and tumorous tissues, a technique for following DNA synthesis as reflected in the incorporation of tritiated thymidine (3H-TdR) into DNA was compared to the conventional radioautographic technique of the labeling index (LI) and to the functional kinetic technique of granulocyte colony formation in vitro. Alterations in DNA synthesis induced by a single dose of cyclophosphamide in normal and tumorous tissues in vivo paralleled in many respects the changes seen when the more time-consuming techniques of the LI or granulocyte colony formation were employed. However, the recovery of granulocyte colony formation after cyclophosphamide therapy lagged behind the recovery of DNA synthesis in the bone marrow, obscuring a kinetic event of potential therapeutic significance. The determination of DNA synthesis simultaneously in normal and tumorous tissues in vivo was easy to perform and supplied therapeutically pertinent results comparatively quickly.


Palladium-catalyzed thiocarbonylation of aryl, vinyl, and benzyl bromides.
A catalytic protocol for synthesis of thioesters from aryl, vinyl, and benzyl bromides as well as benzyl chlorides was developed using only stoichiometric amounts of carbon monoxide, produced from a solid CO precursor inside a two-chamber system. As a catalytic system, the combination of bis(benzonitrile) palladium(II) chloride and Xantphos furnished the highest yields of the desired compounds, along with the weak base, NaOAc, in anisole at 120 °C. The choice of catalytic system as well as solvent turned out to be important in order to ensure a high chemoselectivity in the reaction. Both electron-rich and electron-deficient aryl bromides worked well in this reaction. Addition of 1 equiv of sodium iodide to the reaction improved the chemoselectivity with the electron-deficient aryl bromides. The thiol scope included both aryl and alkyl thiols, including 2-mercaptobenzophenones, whereby a thiocarbonylation followed by a subsequent McMurry coupling yielded differently substituted benzothiophenes. It was demonstrated that the methodology could be applied for (13)C introduction into the thiophene ring.


Lower Extremity Pediatric Tissue Expansion: A Single Surgeon's 16-Year Experience.
Tissue expansion is a well-established approach to soft tissue reconstruction in the pediatric population for lower extremity pathologies. Unfortunately, complication rates range from 19% to 40% in literature, including infection and implant extrusion, leading to delays in reconstruction. These challenges have prompted investigation toward categorizing risk factors for lower extremity tissue expander placement. A retrospective study of pediatric patients who underwent lower extremity tissue expander placement by the senior author (R.J.R.) was performed over a 16-year period. Patient charts were reviewed to categorize baseline and operative characteristics. Primary outcome variables were surgical-site infection, expander extrusion, and expander deflation. Univariate and multivariate logistic regressions were performed (α < 0.05). There were 59 tissue expanders in our cohort. The overall complication rate was 27.1% with a 77.2% successful reconstruction rate. Greater number of expanders placed during 1 operation is associated with 2.5 increased odds of having any complication and is associated with 0.4 decreased odds of having a successful reconstruction. Incisions made in scar tissue for expander placement appear to be associated with a greater than 7 times increased odds of readmission. Reconstruction of soft tissue pathologies using lower extremity tissue expanders in the pediatric population is an effective yet challenging technique. This study identified that the number of expanders inserted during 1 operation, incisions made over scar tissue, and expanders placed in the anterior thigh were correlated with having a negative impact on reconstructive outcomes. Extra care should be taken with patients who require multiple expanders during 1 operation and with choosing the location and incision of expander placement.


Epstein-Barr virus immortalization and latency.
Epstein-Barr virus infects human B lymphocytes. The interaction between the virus and these cells has been the subject of investigation for over three decades. Recent in vitro and in vivo studies, reviewed here, are revealing the mechanisms by which EBV induces and controls proliferation through the expression of six viral nuclear proteins and two plasma membrane proteins. This genetic program is referred to as immortalization and it is suggested that the purpose of immortalization is to use the innate proliferative potential of B cells to inflate the numbers of infected cells prior to virus production and cell lysis. Latency, on the other hand, has only been detected in situ in latently infected humans. It is characterized by the presence of very low numbers of viral episomes that appear to express the RNA for only one protein (Latent Membrane Protein 2) in B cells that bear the markers of a non-activated resting memory subset. Two models are proposed for the mechanism that establishes this state. The differences between immortalization and latency are highlighted in this review and it is suggested that many of the functions currently attributed to latency are actually features of immortalization. An appreciation of this distinction may assist the discussion of the nature of the interaction between the virus and the host in EBV-associated lymphoproliferative diseases and tumors.


Diagnostic markers for diseases: SELDI-TOF profiling of pig sera for PRRS.
Porcine reproductive and respiratory syndrome (PRRS) is a highly infectious viral disease causing severe losses to the pig industry. Most weaning piglets are likely to be exposed to the infection and show at least asymptomatic PRRS viremia strongly related to productive performance. The aims of this study were to set up experimental conditions for pig sera proteomic profiling and to identify biomarkers that differentiate weaning asymptomatic piglets positive to PRRS viremia from negative controls (PCR tested) with potential predictive value for the subsequent occurrence of clinical PRRS. Protein profiles were generated by SELDI-TOF MS using the Bio-Rad Chips WCX, IMAC30 and H50. The discovery phase revealed that a consistent number of highly significant protein peaks can be detected by the WCX and IMAC30 surfaces; however none of these peaks were statistically confirmed by the subsequent validation phase, highlighting that serum concentration of the contaminant and most abundant proteins is a crucial parameterfor SELDI-TOF MS studies. Current protocols are being furtheroptimized and adapted to pig sera to reduce the unfavourable effects of the most abundant proteins and to increase the number of potential detectable biomarkers. Furthermore, proteomic fingerprint profiling has been shown to be a promising diagnostic tool that, in the future, may be useful to provide also insights into the mechanisms of early viral infection in vivo.


TACT: Transcriptome Auto-annotation Conducting Tool of H-InvDB.
Transcriptome Auto-annotation Conducting Tool (TACT) is a newly developed web-based automated tool for conducting functional annotation of transcripts by the integration of sequence similarity searches and functional motif predictions. We developed the TACT system by integrating two kinds of similarity searches, FASTY and BLASTX, against protein sequence databases, UniProtKB (Swiss-Prot/TrEMBL) and RefSeq, and a unified motif prediction program, InterProScan, into the ORF-prediction pipeline originally designed for the 'H-Invitational' human transcriptome annotation project. This system successively applies these constituent programs to an mRNA sequence in order to predict the most plausible ORF and the function of the protein encoded. In this study, we applied the TACT system to 19 574 non-redundant human transcripts registered in H-InvDB and evaluated its predictive power by the degree of agreement with human-curated functional annotation in H-InvDB. As a result, the TACT system could assign functional description to 12 559 transcripts (64.2%), the remainder being hypothetical proteins. Furthermore, the overall agreement of functional annotation with H-InvDB, including those transcripts annotated as hypothetical proteins, was 83.9% (16 432/19 574). These results show that the TACT system is useful for functional annotation and that the prediction of ORFs and protein functions is highly accurate and close to the results of human curation. TACT is freely available at http://www.jbirc.aist.go.jp/tact/.


[Nutrition and bone metabolism (author's transl)].
Calcium and phosphate metabolism as well as those substances which are essential in regulating this metabolism (parathyroid hormone, thyrocalcitonin and cholecalciferol) are briefly discussed. Of three known forms of bone disease (nutritional secondary hyperparathyroidism, rickets and hypervitaminosis A), the clinical symptoms, radiological changes, (histo)pathological findings, therapeutic procedures as well as the aetiological, pathogenic and pathophysiological features will be reviewed.


Incidence of first ever stroke during Hajj ceremony.
The Hajj Ceremony, the largest annual gathering in the world, is the most important life event for any Muslim. This study was designed to evaluate the incidence of stroke among Iranian pilgrims during the Hajj ceremony. We ascertained all cases of stroke occurring in a population of 92,974 Iranian pilgrims between November 27, 2007 and January 12, 2008. Incidence and risk factors of the first ever stroke in Hajj pilgrims were compared, within the same time frame, to those of the Mashhad residents, the second largest city in Iran. Data for the latter group were extracted from the Mashhad Stroke Incidence Study (MSIS) database. During the study period, 17 first-ever strokes occurred in the Hajj pilgrims and 40 first-ever stroke strokes occurred in the MSIS group. Overall, the adjusted incidence rate of first ever stroke in the Hajj cohort was lower than that of the MSIS population (9 vs. 16 per 100,000). For age- and gender-specific subgroups, the Hajj stroke crude rates were in general similar to or lower than the general population of Mashhad, Iran, with the exception of women aged 35 to 44 years and aged >75 years who were at greater risk of having first-ever stroke than the non-pilgrims of the same age. The first ever stroke rate among Iranian Hajj pilgrims was lower than that of the general population in Mashhad, Iran, except for females 35-44 or more than 75 years old. The number of events occurring during the Hajj suggests that Islamic countries should consider designing preventive and screening programs for pilgrims.


[The rapid diagnosis of salmonellosis by a determination of the antigen in the biosubstrates of patients using the coagglutination reaction].
The diagnostic potential of the coagglutination test was checked with the aim of improving the laboratory diagnosis of Salmonella infections by the detection of Salmonella specific antigen in different biological materials (feces, urine, saliva and immune complexes in blood sera). The study of all specimens resulted in the confirmation of the diagnosis in 78% of patients, often during the first days of the disease. The proportion of nonspecific reactions, as shown in the control groups of healthy donors and patients with dysentery and other acute enteric infections, did not exceed 5%.


Involucrin-claudin-6 tail deletion mutant (CDelta206) transgenic mice: a model of delayed epidermal permeability barrier formation and repair.
Preterm birth is a major global health problem that results in a large number of infant deaths, many of which are attributable to the complications of an immature epidermal permeability barrier (EPB), for which there is currently no effective therapeutic option. The mammalian EPB is formed during development and is essential for survival as it maintains thermoregulation and hydration, and provides a defense against infection. Using transgenic mouse technology, we have demonstrated the importance of claudin (Cldn)-containing tight junctions (TJs) in epidermal differentiation and, in particular, that epidermal suprabasal overexpression of Cldn6 results in an EPB-deficient phenotype that phenocopies the dysfunctional EPB of premature human infants. In this study, we used the same approach to target a Cldn6 tail deletion mutant to the epidermis of mice [involucrin (Inv)-Cldn6-CDelta206 transgenic mice]. The Inv-Cldn6-CDelta206 transgenic mice displayed a developmental delay in EPB formation, as shown by the expression of keratins and Cldns, and by X-Gal penetration assays. Trans-epidermal water loss measurements and immunolocalization studies indicated that the epidermal differentiation program was also perturbed in postnatal Inv-Cldn6-CDelta206 transgenic mice resulting in a delayed maturation. Notably, however, expression/localization of epidermal differentiation and maturation markers, including Cldns, indicated that the transgenic epidermis matured and normalized by postnatal day 10, which is 3 days after the wild-type epidermis. Our results suggest that activation of the extracellular signal-regulated kinase 1/2 (Erk1/2) pathway and Cldn1 phosphorylation are associated with the repair and maturation of the skin barrier processes. These studies provide additional support for the crucial role of Cldns in epidermal differentiation, maturation and the formation of the EPB, and describe a novel animal model for evaluating postnatal epidermal maturation and therapies that may accelerate the process.


Preferential inhibition of inducible nitric oxide synthase in intact cells by the 4-amino analogue of tetrahydrobiopterin.
In the present study we demonstrate that the 4-amino analogue of tetrahydrobiopterin, 2,4-diamino-5,6,7,8-tetrahydro-6-(l-erythro-1, 2-dihydroxypropyl)pteridine (4-amino-H4biopterin) binds with high affinity to recombinant endothelial NO synthase and concomitantly inhibits enzyme activity [IC50 = 14.8 +/- 7.5 microm in the presence of added 5,6,7,8-tetrahydro-l-erythrobiopterin (H4biopterin) 10 microm] as efficiently as previously shown for inducible NO synthase [Mayer, B., Wu, C.Q., Gorren, A.C.F., Pfeiffer, S., Schmidt, K., Clark, P., Stuehr, D.J. & Werner, E.R. (1997) Biochemistry 36, 8422-8427]. In cultured porcine endothelial cells, however, 4-amino-H4biopterin was less effective in inhibiting NO formation (IC50 = 420 +/- 36 microm) as compared with inhibition of the inducible isoform in murine fibroblasts (IC50 = 15 +/- 4.9 microm) and in human DLD-1 adenocarcinoma cells (IC50 = 55 +/- 10.3 microm). In all cells investigated, the inhibitory effect of 4-amino-H4biopterin was markedly enhanced by depletion of intracellular H4biopterin and could be overcome by increasing intracellular H4biopterin concentrations. Endothelial cells contained lower amounts of H4biopterin [5.2 +/- 0.3 pmol.(mg protein)-1] than fibroblasts [19.4 +/- 2.7 pmol.(mg protein)-1] and DLD-1 cells [8.3 +/- 1.1 pmol.(mg protein)-1], so that the selectivity of 4-amino-H4biopterin towards inducible NO synthase was not explained by differences in the H4biopterin levels. Because 4-amino-H4biopterin did not suppress expression of NO synthase in cytokine-treated cells, we suggest that high-affinity binding of the inhibitor during protein expression may be responsible for the preferential inhibition of the inducible isozyme in intact cells.


Antidepressant-like effect of geniposide on chronic unpredictable mild stress-induced depressive rats by regulating the hypothalamus-pituitary-adrenal axis.
Geniposide as the major active component of Gardenia jasminoides Ellis has neuroprotective activity. This study elucidated the potential antidepressant-like effect of geniposide and its related mechanisms using a depression rat model induced by 3 consecutive weeks of chronic unpredictable mild stress (CUMS). Sucrose preference test, open field test (OFT) and forced swimming test (FST) were applied to evaluate the antidepressant effect of geniposide. Adrenocorticotropic hormone (ACTH) and corticosterone (CORT) serum levels, adrenal gland index and hypothalamic corticotrophin-releasing hormone (CRH) mRNA expression were measured to assess the activity of hypothalamus-pituitary-adrenal (HPA) axis. Hypothalamic glucocorticoid receptor α (GRα) mRNA expression and GRα protein expression in hypothalamic paraventricular nucleus (PVN) were also determined by real-time PCR and immunohistochemistry, respectively. We found that geniposide (25, 50, 100mg/kg) treatment reversed the CUMS-induced behavioral abnormalities, as suggested by increased sucrose intake, improved crossing and rearing behavior in OFT, shortened immobility and prolonged swimming time in FST. Additionally, geniposide treatment normalized the CUMS-induced hyperactivity of HPA axis, as evidenced by reduced CORT serum level, adrenal gland index and hypothalamic CRH mRNA expression, with no significant effect on ACTH serum level. Moreover, geniposide treatment upregulated the hypothalamic GRα mRNA level and GRα protein expression in PVN, suggesting geniposide could recover the impaired GRα negative feedback on CRH expression and HPA axis. These aforementioned therapeutic effects of geniposide were essentially similar to fluoxetine. Our results indicated that geniposide possessed potent antidepressant-like properties that may be mediated by its effects on the HPA axis.


Transgenic neuronal expression of proopiomelanocortin attenuates hyperphagic response to fasting and reverses metabolic impairments in leptin-deficient obese mice.
Hypothalamic proopiomelanocortin (POMC) gene expression is reduced in many forms of obesity and diabetes, particularly in those attributable to deficiencies in leptin or its receptor. To assess the functional significance of POMC in mediating metabolic phenotypes associated with leptin deficiency, leptin-deficient mice bearing a transgene expressing the POMC gene under control of the neuron-specific enolase promoter were produced. The POMC transgene attenuated fasting-induced hyperphagia in wild-type mice. Furthermore, the POMC transgene partially reversed obesity, hyperphagia, and hypothermia and effectively normalized hyperglycemia, glucosuria, glucose intolerance, and insulin resistance in leptin-deficient mice. Effects of the POMC transgene on glucose homeostasis were independent of the partial correction of hyperphagia and obesity. Furthermore, the POMC transgene normalized the profile of hepatic and adipose gene expression associated with gluconeogenesis, glucose output, and insulin sensitivity. These results indicate that central POMC is a key modulator of glucose homeostasis and that agonists of POMC products may provide effective therapy in treating impairments in glucose homeostasis when hypothalamic POMC expression is reduced, as occurs with leptin deficiency, hypothalamic damage, and aging.


Aerosol therapy of asthma: principles and applications.
Aerosols are the mainstay of therapy of reversible airflow obstruction. Currently available aerosols include bronchodilators such as beta-adrenoceptor agonists and anticholinergic agents which may be used alone or together with prophylactic, anti-inflammatory medications such as sodium cromoglycate and steroids. These agents are characterized by primarily topical activity and thus result in minimal systemic effects. By combining these medications in optimum doses, most patients can be readily managed. In patients with severe airway hyperreactivity, aerosols may need to be combined with oral sustained-release theophylline compounds and/or systemic steroids, the latter being used in minimum doses and if possible on alternate days to minimize adrenal suppression and steroid-related systemic complications. In recent years, an improved understanding of aerosol physics, pharmacology and airway physiology has led to greatly improved aerosol therapy with increasing emphasis on metered dose inhalers as the delivery system of choice while nebulizers are used less frequently except in hospital and pediatric applications. The use of intermittent positive-pressure devices for aerosol delivery has decreased considerably as physicians recognize that simpler delivery methods work equally well at much lower cost. While aerosol therapy techniques are now well worked out, research is continuing on methods of improving the efficiency of aerosol delivery and in the development of newer pharmacological agents such as longer-acting adrenoceptor agonists, calcium channel blockers, antagonists of mediators derived from arachidonic acid as well as higher dose aerosolized steroids.


Fast saccades toward numbers: simple number comparisons can be made in as little as 230 ms.
Visual psychophysicists have recently developed tools to measure the maximal speed at which the brain can accurately carry out different types of computations (H. Kirchner & S. J. Thorpe, 2006). We use this methodology to measure the maximal speed with which individuals can make magnitude comparisons between two single-digit numbers. We find that individuals make such comparisons with high accuracy in 306 ms on average and are able to perform above chance in as little as 230 ms. We also find that maximal speeds are similar for "larger than" and "smaller than" number comparisons and in a control task that simply requires subjects to identify the number in a number-letter pair. The results suggest that the brain contains dedicated processes involved in implementing basic number comparisons that can be deployed in parallel with processes involved in low-level visual processing.


Effect of a uniform partial denervation of the periphery on the peripheral and central vibrissal system in guinea pigs.
In some rodents, somatotopically organized architectonic patterns corresponding precisely to the arrangement of the vibrissae on the face are found in each of the central stations of the trigeminal (V) pathway. Two lines of evidence indicate that these architectonic patterns reflect the level of peripheral innervation. First, in normal mice, the sizes of the individual units within the cortical representation are proportional to the number of fibers supplying the corresponding vibrissal follicles. Second, complete surgical denervation of groups of vibrissae can severely attenuate the sizes of and alter the patterns of their central representations. However, previous studies in this system do not distinguish the effects of the absolute and the relative levels of peripheral innervation on central representations. To address this question, we have studied guinea pigs in which all vibrissae are partially deafferentated before birth by fetal exposure to antibodies against NGF. This approach reduces the absolute level of peripheral innervation in a graded way, and does so uniformly, without changing the pattern of vibrissal innervation. In the most severely affected animals, only 18% of the normal number of V ganglion neurons survive. The effect of this loss on the V system was assessed by comparing the peripheral and central components of the vibrissal system in normal and NGF-deprived animals. Peripheral fibers from the V ganglion neurons, including those to the vibrissae, are less reduced in number (50%) than expected. The number of peripheral fiber fascicles is also decreased. In contrast, neither the patterns nor the areas of the central representations in medulla and cortex differ from those of normal animals. We conclude that 18% of the normal number of V ganglion cells is sufficient to establish normal central architectonic patterns and the size of the central vibrissal representations is independent of the absolute magnitude of peripheral innervation. These observations are of relevance to understanding the role of NGF on the morphogenesis of central somatosensory pathways, the effects of "mismatches" between peripheral innervation, and the development of projections to central targets in the mammalian brain, and provide new data for understanding competitive interactions in the developing central and peripheral trigeminal system.


Membranoproliferative glomerulonephritis associated with low-grade B cell lymphoma presenting in the kidney.
Low-grade B cell lymphoma of mucosa-associated tissue type (MALToma) rarely may involve the kidney. Membranoproliferative glomerulonephritis (MPGN) is an uncommon complication of B cell lymphoma and may be related to cryoglobulin and/or immunoglobulin synthesis by a secretory B cell clone. We report 2 patients with the novel renal biopsy findings of coexistent MALToma and MPGN. Both subjects presented with nephrotic proteinuria and renal insufficiency. One patient had a serum M protein (IgG K) but neither individual had any other clinical or serologic evidence of systemic disease, including hematolymphoid malignancy, autoimmune disease, cryoglobulinemia, or hepatitis C viral infection. Both renal biopsies demonstrated MPGN type I with immunoglobulin deposits that in 1 case showed light chain restriction (IgM K). Electron microscopy disclosed corresponding glomerular electron dense deposits in subendothelial locations. Both biopsies also contained atypical interstitial lymphoid infiltrates comprising marginal zone (centro-cyte-like) cells that infiltrated tubules and showed extra-capsular extension. Immunostains demonstrated a predominantly B cell population that lacked expression of CD5 and cycline D1, and gene rearrangement studies confirmed the presence of a monoclonal B cell population in both cases. These findings indicate that low-grade B cell lymphoma in the kidney may be an unexpected finding in patients with nephrotic syndrome related to MPGN. Immunophenotypic and gene rearrangement studies are important ancillary tools for the evaluation of atypical lymphoid infiltrates in kidney biopsies.


Coccidioidomycosis: a persistent threat to deployed populations.
The world-wide deployability of aircrew members exposes them to a peculiar array of medical problems. Non-immune populations, upon deployment to endemic areas, may present to the deployed flight surgeon with acute, poorly recognizable syndromes, such as acute coccidioidomycosis. Alternatively, the acquisition of the chronic progressive form of coccidioidomycosis in endemic areas may be a problem for the flight surgeon, who may be faced with a poorly recognizable syndrome which first manifests itself weeks to months after the crewmember's return from deployment. We describe three cases of coccidioidomycosis in service members that highlight the difficulty in the diagnosis of this disease. These cases prompted an epidemiologic survey of recent cases of coccidioidomycosis among Air Force beneficiaries, presented here, to better define the impact of this disease on personnel assigned to endemic areas. A brief discussion of the epidemiologic and clinical features of the disease and of the aeromedical disposition of the aviator is presented.


[Use of cryoextraction in different types of airway obstruction].
With the development of mechanically stable cryoprobes, direct endoscopic removal of tissue from the bronchial system with cryotherapy is now possible. Tissue is frozen to the tip of the probe and removed by pulling on the probe together with the fiberscope. This procedure, also referred to as cryoextraction, additionally provides good quality biopsy specimens for histopathological examination. We report here on four patients with different forms of bronchial obstruction, in whom cryoextraction was performed for rapid recanalization. In addition to the immediate effect, the procedure has few complications and is cost-effective. Indications were mucoid impaction, foreign body aspiration, removal of suprastomal granulation tissue prior to decannulation after prolonged mechanical ventilation, and obstruction of the trachea and the main bronchi by a carcinosarcoma. In the latter case, the diagnosis was additionally based on a specimen obtained by cryobiopsy.


Potential Protective Effects of Spirulina (Spirulina platensis) against In Vitro Toxicity Induced by Heavy Metals (Cadmium, Mercury, and Lead) on SH-SY5Y Neuroblastoma Cells.
Spirulina, a filamentous microalga, is used all over the world as a nutraceutical dietary supplement. Recent studies have focused on examining its chelating activity and antioxidant properties, especially as a candidate for protection against neurotoxicity caused by heavy metals. The MTT test and LDH assay were used to examine the viability of the SH-SY5Y cells for 24, 48, and 72 h, to Cd, Hg, and Pb, individually or in combination with Spirulina, and the effects of necrotic cell death. In comparison to the control group, the viability of SH-SY5Y cells decreased after 24 h of exposure, with Cd being more toxic than Hg and Pb being less lethal. The effects of heavy metal toxicity on cell survival were ranked in order after 72 h under identical experimental circumstances as follows: Hg, Pb, and Cd. The viability of the cells was then tested after being exposed to Spirulina at doses of 5 at 50 (%v/v) for 24, 48, and 72 h, respectively. SH-SY5Y cells that had been treated with mixtures of heavy metals and Spirulina underwent the same assay. Cell viability is considerably increased by using Spirulina treatments at the prescribed periods and doses. Instead, the same procedure, when applied to SH-SY5Y cells, caused the release of LDH, which is consistent with the reduction in cell viability. We demonstrated for the first time, considering all the available data, that Spirulina 5, 25, and 50 (%v/v) enhanced the number of viable SH-SY5Y cells utilized as a model system for brain cells. Overall, the data from the present study provide a first insight into the promising positive role of Spirulina against the potentially toxic effects of metals.