Lipoprotein-associated phospholipase A(2): a promising vascular-specific marker for screening cardiovascular risk?
Atherosclerosis is an inflammatory disease that can be assessed by circulating biomarkers. Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is an enzyme produced in atherosclerotic plaque and is bound to low density lipoprotein and high-density lipoprotein cholesterol. It has a role in the pathogenesis of atherosclerosis by promoting vascular inflammation. It is emerging as a vascular-specific marker and predictor of risk for cardiovascular disease (CVD) events. Increasing evidence from many prospective epidemiologic studies have shown that elevated levels of Lp-PLA(2) are associated with future CVD events. Measurement of Lp-PLA(2) in individuals may provide clinically relevant information about their future risk of CVD events. Pharmacologic therapies and/or risk factor modification could be initiated after identification of individuals at risk for CVD. This review provides an overview of the pathophysiology, epidemiologic evidence, and clinical utility of Lp-PLA(2).


Two-dimensional square and hexagonal oxide quasicrystal approximants in SrTiO3 films grown on Pt(111)/Al2O3(0001).
The formation of two-dimensional oxide dodecagonal quasicrystals as well as related complex approximant phases was recently reported in thin films derived from BaTiO3 or SrTiO3 perovskites deposited on (111)-oriented Pt single crystals. Here, we use an all-thin-film approach in which the single crystal is replaced by a 10 nm thick Pt(111) buffer layer grown by molecular beam epitaxy on an Al2O3(0001) substrate. An ultra-thin film of SrTiO3 was subsequently deposited by pulsed laser deposition. The film stacking and structure are fully characterized by diffraction and microscopy techniques. We report the discovery of two new complex phases obtained by reduction of this system through high temperature annealing under ultrahigh vacuum conditions. The formation of a new large square approximant with a lattice parameter equal to 44.4 Å is evidenced by low-energy electron diffraction and scanning tunneling microscopy (STM). Additionally, a new 2D hexagonal approximant phase with a lattice parameter of 28 Å has been observed depending on the preparation conditions. Both phases can be described by two different tilings constructed with the same basic square, triangle and rhombus tiles possessing a common edge length of about 6.7 Å. Using the tiling built from high resolution STM images, we propose an atomic model for each approximant which accounts for the experimental observations. Indeed, the STM images simulated using these models are found to be in excellent agreement with the experimental ones, the bright protrusions being attributed to the topmost Sr atoms. In addition our theoretical approach shows that the adhesion of the oxide layer is rather strong (-0.30 eV Å-2). This is attributed to charge transfer, from the most electropositive elements (Sr and Ti) to the most electronegative ones (Pt and O), and to hybridization with Pt-states. Density of states calculations indicate differences in the electronic structure of the two approximants, suggesting different chemical and physical properties. This all-thin-film approach may be useful to explore the formation of complex two-dimensional oxide phases in other metal-oxide combinations.


Novel Solutions to Student Problems: A Phenomenological Exploration of a Single Session Approach to Art Therapy With Creative Arts University Students.
Within the Australian university context, research has uncovered increasing levels of psychological distress, in the form of stress, anxiety and depression. Higher rates of psychological distress have been reported in undergraduate students specifically enrolled in creative arts programs. Despite these increasing levels of psychological distress, university students are reluctant to engage with mental health and wellbeing supports. To explore ways to meet the mental health and wellbeing needs of creative arts university students, the Creative Arts and Music Therapy Research Unit at The University of Melbourne commenced a project exploring the benefits and pitfalls of a brief creative arts therapies approach for students attending a campus based wellbeing clinic. This exploratory research study formed the art therapy component of this much broader research endeavor. Creative arts students in this research study were invited to participate in a single session art therapy encounter that involved the visual exploration of the miracle question, asking students to visually depict "what the problem looks like and how it will look when the problem is resolved or you feel like you can cope with it better?" The descriptive findings of this exploratory research study revealed how the combination of art therapy used within a single session framework was able to afford students a novel means to externalize problems, leading students to forming a less internalized view of the self.


Effect of Dual Modification on the Spectroscopic, Calorimetric, Viscosimetric and Morphological Characteristics of Corn Starch.
The effect of dual modification of corn starch, including hydrolysis and succinylation, were evaluated through peak viscosity (PV) analysis, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy. This dual modification was shown to increase the reaction efficiency (RE) and degree substitution (DS) compared with starches that were not subjected to acid hydrolysis pretreatment with a 44% and 45% increase respectively. After acid hydrolysis pretreatment, the surface of the corn starch granules exhibited exo-erosion and whitish points due to the accumulation of succinyl groups. The peak viscosity was reduced significantly with the acid hydrolysis pretreatment (between 3 and 3.5-fold decrease), which decreased the pasting temperature and peak time to 20 °C and 100 s respectively. In addition, the dual modification of corn starch altered certain thermal properties, including a reduction in the enthalpy of gelatinization (ΔH) and a higher range of gelatinization (around 6 °C), which may effectively improve industrial applications. Modifications on the FTIR spectra indicated that the dual modification affected the starch crystallinity, while the Raman spectra revealed that the dual modification disrupted the short-range molecular order in the starch. Rearrangement and molecular destabilization of the starch components promoted their granular amphiphilic properties.


Muscular strength is associated with self-esteem in college men but not women.
Muscular strength is a well-known predictor of morbidity and mortality. Similarly, self-esteem is a predictor of health and well-being. The relationship between these two variables, however, is currently unknown. This study examined the cross-sectional relationship between maximal muscular strength (i.e. handgrip and one-repetition-maximum (1-RM) squat) and global self-esteem in 126 college students. Significant correlations were found between both measures of muscular strength and self-esteem. Further analyses revealed that these relationships were only significant for men. Based on these results, additional research is needed to further explore the relationship between muscular strength and self-esteem, especially in other demographic groups and longitudinally.


Biological effects of the aqueous extract of Bridelia grandis stem bark on normal and neoplastic human cells: an in vitro preliminary evaluation.
The aim of the work is to investigate the effects of Bridelia grandis (Pierre ex Hutch) stem bark water extract on human HeLa cancer cells and normal monocytes treated in vitro, evaluating the morphological modifications with light and electron microscopy. The phytocomplex obtained from B. grandis caused a significant decrease in the mitotic index of both HeLa cancer cells and normal monocytes. In addition, a reduction of the typical aneuploid-polyploid pattern has been observed in HeLa cells after treatment. Various alterations at fine structural level, both in neoplastic (HeLa cells) and normal (monocytes) cells have been observed. In particular, electron-dense cells containing condensed mitochondria, autophagic vacuoles and dense spherical cytoplasmic inclusions have been observed. The results show that B. grandis water extracts have an antiproliferative effect on human cells, with a different effect on neoplastic and normal cells. The antiproliferative effect is accompanied by the appearance of various subcellular alterations. The morphological alterations observed are likely to represent the condition of 'dark cell' as a possible preliminary phase towards the autophagic and/or apoptotic cell death.


Antiphospholipid antibodies in a large population-based cohort: genome-wide associations and effects on monocyte gene expression.
The antiphospholipid syndrome (APS) is characterised by venous and/or arterial thrombosis and pregnancy morbidity in women combined with the persistent presence of antiphospholipid antibodies (aPL). We aimed to identify genetic factors associated with the presence of aPL in a population based cohort. Furthermore, we wanted to clarify if the presence of aPL affects gene expression in circulating monocytes. Titres of IgG and IgM against cardiolipin, β2glycoprotein 1 (anti-β2GPI), and IgG against domain 1 of β2GPI (anti-domain 1) were determined in approx. 5,000 individuals from the Gutenberg Health Study (GHS) a population based cohort of German descent. Genotyping was conducted on Affymetrix Genome-Wide Human SNP 6.0 arrays. Monocyte gene expression was determined in a subgroup of 1,279 individuals by using the Illumina HT-12 v3 BeadChip. Gene expression data were confirmed in vitro and ex vivo by qRT-PCR. Genome wide analysis revealed significant associations of anti-β2GPI IgG and APOH on chromosome 17, which had been previously identified by candidate gene approaches, and of anti-domain1 and MACROD2 on chromosome 20 which has been listed in a previous GWAS as a suggestive locus associated with the occurrence of anti-β2GPI antibodies. Expression analysis confirmed increased expression of TNFα in monocytes and identified and confirmed neuron navigator 3 (NAV3) as a novel gene induced by aPL. In conclusion, MACROD2 represents a novel genetic locus associated with aPL. Furthermore, we show that aPL induce the expression of NAV3 in monocytes and endothelial cells. This will stimulate further research into the role of these genes in the APS.


Properties, synthesis, and growth mechanisms of carbon nanotubes with special focus on thermal chemical vapor deposition.
Carbon nanotubes (CNTs) have been extensively investigated in the last decade because their superior properties could benefit many applications. However, CNTs have not yet made a major leap into industry, especially for electronic devices, because of fabrication challenges. This review provides an overview of state-of-the-art of CNT synthesis techniques and illustrates their major technical difficulties. It also charts possible in situ analyses and new reactor designs that might enable commercialization. After a brief description of the CNT properties and of the various techniques used to synthesize substrate-free CNTs, the bulk of this review analyzes chemical vapor deposition (CVD). This technique receives special attention since it allows CNTs to be grown in predefined locations, provides a certain degree of control of the types of CNTs grown, and may have the highest chance to succeed commercially. Understanding the primary growth mechanisms at play during CVD is critical for controlling the properties of the CNTs grown and remains the major hurdle to overcome. Various factors that influence CNT growth receive a special focus: choice of catalyst and substrate materials, source gases, and process parameters. This review illustrates important considerations for in situ characterization and new reactor designs that may enable researchers to better understand the physical growth mechanisms and to optimize the synthesis of CNTs, thus contributing to make carbon nanotubes a manufacturing reality.


Role of thiamine in Huntington's disease pathogenesis: In vitro studies.
Oxidative stress accompanies neurodegeneration and also causes abnormalities in thiaminedependent processes. These processes have been reported to be diminished in the brains of patients with several neurodegenerative diseases. The aim of this work was to conduct a comparative analysis of the impact of supplemented thiamine on the viability of human B lymphocytes with CAG abnormal expanded huntingtin gene (mHTT) (GM13509) and control, B lymphocytes without mHTT (GM14467) through the following studies: determination of the supplemented thiamine concentrations, which are effective for cell growth stimulation after incubation in thiamine deficit conditions; determination of cell capability to intake the exogenous thiamine; evaluation of exogenous thiamine influence on the profile of the genes related to thiamine and energy metabolism; determination of ATP synthesis and activities of thiamine-dependent enzymes, KGDHC and BCKDHC in the intact cells and upon the exogenous thiamine. The following methods were used: EZ4U test for cell growth analysis; HPLC for determination of thiamine intake and ATP synthesis, qRT-PCR for evaluation of the gene profiles and spectrophotometric method for KGDHC and BCKDHC activities determination. Maximal cell growth stimulation was observed at 2.5 mM in GM14467 up to 135% of the control culture and at 5.0 mM in GM13509 cells up to 165% of the control culture. Native levels of total ATP and KGDHC and BCKDHC activities in both cell types were comparable and did not changed upon thiamine deficit or supplementation. GM13509 cells showed more of an increase in growth stimulation upon thiamine supplementation than GM14467 cells and this effect was reflected in the increase of intracellular thiamine concentration. The above results and reported changes in expression of GAPDH, IDH1 and SLC19A3 genes observed upon thiamine deficit conditions suggest that intracellular thiamine status and energy metabolism can have a role in HD pathogenesis.


Breast-feeding rate has dropped to 12.7% in Shanghai.
Only 1 baby in 8 is lucky enough to be fed at the breast in Shanghai. According to the Conference on the Promotion of Breastfeeding in China held on November 11, 1991, the breastfeeding rate in Shanghai has kept dropping with years. The rate is 32% is 1979, 23% in 1984, and only 12.7% in 1991 according to the sample survey. Although breastfeeding has been put on the agenda of the work of maternal and child care since the late 1970s and much work has been done by the maternal and child care workers on this, the trend of dropping of the breastfeeding rate is still hard to be reversed. Specialists in the field think the reason behind this is that many parents as well as grandparents are afraid that their only baby cannot be fully fed at the breast, and they only see fast weight gaining as a sign of healthiness. In addition, the marketing and gifting of the overseas dried-milk businessmen is also dedicated to the dropping of the breastfeeding rate. So now some regulations have been formulated in Shanghai to prohibit businessmen from marketing and gifting dried milk to hospitals.


Molecular expression analysis of beta-naphthoflavone-induced hepatocellular tumors in rats.
The present study was performed to characterize molecular expression levels of preneoplastic and neoplastic lesions induced by beta-naphthoflavone (BNF), an aryl hydrocarbon receptor (AhR) agonist in rat hepatocarcinogenesis. Male F344 rats were initiated with an intraperitoneal injection of 200 mg/kg N-diethylnitrosamine, and two weeks later, they were fed a diet containing 0% or 1% BNF for twenty-eight weeks. All animals were subjected to a two-thirds partial hepatectomy at week 3 and sacrificed at week 30. Histopathologically, BNF increased the incidence and multiplicity of altered foci (1.7-fold and 3.3-fold) and hepatocellular adenomas (HCAs) (4.0-fold and 4.7-fold). Immunohistochemically, BNF increased the number of proliferating cell nuclear antigen (PCNA)-positive cells in altered foci (2.3-fold) and HCAs (6.7-fold) compared with the surrounding tissue and decreased the staining of cell cycle regulators (P21, C/EBPalpha). In addition, loss of reactivity for AhR-regulated (CYP1A1, CYP1B1) molecules and increased reactivity of Nrf-2-regulated (AKR7, GPX2) molecules were also observed in proliferative lesions. Furthermore, increased staining of histone deacetylase (HDAC1) in the nucleus was prominent in HCAs. The differential expression patterns were confirmed at mRNA levels by real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. These results suggest that enhanced cell proliferation and protection against oxidative stress play an important role in BNF-induced hepatocarcinogenesis in rats.


Autoradiographic studies of central alpha 2A- and alpha 2C-adrenoceptors in the rat using [3H]MK912 and subtype-selective drugs.
In the present study we examined the distribution of alpha 2A- and alpha 2C-adrenoceptors in tissue slices from the rat cervical spinal cord and from brain slices collected at the level of the striatum. To differentiate between alpha 2A- and alpha 2C-adrenoceptors, the slices were incubated with [3H]MK912 in the presence of graded concentrations of the alpha 2A-selective drug, BRL44408, or the alpha 2C-selective drug, spiroxatrine. Computer analysis of the autoradiograms indicated that 0.4 nM [3H]MK912 plus 185 nM BRL44408 selectively labeled alpha 2C-adrenoceptors, while 0.4 nM [3H]MK912 plus 220 nM spiroxatrine selectively labeled alpha 2A-adrenoceptors. Using this approach, alpha 2C-adrenoceptors were detected in the striatum, while alpha 2A-adrenoceptors predominated in the cortical layers 1-4, the spinal cord distal dorsal horn, the septum and the endopiriform nucleus.


Positive FAPI PET/CT in a Bilateral Mammary Angiosarcoma Patient With Less Impressive FDG PET/CT Images.
Bilateral primary angiosarcoma of the breast is an extremely rare disease. We describe a case with bilateral angiosarcoma of the breast well visualized on 68Ga-FAPI PET/CT in a 30-year-old woman with a history of right breast-conserving surgery. However, the lesions are less impressive on 18F-FDG PET/CT. No additional findings including lymph node and distant metastases were noted. The patient underwent bilateral mastectomy, and histopathology revealed well-differentiated angiosarcoma that involved bilateral breast parenchyma and the left nipple. Our case illustrates that 68Ga-FAPI PET/CT can be more sensitive in detecting well-differentiated angiosarcoma of the breast.


Diffuse cavernous hemangioma of the rectum (DCHR)--diagnosis and treatment--case report and review of available literature.
Diffuse cavernous hemangioma of the rectum (DCHR) is a rare benign tumor of vascular origin. Approximately 200 such cases have been reported in the literature. Here we present a case of a 49-year old female patient who underwent a surgical procedure due to a mass of the rectum with a history of recurrent, painless gastrointestinal bleeding and anemia in whom DCHR was diagnosed postoperatively. This paper is intended as a metaanalysis of available diagnostic and therapeutic methods to be used in patients with DCHR.


Conversion reaction presenting as acute spinal cord injury.
A conversion reaction, presenting as an acute spinal-cord injury, was diagnosed in 9.8% of new patients admitted to a spinal injury unit during the period from November 1982 to November 1983. There were eight men and one woman (mean age, 23 +/- 8 years). A history of previous episodes of conversion reaction was common in this group. Some patients had evidence of previous vertebral damage. Diagnosis of this condition is difficult, even for those who are familiar with patterns of spinal-cord injuries. All patients were walking independently and had no sensory deficits at the time of discharge from hospital.


Three-dimensional texture analysis of renal cell carcinoma cell nuclei for computerized automatic grading.
The extraction of important features in cancer cell image analysis is a key process in grading renal cell carcinoma. In this study, we analyzed the three-dimensional chromatin texture of cell nuclei based on digital image cytometry. Individual images of 2,423 cell nuclei were extracted from 80 renal cell carcinomas (RCCs) using confocal laser scanning microscopy (CLSM). First, we applied the 3D texture mapping method to render the volume of entire tissue sections. Then, we determined the chromatin texture quantitatively by calculating 3D gray level co-occurrence matrices and 3D run length matrices. Finally, to demonstrate the suitability of 3D texture features for classification, we performed a discriminant analysis. In addition, we conducted a principal component analysis to obtain optimized texture features. Automatic grading of cell nuclei using 3D texture features had an accuracy of 78.30%. Combining 3D textural and 3D morphological features improved the accuracy to 82.19%.


Decaffeination and measurement of caffeine content by addicted Escherichia coli with a refactored N-demethylation operon from Pseudomonas putida CBB5.
The widespread use of caffeine (1,3,7-trimethylxanthine) and other methylxanthines in beverages and pharmaceuticals has led to significant environmental pollution. We have developed a portable caffeine degradation operon by refactoring the alkylxanthine degradation (Alx) gene cluster from Pseudomonas putida CBB5 to function in Escherichia coli. In the process, we discovered that adding a glutathione S-transferase from Janthinobacterium sp. Marseille was necessary to achieve N 7 -demethylation activity. E. coli cells with the synthetic operon degrade caffeine to the guanine precursor, xanthine. Cells deficient in de novo guanine biosynthesis that contain the refactored operon are ″addicted″ to caffeine: their growth density is limited by the availability of caffeine or other xanthines. We show that the addicted strain can be used as a biosensor to measure the caffeine content of common beverages. The synthetic N-demethylation operon could be useful for reclaiming nutrient-rich byproducts of coffee bean processing and for the cost-effective bioproduction of methylxanthine drugs.


Pharmaceutical development of a parenteral lyophilised dosage form for the novel anticancer agent C1311.
C1311 (5-[[2-(diethylamino)ethyl]amino]-8-hydroxyimidazo [4,5,1-de]-acridin-6-one-dihydrochloride trihydrate) is the lead compound from the group of imidazoacridinones, a novel group of rationally designed anticancer agents. C1311 shows significant cytotoxic activity in vitro and in vivo toward a range of colon tumours. The aim of the present study is to develop a sterile and stable, injectable pharmaceutical product for C1311 to be used in phase I clinical trials. C1311 drug substance was structurally and analytically characterised by chromatographic, spectrometric, and diffraction techniques. C1311 was freely soluble in water, and its stability was investigated in several liquid and lyophilised formulations with or without the use of buffering, tonicity, and bulking agents. The final product, containing 100 mg/vial C1311 (as anhydrous free base), was stable for at least 3 months under accelerated storage conditions and at the designated long-term storage condition of 5 +/- 3 degrees C in the dark. The drug is currently used in phase I clinical trials.


The regulatory factors controlling murine erythropoiesis in vitro.
In the mouse, control of the proliferation of erythroid precursors is complex and probably based on the sequential expression of membrane receptors for different regulator molecules during erythroid differentiation. Purified GM-CSF, G-CSF and multi-CSF are all able to initiate proliferation of the earlier erythroid precursors, the multi-CFC and BFU-E, and multi-CSF can stimulate the proliferation of erythroid precursors through all subsequent stages of differentiation. Erythropoietin acts as proliferative stimulus only for the terminal stages of erythropoiesis. There is as yet no evidence for the existence of a distinct BPA regulator with proliferative effects restricted to early erythroid precursors.


Primary chronic daily headache: clinical and pharmacological aspects. A clinic-based study in Oman.
This study on primary chronic daily headache was based on the 1996 proposed revision of the diagnostic criteria of the International Headache Society (IHS). To investigate the relative frequency, clinical characteristics, and associated features of primary chronic daily headache in Omani patients, 171 patients visiting the Neurology Clinic at Sultan Qaboos University Hospital were evaluated. Forty-five percent was diagnosed as suffering from primary chronic daily headache (female to male ratio, 1.7:1). Sixty-two percent suffered from transformed migraine and 34% from chronic tension-type headache. The average age across sexes was 32.3 +/- 12.3 years. A dull heavy feeling in the head was reported by 58% of patients and was associated in less than one third with associated features characteristic of migraine. All headache types shared the same trigger factors. All patients were taking medication, predominantly analgesics, at the time of their first visit. We concluded that primary chronic daily headache is very common with the relative frequency of transformed migraine being similar to that found in Mediterranean studies. Also in Oman, chronic use/overuse of analgesics and nonsteroidal anti-inflammatory drugs is a problem that coexists with primary chronic daily headache. Finally, the proposed revised IHS criteria are highly recommended as a standard classification system for this type of headache.


Treg Cells and Epigenetic Regulation.
The discovery of the epigenetic regulation of Treg cells, a cell population with fundamental immunoregulatory properties, has shed considerable insights into an understanding of the role of these cells in health and disease. Research over the past several years has shown that the interaction of Treg cells with the gut microbiota are critical not only for the development of Treg function in health but also for abnormalities of Treg function that play a critical role in the pathogenesis of human diseases such as the allergic diseases, the autoimmune disorders, and cancer. The equilibrium between phenotypic plasticity and stability of Treg cells is defined by the fine-tuned transcriptional and epigenetic events required to ensure stable expression of Foxp3 in Treg cells. In this chapter, we discuss the molecular events that control Foxp3 gene expression and address the importance of DNA methylation as an important molecular switch that regulates the genetic expression of Treg induction and the possible implications of these findings for the treatment of human diseases characterized by abnormalities of Treg cell function.


Digital imaging of haematological morphology.
Microscopic images of haematological cells are now routinely photographed using digital cameras. Advances in technology mean that the quality of such digital images can now approach that viewed through a microscope. At the same time there is an emerging appreciation that such images can be used in many roles: digital images are now being used to construct digital 'virtual slides', or are being employed together with cell recognition systems for morphological screening. Additionally, an Internet-based viewing systems allow access to on-line annotation, as well as real-time data gathering and feedback. The process of viewing digital images differs from the viewing of glass slides through a microscope; however, such images can provide diagnostic equivalence, and have an emerging role in areas such as education, quality control and continuing professional development. This review explores some of the present strengths, weaknesses and future applications of digital imaging in haematology.


[Radical treatment and prognosis: results of a multicenter surgical-pathologic TNM stomach cancer study].
A multicentre observational study with 22 departments of surgery and 14 departments of pathology was started on April 1st, 1982 to validate TNM classification for patients with gastric carcinoma. 1417 patients with histologically proven gastric cancer entered the study. Follow-up is now 3-5 years. A subgroup of 138 patients (age under 65 years, no metastases M 0, curative resection R 0, only T 2-T 4-carcinomas and tumors situated only in the middle and lower third of the stomach) was studied. 65 had subtotal, 73 total resection. The 3 to 5 years cumulative survival time was at least not worse with subtotal in comparison to total resection.


[Changes in pulmonary function in SARS patients during the three-year convalescent period].
To investigate the patterns of pulmonary function in severe acute respiratory syndrome (SARS) patients during three-year convalescent period, and to investigate the changes and the medium and long term effects on pulmonary function of SARS patients. Pulmonary function tests were conducted for four times in 37 SARS patients during three-year convalescent period. They were discharged from hospital within one month, three months, one year and three years respectively. At the same time, pulmonary function of 15 healthy persons was examined as controls to be used for comparison. Compared with the healthy controls, there were significant decrease in forced vital capacity (FVC), vital capacity (VC), one second forced expiratory volume (FEV1), 25%-75% forced expiratory flow (25%-75%FEF) of SARS patients within three years after their discharge from hospital (all P<0.05). There were no significant differences in FEV1/FVC, total lung capacity (TLC), diffusing capacity of the lung for carbon monoxide (DLCO) between the two groups (all P>0.05). Abnormality rate of DLCO in the SARS patients within three years after discharge was significantly decreased (32.4% vs. 5.4%,P<0.05), but no significant differences in FVC, VC, 25%-75%FEF among SARS patients discharged one month and three years from the hospital (all P>0.05). Ten SARS patients showed lower FVC (mild degree in 9 cases, severe degree in 1 case) three years later, 2 patients showed mildly lowered DLCO. The lung diffusion function of the SARS patients had recovered after they were discharged from hospital within three years, but in 20%-30% patients there is still mild or moderate restrictive ventilation function abnormality and small airway function impairment. The lung functions of most patients have recovered gradually, but in a minority of patients they may be impaired.


Nanostructured lipid carriers loaded with an association of minoxidil and latanoprost for targeted topical therapy of alopecia.
Alopecia is a condition associated with different etiologies, ranging from hormonal changes to chemotherapy, that affects over 80 million people in the USA. Nevertheless, there are currently few FDA-approved drugs for topical treatment, and existing formulations still present skin irritation issues, compromising treatment adherence. This work aimed to develop a safe formulation based on nanostructured lipid carriers (NLC) that entrap an association of minoxidil and latanoprost and target drug delivery to the hair follicles. To do so, thermal techniques combined with FTIR were used to assess the chemical compatibility of the proposed drug association. Then, NLC with 393.5 ± 36.0 nm (PdI < 0.4) and +22.5 ± 0.2 mV zeta potential were produced and shown to entrap 86.9% of minoxidil and 99.9% of latanoprost efficiently. In vitro, the free drug combination was indicated to exert positive effects over human primary epidermal keratinocytes, supporting cell proliferation, migration and inducing the mRNA expression of MKI67 proliferation marker and VEGF - a possible effector for minoxidil-mediated hair growth. Interestingly, such a favorable drug combination profile was optimized when delivered using our NLC. Furthermore, according to the HET-CAM and reconstructed human epidermis assays, the nanoformulation was well tolerated. Finally, drug penetration was evaluated in vitro using porcine skin. Such experiments indicated that the NLC could be deposited preferentially into the hair follicles, causing a considerable increase in the penetration of the two drugs in such structures, compared to the control (composed of the free compounds) and generating a target-effect of approximately 50% for both drugs. In summary, present results suggest that hair follicle-targeted delivery of the minoxidil and latanoprost combination is a promising alternative to treat alopecia.


Inhibitory effects of short hairpin RNA against caspase-8 on apoptosis of murine hepatoma Hepa1-6 cells.
Caspase-8 plays an important role in death-receptor-mediated apoptosis of hepatocytes. We constructed short hairpin RNAs (shRNAs) against caspase-8 and investigated the effects of caspase-8 targeting shRNAs on apoptosis of Hepa1-6 cells induced by TNF-alpha. Oligonucleotides coding for four shRNAs against caspase-8 were cloned into mammalian expression vector Pgenesil-1 containing U6 promoter, which were then introduced into Hepa1-6 cells using liposome-mediated transfection. Effects of caspase-8-shRNAs on apoptosis of Hepa1-6 cells induced by TNF-alpha were detected by PI apoptosis detection kit. Effects of caspase-8-shRNAs on caspase-8 mRNA expression in apoptosis Hepa1-6 cells induced by TNF-alpha were detected by real-time fluorescent RT-PCR. Of the four caspase-8-shRNAs, Pgenesil-caspase-8-1 and Pgenesil-caspase-8-2 were successfully constructed. The apoptosis of Hepa1-6 cells induced by TNF-alpha was significantly inhibited by either Pgenesil-caspase-8-1 or Pgenesil-caspase-8-2 (p < 0.05). Caspase-8 mRNA expression levels in apoptosis Hepa1-6 cells induced by TNF-alpha were significantly decreased by either Pgenesil-caspase-8-1 or Pgenesil-caspase-8-2 (p < 0.05). This study suggested that shRNAs against caspase-8 could effectively inhibit apoptosis of Hepa1-6 cells induced by TNF-alpha by suppressing caspase-8 mRNA expression.


Success and failure of virus-specific T cell responses in hepatitis C virus infection.
Hepatitis C virus (HCV) infection is only cleared in a minority of infected individuals, the majority of patients develop chronic infection. Chronic HCV infection potentially leads to liver fibrosis, cirrhosis and finally hepatocellular carcinoma. The host immune response is an important determinant in the outcome of HCV infection. Innate as well as adaptive cellular and humoral immune responses mediate important antiviral actions; however, virus-specific T cell responses appear to be most critical. Indeed, strong and multispecific CD4+ as well as CD8+ T cell responses are required for viral clearance. Interestingly, individuals who express certain HLA alleles (which are important for antigen presentation to CD4+ and CD8+ T cells) have a higher chance to clear the virus. The mechanisms of protection by HLA class I alleles such as HLA-B27 have been characterized recently. In most individuals, however, the HCV-specific immune response fails to clear the virus. Several mechanisms underlying this HCV-specific T cell failure have been identified. These include viral factors such as viral escape mutations and immunological factors such as the expression of inhibitory receptors, which lead to CD8+ T cell dysfunction. An in-depth understanding of the determinants of success or failure of the HCV-specific T cell response is critical for the development of prophylactic as well as therapeutic vaccination regimes against HCV. Here, we will discuss the virological and immunological determinants of HCV clearance and persistence.


Comparative assessment of microalgal growth kinetic models based on light intensity and biomass concentration.
The comprehensive evaluation and validation of mathematical models for microalgal growth dynamics are essential for improving cultivation efficiency and optimising photobioreactor design. A considerable gap in comprehending the relation between microalgal growth, light intensity and biomass concentration arises since many studies focus solely on associating one of these factors. This paper compares microalgal growth kinetic models, specifically focusing on the combined impact of light intensity and biomass concentration. Considering a dataset (experimental results and literature values) concerning Chlorella vulgaris, nine kinetic models were assessed. Bannister and Grima models presented the best fitting performance to experimental data (RMSE ≤ 0.050 d-1; R2≥0.804; d2≥0.943). Cultivation conditions conducting photoinhibition were identified in some kinetic models. After testing these models on independent datasets, Bannister and Grima models presented superior predictive performance (RMSE = 0.022-0.023 d-1; R2 = 0.878-0.884; d2: 0.976-0.975). The models provide valuable tools for predicting microalgal growth and optimising operational parameters, reducing the need for time-consuming and costly experiments.


Effects of exercise on electroencephalographic mean frequency in depressed elderly subjects.
An analysis of cortical activity by electroencephalogram (EEG) may show different patterns in depressed and normal individuals. Our study aimed (1) to compare the total mean frequency (TMF) and mean EEG frequency per bands (MFB) in depressed and healthy elderly individuals, and (2) to verify the effect of exercise as an additional treatment for major depressive disorder (MDD; according to DSM-IV) via an assessment of depressive symptoms, TMF and MFB. This research was divided into 2 studies. In study 1, we assessed 45 elderly individuals (13 normal and 32 depressive) in a cross-sectional design to search for differences in TMF and MFB. In study 2, we conducted a longitudinal study to assess the effect of exercise on MDD. Twenty depressed elderly persons who were on clinical treatment were allocated to a control group or an exercise group. Subjects were assessed at baseline and after 6 months with the Hamilton Depression Rating Scale, also assessing the TMF and MFB. Our cross-sectional study showed that a lower mean frequency (MF) in posterior areas is related to depression in the elderly. Moreover, in our longitudinal study, we observed that depressed elderly individuals showed a better response to treatment and an increase in MF after physical training. The interaction between exercise and pharmacological treatment may increase the TMF in posterior areas of depressed elderly individuals after 6 months.


Nicotinic Agonist Inhibits Cardiomyocyte Apoptosis in CVB3-Induced Myocarditis via α3β4-nAChR/PI3K/Akt-Dependent Survivin Upregulation.
Cardiomyocyte apoptosis is critical for the development of coxsackievirus B3- (CVB3-) induced myocarditis, which is a common cardiac disease that may result in heart failure or even sudden death. Previous studies have associated CVB3-induced apoptosis with the downregulation of antiapoptotic proteins. Here, attempts were made to examine whether nicotinic acetylcholine receptors (nAChRs), especially α3β4-nAChRs, were a novel therapeutic antiapoptotic target via the activation of survivin, a strong antiapoptotic protein, in viral myocarditis (VMC). In the present study, we demonstrated that nAChRs, α3β4-nAChR subunits in particular, were present and upregulated in CVB3-infected neonatal rat cardiomyocytes (NRC) and H9c2 cells by RT-qPCR. The function of α3β4-nAChRs was next examined using its specific blocker α-CTX AuIB in vitro. The results of the TUNEL assay and western blot experiments showed that the block of α3β4-nAChRs abrogated nicotine-mediated protection of NRC from CVB3-induced apoptosis, and this effect displayed a substantial correlation with the protein expressions of pAkt, survivin, and Cleaved Caspase-3. Hence, the involvement of the PI3K/Akt pathway was further verified by LY294002, a selective inhibitor of PI3K. As a result, nicotine-mediated induction of pAkt and survivin was abolished by LY294002; meanwhile, apoptotic NRC were increased accompanied by an increase of Cleaved Caspase-3 expression. Regarding CVB3-infected BALB/c mice, the α-CTX AuIB- and LY294002-treated groups had a lower survival rate, deteriorative ventricular systolic function, and more severe inflammation than the nicotine-treated group and the modulation of pAkt, survivin, and Cleaved Caspase-3 protein expressions was similar to that in CVB3-infected NRC. In addition, we found that a nicotinic agonist reduced CVB3 replication in a dose-dependent manner in vitro, which indicates that nAChR activation may serve as a possible protection mechanism of CVB3-induced myocarditis. Our study demonstrated that α3β4-nAChR subunits are essential in the nicotine-mediated antiapoptotic effect of protecting cardiomyocytes from CVB3-induced apoptosis in vivo and in vitro. This protection correlated with the PI3K/Akt pathway and the inducement of the antiapoptotic protein survivin. A combination of these mechanisms serves as a novel protective response to treat viral myocarditis.


[A study on early activation of T lymphocytes and soluble tumor necrosis factor receptor in patients with myelodysplastic syndrome].
To investigate the expression of T cell early activation marker (CD(69)) on CD(4)(+) and CD(8)(+) lymphocytes in peripheral blood and the levels of soluble tumor necrosis factor receptor 1 (sTNF-R1) and sTNF-R2 in serum and bone marrow in patients with myelodysplastic syndrome (MDS). Whole blood cell culture procedure was applied to activate T lymphocytes with phytohemagglutinin (PHA) (20 mg/L) in vitro. The expression rates of CD(69) on CD(4)(+) and CD(8)(+) lymphocytes at 0 h and 4 h after culture were analyzed with two-color flow cytometry. The levels of sTNF-R1 and sTNF-R2 in serum and bone marrow were measured with ELISA. There was an increase in the expression rates of CD(69) on CD(4)(+) and CD(8)(+) cells in RA and RAS patients (8.32% and 9.88% respectively) and in the expression rate of CD(69) on CD(8)(+) cells in RAEB and RAEB-T patients (7.92%) before PHA stimulation. CD(69) expression on CD(4)(+) and CD(8)(+) cells were significantly elevated in MDS patients after PHA stimulation (53.46% and 51.63% in RA + RAS; 42.93% and 41.96% in RAEB and RAEB-T) and the expression rate on CD(4)(+) cells was similar to that on CD(8)(+) cells. The levels of the two sTNF-R in MDS patients were elevated. sTNF-R1 in RA and RAS (1.58 +/- 0.68) micro g/L (PB), (2.10 +/- 0.26) micro g/L (BM); sTNF-R2 in RA and RAS (1.41 +/- 0.50) micro g/L (PB), (1.95 +/- 0.64) micro g/L (BM); sTNF-R1 in RAEB and RAEB-T (2.62 +/- 2.55) micro g/L (PB), (3.12 +/- 0.67) micro g/L (BM); sTNF-R2 in RAEB and RAEB-T (1.96 +/- 0.56) micro g/L (PB), (3.09 +/- 0.62) micro g/L (BM). The levels of sTNF-R2 in serum positively correlated to the expression rate of CD(69) on CD(8)(+) cells before PHA stimulation. It is indicate that the increased early activation and activated potentials of T lymphocytes, along with abnormally elevated immunologically active molecules play an important role in immune pathogenesis of patients with MDS.


Communicating scientific uncertainty in a rapidly evolving situation: a framing analysis of Canadian coverage in early days of COVID-19.
The COVID-19 pandemic brought the production of scientific knowledge onto the public agenda in real-time. News media and commentators analysed the successes and failures of the pandemic response in real-time, bringing the process of scientific inquiry, which is also fraught with uncertainty, onto the public agenda. We examine how Canadian newspapers framed scientific uncertainty in their initial coverage of the COVID-19 pandemic and how journalists made sense of the scientific process. We conducted a framing analysis of 1143 news stories and opinion during the first two waves of the COVID-19 pandemic. Using a qualitative analysis software, our analysis focused, first, on how scientific uncertainty was framed in hard news and opinion discourse (editorial, op-ed). Second, we compared how specialist health and science reporters discussed scientific evidence versus non-specialist reporters in hard news and columns. Uncertainty emerged as a "master frame" across the sample, and four additional framing strategies were used by reporters and commentators when covering the pandemic: (1), evidence -focusing on presence or absence of it-; (2) transparency and leadership -focusing on the pandemic response-; (3) duelling experts - highlighting disagreement among experts or criticizing public health decisions for not adhering to expert recommendations-; and (4) mixed messaging -criticizing public health communication efforts. While specialist journalists understood that scientific knowledge evolves and the process is fraught with uncertainty, non-specialist reporters and commentators expressed frustration over changing public health guidelines, leading to the politicization of the pandemic response and condemnation of elected officials' decisions. Managing scientific uncertainty in evolving science-policy situations requires timely and clear communication. Public health officials and political leaders need to provide clear and consistent messages and access to data regarding infection prevention guidelines. Public health officials should quickly engage in communication course corrections if original messages are missing the intended mark, and clearly explain the shift. Finally, public health communicators should be aware of and more responsive to a variety of media reporters, who will bring different interpretative frames to their reporting. More care and effort are needed in these communication engagements to minimize inconsistencies, uncertainty, and politicization.


Blood donation in Guangdong Province, China, from 2006-2014.
To explore the trends in blood collection from 2006 to 2014 in Guangdong, China. Although the Blood Donation Law of the People's Republic of China was implemented in 1998, voluntary non-remunerated blood donation (VNRBD) has been promoted fully for only a decade. The provincial and local governments of Guangdong, one of the most well-developed provinces in China, have promoted blood donation by various means. Official data on blood donation from 2006 to 2014, including the number of blood donations and the family replacement/mutual blood donation (FRMBD) rate, were collected from all blood collection and supply institutions in Guangdong. These data were analysed to explore trends in blood donation in Guangdong Province, and to detect differences among the province's four regions. The number of blood donations in Guangdong increased by 38·23% from 2006 to 2014; overall, the rate increased annually, although it fluctuated in the eastern region. Family replacement/mutual whole blood and platelet donation rates decreased dramatically from 2006 to 2014 (from 39·99% to 20·16% and from 64·15% to 26·51%, respectively), but remained high. Marked disparities in blood donation development were detected among the four regions. With nearly a decade of efforts, blood donation in Guangdong has developed rapidly and sustainably. All blood collection and supply institutions must strengthen efforts to improve awareness of blood donation among the population, retain repeat and regular donors and reduce the rate of FRMBD in favor of the development of VNRBD.


Blunt abdominal trauma: does the use of a second-generation sonographic contrast agent help to detect solid organ injuries?
The objective of our study was to prospectively evaluate whether a second-generation sonography contrast agent (SonoVue) can improve the conspicuity of solid organ injuries (liver; spleen; or kidney, including adrenal glands) in patients with blunt abdominal trauma. Two hundred ten consecutive hemodynamically stable trauma patients underwent both abdominal sonography and CT at admission. The presence of solid organ injuries and the quality of sonography examinations were recorded. Patients with false-negative sonography findings for solid organ injuries in comparison with CT results underwent control sonography. If a solid organ injury was still undetectable, contrast-enhanced sonography was performed. Findings of admission, control, and contrast-enhanced sonograms were compared with CT results for their ability to depict solid organ injuries. Contrast-enhanced sonography was also performed in patients in whom a vascular injury (pseudoaneurysm) was shown on admission or control CT. CT findings were positive for 88 solid organ injuries in 71 (34%) of the 210 patients. Admission, control, and contrast-enhanced sonograms had a detection rate for solid organ injury of 40% (35/88), 57% (50/88), and 80% (70/88), respectively. The improvement in the detection rate between control and contrast-enhanced sonography was statistically significant (p = 0.001). After exclusion of low-quality examinations, contrast-enhanced sonography still missed 18% of solid organ injuries. Five vascular liver (n = 1) and spleen (n = 4) injuries (pseudoaneurysms) were detected on CT; all were visible on contrast-enhanced sonography. Contrast-enhanced sonography misses a large percentage of solid organ injuries and cannot be recommended to replace CT in the triage of hemodynamically stable trauma patients. However, contrast-enhanced sonography could play a role in the detection of pseudoaneurysms.


Relationship between strain levels and permeability of the plasma membrane in statically stretched myoblasts.
Deep tissue injury (DTI) is a life-threatening type of pressure ulcer which initiates subdermally with muscle necrosis at weight-bearing anatomical locations, where localized elevated tissue strains exist. Though it has been suggested that excessive sustained soft tissue strains might compromise cell viability, which then initiates the DTI, there is no experimental evidence to describe how specifically such a process might take place. Here, we experimentally test the hypothesis that macroscopic tissue deformations translated to cell-level deformations and in particular, to localized tensile strains in the plasma membrane (PM) of cells, increase the permeability of the PM which could disrupt vital transport processes. In order to determine whether PM permeability changes can occur due to static stretching of cells we measured the uptake of fluorescein isothiocyanate (FITC)-labeled Dextran (molecular weight = 4 kDa) by deformed vs. undeformed myoblasts, using a fluorescence-activated cell sorting (FACS) method. These PM permeability changes were then correlated with tensile strains in the PM which correspond to the levels of substrate tensile strain (STS) that were applied in the experiments. The PM strains were evaluated by means of confocal-microscopy-based cell-specific finite element (FE) modeling. The FACS studies demonstrated a statistically significant rise in the uptake of the FITC-labeled Dextran with increasing STS levels in the STS ≤ 12% domain, which thereby indicates a rise in the permeability of the PM of the myoblasts with the extent of the applied cellular deformation. The cell-specific FE modeling simulating the experiments further demonstrated that applying average PM tensile strains which exceed 3%, or, applying peak PM tensile strains over 9%, substantially increases the permeability of the PM of myoblasts to the Dextran. Moreover, the permeability of the PM grew rapidly with any further increase in PM strains, though there were no significant changes in the uptake above average and peak PM tensile strain values of 9 and 26%, respectively. These results provide an experimental basis for studying the theory that cell-level deformation-diffusion relationships may be involved in determining the tolerance of soft tissues to sustained mechanical loading, as relevant to the etiology of DTI.


The spatial pattern of international labour flows from and to Pakistan: a preliminary analysis.
"This paper attempts to identify the international labour flows of Pakistinis focussing on their geographical distribution. Based on a survey conducted by the Overseas Pakistanis Foundation and the Population Census of 1981 the study suggests that less developed districts are characterised by low out-migration and high return-migration. In addition, structural characteristics (e.g., land tenure) may be important in explaining low mobility from some underdeveloped districts...."


Determination of oxytetracycline/oxytetracycline hydrochloride in animal feed, fish feed, and veterinary medicinal products by liquid chromatography with fluorescence detection: collaborative study.
A method for determining oxytetracycline (OTC) in animal feed, fish feed, and veterinary medicinal products at medicated use and contamination levels was collaboratively studied. The method is applicable to the analysis of animal feeds and mineral premixes containing levels > or =2 mg/kg, and fish feed containing levels > or =10 mg/kg. Oxytetracycline hydrochloride (OTC.HCI) is extracted from ground feed material in acid-methanol solution using mechanical agitation. After centrifugation for 5 min at 1230 x g, an aliquot of the extract is diluted with water andlor acid-methanol so that the concentration of OTC.HCI is approximately the same as that in the working standard, and the solutions contain at least 50% water. Injectable veterinary medicinal materials (also called animal remedy materials) are diluted with water andlor extractant to reach the target concentration. The extracts are filtered and analyzed by reversed-phase liquid chromatography with fluorescence detection with excitation at 390 nm and emission at 512 nm. Twenty-eight test samples of medicated feeds, supplements, and drug premixes, including 4 test samples for trace-level analysis, were sent to 17 collaborators in Canada, The Netherlands, and the United States. Results were received from 11 laboratories. The RSDr values (within-laboratory repeatability) ranged from 1.26 to 9.21%; RSDR values (among-laboratory reproducibility) ranged from 2.14 to 12.9%, and HorRat values ranged from 0.54 to 3.02. It is recommended that this method be adopted AOAC Official First Action.


Recurrent immune thrombocytopenia following different vaccines.
A boy developed immune thrombocytopenia 2 weeks after receiving his measles-mumps-rubella and varicella vaccines at 12 months of age. He then had a recurrent episode 1 week after the booster doses of his quadrivalent diphtheria-tetanus-pertussis-polio, pneumococcal and meningococcal group C vaccines at 5 years of age. On both the occasions he required hospitalisation and treatment with intravenous immunoglobulin. He received other vaccines, before and in between, without any adverse events. Future vaccines are to be determined on an individual risk-benefit basis and he will be reviewed at the age of 11 when his next routine immunisations are due.


Acetylcholinesterase inhibitors promote angiogenesis in chick chorioallantoic membrane and inhibit apoptosis of endothelial cells.
Alzheimer's disease (AD) is one of the most common causes of dementia in the elderly. Recently, a great attention has been paid to the possible role of vascular changes in the pathogenesis of AD. Reduced microvascular density and degeneration of the endothelium are of structural cerebrovascular changes in AD. Acetylcholinesterase (AChE) inhibitors are widely used for the improvement of AD symptoms. Until now, however, the effects of AChE inhibitors on vascular changes including angiogenesis and endothelial cell apoptosis are not fully understood. In the present work, the effects of three AChE inhibitors (donepezil, rivastigmine, and galantamine) were tested on H2O2-induced apoptosis in human umbilical vein endothelial cells (HUVECs) and on angiogenesis in chicken chorioallantoic membrane model. Incubation of HUVEC with H2O2 led to a significant decrease in cell viability and an increase in the percentage of apoptotic cells. The tested drugs, at concentrations of 1-100 μ M, significantly inhibited the H2O2-induced toxicity. Also, all donepezil, rivastigmine and galantamine significantly increased the number of vessels in the chorioallantoic membrane when injected into fertilized eggs. In conclusion, AChE inhibitors possess angiogenesis-accelerating properties and have antiapoptotic effects on endothelial cells. These effects of AChE inhibitors may be involved in their beneficial effects on AD.


Cutaneous Rosai-Dorfman disease: histopathological presentation as inflammatory pseudotumor. A literature review.
Purely cutaneous Rosai-Dorfman disease is exceptional. The disease is characterized histologically by large, proliferating histiocytes exhibiting inflammatory cells within their cytoplasm (emperipolesis). We present here a case of purely cutaneous generalized disease in which the routine histopathology was suggestive of an inflammatory pseudotumor. Positivity for S-100 protein, alpha1-antitrypsin, alpha1-antichymotrypsin, lysozyme, Mac387 and CD68 proteins, and negativity for CD1a protein confirmed the diagnosis of Rosai-Dorfman disease. The rarity of this case lies in the presence of conspicuous inflammatory pseudotumor-like histopathologic changes, masking an otherwise typical sinus histiocytosis cell infiltrate. This unusual presentation of the disease requires a high index of suspicion by clinicians and pathologists.


[Parents compliance to perform the voiding cystourethrogram test after urinary tract infection].
Urinary tract infections (UTIs) occur in 3-5% of girls and 1% of boys. UTIs have been considered an important risk factor for the development of renal insufficiency, hypertension or end-stage renal disease. Hence, there is a need for early diagnosis and management of UTI to prevent renal scarring. The guidelines concerning the prophylactic treatment and the imaging studies have changed during the past year. It was believed that reflux predisposed to renal infection (pyelonephritis) by causing renal injury or scarring and that imaging studies in children with UTI were meant to identify anatomic abnormalities that predispose them to infection and renal scaring. Today, this concept is no longer prevalent and there is no recommendation to perform voiding cystourethrogram (VCUG] after every UTI. To examine parents' compliance to perform a VCUG test after hospitalization due to UTI and factors affecting their decision. We performed a retrospective study that included all children who were hospitalized and diagnosed with UTI at "Dana" Children's Medical Center during the period 1/2004 - 12/2005. Hospital file records were collected, and the relevant data were obtained. We collected data regarding the parents' compliance to perform a VCUG according to a telephone interview. A total of 227 children participated in the study; 179 (78.85%) parents agreed to partake in the interview. Overall, 52% of the children didn't perform the VCUG because of concern about exposure to radiation (55.91%), fear and distress from pain during the test (43.03%), fear of irreversible damage to the urinary tract (40.86%), lack of relevant information (35.48%] and the primary pediatrician's recommendation to postpone the test. The remaining 48% conducted the test because of the hospital doctor's recommendation (94.18%), Primary pediatrician recommendation (94.18%) and because of the desire to terminate prophylactic treatment (63.95%). There is a significant correlation between the doctors' recommendation and the extent and clarity of their explanation to the parents' understanding of the importance of the test, to the parents' satisfaction from the explanation and to the compliance to perform the test. There is a need to improve the doctors' explanation regarding the performance of VCUG test post UTI. This will improve the patients' compliance to perform their post discharge recommendation.


Smoked heroin self-administration in rhesus monkeys.
The purpose of the present study was to evaluate behavioral and pharmacological determinants of smoked heroin self-administration. Eight rhesus monkeys were trained to self-administer smoked heroin under a chained fixed-ratio, (FR, 64-1024) for lever presses, FR 5 for inhalations schedule during daily experimental sessions. Demand for heroin was determined by plotting consumption (smoke deliveries) as a function of price which was varied by increasing the FR lever press requirement from 64 to 1024. The heroin demand curve was compared to that obtained with smoked cocaine base. Dose-effect determinations were obtained by varying the unit dose of heroin from 0.025 to 1.6 mg/kg per delivery. Pretreatment with naloxone (0.01-1.0 mg/kg IM, 10 min presession) and substitution tests with the peripherally acting opioid loperamide (0.1 mg/kg per delivery) were also conducted. Deliveries of smoked heroin decreased, but lever responding per delivery increased as the FR increased. Demand for heroin was elastic and comparable to demand for smoked cocaine base. Varying the dose of heroin available for self-administration resulted in an asymptotic dose-effect curve. Naloxone pretreatment produced dose-dependent decreases in heroin self-administration. Substitution of loperamide for heroin produced extinction-like responding within one or two sessions, with the total smoke deliveries decreasing by 80% of heroin levels within 8-15 days. Reinstatement of heroin resulted in a rapid return to baseline levels of self-administration. These data suggest that rhesus monkeys will readily and reliably self-administer heroin via the inhalation route, and behavioral and pharmacological manipulations indicate that smoked heroin functioned as a positive reinforcer.


Mapping of genetic determinants of rubella virus associated with growth in joint tissue.
Rubella virus (RV) strains vary in their abilities to replicate and persist in cell cultures derived from human joint tissue (synovial cells [SC]), and this arthrotropism appears to be linked to their association with joint symptoms in vivo. In order to map the genetic determinants of arthrotropism, an infectious clone of the Cendehill vaccine strain of RV was constructed, as well as two chimeric clones containing cDNAs from both Cendehill and Therien (wild-type) strains. Replacement of the entire structural gene region of Therien in the infectious clone pROBO302 with the corresponding region of Cendehill did not affect growth in SC. A further observation that Cendehill bound equally well to SC and the permissive Vero cell line indicated that restriction was not at the level of receptor binding, a function of the envelope proteins. Mutations that affected growth in joint cells were mapped to two locations in the nonstructural gene region. The first of these (nucleotides 2803 and 6416) resulted in a 10-fold decrease in yield of progeny virus from SC. This region contained five mutations, at nucleotides 2829, 3060, 3164, and 3528 (near the carboxy terminus of P150 where the protease domain is located) and at nucleotide 4350 in p90. Further substitution of the sequence representing nucleotides 1 to 2803 to give a complete Cendehill infectious clone restricted growth in SC by a further 100-fold to less than 10 PFU/ml. This region contains three mutations, at nucleotides 34, 37, and 55, within the 5' stem-loop structure. In conclusion, the Cendehill-specific mutations believed to be determinants of joint cell growth are located in two regions, the 5' nontranslated region and in a sequence that encodes the carboxy-terminal region of p150 extending into the helicase domain of p90.


Directed evolution reveals hidden properties of VMAT, a neurotransmitter transporter.
The vesicular neurotransmitter transporter VMAT2 is responsible for the transport of monoamines into synaptic and storage vesicles. VMAT2 is the target of many psychoactive drugs and is essential for proper neurotransmission and survival. Here we describe a new expression system in Saccharomyces cerevisiae that takes advantage of the polyspecificity of VMAT2. Expression of rVMAT2 confers resistance to acriflavine and to the parkinsonian toxin 1-methyl-4-phenylpyridinium (MPP(+)) by their removal into the yeast vacuole. This expression system allowed identification of a new substrate, acriflavine, and isolation of mutants with modified affinity to tetrabenazine (TBZ), a non-competitive inhibitor of VMAT2 that is used in the treatment of various movement disorders including Tourette syndrome and Huntington chorea. Whereas one type of mutant obtained displayed decreased affinity to TBZ, a second type showed only a slight decrease in the affinity to TBZ, displayed a higher K(m) to the neurotransmitter serotonin, but conferred increased resistance to acriflavine and MPP(+). A protein where both types of mutations were combined (with only three amino acid replacements) lost most of the properties of the neurotransmitter transporter (TBZ-insensitive, no transport of neurotransmitter) but displayed enhanced resistance to the above toxicants. The work described here shows that in the case of rVMAT2, loss of traits acquired in evolution of function (such as serotonin transport and TBZ binding) bring about an improvement in older functions such as resistance to toxic compounds. A process that has taken millions of years of evolution can be reversed by three mutations.


Driving under the influence with blood alcohol concentrations over 0.4 g%.
The aim of this study was to evaluate the characteristics of traffic offenders with unusually high blood-alcohol concentrations (BAC>0.4 g%) when arrested. The BAC that kills one person might be easily tolerated by another, depending on, among other things, the person's age, pattern of drinking, and the development of tolerance. The archives of two forensic laboratories, one in Sweden and the other in Wisconsin (USA), were searched to find traffic offenders with BACs>0.4 g%. The results were compared in relation to the person's age and gender, mean BAC and the weekday and time of day of the arrest. The mean age (±standard deviation) of N=158 Swedish offenders was 45±9.0 y, which was not significantly different from the 43±9.4 y in N=233 Wisconsin drivers (p>0.05). Overall there were more men (78%) than women (22%) arrested with BAC's>0.4 g%, although the proportion of women in Wisconsin (35%) was higher than in Sweden (9%) (p<0.001). The mean (median) and highest BAC did not differ between jurisdictions; 0.429 g% (0.422) and 0.546 g% in Sweden and 0.428 g% (0.421 g%) and 0.526 g% in Wisconsin. In Sweden 40% of the arrests occurred on Fridays and Saturdays, whereas in Wisconsin the arrests of people with such high BAC's were more evenly distributed throughout the week. Forty eight percent of the arrests in Sweden were made between 12 noon and 6 pm compared with 37% in Wisconsin. Neither the mean age of offenders nor their mean BAC seemed to depend on the weekday or time of day of the arrest. Attempting to drive with a BAC above 0.4 g% verifies the development of an appreciable tolerance to ethanol-induced cognitive and psychomotor impairment. Reaching such a high BAC probably requires continuous heavy drinking over several days as opposed to an evening's binge drinking.


The association between obesity and low back pain: a meta-analysis.
This meta-analysis assessed the association between overweight/obesity and low back pain. The authors systematically searched the Medline (National Library of Medicine, Bethesda, Maryland) and Embase (Elsevier, Amsterdam, the Netherlands) databases until May 2009. Ninety-five studies were reviewed and 33 included in the meta-analyses. In cross-sectional studies, obesity was associated with increased prevalence of low back pain in the past 12 months (pooled odds ratio (OR) = 1.33, 95% confidence interval (CI): 1.14, 1.54), seeking care for low back pain (OR = 1.56, 95% CI: 1.46, 1.67), and chronic low back pain (OR = 1.43, 95% CI: 1.28, 1.60). Compared with non-overweight people, overweight people had a higher prevalence of low back pain but a lower prevalence of low back pain compared with obese people. In cohort studies, only obesity was associated with increased incidence of low back pain for > or =1 day in the past 12 months (OR = 1.53, 95% CI: 1.22, 1.92). Results remained consistent after adjusting for publication bias and limiting the analyses to studies that controlled for potential confounders. Findings indicate that overweight and obesity increase the risk of low back pain. Overweight and obesity have the strongest association with seeking care for low back pain and chronic low back pain.


Comparison of single and multiple dwell position methods in MammoSite high dose rate (HDR) brachytherapy planning.
The purpose of this study is to dosimetrically compare two plans generated using single dwell position method (SDPM) and multiple dwell position methods (MDPM) in MammoSite high dose rate (HDR) brachytherapy planning for 19 breast cancer patients. In computed tomography (CT) image-based HDR planning, a surface optimization technique was used in both methods. Following dosimetric parameters were compared for fraction 1 plans: %PTV_EVAL (planning target volume for plan evaluation) coverage, dose homogeneity index (DHI), dose con-formal index (COIN), maximum dose to skin and ipsilateral lung, and breast tissue volume receiving 150% (V150[cc]) and 200% (V200[cc]) of the prescribed dose. In addition, a plan was retrospectively generated for each fraction 2-10 to simulate the clinical situation where the fraction 1 plan was used for fractions 2-10 without modification. In order to create nine derived plans for each method and for each of the 19 patients, the catheter location and contours of target and critical structures were defined on the CT images acquired prior to each fraction 2-10, while using the same dwell-time distribution as used for fraction 1 (original plan). Interfraction dose variations were evaluated for 19 patients by comparing the derived nine plans (each for fractions 2-10) with the original plan (fraction 1) using the same dosimetric parameters used for fraction 1 plan comparison. For the fraction 1 plan comparison, the MDPM resulted in slightly increased %PTV_EVAL coverage, COIN, V150[cc] and V200[cc] values by an average of 1.2%, 0.025, 0.5 cc and 0.7cc, respectively, while slightly decreased DHI, maximum skin and ipsilateral lung dose by an average of 0.003, 3.2 cGy and 5.8 cGy, respectively. For the inter-fraction dose variation comparison, the SDPM resulted in slightly smaller variations in %PTV_EVAL coverage, DHI, maximum skin dose and V150[cc] values by an average of 0.4%, 0.0005, 0.5 cGy and 0.2 cc, respectively, while slightly higher average variations in COIN, maximum ipsilateral lung dose and V200[cc] values by 0.0028, 0.2 cGy and 0.2 cc, respectively. All differences were too small to be clinically significant. Compared to the MDPM, the SDPM combined with a surface optimization technique can generate a clinically comparable fraction 1 treatment plan with a similar interfraction dose variation if a single source is carefully positioned at the center of the balloon catheter.


The incidence of pelvic fractures with traumatic lower limb amputation in modern warfare due to improvised explosive devices.
A frequently-seen injury pattern in current military experience is traumatic lower limb amputation as a result of improvised explosive devices (IEDs). This injury can coexist with fractures involving the pelvic ring. This study aims to assess the frequency of concomitant pelvic fracture in IED-related lower limb amputation. A retrospective analysis of the trauma charts, medical notes, and digital imaging was undertaken for all patients arriving at the Emergency Department at the UK military field hospital in Camp Bastion, Afghanistan, with a traumatic lower limb amputation in the six months between September 2009 and April 2010, in order to determine the incidence of associated pelvic ring fractures. Of 77 consecutive patients with traumatic lower limb amputations, 17 (22%) had an associated pelvic fracture (eleven with displaced pelvic ring fractures, five undisplaced fractures and one acetabular fracture). Unilateral amputees (n = 31) had a 10% incidence of associated pelvic fracture, whilst 30 % of bilateral amputees (n = 46) had a concurrent pelvic fracture. However, in bilateral, trans-femoral amputations (n = 28) the incidence of pelvic fracture was 39%. The study demonstrates a high incidence of pelvic fractures in patients with traumatic lower limb amputations, supporting the routine pre-hospital application of pelvic binders in this patient group.


[Granulomatosis with polyangiitis associated with the cytoplasmic antineutrophil antibodies].
The objective of the present work was to analyze the clinical case of granulomatosis with polyangiitis associated with the presence of cytoplasmic antineutrophil antibodies. We considered the medical records of the patient presenting with this condition. It was shown that systemic vasculitis had a polymorphic clinical picture; its diagnostics and treatment encountered serious difficulties. It is concluded that the development of progressive perforation of the nasal septum and destructive changes in the intranasal and adjacent structures after the endonasal surgical intervention implies the necessity of the detailed analysis of the clinical and laboratory observations with the subsequent counselling by a rheumatologist and oncologist for the clarification of the diagnosis and the choice of adequate therapy.


The experience of community first responders in co-producing rural health care: in the liminal gap between citizen and professional.
The involvement of community first responders (CFRs) in medical emergencies in Scotland, and particularly in remote and rural areas, has expanded rapidly in recent years in response to geographical and organisational challenges of emergency medical service access. In 2013 there were over 120 active or developing schemes in a wide variety of settings. Community first responders are volunteers trained in First Person on the Scene (FPOS) first aid, administered prior to the arrival of an ambulance. Although there is limited literature which describes the role of first response, little academic literature has been published about the complexities of their specific role in both the community and organisational contexts. Here we reflect on data from two mixed-methods studies into the role of CFRs in Scotland. We highlight findings that explore the liminal and complex role of the first responder as both 'practitioner' and community member, and how this contributes to a sense of communitas within the study areas. The rural context encompasses additional complexity in relation to the role of emergency care volunteer, having the highest levels of volunteering and this paper questions assumptions that rural areas, are more accepting of volunteerism. Complexities arising from the experience of blurred voluntary/practitioner boundaries emerge as a key feature of voluntary participation in medical emergencies in this setting.


Stride frequency or length? A phylogenetic approach to understand how animals regulate locomotor speed.
Speed regulation in animals involves stride frequency and stride length. While the relationship between these variables has been well documented, it remains unresolved whether animals primarily modify stride frequency or stride length to increase speed. In this study, we explored the interrelationships between these three variables across a sample of 103 tetrapods and assessed whether speed regulation strategy is influenced by mechanical, allometric, phylogenetic or ecological factors. We observed that crouched terrestrial species tend to regulate speed through stride frequency. Such a strategy is energetically costly, but results in greater locomotor maneuverability and greater stability. In contrast, regulating speed through stride length is closely tied to larger arboreal animals with relatively extended limbs. Such movements reduce substrate oscillations on thin arboreal supports and/or helps to reduce swing phase costs. The slope of speed on frequency is lower in small crouched animals than in large-bodied erect species. As a result, substantially more rapid limb movements are matched with only small speed increases in crouched, small-bodied animals. Furthermore, the slope of speed on stride length was inversely proportional to body mass. As such, small changes in stride length can result in relatively rapid speed increases for small-bodied species. These results are somewhat counterintuitive, in that larger species, which have longer limbs and take longer strides, do not appear to gain as much speed increase out of lengthening their stride. Conversely, smaller species that cycle their limbs rapidly do not gain as much speed out of increasing stride frequency as do larger species.


Effects of simvastatin on high-sensitivity C-reactive protein and serum albumin in hemodialysis patients.
A 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor is recommended in hemodialysis (HD) patients with hypercholesterolemia to improve their lipid profiles. We evaluated effects of simvastatin on markers for inflammation, oxidative stress, and coagulation in HD patients. Sixty-two maintenance HD patients with serum cholesterol levels of 200 mg/dL or greater were randomly assigned to the treatment group (n = 31; 8 men, 23 women; age, 63 +/- 11 years) and administered simvastatin, 20 mg/d, for 8 weeks or to the control group (n = 31; 10 men, 21 women; age, 60 +/- 12 years). We measured cholesterol, albumin, high-sensitivity C-reactive protein (hs-CRP), malondialdehyde (MDA; an index of lipid peroxidation), and D-dimer (a marker of intravascular coagulation) in blood at baseline and again at 8 weeks. Fifty-eight of 62 patients completed the study. In the control group, total cholesterol, serum albumin, hs-CRP, MDA, and D-dimer levels did not change. In the treatment group, simvastatin administration for 8 weeks significantly reduced total cholesterol levels from 232 +/- 25 to 165 +/- 39 mg/dL (P < 0.001) and hs-CRP levels from a median of 0.23 mg/dL (range, 0.05 to 1.63 mg/dL) to 0.12 mg/dL (range, <0.006 to 1.45 mg/dL; P < 0.01), whereas it increased serum albumin levels from 3.4 +/- 0.3 to 3.6 +/- 0.4 g/dL (P < 0.001). Administration of simvastatin did not affect MDA and D-dimer levels. These results suggest that in addition to the lipid-lowering effect, simvastatin had an antiinflammatory effect in HD patients. Considering that atherosclerosis is inflammation of the vascular wall, simvastatin may have a beneficial effect on cardiovascular disease, in part because it alleviates inflammation.


A Computational and Experimental View of Hydrogen Bonding in Glycerol Water Clusters.
Polyol-water clusters provide a template to probe ionization and solvation processes of paramount interest in atmospheric and interstellar chemistry. We generate glycerol water clusters in a continuous supersonic jet expansion and interrogate the neutral species with synchrotron-based tunable vacuum ultraviolet photoionization mass spectrometry. A series of glycerol fragments (m/z 44, 61, 62, and 74) clustered with water are observed. A judicious combination of backing pressure, nozzle temperature, and water vapor pressure allows for tuning the mol % of glycerol. The recorded appearance energies of the water cluster series m/z 62 and 74 are similar to that observed in pure glycerol, while the m/z 61 series shows a dependence on cluster composition. Furthermore, this series also tracks the water concentration of the beam. Theoretical calculations on neutral and ionized clusters visualize the hydrogen bond network in these water clusters and provide an assessment of the number of glycerol-glycerol, glycerol-water, and water-water hydrogen bonds in the cluster, as well as their interaction energies. This method of bond counting and interaction energy assessment explains the changes in the mass spectrum as a function of mol % and offers a glimpse of the disruption of the hydrogen bond network in glycerol-water clusters. The calculations also reveal interesting barrierless chemical processes in photoionized glycerol water clusters that are either activated or do not occur without the presence of water. Examples include spontaneous intramolecular proton transfer within glycerol to form a distonic ion, nonactivated breaking of a C-C bond, and spontaneous proton transfer from glycerol to water. These results appear relevant to radiation-induced chemical processing of alcohol-water ices in the interstellar medium.


[Jiangxi planned parenthood].
Jiangxi scored outstanding achievements in planned parenthood in 1980. The natural population growth rate fell from 13.74/1000 in 1979 to below 10/1000. In Yichun Prefecture the growth rate fell to 7.51/1000. Over 70,000 couples with only 1 child took out single-child certificates.


Failed antireflux surgery: surgical outcome of laparoscopic refundoplication in the elderly.
Antireflux surgery has a failure rate between 5 and 20%. Laparoscopic redo-surgery is feasible, but little is known about the surgical outcome in elderly patients. The aim of this prospective study was to evaluate early surgical experience and outcome, including quality of life, after laparoscopic refundoplication in patients older than 65 years. Eleven patients, mean age of 71 years (range: 65-78), underwent laparoscopic redo-surgery. Six patients had the former antireflux procedure performed by the open technique, one having had it twice, one had both laparoscopic and open antireflux procedures, and in 4 the primary intervention was performed laparoscopically. Quality of life was evaluated by using the Gastrointestinal Quality of Life Index. All patients were evaluated prior to surgery, and at 3 months and 12 months after laparoscopic refundoplication, as well as with esophageal manometry and 24-hour pH-monitoring. Redo-procedures were completed laparoscopically in 10 patients. In one patient conversion to an open laparotomy was necessary because of severe bleeding from the spleen. One patient had an injury to the gastric wall, successfully managed laparoscopically. Postoperatively, one patient had moderate dysphagia for a period of two months, another had epigastric pain for the same period. Esophageal manometry and 24-hour pH-monitoring showed normal values in all patients after redo-surgery. Prior to redo-surgery, the mean Gastrointestinal Quality of Life Index was 85.2 points. Three months (mean: 119.8 points) and one year (mean: 119.2 points) after laparoscopic reoperation the general score increased significantly (P < 0.01) and attained the equivalent level of comparable healthy individuals (118.7 points). Laparoscopic refundoplication in the elderly patient is feasible, safe and an effective treatment after failed antireflux surgery. Older patients with failed antireflux surgery have poor quality of life. Laparoscopic redo-surgery improves quality of life significantly to the level of healthy individuals and normalizes objective outcome criteria without any long-term restrictions in daily life.


Antigen 85 variation across lineages of Mycobacterium tuberculosis-implications for vaccine and biomarker success.
Mycobacterium tuberculosis secretes several hundred proteins; many of which elicit immune responses. As a result, many of these proteins have been explored for their potential as diagnostic and vaccine candidates. Of these, the Antigen 85 complex proteins, represented by Antigen85 A, B, and C, are the most studied from the mycobacterial secretome. However, vaccine constructs exploiting Antigen 85 as the sole antigen repertoire have not experienced the pre-clinical and clinical trials success originally anticipated. Anecdotal and biochemical evidence suggests that differences in protein abundance may explain this phenomenon. Here, biochemical, molecular, and mass spectrometry approaches were used to quantify Antigen 85 among six M. tuberculosis strains from four phylogenetically distinct clades. Our data demonstrates that the greatest variation in Antigen 85 is ascribed to protein quantities, whereas few transcriptional differences were found. In addition, the ratio of Antigen 85 A, to B, to C is conserved within clades and phylogenetic neighbors. In contrast, no such relationship between individual protein quantities was observed, and in the case of Antigen85 B, this variation even extends within biological replicates of individual isolates. The relevance of Antigen 85 protein quantities and vaccine efficacy remains to be defined. Absolute quantitation via multiple reaction monitoring mass spectrometry was used to determine the exact molar concentrations of Antigen 85A, B, and C; three key immunodominant proteins present in M. tuberculosis. Further, the concentration of these three proteins was compared among various clades of M. tuberculosis, and demonstrated differences in abundance of two of the three proteins. These proteins have been identified as key antigens in multiple vaccine and diagnostic platforms, thus the potential relevance of their abundance in various M. tuberculosis clades to the successful outcome of these interventions is discussed. This article is part of a Special Issue entitled: Trends in Microbial Proteomics.


Structure-function studies of a plant tyrosyl-DNA phosphodiesterase provide novel insights into DNA repair mechanisms of Arabidopsis thaliana.
TDP1 (tyrosyl-DNA phosphodiesterase 1), a member of the PLD (phospholipase D) superfamily, catalyses the hydrolysis of a phosphodiester bond between a tyrosine residue and the 3'-phosphate of DNA. We have previously identified and characterized the AtTDP gene in Arabidopsis thaliana, an orthologue of yeast and human TDP1 genes. Sequence alignment of TDP1 orthologues revealed that AtTDP has both a conserved C-terminal TDP domain and, uniquely, an N-terminal SMAD/FHA (forkhead-associated) domain. To help understand the function of this novel enzyme, we analysed the substrate saturation kinetics of full-length AtTDP compared with a truncated AtTDP mutant lacking the N-terminal FHA domain. The recombinant AtTDP protein hydrolysed a single-stranded DNA substrate with Km and kcat/Km values of 703±137 nM and (1.5±0.04)×10(9) M(-1)·min(-1) respectively. The AtTDP-(Δ1-122) protein (TDP domain) showed kinetic parameters that were equivalent to those of the full-length AtTDP protein. A basic amino acid sequence (RKKVKP) within the AtTDP-(Δ123-605) protein (FHA domain) was necessary for nuclear localization of AtTDP. Analysis of active-site mutations showed that a histidine and a lysine residue in each of the HKD motifs were critical for enzyme activity. Vanadates, inhibitors of phosphoryl transfer reactions, inhibited AtTDP enzymatic activity and retarded the growth of an Arabidopsis tdp mutant. Finally, we showed that expression of the AtTDP gene could complement a yeast tdp1Δrad1Δ mutant, rescuing the growth inhibitory effects of vanadate analogues and CPT (camptothecin). Taken together, the results of the present study demonstrate the structure-based function of AtTDP through which AtTDP can repair DNA strand breaks in plants.


Leptospirosis: Increasing importance in developing countries.
Leptospirosis is a zoonotic disease caused by the pathogenic helical spirochetes, Leptospira. Symptoms include sudden-onset fever, severe headaches, muscle pain, nausea and chills. Leptospirosis is endemic in developing countries such as Malaysia, India, Sri Lanka, and Brazil where thousands of cases are reported annually. The disease risk factors include the high population of reservoirs, environmental factors, recreational factors, and occupational factors. To end the endemicity of leptospirosis, these factors need to be tackled. The management of leptospirosis needs to be refined. Early diagnosis remains a challenge due to a lack of clinical suspicion among physicians, its non-specific symptoms and a limited availability of rapid point-of-care diagnostic tests. The purpose of this review is to provide insight into the status of leptospirosis in developing countries focusing on the risk factors and to propose methods for the improved management of the disease.


Regional variations in medical spending and utilization: a longitudinal analysis of US Medicare population.
One of the perceived symptoms of US Medicare inefficiency is the existence of the dramatic variation in spending and utilization in different areas of the country. This study uses the Continuous Medicare History Sample, a large longitudinal 5% sample of all Medicare beneficiaries from 1974 to 2003, to study the issue. We show that the spending and utilization disparities are significant at the aggregate state level. More importantly, the variation shows signs of narrowing over time, particularly in the earlier years of the sample period and in some cases following major reforms. However, it remains significant even after an array of demographic, demand and supply side factors are controlled for.


Computer-assisted modelling and optimisation of reversed-phase high-temperature liquid chromatographic (RP-HTLC) separations.
The use of high temperatures (above 100 °C) in reversed-phase liquid chromatography (RP-HTLC) has opened up novel and enhanced applications for this essential separation technique. Although the favourable effects of temperature on LC have been extensively studied both theoretically and practically, its potential application to method development has barely been investigated. These favourable effects include enhanced speed, efficiency, resolution and detectability, as well as changes in selectivity, especially for polar and ionisable compounds, and the emergence of new options such as temperature programming and the concomitant use of solvent and temperature gradients, green separations, and so on. The recent availability of silica-based columns that routinely support high temperatures in addition to more conventional temperature-resistant columns (based on graphitised carbon, polymers and zirconium oxide) and dedicated column ovens that allow accurate temperature control up to 200 °C makes it possible to conceive of RP-HTLC as a routine separation technique in the modern analytical laboratory. On the other hand, the addition of temperature as a new optimisable parameter to RPLC adds further complexity to method development. Thus, new computer-assisted optimisation tools that extend the capabilities of current computer-assisted tools are being specifically developed for this type of separation. A new specially developed computer-assisted method development (CAMD) tool is presented herein, and its efficiency is demonstrated. This CAMD is based on the development of a rugged retention model for peaks, allowing the simulation of any kind of RP-HTLC separation, including isocratic, linear, curved, multilinear and stepwise gradients of solvent composition concomitant with constant, linear and multilinear temperature gradients. Both the retention models and the unattended optimisation of separations are driven by evolutionary algorithms, thus providing negligible-cost, rapid, highly efficient, and user-friendly optimisation processes.


Oxidation of formic acid and carbon monoxide on gold electrodes studied by surface-enhanced Raman spectroscopy and DFT.
The oxidation of formic acid and carbon monoxide was studied at a gold electrode by a combination of electrochemistry, in situ surface-enhanced Raman spectroscopy (SERS), differential electrochemical mass spectrometry, and first-principles DFT calculations. Comparison of the SERS results and the (field-dependent) DFT calculations strongly suggests that the relevant surface-bonded intermediate during oxidation of formic acid on gold is formate HCOO- ad*. Formate reacts to form carbon dioxide via two pathways: at low potentials, with a nearby water to produce carbon dioxide and a hydronium ion; at higher potentials, with surface-bonded hydroxyl (or oxide) to give carbon dioxide and water. In the former pathway, the rate-determining step is probably related to the reaction of surface-bonded formate with water, as measurements of the reaction order imply a surface almost completely saturated with adsorbate. The potential dependence of the rate of the low-potential pathway is presumably governed by the potential dependence of formate coverage. There is no evidence for CO formation on gold during oxidation of formic acid. The oxidation of carbon monoxide must involve the carboxyhydroxyl intermediate, but SERS measurements do not reveal this intermediate during CO oxidation, most likely because of its low surface coverage, as it is formed after the rate-determining step. Based on inconclusive spectroscopic evidence for the formation of surface-bonded OH at potentials substantially below the surface oxidation region, the question whether surface-bonded carbon monoxide reacts with surface hydroxyl or with water to form carboxyhydroxyl and carbon dioxide remains open. The SERS measurements show the existence of both atop and bridge-bonded CO on gold from two distinguishable low-frequency modes that agree very well with DFT calculations.


Analysis of cortical shape in children with simplex autism.
We used surface-based morphometry to test for differences in cortical shape between children with simplex autism (n = 34, mean age 11.4 years) and typical children (n = 32, mean age 11.3 years). This entailed testing for group differences in sulcal depth and in 3D coordinates after registering cortical midthickness surfaces to an atlas target using 2 independent registration methods. We identified bilateral differences in sulcal depth in restricted portions of the anterior-insula and frontal-operculum (aI/fO) and in the temporoparietal junction (TPJ). The aI/fO depth differences are associated with and likely to be caused by a shape difference in the inferior frontal gyrus in children with simplex autism. Comparisons of average midthickness surfaces of children with simplex autism and those of typical children suggest that the significant sulcal depth differences represent local peaks in a larger pattern of regional differences that are below statistical significance when using coordinate-based analysis methods. Cortical regions that are statistically significant before correction for multiple measures are peaks of more extended, albeit subtle regional differences that may guide hypothesis generation for studies using other imaging modalities.


Folliculin directs the formation of a Rab34-RILP complex to control the nutrient-dependent dynamic distribution of lysosomes.
The spatial distribution of lysosomes is important for their function and is, in part, controlled by cellular nutrient status. Here, we show that the lysosome associated Birt-Hoge-Dubé (BHD) syndrome renal tumour suppressor folliculin (FLCN) regulates this process. FLCN promotes the peri-nuclear clustering of lysosomes following serum and amino acid withdrawal and is supported by the predominantly Golgi-associated small GTPase Rab34. Rab34-positive peri-nuclear membranes contact lysosomes and cause a reduction in lysosome motility and knockdown of FLCN inhibits Rab34-induced peri-nuclear lysosome clustering. FLCN interacts directly via its C-terminal DENN domain with the Rab34 effector RILP Using purified recombinant proteins, we show that the FLCN-DENN domain does not act as a GEF for Rab34, but rather, loads active Rab34 onto RILP We propose a model whereby starvation-induced FLCN association with lysosomes drives the formation of contact sites between lysosomes and Rab34-positive peri-nuclear membranes that restrict lysosome motility and thus promote their retention in this region of the cell.


Describing the characteristics and healthcare use of high-cost acute care users at the end of life: a pan-Canadian population-based study.
A minority of individuals use a large portion of health system resources, incurring considerable costs, especially in acute-care hospitals where a significant proportion of deaths occur. We sought to describe and contrast the characteristics, acute-care use and cost in the last year of life among high users and non-high users who died in hospitals across Canada. We conducted a population-based retrospective-cohort study of Canadian adults aged ≥18 who died in hospitals across Canada between fiscal years 2011/12-2014/15. High users were defined as patients within the top 10% of highest cumulative acute-care costs in each fiscal year. Patients were categorized as: persistent high users (high-cost in death year and year prior), non-persistent high users (high-cost in death year only) and non-high users (never high-cost). Discharge abstracts were used to measure characteristics and acute-care use, including number of hospitalizations, admissions to intensive-care-unit (ICU), and alternate-level-of-care (ALC). We identified 191,310 decedents, among which 6% were persistent high users, 41% were non-persistent high users, and 46% were non-high users. A larger proportion of high users were male, younger, and had multimorbidity than non-high users. In the last year of life, persistent high users had multiple hospitalizations more often than other groups. Twenty-eight percent of persistent high users had ≥2 ICU admissions, compared to 8% of non-persistent high users and only 1% of non-high users. Eleven percent of persistent high users had ≥2 ALC admissions, compared to only 2% of non-persistent high users and < 1% of non-high users. High users received an in-hospital intervention more often than non-high users (36% vs. 19%). Despite representing only 47% of the cohort, persistent and non-persistent high users accounted for 83% of acute-care costs. High users - persistent and non-persistent - are medically complex and use a disproportionate amount of acute-care resources at the end of life. A greater understanding of the characteristics and circumstances that lead to persistently high use of inpatient services may help inform strategies to prevent hospitalizations and off-set current healthcare costs while improving patient outcomes.