The epidemiology of ischaemic heart disease in the different ethnic populations in Johannesburg.
From an examination of death certificates of members of different ethnic populations in Johannesburg who have died of ischaemic heart disease (IHD), age-specific mortality rates have been calculated. The period examined in the case of Whites was 1971; in the case of Indians and Coloureds it was 1968-1971 (because of the smaller sizes of the populations); and in the case of Blacks 1968-1971 (because of the small number dying from IHD). Notable findings included: (i) an extremely high mortality rate among Whites (especially among the Jewish sector, undoubtedly the highest extant); (ii) an equally high or higher rate among Indians, a feature which also prevails in Indian immigrant populations elsewhere; (iii) a lower rate among Coloureds; and (iv) an extremely low rate among Blacks, as is also the case among other Black African populations. Comparative information is provided on IHD in other populations.


Hax-1 is rapidly degraded by the proteasome dependent on its PEST sequence.
HS-1-associated protein X-1 (Hax-1), is a multifunctional protein that has sequence homology to Bcl-2 family members. HAX-1 knockout animals reveal that it plays an essential protective role in the central nervous system against various stresses. Homozygous mutations in the HAX-1 gene are associated with autosomal recessive forms of severe congenital neutropenia along with neurological symptoms. The protein level of Hax-1 has been shown to be regulated by cellular protease cleavage or by transcriptional suppression upon stimulation. Here, we report a novel post-translational mechanism for regulation of Hax-1 levels in mammalian cells. We identified that PEST sequence, a sequence rich in proline, glutamic acid, serine and threonine, is responsible for its poly-ubiquitination and rapid degradation. Hax-1 is conjugated by K48-linked ubiquitin chains and undergoes a fast turnover by the proteasome system. A deletion mutant of Hax-1 that lacks the PEST sequence is more resistant to the proteasomal degradation and exerts more protective effects against apoptotic stimuli than wild type Hax-1. Our data indicate that Hax-1 is a short-lived protein and that its PEST sequence dependent fast degradation by the proteasome may contribute to the rapid cellular responses upon different stimulations.


Synchronization of carbohydrate and protein sources on fermentation and passage rates in dairy cows.
Four ruminally cannulated Holstein cows in midlactation were randomly assigned to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments to evaluate two nonstructural carbohydrate sources (corn or barley) with two sources of ruminally undegradable protein (soybean meal or extruded soybean meal) on milk production, ruminal fermentation, and digesta passage rates. Milk production (25.1, 27.5, 23.8, and 23.5 kg/d for the corn and soybean meal, corn and extruded soybean meal, barley and soybean meal, and barley and extruded soybean meal, respectively) and dry matter intake per unit of body weight (3.9, 4.1, 3.7, and 3.7%) were greater for cows fed corn than for cows fed barley and were similar for cows fed soybean meal or extruded soybean meal. Concentrations of ruminal NH3-N were greater for cows fed the corn and soybean meal diet than for cows fed other diets (15.0, 10.4, 9.0, and 11.3 mg/dl). Rumen volatile fatty acid concentrations were greater for cows fed corn than barley (133, 139, 121, and 118 mumol/ml). Fractional passage rates of solids from the rumen were greater for cows fed the barley and soybean meal diet than cows fed the corn and soybean meal diet (3.4, 3.9, 4.2, and 3.8%/h), and ruminal liquid dilution rates were similar for cows fed all diets (11.2, 11.0, 11.1, and 11.9%/h). The attempt to synchronize ruminal nonstructural carbohydrate and crude protein degradability produced minimal benefits for midlactation dairy cows.


Postendocytic sorting of constitutively internalized dopamine transporter in cell lines and dopaminergic neurons.
The dopamine transporter (DAT) mediates reuptake of released dopamine and is the target for psychostimulants, such as cocaine and amphetamine. DAT undergoes marked constitutive endocytosis, but little is known about the fate and sorting of the endocytosed transporter. To study DAT sorting in cells lines, we fused the one-transmembrane segment protein Tac to DAT, thereby generating a transporter (TacDAT) with an extracellular antibody epitope suited for trafficking studies. TacDAT was functional and endocytosed constitutively in HEK293 cells. According to an ELISA-based assay, TacDAT intracellular accumulation was increased by the lysosomal protease inhibitor leupeptin and by monensin, an inhibitor of lysosomal degradation and recycling. Monensin also reduced TacDAT surface expression consistent with partial recycling. In both HEK293 cells and in the dopaminergic cell line 1Rb3An27, constitutively internalized TacDAT displayed primary co-localization with the late endosomal marker Rab7, less co-localization with the "short loop" recycling marker Rab4, and little co-localization with the marker of "long loop" recycling endosomes, Rab11. Removal by mutation of N-terminal ubiquitination sites did not affect this sorting pattern. The sorting pattern was distinct from a bona fide recycling membrane protein, the beta(2)-adrenergic receptor, that co-localized primarily with Rab11 and Rab4. Constitutively internalized wild type DAT probed with the fluorescently tagged cocaine analogue JHC 1-64, exhibited the same co-localization pattern as TacDAT in 1Rb3An27 cells and in cultured midbrain dopaminergic neurons. We conclude that DAT is constitutively internalized and sorted in a ubiquitination-independent manner to late endosomes/lysosomes and in part to a Rab4 positive short loop recycling pathway.


Construction of the multidecker anthracene-silver(I) system by intramolecular pi-pi interactions.
As a continuation of the study on the construction of the multidecker aromatic compounds with pi-pi interactions, the anthracene derivatives 9,10-dimethylanthracene (dman), 9,10-diphenylanthracene (dpan), and 7,12-dimethylbenz[a]anthracene (dmban) have been studied toward complexation with the silver(I) ion. The crystal structures of [Ag(dman)(0.5)(p-xylene)(ClO(4))], [Ag(2)(dpan)(0.5)(C(6)H(6))(0.5)(CF(3)SO(3))(2)], and [Ag(2)(dmban)(2)(ClO(4))(2)](p-xylene), together with the metal-free ligand dpan, have been determined by single-crystal X-ray diffraction. These pi-electron-rich aromatic compounds are found to have great promise as an approach to the effective self-assembly of high nuclearity in a multilayer fashion.


Comparative biodistribution of 12 ¹¹¹In-labelled gastrin/CCK2 receptor-targeting peptides.
Cholecystokinin 2 (CCK-2) receptor overexpression has been demonstrated in various tumours such as medullary thyroid carcinomas and small-cell lung cancers. Due to this high expression, CCK-2 receptors might be suitable targets for radionuclide imaging and/or radionuclide therapy. Several CCK-2 receptor-binding radiopeptides have been developed and some have been tested in patients. Here we aimed to compare the in vivo tumour targeting properties of 12 (111)In-labelled 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-conjugated gastrin/CCK2 receptor-binding peptides. Two CCK8-based peptides and ten gastrin-based peptide analogues were tested. All peptides were conjugated with DOTA and labelled with (111)In. Biodistribution studies were performed in mice with subcutaneous CCK2/gastrin receptor-expressing tumours and with receptor-negative tumours contralaterally. Biodistribution was studied by counting dissected tissues at 1 and 4 h after injection. Both the CCK analogues displayed relatively low tumour uptake (approximately 2.5%ID/g) as compared to minigastrin analogues. Two linear minigastrin peptides (MG0 and sargastrin) displayed moderate tumour uptake at both 1 and 4 h after injection, but also very high kidney uptake (both higher than 48%ID/g). The linear MG11, lacking the penta-Glu sequence, showed lower tumour uptake and also low kidney uptake. Varying the N-terminal Glu residues in the minigastrin analogues led to improved tumour targeting properties, with PP-F11 displaying the optimal biodistribution. Besides the monomeric linear peptides, a cyclized peptide and a divalent peptide were tested. Based on these studies, optimal peptides for peptide receptor radionuclide targeting of CCK2/gastrin receptor-expressing tumours were the linear minigastrin analogue with six D-Glu residues (PP-F11), the divalent analogue MGD5 and the cyclic peptide cyclo-MG1. These peptides combined high tumour uptake with low kidney retention, and may therefore be good candidates for future clinical studies.


Validation of in vitro labeling method for human use of heat-damage red blood cells to detect splenic tissue and hemocateretic function.
Selective imaging of the splenic tissue is obtained with heat-damaged, or heat-denatured, red blood cells (RBCs) of the patient labeled with 99mTc in a variety of clinical scenarios. Aim of the study was to validate the process used for labelling heat-damaged red blood cells "totally in vitro", after blood sample collection, before re-inject labeled RBCs to the patient. Moreover, we assessed efficacy of the staff training programme in order to guarantee repeatibility and method standardization in the clinical routine. The validation process of the labeling procedure was performed in three different patients during three consecutive days. After collection of a blood sample using a heparinized syringe, we isolated erythrocytes from other blood components by centrifugation and washing steps. Then, we added the stannous pyrophosphate (PYP) to the erythrocytes pellet, after pH control. The 'pretinning' of RBCs was necessary to reduce Tc-99m once pertechnetate was entered them. After the labeling reaction with 130 MBq of 99mTc-pertechnetate, the erythrocytes were denatured in a water bath at a temperature of 49°-50°C, for 10 min. Radioactivity of blood aliquotes was measured with a dose calibrator and labelling efficiency (LE%) was determined. The labelling purity was measured using a gamma counter and calculated using the formula: counts of pellet/counts of pellet+(counts of surnatant)*100.Training program was evaluated using a Learning Questionnaire (LQ). with a grading score from 6 ("") to 1 ("nothing") for each operator (n=3). We didn't observed the presence of macroaggregates during the entire process, until the final sample. The labelling efficiency resulted at very high values in the three consecutive measured aliquotes (mean value 73.67%) as well as the labelling purity (>95.22%). In our instituion, we use splenic imaging with labelled heat-damaged RBCs to detect ectopic spleen, splenosis, extramedullary hematopoiesis. We performed 3 procedures with heat-damaged labeled RBCs with a mean labelling efficiency 73.67%.Training and learning programmes were scored by key objective areas with a mean value of 5. Our in vitro labeling process of heat-damaged RBCs is simple and safe, providing a useful technique easy to implement in clinical routine for splenic imaging Learning outcome of the training programme was scored as effective by all the operators with evidence of high-efficiency-reproducible procedure mantained over time.


Routine opioid outcome monitoring in community pharmacy: Pilot implementation study protocol.
Increases in opioid use and related harms such as mortality are occurring in many high income countries. Community pharmacists are often in contact with patients at risk of opioid-related harm and represent an ideal point for intervention. Best practice in monitoring opioid-related outcomes involves assessing analgesia, pain functioning, mood, risks and harms associated with opioid use. Community pharmacists are well-placed to undertake these tasks. Our pilot study will test the implementation of a computer-facilitated screening and brief intervention (SBI). The SBI will support pharmacist identification of opioid-related problems and provide capacity for brief intervention including verbal reinforcement of tailored information sheets, supply of naloxone and referral back to the opioid prescriber. The SBI utilises software that embeds study procedures into dispensing workflow and assesses opioid outcomes with domains aligned with a widely accepted clinical framework. We will recruit and train 75 pharmacists from 25 pharmacies to deliver the Routine Opioid Outcome Monitoring (ROOM) SBI. Pharmacists will complete the SBI with up to 500 patients in total (20 per pharmacy). Data will be collected on pharmacists' knowledge and confidence through pre- and post-intervention online surveys. Data on feasibility, acceptability and implementation outcomes, including naloxone supply, will also be collected. Our study will examine changes in pharmacists' knowledge and confidence to deliver the SBI. Through the implementation pilot, we will establish the feasibility and acceptability of a pharmacist SBI that aims to improve monitoring and clinical management of patients who are prescribed opioids.


The supraorbital keyhole approach with eyebrow incisions for treating lesions in the anterior fossa and sellar region.
Keyhole surgery has developed since the 1990s as a less invasive therapeutic strategy for intracranial lesions, initially for the treatment of intracranial aneurysms. The purpose of this study was to describe and evaluate the results of surgical treatment of lesions in the anterior fossa and sellar region via a supraorbital keyhole approach using eyebrow incisions. Between April 1994 and July 2003, 54 patients with lesions in the anterior fossa and sellar region were operated on via the supraorbital keyhole approach. The surgical results were studied retrospectively and compared with that of patients with lesions at the same locations but treated via a conventional subfrontal approach. No significant difference in curative effect was found between the conventional subfrontal approach and the supraorbital keyhole approach. However, the supraorbital approach required a much smaller skin incision, causing less surgical trauma, while achieving excellent surgical exposure and good recovery. The supraorbital keyhole approach using an eyebrow incision is safe, effective, and both suitable and convenient for treating lesions in the anterior fossa and sellar region, with almost no adverse consequences on the facial features of patients.


An economic evaluation of an augmented cognitive behavioural intervention vs. computerized cognitive training for post-stroke depressive symptoms.
Stroke survivors encounter emotional problems in the chronic phase after stroke. Post-stroke depressive symptoms have major impact on health-related quality of life (HRQol) and lead to increased hospitalization and therefore substantial healthcare costs. We present a cost-effectiveness and cost-utility evaluation of a cognitive behavioural therapy augmented with occupational and movement therapy to support patients with a stroke with depressive symptoms in goal-setting and goal attainment (augmented CBT) in comparison with a computerized cognitive training program (CogniPlus) as a control intervention. A trial-based economic evaluation was conducted from a societal perspective with a time horizon of 12 months. Stroke patients (aged 18+ years) with signs of depression (Hospital Anxiety and Depression Scale (HADS) - subscale depression > 7) were eligible to participate. Primary outcomes were the HADS and Quality Adjusted Life Years (QALYs) based on the three-level five-dimensional EuroQol (EQ-5D-3 L). Missing data were handled through mean imputation (costs) and multiple imputation (HADS and EuroQol), and costs were bootstrapped. Sensitivity analyses were performed to test robustness of baseline assumptions. Sixty-one patients were included. The average total societal costs were not significantly different between the control group (€9,998.3) and the augmented CBT group (€8,063.7), with a 95 % confidence interval (-5,284, 1,796). The augmented CBT intervention was less costly and less effective from a societal perspective on the HADS, and less costly and slightly more effective in QALYs, in comparison with the control treatment. The cost-effectiveness and cost-utility analyses provided greater effects and fewer costs for the augmented CBT group, and fewer effects and costs for the HADS. Based on a willingness to pay (WTP) level of €40,000 per QALY, the augmented CBT intervention had a 76 % chance of being cost-effective. Sensitivity analyses showed robustness of results. The stroke-specific augmented CBT intervention did not show convincing cost-effectiveness results. In addition to other literature, this study provided new insights into the potential cost-effectiveness of an adjusted cognitive behavioural therapy intervention. However, as our study showed a 76 % chance of being cost-effective for one outcome measure (QALY) and did not provide convincing cost-effectiveness results on the HADS we recommend further research in a larger population.


Polyoxin D inhibits colloidal gold-wheat germ agglutinin labelling of chitin in dimorphic forms of Candida albicans.
Yeasts and mycelia of the pathogen Candida albicans grown in the presence of polyoxin D, a competitive inhibitor of chitin synthase, formed chains of swollen bulbous cells as observed by fluorescence microscopy. Wheat germ agglutinin (WGA) complexed to colloidal gold (Au) was used as a specific label at the ultrastructural level to visualize chitin in walls of control and polyoxin-treated cells. In control cells, Au-WGA labelling was preferentially localized in the innermost wall layers and was predominant at bud scars and septa. After 4.5 h in 4 mM-polyoxin D, budding in yeasts and lateral wall growth in mycelia continued, but primary septa failed to form and no Au-WGA labelling was detected in the walls. These results demonstrated that the morphological alterations caused by polyoxin D were due to the absence of chitin, a wall component important for formation of primary septa and for maintenance of structural integrity during morphogenesis.


The quality of bioethics debate: implications for clinical ethics committees.
Bioethicists have recently expressed concern over a lack of quality control within the field. This apprehension focuses on bioethics expanding in ways that obscure its distinctive ethical remit and the specialist reasoning skills it requires. This thesis about the quality and conduct of bioethics may have particular relevance for clinical ethics. As one of the youngest offshoots of bioethics, the field focuses on the ethical issues that arise specifically in a clinical context. However, non-ethics specialists are increasingly involved in this field. This means that clinical ethics could be especially vulnerable to the quality control concerns articulated within bioethics. The growing public profile of clinical ethics means that concerns over quality in this area warrant specific attention by those concerned with declining standards in bioethics and those working in clinical ethics.


Dermoscopy of skin metastases from breast cancer: two case reports.
Cutaneous metastatic breast cancer is the most common cutaneous metastatic malignancy in women. The assessment of cutaneous metastatic disease can be perplexing because the clinical presentation appears similar to other skin malignancies like angiosarcoma or melanoma, or benign diseases like cellulitis and lymphedema. To date, only a limited number of dermoscopic images of cutaneous metastatic solid tumors, especially breast cancer, have been published. The authors report two Moroccan cases highlighting dermoscopy as a quick tool to recognize skin metastasis of breast cancer in two different clinical presentations. A 51-year-old Moroccan woman presented with nodules of various sizes on and around a mastectomy scar, and a 65-year-old Moroccan woman presented with cellulitis-like lesions on her chest wall and her back. Dermoscopic features were similar in the two cases with findings of yellow central areas, polymorphic vessels, whitish bright lines, whitish structureless areas, and linear irregular fissure-like depressions on a pink-orange background. The recognition of dermoscopic patterns of cutaneous metastasis of breast cancer is not only useful to facilitate diagnosis at an early stage and to rule out other differentials, especially in difficult presentations such as cellulitis-like lesions or lymphedema, but it may also be used by physicians in monitoring mastectomy scars.


Activation of protein kinase R is required for induction of stress granules by respiratory syncytial virus but dispensable for viral replication.
We performed experiments to determine the effect of PKR activation on respiratory syncytial virus (RSV) replication. We first determined that RSV infection activates PKR which induces the phosphorylation of eIF2α, resulting in the formation of host stress granules. We used RNA interference to decrease endogenous PKR levels. RSV replication was not altered in cells deficient for PKR expression. However, RSV-mediated stress granule formation was significantly reduced in PKR-knockdown cells. As an alternative method to block PKR activation, we used treatment with the kinase inhibitor 2-aminopurine (2-AP). We observed that 2-AP treatment significantly reduced viral replication. We also treated PKR-knockdown cells with 2-AP and inoculated with RSV. Under these conditions, 2-AP treatment diminished viral replication in the absence of PKR expression. These results suggest that PKR activation has a minimal effect on RSV replication and that the antiviral effect of 2-AP during RSV infection likely occurs via a PKR-independent mechanism.


The consistency of the posterior probability of linkage.
When searching for trait loci along the genome, properly incorporating prior genomic information into the analysis will almost certainly increase the chance of success. Recently, we devised a method that utilizes such prior information in the mapping of trait genes for complex disorders (Vieland, 1998; Wang el al. 1999; Vieland et al. 2000). This method uses the posterior probability of linkage (PPL) based on the admixture model as a measure of linkage information. In this paper, we study the consistency of the PPL. It is shown that, as the number of pedigrees increases, the PPL converges in probability to 1 when there is linkage between the marker and a trait locus, and converges to 0 otherwise. This conclusion is shown to be true for general pedigrees and trait models, even, when the likelihood functions are based on misspecified trait models. As part of the effort to prove this conclusion, it is shown that when there is no linkage, the maximum likelihood estimator of the recombination fraction in the admixture model is asymptotically 0.5, even when the admixture model misrepresents the true model.


Surgical pathology of the thymus: 20 years' experience.
Fifty five patients underwent thymic surgery at Papworth Hospital from April 1964 to March 1984. The number presenting and the percentage with symptoms annually remained unchanged during this period. Forty four of these 55 patients had tumours. Twenty eight had thymomas (18% of whom had myasthenia gravis and 7% red cell aplasia), nine Hodgkin's disease, four germ cell tumours, and three secondary carcinomatous tumours. Five tumours were cystic. Six further patients had non-tumourous cystic lesions (four simple, one foregut, one lymphangiectatic). The remaining five patients had follicular hyperplasia; all of these had myasthenia gravis. Complete excision was performed in 41 of the 55 patients. So far survival is 100% in those with benign lesions other than benign thymomas, where the survival was 70% at five years. Those with malignant thymomas had a 60% survival rate at five years and those with Hodgkin's disease 29%.


Oculopharyngeal muscular dystrophy among Bulgarian Jews: a new cluster?
Oculopharyngeal muscular dystrophy (OPMD) produced by the (GCG)13 expansion mutation in the PABPN1 gene is frequent among Uzbek Jews in Israel. To describe the phenotypic and genotypic features in five Bulgarian Jewish patients, from different families, with autosomal dominant OPMD. We performed clinical follow-up, electrodiagnostic tests and mutation detection. Blood samples were obtained after informed consent and DNA was extracted; measurement of GCG repeats in both PABPN1 alleles and sequencing of OPMD mutations were performed according to standard techniques. We identified five patients (four females), aged 58 to 71 years, with bilateral ptosis, dysphagia, dysphonia (n = 3) and myopathic motor units by electromyography. In all patients we noticed proximal weakness of the upper limbs with winging scapulae in three of them. All cases shared the (GCG)13-(GCG)10 PABPN1 genotype. OPMD among Bulgarian Jews is produced by a (GCG)13 expansion, identical to the mutation in Uzbek Jews and French Canadians. In addition to the classical neurological and neuro-ophthalmological features, early shoulder girdle weakness is common in Bulgarian Jewish patients; this is an unusual feature during the early stages of OPMD produced by the same mutation in other populations. We suggest that besides the disease-producing GCG expansion, additional ethnicity-related genetic factors may influence the OPMD phenotype. OPMD is a rare disease, and the identification of five affected families in the rather small Bulgarian Jewish community in Israel probably represents a new cluster; future haplotype studies may elucidate whether a founder effect occurred.


Novel facets of glioma invasion.
Malignant gliomas including Glioblastoma (GBM) are characterized by extensive diffuse tumor cell infiltration throughout the brain, which represents a major challenge in clinical disease management. While surgical resection is beneficial for patient outcome, it is well recognized that tumor cells at the invasive front or beyond stay behind and constitute a major source of tumor recurrence. Invasive glioma cells also represent a difficult therapeutic target since they are localized within normal functional brain areas with an intact blood brain barrier (BBB), thereby excluding most systemic drug treatments. Cell movement is mediated via the actin cytoskeleton where corresponding membrane protrusions play essential roles. This review provides an overview of the various paths of glioma cell invasion and underlines the specific aspects of the brain microenvironment. We highlight recent insight into tumor microtubes, neuro-glioma synapses and tumor metabolism which can regulate collective invasion processes. We also focus on the deregulation of actin cytoskeleton-related components in the context of glioma invasion, a deregulation that may be controlled by genomic alterations in tumor cells as well as by various external factors, including extracellular matrix (ECM) components and non-malignant stromal cells. Finally we critically assess the challenges and opportunities for therapeutically targeting glioma cell invasion.


Individualized care in patients with chronic congestive heart failure.
Based on our experience with the prospective follow-up study in a group of patients (n = 21) with congestive heart failure (CHF) (NYHA III n = 10, NYHA IV n = 11) and left ventricular ejection fraction below 30%, who were referred to our ordinary care unit by their family physicians after an initial work-up, we investigated the impact of a special treatment program in 25 patients with similar CHF despite therapy (60 +/- 4 years, NYHA III n = 11, NYHA IV n = 14, LVEF less than 30%, 17 +/- 3%). The program focused on three issues: (1) individualized medical therapy of CHF, (2) antiarrhythmic treatment and close follow-up visits, and (3) continuous education of patients and physicians in order to improve treatment compliance and the early management of complications. Medical treatment was based on diuretic and vasodilator therapy in all the patients, while positive inotropic substances including digoxin were given selectively. Vasodilator treatment was started with prazosin in 22 patients and with ACE inhibitors in three patients with low serum sodium. Fifty-five percent of the patients on prazosin had to be changed over to ACE inhibitors. Amiodarone was used as first line drug to treat symptomatic ventricular tachycardia in two patients and two survivors of ventricular fibrillation.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of potentiality of traditional hygienic practices for the mitigation of the 2019-2020 Corona Pandemic.
The coronavirus outbreak threatens global well-being, costing thousands of lives and an unstable economy. Traditional hand-washing (HaWa) with clean water and hand-washing sanitizers (HaWaSas) are among WHO recommended practices against the outbreaks. HaWaSa and HaWa lacked proper quality monitoring evaluation that may pose health risks. The current study investigates the potential of traditional hygienic practices adopted to mitigate the 2019-2020 Pandemic. It involved assessing the effectiveness of 15 HaWaSa collected from the Dodoma market. Thirty volunteered participants were subjected to different hygienic practices involving HaWa and HaWaSa. The obtained hand-swabbed samples were subjected to the streak plate technique for inoculation, followed by counting colonies. The results revealed that, among the collected HaWaSa brands, only 33.3% were certified by the Tanzania Bureau of Standards (TBS). In the setting under no hand-wash, each client had a different number of colony counts. HaWa did not eliminate all colonies since water acted as a secondary source while only one sanitizer proved effective. The p-values obtained in either setting were greater than 0.05, indicating that each variable was independent. Despite global awareness and fighting against the Coronavirus Disease 2019 (Covid-19) outbreak, quality monitoring of all diagnostic tools, preventive HaWaSa, and related drugs should be prioritized for human safeguard.


Early failure of articular surface replacement XL total hip arthroplasty.
The ASR (articular surface replacement) XL (DePuy, Warsaw, Ind) metal-on-metal hip arthroplasty offers the advantage of stability and increased motion. However, an alarming number of early failures prompted the evaluation of patients treated with this system. A prospective study of patients who underwent arthroplasty with the ASR XL system was performed. Patients with 2-year follow-up or any revision were included. Failure rates, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores, and radiographs were evaluated. Ninety-five patients (105 hips) were included. There were 16 revisions. Thirteen (12%) were aseptic acetabular failures. Eight were revised for aseptic loosening; 4, for metallosis; 1, for malposition; 2, for infection; and 1, for periprosthetic fracture. Mean time to revision was 1.6 years (0.18-3.4 years). The ASR XL with a revision rate of 12% is the second reported 1 piece metal-on-metal system with a significant failure rate at early follow-up. This particular class of implants has inherent design flaws that lead to early failure.


Physiological effects of fenpropimorph on wild-type Saccharomyces cerevisiae and fenpropimorph-resistant mutants.
Fenpropimorph-resistant mutants of Saccharomyces cerevisiae were isolated by a gradient selection procedure. The mutants were cross-resistant to other morpholines (fenpropidin, dodemorph, tridemorph) and 15-azasterol, but were susceptible to azoles (miconazole, clotrimazole, ketoconazole) and nystatin. In the absence of fenpropimorph, the major sterol produced by the mutants and the parental strain was ergosterol. In the presence of fenpropimorph, ignosterol (ergosta-8,14-dien-3 beta-ol) was the major sterol produced by the mutants and the parental strain. The resistance to fenpropimorph involves two recessive genes, each of which allows a semiresistance, when they are isolated apart from one another. Strain JR4 (erg3 erg11), which produces 14-methylfecosterol [14 alpha-methyl-ergosta-8,24(28)-dien- 3-beta-ol) as the major sterol in the presence or absence of fenpropimorph, was also found to be resistant to the drug. The growth inhibitory effect of fenpropimorph on wild-type cells appears to be linked to the production of ignosterol. The uptake of exogenous sterol by wild-type cells was greatly enhanced in the presence of fenpropimorph. The growth inhibition caused by fenpropimorph could only be overcome with bulk levels of exogenous C-5,6-unsaturated sterols.


Mass spectrometric metabolomic imaging of biofilms on corroding steel surfaces using laser ablation and solvent capture by aspiration.
Ambient laser ablation and solvent capture by aspiration (LASCA) mass spectrometric imaging was combined with metabolomics high-performance liquid chromatography (HPLC) mass spectrometry analysis and light profilometry to investigate the correlation between chemical composition of marine bacterial biofilms on surfaces of 1018 carbon steel and corrosion damage of steel underneath the biofilms. Pure cultures of Marinobacter sp. or a wild population of bacteria present in coastal seawater served as sources of biofilms. Profilometry data of biofilm-free surfaces demonstrated heterogeneous distributions of corrosion damage. LASCA data were correlated with areas on the coupons varying in the level of corrosion attack, to reveal differences in chemical composition within biofilm regions associated with corroding and corrosion-free zones. Putative identification of selected compounds was carried out based on HPLC results and subsequent database searches. This is the first report of successful ambient chemical and metabolomic imaging of marine biofilms on corroding metallic materials. The metabolic analysis of such biofilms is challenging due to the presence in the biofilm of large amounts of corrosion products. However, by using the LASCA imaging interface, images of more than 1000 ions (potential metabolites) are generated, revealing striking heterogeneities within the biofilm. In the two model systems studied here, it is found that some of the patterns observed in selected ion images closely correlate with the occurrence and extent of corrosion in the carbon steel substrate as revealed by profilometry, while others do not. This approach toward the study of microbially influenced corrosion (MIC) holds great promise for approaching a fundamental understanding of the mechanisms involved in MIC.


[Prediction of progression in patients with mild cognitive impairment using IMP-SPECT].
To examine the difference in functional brain imaging between mild cognitive impairment (MCI) and normal aging, we measured rCBF on functional brain imaging using 123I-IMP single photon emission computed tomography (IMP-SPECT) in 19 MCI patients who progressed to develop AD on follow-up and 23 probable Alzheimer's disease (AD) patients as well as 15 age-matched normal subjects. Baseline MMSE score was 25.3 (SD 1.2) in the MCI group and 17.5 (SD 3.3) in the AD group. The regions of interest (ROI) in the posterior cingulate gyrus, frontal, temporal and parietal cortices were drawn on the image of IMP-SPECT with reference to an individual MRI image. The rCBF ratio was calculated using ROI value in the cerebellum as a reference. Voxel-based analysis was also preformed using statistical parametric mapping (SPM). The rCBF ratio in the posterior cingulate gyrus was significantly reduced in the MCI group (mean 0.956, SD 0.080) and the AD group (mean 0.833, SD 0.118) compared to that in the normal group (mean 1.083, SD 0.084). In the frontal, temporal and parietal cortices, the rCBF ratio was significantly reduced only in the AD group compared to the normal group. At a fixed specificity of 80%, the diagnostic sensitivity in the discrimination between MCI patients and normal subjects was 80.5% when using rCBF ratio in posterior cingulate gyrus. In the SPM analysis, significant reduction of the rCBF in MCI group was observed only in the posterior cingulate gyrus, compared with normal subject group. Our results suggest that MCI patients presenting with a posterior cingulate hypoperfusion are at higher risk for transition from MCI to clinically recognizable AD.


Gallium-67 lung scintigraphy in patients with acquired immune deficiency syndrome (AIDS).
Fifteen 67Ga lung scans were obtained from 11 men with AIDS to detect opportunistic lung infection. Results were compared with clinical findings, chest radiographs, CO-transfer and transbronchial biopsies or BAL. Clinical symptoms were least helpful in diagnosing pneumonia. Chest radiographs were normal in six of eight normal 67Ga studies and abnormal in four of seven abnormal scans. In four cases the X-ray showed equivocal abnormalities, twice corresponding to a normal and twice to an abnormal scan. Once the X-ray was normal, but the scan showed diffuse abnormalities. CO-transfer in patients with a normal scan was higher (55-68%) than in patients with an abnormal scan (22-44%). Biopsy or BAL revealed P. carinii in five of six patients. Cytomegalo-virus was isolated once. All six patients had abnormal 67Ga scans. 67Ga lung scintigraphy is a sensitive indicator of active lung disease in AIDS, especially when chest X-rays are normal or equivocal.


Sepsis in Older Adults in Long-Term Care Facilities: Challenges in Diagnosis and Management.
Despite the current understanding of the pathophysiology of sepsis and advances in its treatment, the rate of sepsis is increasing globally. Sepsis is a common cause of hospitalization in older adults, and infections are among the most common diagnoses among residents transferred to the hospital from long-term care facilities (LTCFs). LTCFs and hospitals are facing financial and regulatory requirements to reduce potentially preventable emergency department visits, hospitalizations, and hospital readmissions due to infections and other causes. In addition, the human and financial costs of these events are substantial. Current criteria for early identification of sepsis have low sensitivity and specificity among LTCF residents. Early diagnosis must focus on changes in clinical, mental, and functional status, and vital signs including pulse oximetry. Laboratory data can increase the suspicion of sepsis, but the availability of testing and timing of results limits its usefulness in most LTCFs.While new diagnostic criteria for sepsis are being developed and validated in the LTCF setting, clinical practice and decision support tools are available to guide management. Most LTFCs do not have the capabilities to manage sepsis based on current guidelines despite availability of qualified nursing staff and clinicians. Thus excluding circumstances in which a resident's desire is palliative or hospice care without transfer to a hospital, most LTCFs will continue to transfer residents with severe infections at risk for evolving into sepsis to an acute hospital setting. J Am Geriatr Soc 67:2234-2239, 2019.


Inflammation-associated insulin resistance: differential effects in rheumatoid arthritis and systemic lupus erythematosus define potential mechanisms.
Insulin resistance is increased by inflammation, but the mechanisms are unclear. The present study was undertaken to test the hypothesis that decreased insulin sensitivity is differentially associated with mediators of inflammation by studying 2 chronic inflammatory diseases of different pathogenesis, systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). We measured fasting insulin, glucose, and lipid levels, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), and coronary artery calcification in 103 patients with SLE and in 124 patients with RA. Insulin sensitivity was measured using the homeostasis model assessment (HOMA) index. The HOMA value was higher in RA patients (median 2.05 [interquartile range (IQR) 1.05-3.54]) than in SLE patients (1.40 [0.78-2.59]) (P = 0.007). CRP and ESR did not differ significantly in RA and SLE patients. Body mass index (BMI) was significantly correlated with the HOMA index in both RA (rho = 0.20) and SLE (rho = 0.54), independently of age, sex, race, and current use of corticosteroids. In RA patients, the HOMA index was also significantly positively correlated with IL-6 (rho = 0.63), TNFalpha (rho = 0.50), CRP (rho = 0.29), ESR (rho = 0.26), coronary calcification (rho = 0.26), and Disease Activity Score in 28 joints (rho = 0.21); associations adjusted for age, sex, race, BMI, and current use of corticosteroids remained significant (P < 0.05). In SLE patients, the HOMA index was also significantly correlated with ESR (rho = 0.35) and CRP (rho = 0.25), but not with other variables. The association between the ESR and the HOMA value in patients with SLE remained significant after adjustment for confounding covariates (P = 0.008). In multivariable models, the major contributing factors to the HOMA index were the BMI in SLE patients, and IL-6 and TNFalpha levels in RA patients. The pathogenesis of insulin resistance and its contribution to atherogenesis varies in different inflammatory settings.


A Follow-Up Study from a Multisite, Randomized Controlled Trial for Traumatized Children Receiving TF-CBT.
Trauma-focused cognitive behavioral therapy (TF-CBT) is the treatment of choice for traumatized youth, however, follow-up studies are scarce, and treatment effects for co-occurring depression show mixed findings. The aims of this study were to examine whether treatment effects of TF-CBT are maintained at 18 month follow-up and whether degree of co-occurring depression influences treatment effects. As rapid improvement in psychological functioning is warranted for youth, we also investigated whether the symptom trajectory was different for TF-CBT compared to therapy as usual (TAU). The sample consisted of 156 youth (M age = 15.05, 79.50% girls) randomly assigned to TF-CBT or TAU. The youth were assessed for posttraumatic stress symptoms (PTSS), depression, anxiety and general mental health symptoms. Mixed effects analyses followed the symptom courses over 5 time points. Youth receiving TF-CBT maintained their symptom improvement at 18 months follow-up with scores below clinical cut-of on all symptom measures. The most depressed youth had also a significant decline in symptoms that were maintained at follow-up. Symptom trajectories differed as the TF-CBT group reported a more rapid symptom reduction compared to the TAU condition. In the TAU condition, participants received 1.5 times the number of treatment sessions compared to the TF-CBT participants. After 18 months the groups were significantly different on general mental health symptoms only. In conclusion, youth receiving TF-CBT experienced more efficient improvement in trauma related symptoms than youth receiving TAU and these improvements were maintained after 18 months. Also youth experiencing serious co-occurring depression benefitted from TF-CBT.


Facial Swelling and Shortness of Breath Are Not Always an Allergic Reaction: It Could Be Acute Superior Vena Cava Syndrome.
Historically, it has been found that malignancy is associated with superior vena cava (SVC) syndrome. The past decade has seen more cases of thrombogenic and stenotic SVC syndrome due to increased use of pacemakers and indwelling central lines. As compared to the slowly progressing obstruction in malignancy, rapid thrombogenesis rate and a lack of venous collateral sequelae lead to more acute sequelae in these patients. It is important to timely assess patients presented with an acute process of SVC syndrome in the emergency room. Diagnosis can quickly be made by using computed tomography angiography (CTA) or magnetic resonance angiography (MRA) modalities. The underlying cause of the syndrome is the focus of the treatment. Anticoagulation is the basis of the treatment in the case of thrombogenic catheter-associated SVC syndrome. In order to promptly manage symptoms, it was observed that balloon angioplasty with stenting and thrombolytics proved to be beneficial. Herein we are describing a 68-year-old female with past medical history of colon cancer with liver metastasis on chemotherapy via port, presented to the emergency room with acute shortness of breath and facial and neck swelling, and was found to have acute superior vena cava syndrome.


Learning Temporal Quantum Tomography.
Quantifying and verifying the control level in preparing a quantum state are central challenges in building quantum devices. The quantum state is characterized from experimental measurements, using a procedure known as tomography, which requires a vast number of resources. However, tomography for a quantum device with temporal processing, which is fundamentally different from standard tomography, has not been formulated. We develop a practical and approximate tomography method using a recurrent machine learning framework for this intriguing situation. The method is based on repeated quantum interactions between a system called quantum reservoir with a stream of quantum states. Measurement data from the reservoir are connected to a linear readout to train a recurrent relation between quantum channels applied to the input stream. We demonstrate our algorithms for representative quantum learning tasks, followed by the proposal of a quantum memory capacity to evaluate the temporal processing ability of near-term quantum devices.


An index for evaluating difficulty of Chewing Index for chewable tablets.
Chewing difficulty index, a potential measure of difficulty in chewing the chewable tablets, has been described herein as the product of tablet thickness and tablet hardness measured under the diametral loading. The proposed index was evaluated by measuring the dimensions and mechanical strength of commercial and in-house prepared chewable tablets. Data collected on tablets with different thickness but same hardness or tensile strength suggests that the proposed index provides a good assessment of the force needed to chew the chewable tablets. Influence of brief exposure to salivary fluid during chewing on the mechanical strength of the chewable tablets was also evaluated. Thirty seconds exposure to the simulated salivary fluid was also found to significantly reduce (p < 0.05) the hardness and the chewing difficulty index of a number of evaluated chewable tablet drug products.


Multimodal Magnetic Resonance Imaging Data Fusion Reveals Distinct Patterns of Abnormal Brain Structure and Function in Catatonia.
Catatonia is a nosologically unspecific syndrome, which subsumes a plethora of mostly complex affective, motor, and behavioral phenomena. Although catatonia frequently occurs in schizophrenia spectrum disorders (SSD), specific patterns of abnormal brain structure and function underlying catatonia are unclear at present. Here, we used a multivariate data fusion technique for multimodal magnetic resonance imaging (MRI) data to investigate patterns of aberrant intrinsic neural activity (INA) and gray matter volume (GMV) in SSD patients with and without catatonia. Resting-state functional MRI and structural MRI data were collected from 87 right-handed SSD patients. Catatonic symptoms were examined on the Northoff Catatonia Rating Scale (NCRS). A multivariate analysis approach was used to examine co-altered patterns of INA and GMV. Following a categorical approach, we found predominantly frontothalamic and corticostriatal abnormalities in SSD patients with catatonia (NCRS total score ≥ 3; n = 24) when compared to SSD patients without catatonia (NCRS total score = 0; n = 22) matched for age, gender, education, and medication. Corticostriatal network was associated with NCRS affective scores. Following a dimensional approach, 33 SSD patients with catatonia according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision were identified. NCRS behavioral scores were associated with a joint structural and functional system that predominantly included cerebellar and prefrontal/cortical motor regions. NCRS affective scores were associated with frontoparietal INA. This study provides novel neuromechanistic insights into catatonia in SSD suggesting co-altered structure/function-interactions in neural systems subserving coordinated visuospatial functions and motor behavior.


Sleep problems and depression among 237 023 community-dwelling adults in 46 low- and middle-income countries.
Sleep problems are considered a core symptom of depression. However, there is little information about the comorbidity of sleep problems and depression in low- and middle-income countries (LMICs), and whether sleep problems with depression confer additional risk for decrements in health compared to sleep problems alone. This study thus examined the association between sleep problems and depression and whether sleep problems with depression are associated with an increased risk for poorer health in 46 LMICs. Cross-sectional, community-based data from 237 023 adults aged ≥18 years from the World Health Survey (WHS) 2002-2004 were analyzed. Information on sleep problems (severe/extreme) and International Classification of Diseases 10th Revision depression/depression subtypes was collected. Multivariable logistic (binary and multinomial) and linear regression analyses were performed. Sleep problems were associated with subsyndromal depression (odds ratio [OR]: 2.23, 95% confidence interval [CI]: 1.84-2.70), brief depressive episode (OR = 2.48, 95% CI = 2.09-2.95) and depressive episode (OR = 3.61, 95% CI = 3.24-4.03). Sleep problems with depression (vs. sleep problems alone) conferred additional risk for anxiety, perceived stress and decrements in health in the domains of mobility, self-care, pain, cognition, and interpersonal activities. Clinicians should be aware that the co-occurrence of sleep problems and depression is associated with a variety of adverse health outcomes in LMICs. Detecting this co-occurrence may be important for treatment planning.


The relative efficiency of modular and non-modular networks of different size.
Most biological networks are modular but previous work with small model networks has indicated that modularity does not necessarily lead to increased functional efficiency. Most biological networks are large, however, and here we examine the relative functional efficiency of modular and non-modular neural networks at a range of sizes. We conduct a detailed analysis of efficiency in networks of two size classes: 'small' and 'large', and a less detailed analysis across a range of network sizes. The former analysis reveals that while the modular network is less efficient than one of the two non-modular networks considered when networks are small, it is usually equally or more efficient than both non-modular networks when networks are large. The latter analysis shows that in networks of small to intermediate size, modular networks are much more efficient that non-modular networks of the same (low) connective density. If connective density must be kept low to reduce energy needs for example, this could promote modularity. We have shown how relative functionality/performance scales with network size, but the precise nature of evolutionary relationship between network size and prevalence of modularity will depend on the costs of connectivity.


Graduate Midwifery Education in Uganda Aiming to Improve Maternal and Newborn Health Outcomes.
Maternal and newborn health outcomes in Uganda have remained poor. The major challenge affecting the implementation of maternal and newborn interventions includes a shortage of skilled midwives. In 2013, Lira University, a Ugandan Public University, in partnership with Seed Global Health, started the first Bachelor of Science in Midwifery (BScM) in Uganda with a vision to develop a Master of Science in Midwifery (MScM) in the future. Evaluate results of Lira University's Bachelors in Midwifery program to help inform the development of a Masters in Midwifery program, which would expand midwifery competencies in surgical obstetric and newborn care. Lira University and Ministry of Health records provided data on curriculum content, student enrollment and internships. The internship reports of the graduate midwives were reviewed to collect data on their employment and scope of practice. Interviews were also conducted with the graduates to confirm the added skills they were able to apply and their outcomes. The critical competences incorporated into the Bachelor in Midwifery curriculum included competences to care for pre- and post-operative caesarian section patients or assist in a caesarean section, newborn care (e.g. resuscitation from birth asphyxia), anesthesia, and theatre techniques, among others. Overall, 356 students (40.2% male, 59.8% female) enrolled in the BScM program over the period 2013-2018. Annual data shows an increasing trend in enrollment. Of the 32 graduates in January 2019, 87.6% were employed in maternal and newborn healthcare facilities, and 12.4% were employed in midwifery private practice. Follow-up interviews revealed that the graduate midwives reported positive maternal and newborn outcomes and the ability to practice advanced obstetrics and newborn care skills they acquired from the training. There is growing interest in a graduate midwifery education program in Uganda for both male and female students. The retention of the graduate midwives in healthcare facilities gives a renewed hope for mothers and newborns, who benefit from their extra obstetrics and newborn care competences in settings where there are neither medical doctors nor obstetricians and gynecologists. Further, larger tracer studies of the graduate midwives to identify the kinds of obstetric surgeries and newborn care services they ably performed and their corresponding maternal and newborn health outcomes is recommended. Also recommended is advocacy for recognition of extra skills of graduate midwives by health authorities in Uganda and the region.


Global importance analysis: An interpretability method to quantify importance of genomic features in deep neural networks.
Deep neural networks have demonstrated improved performance at predicting the sequence specificities of DNA- and RNA-binding proteins compared to previous methods that rely on k-mers and position weight matrices. To gain insights into why a DNN makes a given prediction, model interpretability methods, such as attribution methods, can be employed to identify motif-like representations along a given sequence. Because explanations are given on an individual sequence basis and can vary substantially across sequences, deducing generalizable trends across the dataset and quantifying their effect size remains a challenge. Here we introduce global importance analysis (GIA), a model interpretability method that quantifies the population-level effect size that putative patterns have on model predictions. GIA provides an avenue to quantitatively test hypotheses of putative patterns and their interactions with other patterns, as well as map out specific functions the network has learned. As a case study, we demonstrate the utility of GIA on the computational task of predicting RNA-protein interactions from sequence. We first introduce a convolutional network, we call ResidualBind, and benchmark its performance against previous methods on RNAcompete data. Using GIA, we then demonstrate that in addition to sequence motifs, ResidualBind learns a model that considers the number of motifs, their spacing, and sequence context, such as RNA secondary structure and GC-bias.


Intentional signal in prefrontal cortex generalizes across different sensory modalities.
Biofeedback-EEG training to learn the mental control of an external device (e.g., a cursor on the screen) has been an important paradigm to attempt to understand the involvements of various areas of the brain in the volitional control and the modulation of intentional thought processes. Often the areas to adapt and to monitor progress are selected a priori. Less explored, however, has been the notion of automatically emerging activation in a particular area or subregions within that area recruited above and beyond the rest of the brain. Likewise, the notion of evoking such a signal as an amodal, abstract one remaining robust across different sensory modalities could afford some exploration. Here we develop a simple binary control task in the context of brain-computer interface (BCI) and use a Bayesian sparse probit classification algorithm to automatically uncover brain regional activity that maximizes task performance. We trained and tested 19 participants using the visual modality for instructions and feedback. Across training blocks we quantified coupling of the frontoparietal nodes and selective involvement of visual and auditory regions as a function of the real-time sensory feedback. The testing phase under both forms of sensory feedback revealed automatic recruitment of the prefrontal cortex with a parcellation of higher strength levels in Brodmann's areas 9, 10, and 11 significantly above those in other brain areas. We propose that the prefrontal signal may be a neural correlate of externally driven intended direction and discuss our results in the context of various aspects involved in the cognitive control of our thoughts.


Accurate Physics-Based Registration for the Outcome Validation of Minimal Invasive Interventions and Open Liver Surgeries.
The purpose of this paper is to present an outcome validation tool for tumor radiofrequency (RF) ablation and resection. Intervention assessment tools require an accurate registration of both pre- and postoperative computed tomographies able to handle big deformations. Therefore, a physics-based method is proposed with that purpose. To increase the accuracy both automatically detected internal and surface physical landmarks are incorporated in the registration process. The algorithm has been evaluated in 25 clinical datasets containing RF ablations, resections, and patients with recurrent tumors. The achieved accuracy is 1.2 mm measured as mean internal distance between vessel landmarks and a positive predictive value of 0.95. The quantitative and qualitative results of the outcome validation tool show that in 50% of the cases tumors were only partially covered by the treatment. The use of internal and surface landmarks combined with a physics-based registration method increases the accuracy of the results compared to the accuracy of state of the art methods. An accurate outcome validation tool is important in order to certify that the tumor and its safety margin were fully covered by the treatment. An accurate outcome validation tool can result in a decrease of the tumor recurrence rate.


Analysis of Gut Characteristics and Microbiota Changes with Maternal Supplementation in a Neural Tube Defect Mouse Model.
Adequate nutrient supply is crucial for the proper development of the embryo. Although nutrient supply is determined by maternal diet, the gut microbiota also influences nutrient availability. While currently there is no cure for neural tube defects (NTDs), their prevention is largely amenable to maternal folic acid and inositol supplementation. The gut microbiota also contributes to the production of these nutrients, which are absorbed by the host, but its role in this context remains largely unexplored. In this study, we performed a functional and morphological analysis of the intestinal tract of loop-tail mice (Vangl2 mutants), a mouse model of folate/inositol-resistant NTDs. In addition, we investigated the changes in gut microbiota using 16S rRNA gene sequencing regarding (1) the host genotype; (2) the sample source for metagenomics analysis; (3) the pregnancy status in the gestational window of neural tube closure; (4) folic acid and (5) D-chiro-inositol supplementation. We observed that Vangl2+/Lp mice showed no apparent changes in gastrointestinal transit time or fecal output, yet exhibited increased intestinal length and cecal weight and gut dysbiosis. Moreover, our results showed that the mice supplemented with folic acid and D-chiro-inositol had significant changes in their microbiota composition, which are changes that could have implications for nutrient absorption.


Reoviruslike agent in stools: association with infantile diarrhea and development of serologic tests.
Reoviruslike particles were visualized by electron microscopy in stool filtrates prepared from stools of infants and young children with severe acute gastroenteritis. Patients who had such particles in their stools and whose paired acute and convalescent serums were tested developed an antibody response to the reoviruslike agent, which was measured by immune electron microscopy and by complement fixation. The reoviruslike agent was antigenically related to the epizootic diarrhea of infant mice virus and the Nebraska calf diarrhea virus.


Study on Awareness of Preconception Care and Reproductive Health Behaviors in Pre-honeymooners.
The purpose of this study was 1) to examine levels of awareness on preconception care and pregnancy, and reproductive health promoting behavior among pre-honeymooners; and 2) to explore the relationship between awareness levels of preconception care and pregnancy, and reproductive health promoting behavior. This study was a correlation design with a total of 134 participants (67 couples of pre-honeymooners). Data were collected with questionnaire. Mean score of self-perception of awareness of preconception care and pregnancy was 2.82 out of 4. Content awareness level was 6.83 out of 13, and reproductive health promoting behavior score was 3.02 out of 4, indicating low to mid-level. Women reported higher scores in all variables than those in men. Self-perception awareness and content awareness of care and pregnancy had a weak positive correlation (r=.18, p=.038). Reproductive health promoting behavior was positively related to self-perception awareness of care and pregnancy (r=.33, p<.001) and content awareness of care and pregnancy (r=.23, p=.008). Based on this results, a variety of nursing intervention strategies may need to promote reproductive health behavior such as raising awareness of the preconception care and pregnancy.


Prediction of dropout from respiratory symptoms and airflow limitation in a longitudinal respiratory study.
This study investigated the possibility that employees reporting respiratory symptoms were more likely than asymptomatic workers to dropout of a respiratory study carried out in Norwegian smelters. The study included 3924 employees in 24 Norwegian smelters. They were examined annually using a respiratory questionnaire and spirometry. The employees who did not meet for the follow-up within 18 months prior to the end of the study were considered dropouts. The data were analyzed using Cox regression for time-dependent covariates. The total and the median follow-up times were 16 997 and 4.9 years, respectively. The overall dropout rate was 44.5 [95% confidence interval (95% CI) 41.5-47.8 per 1000 person-years]. The hazard ratio (HR) for dropout was 1.38 (95% CI 1.15-1.66) for the workers reporting any respiratory symptom compared with the asymptomatic workers. The effect was the strongest among the employees who reported dyspnea, and it was stronger regarding symptoms at the last visit than for the baseline symptoms. Similarly, the hazard ratio for dropout for those with an airflow limitation [forced expiratory volume/forced vital capacity below the 5th percentile of the predicted value] was 1.31 (95% CI 1.01-1.69) when they were compared with employees without any airflow limitation. Respiratory symptoms and airflow limitation are important predictors of dropout from a longitudinal respiratory study.


An empowerment-based approach to developing innovative e-health tools for self-management.
E-health is seen as an important technological tool in achieving self-management; however, there is little evidence of how effective e-health is for self-management. Example tools remain experimental and there is limited knowledge yet about the design, use, and effects of this class of tools. By way of introducing a new view on the development of e-health tools dedicated to self-management we aim to contribute to the discussion for further research in this area. Our assumption is that patient empowerment is an important mechanism of e-health self-management and we suggest incorporating it within the development of self-management tools. Important components of empowerment selected from literature are: communication, education and health literacy, information, self-care, decision aids and contact with fellow patients. All components require skills of both patients and the physicians. In this discussion paper we propose how the required skills can be used to specify effective self-management tools.


PPGR: a comprehensive perennial plant genomes and regulation database.
Perennial woody plants hold vital ecological significance, distinguished by their unique traits. While significant progress has been made in their genomic and functional studies, a major challenge persists: the absence of a comprehensive reference platform for collection, integration and in-depth analysis of the vast amount of data. Here, we present PPGR (Resource for Perennial Plant Genomes and Regulation; https://ngdc.cncb.ac.cn/ppgr/) to address this critical gap, by collecting, integrating, analyzing and visualizing genomic, gene regulation and functional data of perennial plants. PPGR currently includes 60 species, 847 million protein-protein/TF (transcription factor)-target interactions, 9016 transcriptome samples under various environmental conditions and genetic backgrounds. Noteworthy is the focus on genes that regulate wood production, seasonal dormancy, terpene biosynthesis and leaf senescence representing a wealth of information derived from experimental data, literature mining, public databases and genomic predictions. Furthermore, PPGR incorporates a range of multi-omics search and analysis tools to facilitate browsing and application of these extensive datasets. PPGR represents a comprehensive and high-quality resource for perennial plants, substantiated by an illustrative case study that demonstrates its capacity in unraveling gene functions and shedding light on potential regulatory processes.


Treatment of Neurogenic Cough with Tramadol: A Pilot Study.
This study employs validated cough assessment tools to prospectively determine the impact of tramadol on cough severity and quality of life in subjects with neurogenic cough. The study was a prospective case series with planned data collection at a tertiary care academic medical center laryngology practice. Sixteen consecutive collected subjects with neurogenic cough prospectively completed pre- and posttreatment validated cough assessment tools, the cough severity index (CSI) and Leicester Cough Questionnaire (LCQ). All subjects in the study reported at least some improvement in their cough symptoms. In a Wilcoxon signed rank test that compared paired results, CSI scores improved from 23 to 14 and LCQ scores improved from 74 to 103 ( P = .003 and P = .005, respectively). This small preliminary assessment suggests that tramadol warrants additional evaluation as a treatment for neurogenic cough.


'One-visit' therapy for pelvic inflammatory disease.
Acute pelvic inflammatory disease is a very common gynaecological disorder, and more often than not it is caused by a mixed infection of aerobic and anaerobic organisms. Routine bacteriological investigation is consequently of little practical value in determining the therapeutic approach. In an open comparative trial, 60 patients were divided into two treatment groups of 30 patients each. Both groups received a combination of penicillins, probenecid, a sulphonamide and an imidazole derivative active against protozoa and anaerobes. Both regimens resulted in virtually the same therapeutic response in patients hospitalized for 6 days. The regimen consisting of ornidazole (as opposed to metronidazole) and the ultralong-acting sulphonamide, sulphadoxine (as opposed to sulphadimidine), can be given in a once-only administration, thereby greatly reducing the possibility of non-compliance in patients treated, as is customary, on an outpatient basis.


Effects of delta-9-tetrahydrocannabinol and (R)-methanandamide on open-field behavior in rats.
This study compared the effects of (R)-methanandamide, an analog of the mammalian brain constituent anandamide, and delta-9-tetrahydrocannabinol on the open-field behavior of male Sprague-Dawley rats. Rats were individually housed with free access to food and water. Animals were treated with 0, 1, 3, and 5.6 mg/kg delta-9-tetrahydrocannabinol given i.p. 30 min pre-session; and 0, 3, 10, and 18 mg/kg (R)-methanandamide, 15 min pre-session. The behavioral categories recorded were ambulation (the number of squares crossed), rearing (the number of times the rat stood erect on its hind-legs), latency (the time in seconds to leave the starting area, the circle in the center of the field), circling (the number of times the animal turned around its vertical axis, 0.5 point given for each 180 degrees turn), grooming (the number of cleaning bouts), urination and defecation (the number of urination spots and fecal boli deposited during the 5 min observation period). Delta-9-tetrahydrocannabinol was more potent than (R)-methanandamide, but otherwise the effects of delta-9-tetrahydrocannabinol and (R)-methanandamide were similar, with one exception; whereas delta-9-tetrahydrocannabinol produced dose-related increases in circling, (R)-methanandamide did not increase circling over the doses examined. The delta-9-tetrahydrocannabinol-induced increase in circling was blocked by the central cannabinoid receptor CB1 antagonist SR 141716. The differential effects with regard to circling may indicate that there are qualitative behavioral differences in the effects of delta-9-tetrahydrocannabinol and (R)-methanandamide.


Observer-based adaptive fuzzy-neural control for unknown nonlinear dynamical systems.
In this paper, an observer-based adaptive fuzzy-neural controller for a class of unknown nonlinear dynamical systems is developed. The observer-based output feedback control law and update law to tune on-line the weighting factors of the adaptive fuzzy-neural controller are derived. The total states of the nonlinear system are not assumed to be available for measurement. Also, the unknown nonlinearities of the nonlinear dynamical systems are not restricted to the system output only. The overall adaptive scheme guarantees that all signals involved are bounded. Simulation results demonstrate the applicability of the proposed method in order to achieve desired performance.


Resting energy expenditure in Parkinson's disease patients under dopaminergic treatment.
Background: Weight homeostasis is complex in Parkinson's disease (PD) and body weight changes substantially throughout the course of the disease. We designed a case-control study to (i) investigate whether PD is associated with changes in resting energy expenditure (REE), (ii) to assess how accurately REE could be predicted for individuals with PD utilizing the equations constructed for healthy individuals, and (iii) to eventually construct a new equation.Materials & Methods: Measured REE (mREE) was compared between 122 PD patients and 122 gender and body mass index (BMI)-matched controls. The accuracy of estimated REE by 5 common equations (Harris/Benedict-1919, Roza/Shizgal-1984, Mifflin St. Jeor, WHO/FAO and aggregate formula) was investigated in PD using Bland-Altman analysis and reported as the frequency of accurate predictions (±10%). Concordance correlation coefficients (CCC) were also calculated. Then, we regressed a new REE equation - using gender, age, weight, height and Hoehn-Yahr stage - and validated it in an independent sample (N = 100).Results: No significant difference in mREE was recorded between the whole PD sample and healthy controls. However, mREE was increased in patients with BMI ≥ 30 kg/m2 and Hoehn-Yahr stage ≥ 3. Limited accuracy was present in the available REE equations (accurate prediction [±10%] frequency, <60% for all). For the new equation, the proportion of accurate prediction was 67.0% (overestimation, 24.0%) and CCC was 0.77.Conclusion: PD patients are not commonly characterized by an increase in REE. This is limited to patients suffering from obesity and more severe disease. Common REE equations appear to be inaccurate. The new predictive equation proposed in this study provided better REE estimates.


Beneficial effects of softened fabrics on atopic skin.
There is general concern about the possible cutaneous adverse effects of wearing garments treated with household laundry products, particularly on atopic skin. Our objective was to compare softened and non- softened fabrics in a forearm wet and dry test, under conditions simulating real-life conditions. Twenty atopic volunteers entered a single-blind 12-day (3 sessions per day) forearm wetting and drying test. Cotton fabrics were machine washed and liquid fabric conditioner was added or not to the final rinse. To simulate conditions of skin damage, a dilute solution of sodium lauryl sulphate was applied under occlusion to the forearm of each volunteer before the start of the study. Skin effects were evaluated by visual grading (redness, dryness and smoothness), squamometry and in vivo instrumental measurements (capacitance, transepidermal water loss and colorimetry). Rubbing of atopic skin with fabrics generally resulted in discrete to moderate alterations of the structure of the stratum corneum. Both for control and pre-irritated skin, all measured parameters indicated that softened fabric was less aggressive to the skin than unsoftened fabric. In the case of pre-irritated skin, the recovery of the skin was significantly faster when rubbed with softened than with unsoftened fabrics. In conclusion, softened fabrics help mitigate the skin condition in atopic patients.


Molecular interplay between Δ5/Δ6 desaturases and long-chain fatty acids in the pathogenesis of non-alcoholic steatohepatitis.
The mechanisms underlying non-alcoholic steatohepatitis (NASH) are not completely elucidated. In the current study we integrated gene expression profiling of liver biopsies from NASH patients with translational studies in mouse models of steatohepatitis and pharmacological interventions in isolated hepatocytes to identify new molecular targets in NASH. Using oligonucleotide microarray analysis we identified a significant enrichment of genes involved in the multi-step catalysis of long-chain polyunsaturated fatty acids, namely, Δ-5 desaturase (Δ5D) and Δ6D in NASH. Increased expression of Δ5D and Δ6D at both mRNA and protein level were confirmed in livers from mice with high-fat diet-induced obesity and NASH. Gas chromatography analysis revealed impaired desaturation fluxes toward the ω-6 and ω-3 pathways resulting in increased ω-6 to ω-3 ratio and reduced ω-3 index in human and mouse fatty livers. Restoration of hepatic ω-3 content in transgenic fat-1 mice expressing an ω-3 desaturase, which allows the endogenous conversion of ω-6 into ω-3 fatty acids, produced a significant reduction in hepatic insulin resistance, steatosis, macrophage infiltration, necroinflammation and lipid peroxidation, accompanied by attenuated expression of genes involved in inflammation, fatty acid uptake and lipogenesis. These results were mostly reproduced by feeding obese mice with an exogenous ω-3-enriched diet. A combined Δ5D/Δ6D inhibitor, CP-24879, significantly reduced intracellular lipid accumulation and inflammatory injury in hepatocytes. Interestingly, CP-24879 exhibited superior antisteatotic and anti-inflammatory actions in fat-1 and ω-3-treated hepatocytes. These findings indicate that impaired hepatic fatty acid desaturation and unbalanced ω-6 to ω-3 ratio play a role in the pathogenesis of NASH.


Expression profile analysis of mycotoxin-related genes in cartilage with endemic osteochondropathy Kashin-Beck Disease.
Kashin-Beck Disease (KBD) is an endemic osteochondropathy. Mycotoxins are believed to play an important role in the pathogenesis of KBD. Because the molecular mechanism of mycotoxin-induced cartilage lesions remains unclear, there is not effective treatment for KBD now. To identify key genes involved in the mycotoxin-induced cartilage lesions, we compared the expression profiles of mycotoxin-related genes (MRG) between KBD cartilage and healthy cartilage. Total RNA was isolated from cartilage samples, following by being amplified, labeled and hybridized to Agilent human whole genome microarray chip. qRT-PCR was conducted to validate the microarray data. 1,167 MRG were derived from the environmentally related genomic database Toxicogenomics. The microarray data of MRG was subjected to single gene and gene ontology (GO) expression analysis for identifying differently expressed genes and GO. We identified 7 up-regulated MRG and 2 down-regulated MRG in KBD cartilage, involved in collagen, apoptosis, metabolism and growth & development. GO expression analysis found that 4 apoptosis-related GO and 5 growth & development-related GO were significantly up-regulated in KBD cartilage. Based on the results of previous and our studies, we suggest that mycotoxins might contribute to the development of KBD through dysfunction of MRG involved in collagen, apoptosis and growth & development in cartilage.


[A follow up study of total IgM, IgG, nucleoprotein and spike protein antibodies against severe acute respiratory syndrome (SARS) coronavirus in patients with SARS].
To survey the dynamic changing and persistence of the special antibodies, including total IgM, IgG, nucleocapsid protein and spike protein antibodies, against severe acute respiratory syndrome coronavirus (SARS-CoV) in patients with SARS. 146 cases, all clinically diagnosed as SARS with positive SARS-CoV IgG, were followed up. 362 serum samples were collected from the onset of the disease to 660 days afterward. Total IgM and IgG against SARS-CoV were tested with commercial ELISA kits. For recombinant nucleoprotein and spike protein, we developed an ELISA to test these two antibodies. Within 20 days of the onset, the positive rate of anti-SARS-CoV IgM was 46.5% (20/43); it reached a peak after 21 - 40 days (80.6%, 25/31). Then, the positive rate of IgM went down gradually to 8.2% (6/73) until 550 days after the onset. The patient's IgG positive rate was lower (34.9%, 15/43) than that of IgM within 20 days of the onset. Then it went up rapidly to 100%. It remained positive (98.6%, 70/71) until 600 - 660 days after the onset. When N-IgG and S-IgG were tested 40 days after the onset of the disease at three different times, the positive rate of N-IgG (92.5%, 37/40) was higher than that of S-IgG (67.5%, 27/40), but the two structure protein antibodies were always lower than the total IgG. In SARS patients with definite clinical and etiological diagnosis, the highest positive rate of the antibodies against SARS-CoV was found at 21 - 40 days after the onset. IgM disappeared almost 500 days (91.8%) after the onset. Total IgG positive rate could reach 100% and 98.6% and the positivity might persist nearly two years. So it is speculated that the total IgG antibody may be positive 3 to 5 years after infection, but it seems that N-IgG and S-IgG keep positive shorter in time than total IgG antibody.


Continuous warfarin administration versus heparin bridging therapy in post colorectal polypectomy haemorrhage: a study protocol for a multicentre randomised controlled trial (WHICH study).
Endoscopic removal of colorectal adenoma is considered an effective treatment for reducing the mortality rates associated with colorectal cancer. Warfarin, a type of anticoagulant, is widely used for the treatment and prevention of thromboembolism; however, bleeding may increase with its administration after polypectomy. In recent times, a high incidence of bleeding after endoscopic polypectomy has been reported in patients receiving heparin bridge therapy. However, previous studies have not compared the bleeding rate after endoscopic colorectal polypectomy between patients who continued with anticoagulant therapy and those who received heparin bridge therapy. We hypothesised that endoscopic colorectal polypectomy under the novel treatment with continuous warfarin is not inferior to endoscopic colorectal polypectomy under standard treatment with heparin bridge therapy with respect to the rate of postoperative bleeding. This study aims to compare the efficacy of endoscopic colorectal polypectomy with continuous warfarin administration and endoscopic colorectal polypectomy with heparin bridge therapy with respect to the rate of postoperative bleeding. We will conduct a prospective multicentre randomised controlled non-inferiority trial of two parallel groups. We will compare patients scheduled to undergo colorectal polypectomy under anticoagulant therapy with warfarin. There will be 2 groups, namely, a standard treatment group (heparin bridge therapy) and the experimental treatment group (continued anticoagulant therapy). The primary outcome measure is the rate of postoperative bleeding. On the contrary, the secondary outcomes include the rate of cumulative bleeding, rate of overt haemorrhage (that does not qualify for the definition of haemorrhage after endoscopic polypectomy), incidence of haemorrhage requiring haemostasis during endoscopic polypectomy, intraoperative bleeding during endoscopic colorectal polypectomy requiring angiography, abdominal surgery and/or blood transfusion, total rate of bleeding, risk factors for postoperative bleeding, length of hospital stay, incidence of thromboembolism, prothrombin time-international ratio (PT-INR) 28 days after the surgery, and incidence of serious adverse events. The results of this randomised controlled trial will provide valuable information for the standardisation of management of anticoagulants in patients scheduled to undergo colorectal polypectomy. UMIN-CTR UMIN000023720 . Registered on 22 August 2016.


The Ouchi illusion as an artifact of biased flow estimation.
A pattern by Ouchi has the surprising property that small motions can cause illusory relative motion between the inset and background regions. The effect can be attained with small retinal motions or a slight jiggling of the paper and is robust over large changes in the patterns, frequencies and boundary shapes. In this paper, we explain that the cause of the illusion lies in the statistical difficulty of integrating local one-dimensional motion signals into two-dimensional image velocity measurements. The estimation of image velocity generally is biased, and for the particular spatial gradient distributions of the Ouchi pattern the bias is highly pronounced, giving rise to a large difference in the velocity estimates in the two regions. The computational model introduced to describe the statistical estimation of image velocity also accounts for the findings of psychophysical studies with variations of the Ouchi pattern and for various findings on the perception of moving plaids. The insight gained from this computational study challenges the current models used to explain biological vision systems and to construct robotic vision systems. Considering the statistical difficulties in image velocity estimation in conjunction with the problem of discontinuity detection in motion fields suggests that theoretically the process of optical flow computations should not be carried out in isolation but in conjunction with the higher level processes of 3D motion estimation, segmentation and shape computation.


A high-definition view of functional genetic variation from natural yeast genomes.
The question of how genetic variation in a population influences phenotypic variation and evolution is of major importance in modern biology. Yet much is still unknown about the relative functional importance of different forms of genome variation and how they are shaped by evolutionary processes. Here we address these questions by population level sequencing of 42 strains from the budding yeast Saccharomyces cerevisiae and its closest relative S. paradoxus. We find that genome content variation, in the form of presence or absence as well as copy number of genetic material, is higher within S. cerevisiae than within S. paradoxus, despite genetic distances as measured in single-nucleotide polymorphisms being vastly smaller within the former species. This genome content variation, as well as loss-of-function variation in the form of premature stop codons and frameshifting indels, is heavily enriched in the subtelomeres, strongly reinforcing the relevance of these regions to functional evolution. Genes affected by these likely functional forms of variation are enriched for functions mediating interaction with the external environment (sugar transport and metabolism, flocculation, metal transport, and metabolism). Our results and analyses provide a comprehensive view of genomic diversity in budding yeast and expose surprising and pronounced differences between the variation within S. cerevisiae and that within S. paradoxus. We also believe that the sequence data and de novo assemblies will constitute a useful resource for further evolutionary and population genomics studies.


Effects of Carcass Weight and Back-fat Thickness on Carcass Properties of Korean Native Pigs.
Our study analyzed the carcass properties of 170 Korean native pigs in relation to carcass weight and back-fat thickness to provide general data for the production and distribution of high quality pig meat. The 70-74 kg group showed highest yield (73.41%). The ≥80 kg group showed the highest thickest back-fat (24.13 mm) (p<0.05). The ≥80 kg group showed the best quality grade (1.00). Back-fat thickness showed significant differences in the weight among groups (p<0.05). The ≥25 mm group showed the highest carcass weight (75.93 kg). The thickest back-fat group (≥25 mm) showed the highest yield (73.03%). There were significant differences in back-fat thickness among groups (p<0.05), and the ≥25 mm group showed the highest thickness back-fat (27.60 mm). We found a strong positive correlation between carcass weight and back-fat thickness (r=0.346) as well as meat quality grade (r=0.739). Back-fat thickness had a relatively strong positive correlation with meat quality grade (r=0.444). Therefore, there are required to manage the breeding through selection of excellent native species for increasing their carcass weight and enhance meat quality.


Prognostic Value of Change in Cardiac Index After Prostacyclin Initiation in Pediatric Pulmonary Hypertension.
Invasive hemodynamic assessment remains the gold standard for the diagnosis of pediatric pulmonary hypertension and for longitudinal assessment of response to therapy. This analysis sought to describe the changes in hemodynamic variables after initiation of prostacyclin therapy and determine which changes bear predictive power of adverse clinical outcomes. A retrospective chart review of established patients at Cincinnati Children's Hospital with pulmonary arterial hypertension (PAH) who required prostacyclin therapy between 2004 and 2018 was performed. The baseline hemodynamic parameters at diagnosis as well as change in those parameters between initial catheterization and post-prostacyclin initiation catheterization were independent variables. Cox proportional hazard regression and recursive partitioning analysis were used to characterize which hemodynamic factors predicted the composite adverse outcome (CAO) defined as death, lung transplantation, or reverse Pott's shunt surgery. During the study period, 29 patients met inclusion criteria in which there were 7 CAOs: 4 deaths, 3 lung transplants, and 2 reverse Pott's shunts. Median time between catheterizations was 86 days and between the initiation of prostacyclin therapy and the second catheterization was 54 days. Cox regression revealed that only baseline pulmonary artery pressure (> 51 mmHg) and a failure to increase cardiac index illustrated statistically significant hazard for occurrence of the CAO (p < 0.01). These criteria significantly dichotomized the population in a Kaplan-Meier analysis into likelihoods of experiencing the CAO. While controlling for other hemodynamic variables, the absence of augmentation of cardiac index after the initiation of prostacyclin therapy is a valuable prognostic indicator of adverse PAH outcomes in pediatrics.


Neuronal apoptosis by glial NO: involvement of inhibition of glyceraldehyde-3-phosphate dehydrogenase.
In primary cocultures of neurons and glial cells prepared from the neonatal rat brain, lipopolysaccharide (LPS) reduced the numbers of neuronal cells but the effects were markedly inhibited by NG-monomethyl-L-arginine, indicating the involvement of NO and LPS-induced NO synthase in neuronal death. LPS stimulated the expression of inducible NOS (iNOS) in preparations of primary cultured microglias/astrocytes, but not in primary cultured neurons. In addition, LPS caused DNA fragmentation only in NG108-15 cells but not in primary cultured astrocytes as well as astrocytes in cocultures of the two cell types, suggesting that NOS induces the apoptosis of neurons but not glial cells. We then examined the NO-induced neuronal death in NG108-15 cells using NO donors. SNP, and NO donor, caused NO-2 accumulation in the reaction medium and lactate dehydrogenase (LDH) leakage from NG108-15 cells. Although SNP stimulated guanylyl cyclase and accumulated cGMP, cGMP analogs did not affect LDH leakage. In addition, SNP induced chromosomal condensation and fragmentation of nuclei in NG108-15 cells. Gel electrophoretic analysis of cellular DNA extracted from SNP-treated cells, confirmed the internucleosomal DNA fragmentation typical of apoptosis in this culture. SNP increased the amount of radioisotopic labeled glyceraldehyde-3 phosphate dehydrogenase (GAPDH) in the presence of [32P]NAD and inhibited the enzyme activity. The results suggested that SNP-induced cell death is partly due to the NO-induced inhibition of GAPDH, perhaps by stimulating the binding of NAD to GAPDH.


Simultaneous Trace Identification and Quantification of Common Types of Microplastics in Environmental Samples by Pyrolysis-Gas Chromatography-Mass Spectrometry.
The content of microplastics (MP) in the environment is constantly growing. Since the environmental relevance, particularly bioavailability, rises with decreasing particle size, the knowledge of the MP proportion in habitats and organisms is of gaining importance. The reliable recognition of MP particles is limited and underlies substantial uncertainties. Therefore spectroscopic methods are necessary to ensure the plastic nature of isolated particles, determine the polymer type and obtain particle count related quantitative data. In this study Curie-Point pyrolysis-gas chromatography-mass spectrometry combined with thermochemolysis is shown to be an excellent analytical tool to simultaneously identify and optionally quantify MP in environmental samples on a polymer specific mass related trace level. The method is independent of any mechanical preselection or particle appearance. For this purpose polymer characteristic pyrolysis products and their indicative fragment ions were used to analyze eight common types of plastics. Further aspects of calibration, recoveries, and potential matrix effects are discussed. The method is exemplarily applied on selected fish samples after an enzymatic-chemically pretreatment. This new approach with mass-related results is complementary to established FT-IR and Raman methods providing particle counts of individual polymer particles.


Polo-like kinase 1, a new therapeutic target in hepatocellular carcinoma.
To investigate the role of polo-like kinase 1 (PLK1) as a therapeutic target for hepatocellular carcinoma (HCC). PLK1 gene expression was evaluated in HCC tissue and HCC cell lines. Gene knockdown with short-interfering RNA (siRNA) was used to study PLK1 gene and protein expression using real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, and cell proliferation using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2(4-sulfophenyl)-2H-tetrazolium (MTS) and bromodeoxyuridine (BrdU) assays. Apoptosis was evaluated using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and caspase-inhibition assay. Huh-7 cells were transplanted into nude mice and co-cultured with PLK1 siRNA or control siRNA, and tumor progression was compared with controls. RT-PCR showed that PLK1 was overexpressed 12-fold in tumor samples compared with controls, and also was overexpressed in Huh-7 cells. siRNA against PLK1 showed a reduction in PLK1 gene and protein expression of up to 96% in Huh-7 cells, and a reduction in cell proliferation by 68% and 92% in MTS and BrdU cell proliferation assays, respectively. There was a 3-fold increase in apoptosis events, and TUNEL staining and caspase-3 assays suggested that this was caspase-independent. The pan-caspase inhibitor Z-VAD-FMK was unable to rescue the apoptotic cells. Immnofluorescence co-localized endonuclease-G to fragmented chromosomes, implicating it in apoptosis. Huh-7 cells transplanted subcutaneously into nude mice showed tumor regression in siPLK1-treated mice, but not in controls. Knockdown of PLK1 overexpression in HCC was shown to be a potential therapeutic target, leading to apoptosis through the endonuclease-G pathway.


POODLE-I: disordered region prediction by integrating POODLE series and structural information predictors based on a workflow approach.
Under physiological conditions, many proteins that include a region lacking well-defined three-dimensional structures have been identified, especially in eukaryotes. These regions often play an important biological cellular role, although they cannot form a stable structure. Therefore, they are biologically remarkable phenomena. From an industrial perspective, they can provide useful information for determining three-dimensional structures or designing drugs. For these reasons, disordered regions have attracted a great deal of attention in recent years. Their accurate prediction is therefore anticipated to provide annotations that are useful for wide range of applications. POODLE-I (where "I" stands for integration) is a web-based disordered region prediction system. POODLE-I integrates prediction results obtained from three kinds of disordered region predictors (POODLEs) developed from the viewpoint that the characteristics of disordered regions change according to their length. Furthermore, POODLE-I combines that information with predicted structural information by application of a workflow approach. When compared with server teams that showed best performance in CASP8, POODLE-I ranked among the top and exhibited the highest performance in predicting unfolded proteins. POODLE-I is an efficient tool for detecting disordered regions in proteins solely from the amino acid sequence. The application is freely available at http://mbs.cbrc.jp/poodle/poodle-i.html.


Nutritional and metabolic status of HIV-positive patients with lipodystrophy during one year of follow-up.
The aim of this prospective study was to compare changes in lipid metabolism and nutritional status after either 6 and 12 months of follow-up in subjects with lipodystrophy syndrome after traditional lifestyle therapy with or without fibric acid analogue intervention (bezafibrate and clofibrate). Food intake, alterations in body composition and metabolic abnormalities were assessed in subjects with lipodystrophy syndrome at the beginning of the study. The nutritional status and metabolic alterations of the subjects were monitored, and the subjects received nutritional counseling each time they were seen. The subjects were monitored either two times over a period no longer than six months (Group A; n = 18) or three times over a period of at least 12 months (Group B; n = 35). All of the subjects underwent nutrition counseling that was based on behavior modification. The fibric acid analogue was only given to patients with serum triglyceride levels above 400 mg/dL. After six months of follow-up, Group A showed no alterations in the experimental parameters. After twelve months, there was a decrease in serum triglyceride levels (410.4 ± 235.5 vs. 307.7 ± 150.5 mg/dL, p < 0.05) and an increase in both HDLc levels (37.9 ± 36.6 vs. 44.9 ± 27.9 mg/dL, p,0.05) and lean mass (79.9 ± 7.8 vs. 80.3 ± 9.9 %, p < 0.05) in Group B. After one year of follow-up (three sessions of nutritional and medical counseling), the metabolic parameters of the subjects with lipodystrophy improved after traditional lifestyle therapy with or without fibric acid analogue intervention.