Phosphate recovery potential from wastewater by chemical precipitation at batch conditions.
This study aimed to investigate the potential for phosphate recovery, instead of removal, from wastewater at conventional biological treatment processes. In the first part of this study, struvite and apatite phosphate-salts formation potential were investigated using artificially prepared solutions. In the second part, real municipal wastewater was subjected to batch-basis aerobic/anaerobic sequential order operation and supernatant of high phosphate-concentrations released into the liquid medium during the anaerobic phase were withdrawn and subjected to chemical precipitation tests. The formed mixed-liquor suspended solid suspensions were subjected to quiescent settling to ensure the separation of solids. Then, to form chemical bonds between calcium and phosphate ions, calcium was added to the withdrawn supernatants. The ability of apatite formation by itself as well as support-aided apatite formation was tested in supernatants. All the precipitation tests were carried out at orbital incubator at constant 20 degrees C and 25 degrees C temperature which were reported to be the range yielding the highest phosphate precipitation efficiency. In order to evaluate the solid-liquid separation ability of the chemical slurries, both centrifugation at 2000 rpm and membrane filtration at 100 mbar vacuum pressure were tested. The recovered solid phases as well as supernatants were analyzed for phosphate contents and it was revealed that the highest efficiency of phosphate precipitation was at pH of 9.50 for apatite and 9.40 for struvite. The results also suggested that struvite precipitation is more applicable for technical purposes while apatite precipitation is more applicable for economical reasons.


Near-infrared light triggered drug release from mesoporous silica nanoparticles.
Stimuli triggered drug delivery systems enable controlled release of drugs at the optimal space and time, thus achieving optimal therapeutic effects. As one of the most important stimuli used in bioapplications, near-infrared (NIR) light possesses unique advantages such as deep tissue penetration with minimum auto-fluorescence & tissue scattering and high biosafety. Mesoporous silica nanoparticles (MSNs) are one of the most studied nanocarriers; apart from having a high surface area and large pore volume for loading of drugs, they can be easily functionalized with inorganic nanomaterials and stimuli responsive polymers or organic switch molecules, creating possibilities for designing complex stimuli triggered drug delivery systems. Considering the high tissue penetration depth of NIR light and the unique mesoporous structure of MSNs, NIR responsive inorganic nanoparticle functionalized MSNs can be further combined with stimuli responsive materials to form smart "nano-devices" for controlled drug delivery toward tumors, and to date much progress has been made. In this article, recent advances in the design of NIR triggered mesoporous silica drug delivery systems are systematically summarized and some outstanding studies are highlighted. We will also discuss the shortcomings, challenges and opportunities in the field.


Patients with severe-to-profound hearing impairment and simultaneous severe vision impairment: a quality-of-life study.
Patients with severe vision impairment in combination with severe-to-profound hearing loss seem to have a higher risk for effects on QoL, including: mobility, the ability to provide self-care and perform usual activities, and levels of anxiety and depression, compared with patients with only severe-to-profound hearing loss. To study the quality-of-life (QoL) and audiological rehabilitation of the severely vision-impaired patient population among adults with severe-to-profound hearing loss in Sweden. A study of data collected from 543 patients with severe-to-profound hearing loss combined with severe vision impairment among the total of 2319 persons registered in the Swedish Quality Register of Otorhinolaryngology. QoL was measured with the following instruments: EQ5D, PIRS, and HADS. Audiological rehabilitation was described and evaluated. The patients with dual sensory loss were younger, were more likely to live alone, and had a lower level of education than the control group. The QoL of the study group was significantly negatively affected. In total, 89% of the study group had been rehabilitated with hearing aids, while 8% had received rehabilitation with cochlear implants. A total of 32% of the study group had received extended audiological rehabilitation.


Research priority setting in obesity: a systematic review.
Obesity research priority setting, if conducted to a high standard, can help promote policy-relevant and efficient research. Therefore, there is a need to identify existing research priority setting studies conducted in the topic area of obesity and to determine the extent to which they followed good practice principles for research priority setting. Studies examining research priority setting in obesity were identified through searching the MEDLINE, PBSC, CINAHL, PsycINFO databases and the grey literature. The nine common themes of good practice in research priority setting were used as a methodological framework to evaluate the processes of the included studies. These were context, use of a comprehensive approach, inclusiveness, information gathering, planning for implementation, criteria, methods for deciding on priorities, evaluation and transparency. Thirteen articles reporting research prioritisation exercises conducted in different areas of obesity research were included. All studies reported engaging with various stakeholders such as policy makers, researchers and healthcare professionals. Public involvement was included in six studies. Methods of research prioritisation commonly included both Delphi and nominal group techniques and surveys. None of the 13 studies fulfilled all nine of the good practice criteria for research priority setting, with the most common limitations including not using a comprehensive approach and lack of inclusivity and evaluating on their processes. There is a need for research priority setting studies in obesity to involve the public and to evaluate their exercises to ensure they are of high quality.


A biologically realistic model of contrast invariant orientation tuning by thalamocortical synaptic depression.
Simple cells in layer 4 of the primary visual cortex of the cat show contrast-invariant orientation tuning, in which the amplitude of the peak response is proportional to the stimulus contrast but the width of the tuning curve hardly changes with contrast. This study uses a detailed model of spiny stellate cells (SSCs) from cat area 17 to explain this property. The model integrates our experimental data, including morphological and intrinsic membrane properties and the number and spatial distribution of four major synaptic input sources of the SSC: the dorsal lateral geniculate nucleus (dLGN) and three cortical sources. The model also includes synaptic properties of these inputs. The cortical input served as sources of background activity, and visual stimuli was modeled as sinusoidal grating. For all contrasts, strong synaptic depression of the dLGN feedforward afferents compresses the firing rates in response to orthogonal stimuli, keeping these rates at practically the same low level. However, at preferred orientations, despite synaptic depression, firing rate changes as a function of contrast. Thus, when embedded in an active network, strong synaptic depression can explain contrast-invariant orientation tuning of simple cells. This is true also when the dLGN inputs are partially depressed as a result of their spontaneous activity and to some extent also when parameters were fitted to a more moderate level of synaptic depression. The model response is in close agreement with experimental results, in terms of both output spikes and membrane voltage (amplitude and fluctuations), with reasonable exceptions given that recurrent connections were not incorporated.


Exploring Exercise- and Context-Induced Peptide Changes in Mice by Quantitative Mass Spectrometry.
Recent research suggests that exercise may help facilitate abstinence from cocaine addiction, though the mechanisms are not well understood. In mice, wheel running accelerates the extinction of conditioned place preference (CPP) for cocaine, providing an animal model for evaluating potential neurological mechanisms. The objective of this study was to quantify dynamic changes in endogenous peptides in the amygdala and dentate gyrus of the hippocampus in mice exposed to a context paired with the effects of cocaine, and in response to exercise. Male C57BL/6J mice conditioned to cocaine were housed with or without running wheels for 30 days. Following a CPP test and final exposure to either a cocaine- or saline-associated context, peptides were measured in brain tissue extracts using label-free matrix-assisted laser desorption/ionization mass spectrometry (MS) and stable isotopic labeling with liquid chromatography and electrospray ionization MS. CPP in mice was significantly reduced with running, which correlated to decreased myelin basic protein derivatives in the dentate gyrus extracts, possibly reflecting increased unmyelinated granule neuron density. Exposure to a cocaine-paired context increased hemoglobin-derived peptides in runners and decreased an actin-derived peptide in sedentary animals. These results allowed us to characterize a novel set of biomarkers that are responsive to exercise in the hippocampus and in a cocaine-paired context in the amygdala.


Determinants of promoter and enhancer transcription directionality in metazoans.
Divergent transcription from promoters and enhancers is pervasive in many species, but it remains unclear if it is a general feature of all eukaryotic cis regulatory elements. To address this, here we define cis regulatory elements in C. elegans, D. melanogaster and H. sapiens and investigate the determinants of their transcription directionality. In all three species, we find that divergent transcription is initiated from two separate core promoter sequences and promoter regions display competition between histone modifications on the + 1 and -1 nucleosomes. In contrast, promoter directionality, sequence composition surrounding promoters, and positional enrichment of chromatin states, are different across species. Integrative models of H3K4me3 levels and core promoter sequence are highly predictive of promoter and enhancer directionality and support two directional classes, skewed and balanced. The relative importance of features to these models are clearly distinct for promoters and enhancers. Differences in regulatory architecture within and between metazoans are therefore abundant, arguing against a unified eukaryotic model.


Marginal cells of the stria vascularis of gerbils take up glucose via the facilitated transporter GLUT: application of autofluorescence.
Strial marginal cells are known to take up and metabolize glucose as their main source of metabolic energy. The membrane transport mechanisms for glucose uptake into strial marginal cells, however, are largely unknown. Two types of glucose transporters in mammalian cells have been described, the facilitated glucose transporter GLUT and the sodium/glucose cotransporter SGLT. The goal of the present study was to determine which of these represent the main glucose uptake mechanism in strial marginal cells. Glucose uptake into strial marginal cells was assessed by monitoring the cellular concentration of the reduced form of nicotinamide adenine dinucleotide (NADH) fluorometrically. The relation between the autofluorescence from marginal cells and cellular metabolism was verified as follows. The autofluorescence (excitation: 340 nm, emission: 450-490 nm) decreased when oxidative phosphorylation in the mitochondria was uncoupled with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) and increased when cytochrome oxidase was inhibited with cyanide. These effects indicate that the autofluorescence is dependent on the mitochondrial metabolic state, and more specifically on the level of NADH in mitochondria. Glucose removal from the bath solution elicited a 39% decrease in the autofluorescence intensity within 5 min. Similarly, cytochalasin B (10 microM) reduced the fluorescence intensity by 34% in 5 min. In contrast, neither phlorizin (0.1 mM) nor Na+ removal from the bath solution caused any appreciable change in the autofluorescence intensity. These results indicate that glucose depletion affects the metabolic state of the marginal cell within a few minutes, and that marginal cells take up glucose via GLUT, but not via SGLT. Since the excitation and emission wavelengths of several fluorescent dyes used in physiological studies (e.g., Fura-2 and SBFI) are similar to those of NADH, possible effects of autofluorescence on recording signals should always be taken into account when these dyes are utilized.


Intramolecular crosslinking of tropomyosin via disulfide bond formation: evidence for chain register.
Rabbit skeletal muscle tropomyosin can be crosslinked in the native state by the use of 5,5'-dithiobis(2-nitrobenzoate), which forms disulfide bonds between the two subunits. Using polyacrylamide gel electrophoresis in sodium dodecyl sulfate we have shown that this crosslinking goes to completion over a wide range of protein concentration, ionic strength, and reagent concentration. Crosslinks are not formed in denaturing solvents such as sodium dodecyl sulfate and guanidine hydrochloride despite the fact that the same number of SH groups react as in the native state (2.3 +/- 0.2). The sedimentation coefficients of crosslinked and non-crosslinked samples are identical. Thus, crosslinks are formed between corresponding cysteines on different chains of the same molecule. This provides strong evidence for a model of chain interaction with both chains in register. Evidence has also been obtained that rabbit skeletal tropomyosin is composed only of alphaalpha and alphabeta subunits rather than a random mixture of chains.


Uniportal subxiphoid video-assisted thoracoscopic thymectomy.
With a uniportal subxiphoid video-assisted thoracoscopic surgery (VATS) thymectomy, the thymus is not accessed through the ribs, which avoids intercostal nerve damage. Furthermore, compared to a VATS thymectomy via the lateral chest, decreased use of postoperative analgesics (lower doses and shorter duration), decreased blood loss, and shorter surgical duration and hospital stay have been reported. Here we report our surgical method and solutions for a uniportal subxiphoid VATS thymectomy. A uniportal subxiphoid VATS thymectomy is indicated for patients with myasthenia gravis and anterior mediastinal tumors who do not require surgical suturing. The operator stands between the patient's legs and the scopist stands to the right of the patient to operate the camera scope. To begin with, a 3-cm transverse incision was made along Langer's lines 1 cm caudal below the xiphoid process. The CO2 insufflation is performed in the mediastinum at 8 mmHg. The pericardial adipose tissue and thymus are detached from the pericardium in an anterior manner from the bilateral phrenic nerves. The resected thymus is placed in a pouch in the mediastinum and removed from the body through the subxiphoid incision. The subxiphoid approach is highly beneficial for the patient in that it results in superior esthetic outcomes considering that there is no sternotomy and no intercostal nerve damage; thus, it is a surgical procedure that surgeons should learn.


Looking ahead: Anticipatory cueing of attention to objects others will look at.
Seeing a face gaze at an object elicits rapid attention shifts toward the same object. We tested whether gaze cueing is predictive: do people shift their attention toward objects others are merely expected to look at? Participants categorized objects while a face either looked at this object, at another object, or straight ahead. Unbeknownst to participants, one face would only look at drinks and the other at foods. We tested whether attention was drawn toward objects "favored" by a face even when currently looking straight ahead. Indeed, while gaze expectations initially had a disruptive effect, participants did shift attention to the faces' favored objects once learning had been established, as long as emotional expressions had indicated personal relevance of the object to the individual. These data support predictive models of social perception, which assume that predictions can drive perception and action, as if these stimuli were directly perceived.


Primary pancreatic lymphoma: A rare cause of pancreatic mass.
Primary pancreatic lymphoma (PPL) is a rare entity, most likely to be clinically misdiagnosed as pancreatic cancer. The cure rate of PPL is higher compared with that of pancreatic adenocarcinoma. This is the case report of a 57-year-old male patient who was hospitalized with complaints of abdominal pain, weight loss and jaundice. The radiological evaluation revealed a pancreatic head mass and, following endoscopic ultrasound-guided fine-needle aspiration biopsy, the tumor was diagnosed as diffuse large B-cell lymphoma. The final diagnosis was PPL, and the patient went into remission after receiving three cycles of treatment with rituximab, doxorubicin, cyclophosphamide, vincristine and prednisolone (R-CHOP regimen). Therefore, PPL should be considered in the differential diagnosis of pancreatic masses and its management differs from that of other types of pancreatic tumor.


Functional dichotomy of NK cells in organ transplantation.
Evaluation of: Yu G, Xu X, Vu MD, Kilpatrick ED, Li XC. NK cells promote transplant tolerance by killing donor antigen-presenting cells. J. Exp. Med. 203, 1851-1858 (2006). Natural killer (NK) cells have the potential to display different functional activities after transplantation. The traditional view is that NK cells have the capacity to contribute to rejection by facilitating the activation/differentiation of leukocytes that destroy the graft. By contrast, in the article under review, a novel role for NK cells was identified in the setting of costimulation blockade where alloreactive NK cells of recipient origin were found to have the capacity to kill donor-derived antigen-presenting cells, thereby reducing T-cell priming and promoting long-term skin graft acceptance.


14-3-3ε plays a role in cardiac ventricular compaction by regulating the cardiomyocyte cell cycle.
Trabecular myocardium accounts for the majority of the ventricles during early cardiogenesis, but compact myocardium is the primary component at later developmental stages. Elucidation of the genes regulating compact myocardium development is essential to increase our understanding of left ventricular noncompaction (LVNC), a cardiomyopathy characterized by increased ratios of trabecular to compact myocardium. 14-3-3ε is an adapter protein expressed in the lateral plate mesoderm, but its in vivo cardiac functions remain to be defined. Here we show that 14-3-3ε is expressed in the developing mouse heart as well as in cardiomyocytes. 14-3-3ε deletion did not appear to induce compensation by other 14-3-3 isoforms but led to ventricular noncompaction, with features similar to LVNC, resulting from a selective reduction in compact myocardium thickness. Abnormal compaction derived from a 50% decrease in cardiac proliferation as a result of a reduced number of cardiomyocytes in G(2)/M and the accumulation of cardiomyocytes in the G(0)/G(1) phase of the cell cycle. These defects originated from downregulation of cyclin E1 and upregulation of p27(Kip1), possibly through both transcriptional and posttranslational mechanisms. Our work shows that 14-3-3ε regulates cardiogenesis and growth of the compact ventricular myocardium by modulating the cardiomyocyte cell cycle via both cyclin E1 and p27(Kip1). These data are consistent with the long-held view that human LVNC may result from compaction arrest, and they implicate 14-3-3ε as a new candidate gene in congenital human cardiomyopathies.


Sinus node dysfunction requiring permanent pacemaker implantation in a young adult with Klinefelter syndrome.
Klinefelter syndrome is the most common genetic cause of male infertility and affects approximately 1 in 500 live births. Although accompanying cardiac disorder is not a specific feature of Klinefelter syndrome, rarely associated anomalies such as mitral valve prolapse, atrial septal defect, ventricular septal defect, tetralogy of Fallot, patent ductus arteriosus, and hypertrophic obstructive cardiomyopathy have been reported. A clear association between Klinefelter syndrome and arrhythmic disorders has not yet been demonstrated. We report a case of a sinus node dysfunction that required permanent pacemaker implantation in a young adult with Klinefelter syndrome. The patient was consulted to cardiology clinic due to bradycardia. On physical examination, no cardiac abnormality was detected except for bradycardia. Holter results showed sinus arrhythmia with a minimum heart rate of 33 bpm and maximum of 154 Bpm. There were 3612 ventricular premature beats, 30 ventricular pairs, 804 supraventricular premature beats, 7 supraventricular pairs, and 4 supraventricular runs, the longest of which was 5 beats. The patient had defined dizziness and nausea during Holter monitoring. Electrophysiological study (EPS) was planned because existing findings indicated risk of cardiac syncope. Findings of EPS were interpreted as sinus node dysfunction. A permanent pacemaker implantation was performed and the patient has been free of symptoms since. This concomitance should be kept in mind when examining patients with Klinefelter syndrome with bradycardia and/or syncope. It is easily mistaken for epilepsy, which is a commonly encountered abnormality in Klinefelter syndrome.


Carboplatin and paclitaxel with vs without bevacizumab in older patients with advanced non-small cell lung cancer.
A previous randomized trial demonstrated that adding bevacizumab to carboplatin and paclitaxel improved survival in advanced non-small cell lung cancer (NSCLC). However, longer survival was not observed in the subgroup of patients aged 65 years or older. To examine whether adding bevacizumab to carboplatin and paclitaxel chemotherapy is associated with improved survival in older patients with NSCLC. Retrospective cohort study of 4168 Medicare beneficiaries aged 65 years or older with stage IIIB or stage IV non-squamous cell NSCLC diagnosed in 2002-2007 in a Surveillance, Epidemiology, and End Results (SEER) region. Patients were categorized into 3 cohorts based on diagnosis year and type of initial chemotherapy administered within 4 months of diagnosis: (1) diagnosis in 2006-2007 and bevacizumab-carboplatin-paclitaxel therapy; (2) diagnosis in 2006-2007 and carboplatin-paclitaxel therapy; or (3) diagnosis in 2002-2005 and carboplatin-paclitaxel therapy. The associations between carboplatin-paclitaxel with vs without bevacizumab and overall survival were compared using Cox proportional hazards models and propensity score analyses including information about patient characteristics recorded in SEER-Medicare. Overall survival measured from the first date of chemotherapy treatment until death or the censoring date of December 31, 2009. The median survival estimates were 9.7 (interquartile range [IQR], 4.4-18.6) months for bevacizumab-carboplatin-paclitaxel, 8.9 (IQR, 3.5-19.3) months for carboplatin-paclitaxel in 2006-2007, and 8.0 (IQR, 3.7-17.2) months for carboplatin-paclitaxel in 2002-2005. One-year survival probabilities were 39.6% (95% CI, 34.6%-45.4%) for bevacizumab-carboplatin-paclitaxel vs 40.1% (95% CI, 37.4%-43.0%) for carboplatin-paclitaxel in 2006-2007 and 35.6% (95% CI, 33.8%-37.5%) for carboplatin-paclitaxel in 2002-2005. Neither multivariable nor propensity score-adjusted Cox models demonstrated a survival advantage for bevacizumab-carboplatin-paclitaxel compared with carboplatin-paclitaxel cohorts. In propensity score-stratified models, the hazard ratio for overall survival for bevacizumab-carboplatin-paclitaxel compared with carboplatin-paclitaxel in 2006-2007 was 1.01 (95% CI, 0.89-1.16; P = .85) and compared with carboplatin-paclitaxel in 2002-2005 was 0.93 (95% CI, 0.83-1.06; P = .28). The propensity score-weighted model and propensity score-matching model similarly failed to demonstrate a statistically significant superiority for bevacizumab-carboplatin-paclitaxel. Subgroup and sensitivity analyses for key variables did not change these findings. Adding bevacizumab to carboplatin and paclitaxel chemotherapy was not associated with better survival among Medicare patients with advanced NSCLC.


Coping, social support and quality of life over time after myocardial infarction.
This paper describes gender differences in perceived coping, social support and quality of life 1, 4 and 12 months after myocardial infarction. There is a shortage of studies with a longitudinal research design investigating coping, social support and quality of life in women and men after myocardial infarction. A longitudinal, descriptive and comparative design was used for the study, which included 74 women and 97 men. At 12 months, 60 women and 88 men remained. Data were collected using the Jalowiec Coping Scale, a social support questionnaire, the SF-36 Health Survey (health-related quality of life) and the Quality of Life Index-Cardiac version (quality of life). The data were collected during the period 1999-2001. No statistically significant changes over time in coping assessments emerged in the study group, except for fatalistic coping, which diminished over time in men. Women used more evasive coping than men at 4 and 12 months. The perceived efficiency in coping with physical aspects of the heart disease increased. More women than men perceived available support from grandchildren and staff of the church. Health-related quality of life increased in women and men in physical functioning, role-physical, vitality, social functioning, and role-emotional scales. Moreover, an improvement in the mental health scale was evident in women and a reduction in pain in men. No statistically significant gender differences were found for quality of life at any point in time. The findings can be used to inform caregivers that optimistic, self-reliant and confrontational coping were the most frequently used by both women and men over the first year after myocardial infarction, and that confrontational coping has been shown to have positive outcomes in the longer term. Nurses should tell women about the importance of seeking prompt treatment and discuss health problems with caregivers and significant others. Care planning should include family members and significant others so that they can support and encourage patients to cope with problems in daily life.


Formation of pyrene excimers in mesoporous ormosil thin films for visual detection of nitro-explosives.
We report the preparation of mesoporous thin films with bright pyrene excimer emission and their application in visual and rapid detection of nitroaromatic explosive vapors. The fluorescent films were produced by physically encapsulating pyrene molecules in the organically modified silica (ormosil) networks which were prepared via a facile template-free sol-gel method. Formation and stability of pyrene excimer emission were investigated in both porous and nonporous ormosil thin films. Excimer emission was significantly brighter and excimer formation ability was more stable in porous films compared to nonporous films. Rapid and selective quenching was observed in the excimer emission against vapors of nitroaromatic molecules; trinitrotoluene (TNT), dinitrotoluene (DNT), and nitrobenzene (NB). Fluorescence quenching of the films can be easily observed under UV light, enabling the naked-eye detection of nitro-explosives. Furthermore, excimer emission signal can be recovered after quenching and the films can be reused at least five times.


Diagnostic imaging of osteoporosis and sarcopenia: a narrative review.
Osteoporosis and sarcopenia represent two major health problems with an increasing prevalence in the elderly population. The correlation between these diseases has been widely reported, leading to the development of the term "osteosarcopenia" to diagnose those patients suffering from both diseases. Several imaging methods for the diagnosis and management of osteoporosis exist, with dual-energy X-ray absorptiometry (DXA) being the most commonly used for measuring bone mineral density (BMD). Imaging technique other than DXA is represented by conventional radiography, computed tomography (CT) and ultrasound (US). Similarly, the imaging technologies used to detect loss of skeletal muscle mass in sarcopenia include DXA, CT, US and magnetic resonance imaging (MRI). These methods differ in terms of reliability, radiation exposure and costs. CT and MRI represent the gold standard for evaluating body composition (BC), but are costly and time-consuming. DXA remains the most often used technology for studying BC, being quick, widely available and with low radiation exposure.


Pancreatic somatostatinoma diagnosed preoperatively: report of a case.
Somatostatinoma is a rare neoplasm of the pancreas. Preoperative diagnosis is often difficult. We report a 72-year-old woman with a pancreatic head tumor measuring 37 mm in diameter, and enlargement of the lymph nodes on the anterior surface of the pancreatic head and the posterior surface of the horizontal part of the duodenum. Laboratory data showed an elevated plasma somatostatin concentration. Examination of a biopsy specimen of the pancreatic head mass obtained by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) showed histopathological features of a neuroendocrine tumor. Immunohistochemical staining showed that the tumor cells were positive for somatostatin, leading to a preoperative diagnosis of pancreatic somatostatinoma. The patient underwent pylorus-preserving pancreaticoduodenectomy. The plasma somatostatin concentration decreased progressively after surgery. A rare case of pancreatic somatostatinoma with lymph node metastases was presented. Immunohistochemical analysis of a biopsy specimen obtained by EUS-FNA was useful for preoperative diagnosis.


WHO/ILO work-related burden of disease and injury: Protocol for systematic reviews of occupational exposure to solar ultraviolet radiation and of the effect of occupational exposure to solar ultraviolet radiation on cataract.
The World Health Organization (WHO) and the International Labour Organization (ILO) are developing a joint methodology for estimating the national and global work-related burden of disease and injury (WHO/ILO joint methodology), with contributions from a large network of experts. Here, we present the protocol for two systematic reviews of parameters for estimating the number of disability-adjusted life years of cataracts from occupational exposure to solar ultraviolet radiation, to inform the development of the WHO/ILO joint methodology. We aim to systematically review studies on occupational exposure to solar ultraviolet radiation (Systematic Review 1) and systematically review and meta-analyse estimates of the effect of occupational exposure to solar ultraviolet radiation on the development of cataract (Systematic Review 2), applying the Navigation Guide systematic review methodology as an organizing framework and conducting both systematic reviews in tandem and in a harmonized way. Separately for Systematic Reviews 1 and 2, we will search electronic academic databases for potentially relevant records from published and unpublished studies, including Ovid Medline, PubMed, EMBASE, and Web of Sciences. We will also search electronic grey literature databases, Internet search engines and organizational websites; hand search reference list of previous systematic reviews and included study records; and consult additional experts. We will include working-age (≥15 years) workers in WHO and/or ILO Member States, but exclude children (<15 years) and unpaid domestic workers. For Systematic Review 1, we will include quantitative studies on the prevalence of relevant levels of occupational exposure to solar ultraviolet radiation and of the total working time spent outdoors from 1960 to 2018, stratified by sex, age, country and industrial sector or occupation. For Systematic Review 2, we will include randomized controlled trials, cohort studies, case-control studies and other non-randomized intervention studies with an estimate of the effect of any occupational exposure to solar ultraviolet radiation (i.e. ≥30 Jm-2/day of occupational solar UV exposure at the surface of the eye) on the prevalence or incidence of cataract, compared with the theoretical minimum risk exposure level (i.e. <30 Jm-2/day of occupational solar UV exposure at the surface of the eye). At least two review authors will independently screen titles and abstracts against the eligibility criteria at a first stage and full texts of potentially eligible records at a second stage, followed by extraction of data from qualifying studies. At least two review authors will assess risk of bias and the quality of evidence, using the most suited tools currently available. For Systematic Review 2, if feasible, we will combine relative risks using meta-analysis. We will report results using the guidelines for accurate and transparent health estimates reporting (GATHER) for Systematic Review 1 and the preferred reporting items for systematic reviews and meta-analyses guidelines (PRISMA) for Systematic Review 2. PROSPERO registration: CRD42018098897.


Comparison of the SAWNUC model with CLOUD measurements of sulphuric acid-water nucleation.
Binary nucleation of sulphuric acid-water particles is expected to be an important process in the free troposphere at low temperatures. SAWNUC (Sulphuric Acid Water Nucleation) is a model of binary nucleation that is based on laboratory measurements of the binding energies of sulphuric acid and water in charged and neutral clusters. Predictions of SAWNUC are compared for the first time comprehensively with experimental binary nucleation data from the CLOUD chamber at European Organization for Nuclear Research. The experimental measurements span a temperature range of 208-292 K, sulphuric acid concentrations from 1·106 to 1·109 cm-3, and distinguish between ion-induced and neutral nucleation. Good agreement, within a factor of 5, is found between the experimental and modeled formation rates for ion-induced nucleation at 278 K and below and for neutral nucleation at 208 and 223 K. Differences at warm temperatures are attributed to ammonia contamination which was indicated by the presence of ammonia-sulphuric acid clusters, detected by an Atmospheric Pressure Interface Time of Flight (APi-TOF) mass spectrometer. APi-TOF measurements of the sulphuric acid ion cluster distributions ( (H2SO4)i·HSO4- with i = 0, 1, ..., 10) show qualitative agreement with the SAWNUC ion cluster distributions. Remaining differences between the measured and modeled distributions are most likely due to fragmentation in the APi-TOF. The CLOUD results are in good agreement with previously measured cluster binding energies and show the SAWNUC model to be a good representation of ion-induced and neutral binary nucleation of sulphuric acid-water clusters in the middle and upper troposphere.


Quality of life among older people in Sweden receiving help from informal and/or formal helpers at home or in special accommodation.
The present study describes and compares quality of life (QoL) and factors which predict QoL among people aged 75 years and over who receive help with activities of daily living (ADLs) from formal and/or informal helpers. The subjects were living at home or in special accommodation in Sweden. A postal questionnaire was sent to a randomly selected and age-stratified sample of 8500 people. The response rate was 52.8% (n = 4337), and 1247 people [mean age (+/- SD) = 86.4 +/- 5.9 years] received help and indicated who helped them with ADLs. The findings suggest that a greater age, being a woman, being a widow/widower, a higher number of health-related complaints, needing more help with ADLs and a lower QoL were found among those receiving help in special accommodation in comparison with those receiving help at home. The extent of help was highest among those receiving help in special accommodation. Having help with ADLs every day at home indicated having help from both informal and formal helpers, while respondents receiving help from only informal or only formal helpers received the smallest amount of help with ADLs. A need for greater help with ADLs, and a higher number of self-reported diseases and complaints determined low QoL, whilst a social network (contact with more than three people) and a greater age determined high QoL. However, who the helpers were did not have a significant influence on QoL; it was the extent of help with ADLs that influenced QoL negatively and the density of the social network that influenced QoL positively.


Developmentally regulated expression and activity of 17alpha-hydroxylase/C-17,20-lyase cytochrome P450 in rat liver.
We have investigated the developmental pattern of expression and activity of 17alpha-hydroxylase/C-17,20-lyase cytochrome P450 (cytochrome P450c17) in the liver, stomach, duodenum, and testis of rats from day 18 of pregnancy to adulthood. In the male liver, the enzyme became detectable at birth (135 pmol/mg protein x min) at a level comparable to that in the testis (188 pmol/mg protein x min). The activity then increased dramatically, reaching a peak at 8 days (691 pmol/mg protein x min), which was more than 4-fold the testicular levels in rats of the same age or in adults. Thereafter it declined steadily, becoming undetectable from puberty onward. The hepatic peak followed a depression in testicular activity (58 pmol/mg protein x min) on day 6. Northern and immunoblot analyses showed a good temporal correlation between enzyme activity and the occurrence of P450c17 messenger RNA (mRNA) and protein. The same patterns of mRNA and protein occurrence were observed in female rat liver, indicating that the hepatic CYP17 expression is not sexually dimorphic. Sequencing confirmed a complete identity in the coding region between hepatic and gonadal mRNAs. Hepatic P450c17 mRNA, however, was 150-200 bases longer than the gonadal counterparts. No significant expression of mRNAs encoding P450scc and P450arom was observed in liver of either sex at any age. In stomach and duodenum, enzyme activity was much lower (maxima at 25 and 14 pmol/mg protein x min, respectively) than that in liver, but persisted from the time of weaning onward. It is suggested that the hepatic peak in P450c17 activity may serve to convert circulating progestogens into androgens for gonadal aromatization during Sertoli and granulosa cell proliferation.


Optimizing nonpharmacological management following an acute exacerbation of chronic obstructive pulmonary disease.
Though the guidelines for the optimal management of chronic obstructive pulmonary disease (COPD) following an acute exacerbation (AE) are well established, issues associated with poor adherence to nonpharmacological interventions such as self-management advice and pulmonary rehabilitation will impact on hospital readmission rates and health care costs. Systems developed for clinically stable patients with COPD may not be sufficient for those who are post-exacerbation. A redesign of the manner in which such interventions are delivered to patients following an AECOPD is necessary. Addressing two or more components of the chronic care model is effective in reducing health care utilization in patients with COPD, with self-management support contributing a key role. By refining self-management support to incorporate the identification and treatment of psychological symptoms and by providing health care professionals adequate time and training to deliver respiratory-specific advice and self-management strategies, adherence to nonpharmacological therapies following an AE may be enhanced. Furthermore, following up patients in their own homes allows for the tailoring of advice and for the delivery of consistent health care messages which may enable knowledge to be retained. By refining the delivery of nonpharmacological therapies following an AECOPD according to components of the chronic care model, adherence may be improved, resulting in better disease management and possibly reducing health care utilization.


Self-assembled monolayer films of phosphonates for bonding RGD to titanium.
Modification of the implant surface with the Arg-Gly-Asp tripeptide (RGD) putatively facilitates osteoblast attachment for improved implant fixation in the laboratory. We compared the histomorphometric and mechanical performance of titanium implants coated with RGD using a novel interface of self-assembled monolayers of phosphonates (RGD/SAMP) and implants coated with RGD using the more conventional thiolate-gold interface (RGD/thiolate-gold). We hypothesized RGD/SAMP-coated implants would show greater bone ongrowth and implant fixation than RGD/thiolate-gold-coated ones. We implanted an RGD/SAMP-coated implant in one femur and an RGD/thiolate-gold-coated in the contralateral femur of 60 rats. At 2, 4, and 8 weeks after implantation, 10 rats were sacrificed for histologic evaluation and another 10 for biomechanical testing. Bone-implant ongrowth and implant force-to-failure of the two implants were similar at all times. Although RGD/SAMP-coated implants did not show superior bone ongrowth and implant fixation, RGD/SAMP-coated implants have at least equally good histomorphometric and mechanical in vivo performance as RGD/thiolate-gold-coated ones. Additional in vivo characterization of self-assembled monolayer films of phosphonates as interface to bond RGD to titanium is needed to explore its full potential and seems justified based on the results of this study.


Caffeine, cajoling, and other strategies to maximize clinician survey response rates.
An ongoing objective in health services research is to increase response rates to clinician surveys to ensure generalizability of findings. Three HMOs in the Cancer Research Network participated in a primary care clinician survey to better understand organizational characteristics affecting adoption and implementation of breast and cervical cancer screening guidelines. A four-stage data collection strategy was implemented to maximize response. This included careful attention to survey design and layout, extensive piloting, choice of token incentive, use of "local champions," and denominator management. An overall response rate of 91% was attained, ranging from 83 to 100% among the plans (N = 621). Although the response rate after the second stage of data collection met commonly used standards, the authors argue for the four-stage method due to the possibility of differences when comparing early and late responders. This is important when multiple plans with differing structure and internal characteristics are surveyed.


A high-pressure structure in curium linked to magnetism.
Curium lies at the center of the actinide series and has a half-filled shell with seven 5f electrons spatially residing inside its radon core. As a function of pressure, curium exhibits five different crystallographic phases up to 100 gigapascals, of which all but one are also found in the preceding element, americium. We describe here a structure in curium, Cm III, with monoclinic symmetry, space group C2/c, found at intermediate pressures (between 37 and 56 gigapascals). Ab initio electronic structure calculations agree with the observed sequence of structures and establish that it is the spin polarization of curium's 5f electrons that stabilizes Cm III. The results reveal that curium is one of a few elements that has a lattice structure stabilized by magnetism.


Protein kinase C-induced activin A switches adrenocortical steroidogenesis to aldosterone by suppressing CYP17A1 expression.
Functional zonation of the adrenal cortex is a consequence of the zone-specific expression of P450c17 (CYP17A1) and its cofactors. Activin and inhibin peptides are differentially produced within the zones of the adrenal cortex and have been implicated in steroidogenic control. In this study, we investigated whether activin and inhibin can function as intermediates in functional zonation of the human adrenal cortex. Activin A suppressed CYP17A1 expression and P450c17 function in adrenocortical cell lines as well as in primary adrenal cell cultures. Inhibin βA-subunit mRNA and activin A protein levels were found to be increased up to 1,900-fold and 49-fold, respectively, after protein kinase C (PKC) stimulation through PMA or angiotensin II in H295R adrenocortical carcinoma cells. This was confirmed in HAC15 cells and for PMA in primary adrenal cell cultures. Both PMA and Ang II decreased CYP17A1 expression in the adrenocortical cell lines, whereas PMA concurrently suppressed CYP17A1 levels in the primary cultures. Inhibition of activin signaling during PKC stimulation through silencing of the inhibin βA-subunit or blocking of the activin type I receptor opposed the PMA-induced downregulation of CYP17A1 expression and P450c17 function. In contrast, PKA stimulation through adrenocorticotrophin or forskolin increased expression of the inhibin α-subunit and betaglycan, both of which are antagonists of activin action. These data indicate that activin A acts as a PKC-induced paracrine factor involved in the suppression of CYP17A1 in the zona glomerulosa and can thereby contribute to functional adrenocortical zonation.


DNA analysis indicates that Asian elephants are native to Borneo and are therefore a high priority for conservation.
The origin of Borneo's elephants is controversial. Two competing hypotheses argue that they are either indigenous, tracing back to the Pleistocene, or were introduced, descending from elephants imported in the 16th-18th centuries. Taxonomically, they have either been classified as a unique subspecies or placed under the Indian or Sumatran subspecies. If shown to be a unique indigenous population, this would extend the natural species range of the Asian elephant by 1300 km, and therefore Borneo elephants would have much greater conservation importance than if they were a feral population. We compared DNA of Borneo elephants to that of elephants from across the range of the Asian elephant, using a fragment of mitochondrial DNA, including part of the hypervariable d-loop, and five autosomal microsatellite loci. We find that Borneo's elephants are genetically distinct, with molecular divergence indicative of a Pleistocene colonisation of Borneo and subsequent isolation. We reject the hypothesis that Borneo's elephants were introduced. The genetic divergence of Borneo elephants warrants their recognition as a separate evolutionary significant unit. Thus, interbreeding Borneo elephants with those from other populations would be contraindicated in ex situ conservation, and their genetic distinctiveness makes them one of the highest priority populations for Asian elephant conservation.


Pulmonary function in patients with Huntington's disease.
Huntington's disease (HD) is a neurodegenerative disorder characterized by progressive motor, cognitive and psychiatric disturbances. Chest muscle rigidity, respiratory muscle weakness, difficulty in clearing airway secretions and swallowing abnormalities have been described in patients with neurodegenerative disorders including HD. However limited information is available regarding respiratory function in HD patients. The purpose of this study was to investigate pulmonary function of patients with HD in comparison to healthy volunteers, and its association with motor severity. Pulmonary function measures were taken from 18 (11 male, 7 female) manifest HD patients (53 ± 10 years), and 18 (10 male, 8 female) healthy volunteers (52 ± 11 years) with similar anthropometric and life-style characteristics to the recruited HD patients. Motor severity was quantified by the Unified Huntington's Disease Rating Scale-Total Motor Score (UHDRS-TMS). Maximum respiratory pressure was measured on 3 separate days with a week interval to assess test-retest reliability. The test-retest reliability of maximum inspiratory and expiratory pressure measurements was acceptable for both HD patient and control groups (ICC ≥0.92), but the values over 3 days were more variable in the HD group (CV < 11.1%) than in the control group (CV < 7.6%). The HD group showed lower respiratory pressure, forced vital capacity, peak expiratory flow and maximum voluntary ventilation than the control group (p < 0.05). Forced vital capacity, maximum voluntary ventilation and maximum respiratory pressures were negatively (r = -0.57; -0.71) correlated with the UHDRS-TMS (p < 0.05). Pulmonary function is decreased in manifest HD patients, and the magnitude of the decrease is associated with motor severity.


A simple extemporaneous oral suspension of aprepitant yields sufficient pharmacokinetic exposure in children.
Aprepitant is used for the treatment of chemotherapy induced nausea and vomiting. A liquid formulation is needed for treatment of young children. However, the commercial (powder for) suspension was not available worldwide for a prolonged period of time and, therefore, a 10 mg/mL aprepitant oral suspension was extemporarily prepared to prevent suboptimal antiemetic treatment. The current pharmacokinetic study was developed to investigate whether this extemporaneous oral suspension offers an appropriate treatment option. From 49 pediatric patients (0.7-17.9 years) 235 plasma concentrations were collected. Patients were either treated with our extemporaneous oral suspension (n = 26; 53%), commercially available capsules (n = 18; 37%), or the intravenous prodrug formulation of aprepitant (fosaprepitant, n = 5; 10%). Pharmacokinetic analyses were performed using nonlinear mixed effects modelling. A one-compartment model adequately described the pharmacokinetics of aprepitant in children. The bioavailability of the extemporaneous oral suspension was not significantly different to that of the capsules (P = 0.26). The observed bioavailability throughout the total population was 83% (95% CI 69%-97%). The absorption of the extemporaneous oral suspension was 39.4% (95%CI 19.5-57.4%) faster than that of capsules (mean absorption time of 1.78 h (95%CI 1.32-2.35), but was comparable to that of the commercial oral suspension. The median area under the curve after (fos)aprepitant was 22.2 mg/L*h (range 8.9-50.3 mg/L*h) on day 1. Our extemporaneous oral suspension is an adequate alternative for the commercially (un)available oral suspension in young children. An adequate exposure to aprepitant in children was yielded and the bioavailability of the extemporaneous suspension was comparable to capsules.


Stochastic transition states: reaction geometry amidst noise.
Classical transition state theory (TST) is the cornerstone of reaction-rate theory. It postulates a partition of phase space into reactant and product regions, which are separated by a dividing surface that reactive trajectories must cross. In order not to overestimate the reaction rate, the dynamics must be free of recrossings of the dividing surface. This no-recrossing rule is difficult (and sometimes impossible) to enforce, however, when a chemical reaction takes place in a fluctuating environment such as a liquid. High-accuracy approximations to the rate are well known when the solvent forces are treated using stochastic representations, though again, exact no-recrossing surfaces have not been available. To generalize the exact limit of TST to reactive systems driven by noise, we introduce a time-dependent dividing surface that is stochastically moving in phase space, such that it is crossed once and only once by each transition path.


Pathogenic STX3 variants affecting the retinal and intestinal transcripts cause an early-onset severe retinal dystrophy in microvillus inclusion disease subjects.
Biallelic STX3 variants were previously reported in five individuals with the severe congenital enteropathy, microvillus inclusion disease (MVID). Here, we provide a significant extension of the phenotypic spectrum caused by STX3 variants. We report ten individuals of diverse geographic origin with biallelic STX3 loss-of-function variants, identified through exome sequencing, single-nucleotide polymorphism array-based homozygosity mapping, and international collaboration. The evaluated individuals all presented with MVID. Eight individuals also displayed early-onset severe retinal dystrophy, i.e., syndromic-intestinal and retinal-disease. These individuals harbored STX3 variants that affected both the retinal and intestinal STX3 transcripts, whereas STX3 variants affected only the intestinal transcript in individuals with solitary MVID. That STX3 is essential for retinal photoreceptor survival was confirmed by the creation of a rod photoreceptor-specific STX3 knockout mouse model which revealed a time-dependent reduction in the number of rod photoreceptors, thinning of the outer nuclear layer, and the eventual loss of both rod and cone photoreceptors. Together, our results provide a link between STX3 loss-of-function variants and a human retinal dystrophy. Depending on the genomic site of a human loss-of-function STX3 variant, it can cause MVID, the novel intestinal-retinal syndrome reported here or, hypothetically, an isolated retinal dystrophy.


Early induction of interleukin-12 by human monocytes exposed to Cryptococcus neoformans mannoproteins.
Interleukin-12 (IL-12) production by human monocytes stimulated with mannoproteins (MPs) of Cryptococcus neoformans was investigated. The results reported show that secreted or cell-associated MPs induce an early and significant production of IL-12. MPs show different capabilities to quantitatively affect IL-12 production; MP2, an 8. 2-kDa MP purified from the culture supernatant of C. neoformans, appears to be the most potent stimulator. Cytochalasin B inhibits both internalization and IL-12 induction by MP. In addition, a drastic reduction of IL-12 was observed when monocytes were cultured in the absence of normal human serum or treated with soluble mannan. Early production of IL-12 promotes early secretion of gamma interferon by T cells but does not influence the magnitude of the MP-induced lymphoproliferative response. Overall our results identify cryptococcal antigens responsible for rapid and potent induction of IL-12 in monocytes. MPs appear to regulate IL-12 secretion by internalization via the endocytic pathway and by interaction with monocyte receptors or serum factors.


UK endoscopy workload and workforce patterns: is there potential to increase capacity? A BSG analysis of the National Endoscopy Database.
The lack of comprehensive national data on endoscopy activity and workforce hampers strategic planning. The National Endoscopy Database (NED) provides a unique opportunity to address this in the UK. We evaluated NED to inform service planning, exploring opportunities to expand capacity to meet service demands. Data on all procedures between 1 March 2019 and 29 February 2020 were extracted from NED. Endoscopy activity and endoscopist workforce were analysed. 1 639 640 procedures were analysed (oesophagogastroduodenoscopy (OGD) 693 663, colonoscopy 586 464, flexible sigmoidoscopy 335 439 and endoscopic retrograde cholangiopancreatography 23 074) from 407 sites by 4990 endoscopists. 89% of procedures were performed in NHS sites. 17% took place each weekday, 10% on Saturdays and 6% on Sundays. Training procedures accounted for 6% of total activity, over 99% of which took place in NHS sites. Median patient age was younger in the independent sector (IS) (51 vs 60 years, p<0.001). 74% of endoscopists were male. Gastroenterologists and surgeons each comprised one-third of the endoscopist workforce; non-medical endoscopists (NMEs) comprised 12% yet undertook 23% of procedures. Approximately half of endoscopists performing OGD (52%) or colonoscopies (48%) did not meet minimum annual procedure numbers. This comprehensive analysis reveals endoscopy workload and workforce patterns for the first time across both the NHS and the IS in all four UK nations. Half of all endoscopists perform fewer than the recommended minimum annual procedure numbers: a national strategy to address this, along with expansion of the NME workforce, would increase endoscopy capacity, which could be used to exploit latent weekend capacity.


Use of a Multiplex Transcript Method for Analysis of Pseudomonas aeruginosa Gene Expression Profiles in the Cystic Fibrosis Lung.
The discovery of therapies that modulate Pseudomonas aeruginosa virulence or that can eradicate chronic P. aeruginosa lung infections associated with cystic fibrosis (CF) will be advanced by an improved understanding of P. aeruginosa behavior in vivo We demonstrate the use of multiplexed Nanostring technology to monitor relative abundances of P. aeruginosa transcripts across clinical isolates, in serial samples, and for the purposes of comparing microbial physiology in vitro and in vivo The expression of 75 transcripts encoded by genes implicated in CF lung disease was measured in a variety of P. aeruginosa strains as well as RNA serial sputum samples from four P. aeruginosa-colonized subjects with CF collected over 6 months. We present data on reproducibility, the results from different methods of normalization, and demonstrate high concordance between transcript relative abundance data obtained by Nanostring or transcriptome sequencing (RNA-Seq) analysis. Furthermore, we address considerations regarding sequence variation between strains during probe design. Analysis of P. aeruginosa grown in vitro identified transcripts that correlated with the different phenotypes commonly observed in CF clinical isolates. P. aeruginosa transcript profiles in RNA from CF sputum indicated alginate production in vivo, and transcripts involved in quorum-sensing regulation were less abundant in sputum than strains grown in the laboratory. P. aeruginosa gene expression patterns from sputum clustered closely together relative to patterns for laboratory-grown cultures; in contrast, laboratory-grown P. aeruginosa showed much greater transcriptional variation with only loose clustering of strains with different phenotypes. The clustering within and between subjects was surprising in light of differences in inhaled antibiotic and respiratory symptoms, suggesting that the pathways represented by these 75 transcripts are stable in chronic CF P. aeruginosa lung infections.


SUMOylation regulates telomere length homeostasis by targeting Cdc13.
Telomere length homeostasis is an important aspect of telomere biology. Here, we show that SUMOylation limits telomere length and targets multiple telomere proteins in Saccharomyces cerevisiae. A main target is Cdc13, which both positively and negatively regulates telomerase and confers end protection. We demonstrate that Cdc13 SUMOylation restrains telomerase functions by promoting Cdc13 interaction with the telomerase inhibitor Stn1 without affecting end protection. Mutation of the Cdc13 SUMOylation site (cdc13-snm) lengthens telomeres and reduces the Stn1 interaction, whereas Cdc13-SUMO fusion has the opposite effects. cdc13-snm's effect on telomere length is epistatic with stn1, but not with yku70, tel1 or est1 alleles, and is suppressed by Stn1 overexpression. Cdc13 SUMOylation peaks in early-mid S phase, prior to its known Cdk1-mediated phosphorylation, and the two modifications act antagonistically, suggesting that the opposite roles of Cdc13 in telomerase regulation can be separated temporally and regulated by distinct modifications.


Prostate biopsy findings in Indian men: a hospital-based study.
To review prostatic biopsy findings in Indian patients with elevated serum prostate-specific antigen (PSA) attending the Urology department at a tertiary care hospital. A retrospective study of 119 patients, who underwent TRUS-guided prostatic biopsy, was conducted. A total of 119 patients undergoing TRUS-guided prostatic biopsy were evaluated. Age, presentation, PSA, digital rectal examination, number of cores, and final histology were analyzed. Minimum 10 cores biopsies were performed in 109/119 (92%) and 12 cores in 92/119 (77%). Patients were stratified into three groups based on their PSA: 4-10 ng/ml (group I), 10-20 ng/ml (group II), and >20 ng/ml (group III). Unpaired t-test, Chi-square test, and logistic regression were calculated using an Excel (Ver 2007) and online calculators (P < 0.05 significant). Mean age was 67.6 years. Inflammatory pathology (30/119) was common at all PSA levels. In men with negative DRE and PSA > 10 ng/ml, inflammatory pathology was more likely (Chi 4.2798, P = 0.039). Cancer was found in 29/119 biopsies (group I 2/28, group II 3/45, and group III 24/46). Patients with PSA > 20 ng/ml were more likely to show cancer. Precursor lesions were noted in 10/119 (8.4%). On univariate analysis age, PSA, and DRE all showed significant association with histologic cancer but on multiple logistic regression analysis, only PSA (OR 1.03, P = 0.0021) and DRE (OR 8.07, P = 0.0007) were predictive of cancer. Cancer is less common and inflammatory lesions more common at all levels of PSA in our patients. The effect of antibiotics on PSA and biopsy in our patients needs to be explored.


The study of polymorphism within the promoter region of the osteopontin (OPN) gene in sows.
The aim of the present study was to find a possible polymorphism in the promoter region of the osteopontin (OPN) gene, as a potential mutation region, connected with the transcription factor-binding sites or regulatory sequences and to estimate the expression of this gene in ovaries and oviduct of sows. Sixty wbp x pbz sows after the first mating were slaughtered and tissues samples of the ovaries and oviduct were taken. Primer pairs for PCR analysis were designed on the basis of swine osteopontin 5' end sequence driven from GenBank. This study's amplified DNA fragment, spanning 274 bp of the promoter region, was chosen because of its contents of tree specific sites: type II collagen silence sequence (CACCTCC) at -682 (from the transcription initiation site), glucocortIcoid response site (TGTCCT) at -658 and CAAT box -592. This DNA fragment was subjected to a MSSCP analysis. A different MSSCP pattern was shown which indicates that mutation is located in this region. The samples of different conformers were sequenced and the A --> G transitions was identified in two positions -617 and -608. Restriction analysis of the DNA was performed, but unfortunately none of the known restriction enzymes recognized the novel SNPs, which is why the specific primer pairs characteristic for nucleotide A or for nucleotide G were chosen. In the second stage of the presented study the total RNA was extracted from the tissues of the ovaries and oviduct and the complementary DNA (cDNA) was synthesized. The real-time PCR analysis to determined the expression dynamics of the OPN gene in pig tissues was performed.


Measuring disability in multiple sclerosis: is the Community Dependency Index an improvement on the Barthel Index?
The Community Dependency Index (CDI) was developed due to concerns that the Barthel Index (BI) was limited as a measure of physical function in community settings. However, no studies have compared the two rating scales within multiple sclerosis (MS). The aim of this study was to determine whether, in a community-based sample of people with MS, the CDI is a better measure than the BI. BI and CDI data were collected from 90 people with MS. Four measurement properties were compared: scaling assumptions (item mean scores, corrected item-total correlations), acceptability (score distributions, floor/ceiling effects), reliability (Cronbach's alpha) and validity (concurrent, discriminant, group differences, relative validity). Both scales satisfied recommended criteria for scaling assumptions (indicating it was legitimate to report a summed score) and internal consistency reliability (alpha > 0.85). The scales were highly correlated (r = 0.96), indicating they measured the same construct. Both scales demonstrated good group differences validity, but the BI was marginally superior. Notable ceiling effects (BI > CDI) were demonstrated for both scales, particularly in those less disabled. This study sample had relatively minor levels of disability, with over 70% still being independently mobile. In this sample of people with MS, the measurement properties of the BI and CDI examined were very similar, suggesting the CDI does not appear to have achieved its goal of better measurement.


Midwives' counselling of women at specialised fear of childbirth clinics: A qualitative study.
Fear of childbirth is a well-known problem affecting women's wellbeing and health. The prevalence of intense fear varies across countries from 4.8 to 14.8%. During the past 25 years in Sweden women with intense fear of childbirth have been offered counselling at specialised clinics staffed by midwives. Although the counselling demonstrates positive results, the training, education, supervision and organisation differ between clinics. It is still unclear which approaches and practices are the most beneficial. The aim was to explore and describe the counselling of women with intense fear of childbirth from the viewpoint of midwives who provide counselling in specialised fear of childbirth clinics in one region of Sweden. A qualitative study of 13 midwives using focus group interviews and inductive content analysis. The midwives' counselling of women with intense fear of childbirth is described as 'striving to create a safe place for exploring fear of childbirth', comprising the following categories: Providing a reliable relationship; Investigating previous and present fears; and A strong dedication to the women. Although there are no guidelines for the counselling the midwives described similar frameworks. Some approaches were general, while others were specific and related to the individual woman's parity. The midwives achieved professional and personal development through counselling experiences. The findings add to the existing literature on counselling and can be used to inform the development of midwife-led interventions for women with intense fear of childbirth and previous traumatic births, as well as for the formal education of midwives.


Induction of differentiation by pyruvate and DMEM in the human retinal pigment epithelium cell line ARPE-19.
Cultured retinal pigment epithelium (RPE) may become a therapeutic option for transplantation in retinal disease. However maintaining a native RPE phenotype in vitro has proven challenging. The human RPE cell-line ARPE-19 is used widely as an alternative to primary RPE. It is grown in DMEM/F12 medium as standard, but its phenotype is dependent on culture conditions, and many differentiation markers are usually absent. The purpose of this study was to examine how this sensitive phenotype of ARPE-19 can be modulated by growth media with or without the metabolite pyruvate to elucidate better RPE growth conditions. ARPE-19 cells at passages p22 to p28 were cultured on filters for up to 3 months in DMEM/F12 or DMEM media with or without pyruvate and 1% fetal calf serum. Assessment of differentiation was performed using pigmentation, immunocytochemistry, protein/mRNA expression, transepithelial resistance, VEGF secretion, and ultrastructure. Pyruvate, in combination with DMEM, induced dark pigmentation and promoted differentiation markers such as CRALBP and MerTK. Importantly, RPE65 protein was detected by Western blotting and was enhanced by pyruvate, high glucose, and DMEM. ARPE-19 cells maintained in this medium could also phagocytose human photoreceptor outer segments (POS). VEGF secretion was greater in DMEM cultures and was affected by glucose but not by pyruvate. Pigmentation never occurred in DMEM/F12. This study demonstrated important differentiation markers, including pigmentation and Western blots of RPE65 protein, and showed human POS phagocytosis in ARPE-19 cultures using a simple differentiation protocol. The results favor the use of high-glucose DMEM with pyruvate for future RPE differentiation studies.


The expression of ribonucleotide reductase M2 in the carcinogenesis of uterine cervix and its relationship with clinicopathological characteristics and prognosis of cancer patients.
To investigate the implication of ribonucleotide reductase M2 (RRM2) in the carcinogenesis of uterine cervix and its relationship with clinicopathological characteristics and prognosis of cancer patients. The impact of RRM2 on cell viability was investigated in SiHa cervical cancer cells after RRM2 knockdown and the addition of cisplatin, which induces inter- and intra-strand DNA crosslinks. RRM2 immunoreactivity was evaluated by semi-quantitative H score among 29 normal, 30 low-grade dysplasia, 30 high-grade dysplasia and 103 invasive cancer tissue specimens of the uterine cervix, using tissue microarrays. RRM2 was then correlated with the clinicopathological variables of cervical cancer and patient survival. A greater toxic effect on cell viability using cisplatin was reflected by the greater reduction in RRM2 protein expression in SiHa cells. The RRM2 expression in cancer tissues was higher than that in high-grade dysplasia, low-grade dysplasia or normal cervical tissues. RRM2 upregulation was correlated with deep stromal invasion, large tumors and parametrial invasion and predicted poor survival. RRM2 is a new molecular marker for the diagnosis and clinical outcomes of cervical cancer. It is involved in cervical carcinogenesis and predicts poor survival, and may be a potential therapeutic target including in cisplatin treatment.


Risk factors, falls, and fracture of the distal forearm in Manchester, UK.
To determine the risk factors associated with fracture of the distal forearm, and to evaluate the influence of falls on these risks. This was a case-control study. Manchester, UK. The cases were 62 white women aged 45-82 years who had sustained a fracture of the distal forearm and had attended local hospitals. Two control groups were studied - 50 women who had fallen onto the hand but had not sustained a fracture (recruited from the same source as those with fracture) and 116 women randomly selected from primary care age and sex registers in the catchment area of the hospitals. Both cases and controls were sent a letter inviting them to take part in the study. Data were collected by questionnaire completed by an interviewer. Compared with the population control group, those with fracture were more likely to walk at a brisk pace (odds ratio (OR) = 3.5; 95% confidence interval (CI) 1.3, 9.6) though they had undertaken less physical activity at home or work on a daily basis throughout life (OR = 0.4; 95% CI 0.2, 0.9). The risk associated with brisk walking was less marked when the cases were compared with fall controls. Other lifestyle factors including calcium intake, smoking, and alcohol consumption were not associated with fracture. Analysis of gynaecological and hormonal factors suggested that compared with population controls, those with fracture of the distal forearm had had fewer fertile years (OR = 0.4; 95% CI 0.1, 0.9) and were less likely to have used oral contraceptives (OR = 0.3; 95% CI 0.1, 0.9). The data highlight the need for caution when advising middle aged and elderly subjects about exercise. Such advice should be combined with practical information about the prevention of falls. Hormonal factors seem to be additional determinants of fracture. Other lifestyle interventions seem unlikely to play an important part in preventing distal forearm fracture.


A pilot feasibility study of an extension of the acquaintanceship recruitment procedure in recent-onset psychosis.
The acquaintanceship recruitment procedure is an appealing yet infrequently used method to recruit controls, allowing a very close match between patients and controls. We used an extension of the acquaintanceship procedure to investigate the feasibility of this method to recruit controls in a neuropsychological study of recent-onset psychotic patients. Twenty-five recent-onset psychotic patients attending a multidisciplinary program devoted to recent-onset psychoses were contacted, among whom 13 agreed to participate to the study. At the end of the process, only four control participants were assessed. This pilot study suggests that several obstacles prevent the use of this procedure to recruit controls in this research focusing on recent-onset psychotic disorders.


COVID-19 response in South African communities: Screening, testing, tracing and movement modelling.
In South Africa (SA), the first case of COVID-19 was reported on 5 March 2020 from a traveller who had returned from Italy. Increases in COVID-19 cases and deaths necessitated the design and implementation of community screening, testing, and tracing as a control strategy. The SA government's plans to implement community-based screening, testing, contact tracing and movement modelling during the early phases of the COVID-19 pandemic presented both opportunities and challenges. In this article, we present our experiences, opportunities and lessons for community-based COVID-19 response, anchoring these efforts in the primary healthcare system.


Guidelines for the Treatment of Latent Tuberculosis Infection: Recommendations from the National Tuberculosis Controllers Association and CDC, 2020.
Comprehensive guidelines for treatment of latent tuberculosis infection (LTBI) among persons living in the United States were last published in 2000 (American Thoracic Society. CDC targeted tuberculin testing and treatment of latent tuberculosis infection. Am J Respir Crit Care Med 2000;161:S221-47). Since then, several new regimens have been evaluated in clinical trials. To update previous guidelines, the National Tuberculosis Controllers Association (NTCA) and CDC convened a committee to conduct a systematic literature review and make new recommendations for the most effective and least toxic regimens for treatment of LTBI among persons who live in the United States.The systematic literature review included clinical trials of regimens to treat LTBI. Quality of evidence (high, moderate, low, or very low) from clinical trial comparisons was appraised using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) criteria. In addition, a network meta-analysis evaluated regimens that had not been compared directly in clinical trials. The effectiveness outcome was tuberculosis disease; the toxicity outcome was hepatotoxicity. Strong GRADE recommendations required at least moderate evidence of effectiveness and that the desirable consequences outweighed the undesirable consequences in the majority of patients. Conditional GRADE recommendations were made when determination of whether desirable consequences outweighed undesirable consequences was uncertain (e.g., with low-quality evidence).These updated 2020 LTBI treatment guidelines include the NTCA- and CDC-recommended treatment regimens that comprise three preferred rifamycin-based regimens and two alternative monotherapy regimens with daily isoniazid. All recommended treatment regimens are intended for persons infected with Mycobacterium tuberculosis that is presumed to be susceptible to isoniazid or rifampin. These updated guidelines do not apply when evidence is available that the infecting M. tuberculosis strain is resistant to both isoniazid and rifampin; recommendations for treating contacts exposed to multidrug-resistant tuberculosis were published in 2019 (Nahid P, Mase SR Migliori GB, et al. Treatment of drug-resistant tuberculosis. An official ATS/CDC/ERS/IDSA clinical practice guideline. Am J Respir Crit Care Med 2019;200:e93-e142). The three rifamycin-based preferred regimens are 3 months of once-weekly isoniazid plus rifapentine, 4 months of daily rifampin, or 3 months of daily isoniazid plus rifampin. Prescribing providers or pharmacists who are unfamiliar with rifampin and rifapentine might confuse the two drugs. They are not interchangeable, and caution should be taken to ensure that patients receive the correct medication for the intended regimen. Preference for these rifamycin-based regimens was made on the basis of effectiveness, safety, and high treatment completion rates. The two alternative treatment regimens are daily isoniazid for 6 or 9 months; isoniazid monotherapy is efficacious but has higher toxicity risk and lower treatment completion rates than shorter rifamycin-based regimens.In summary, short-course (3- to 4-month) rifamycin-based treatment regimens are preferred over longer-course (6-9 month) isoniazid monotherapy for treatment of LTBI. These updated guidelines can be used by clinicians, public health officials, policymakers, health care organizations, and other state and local stakeholders who might need to adapt them to fit individual clinical circumstances.


Measuring and monitoring ICP in Neurocritical Care: results from a national practice survey.
The use of intracranial pressure (ICP) monitors is nearly synonymous with Neurocritical Care. Recent studies in nursing literature have report high levels of practice variance associated with ICP monitoring and treatment. There are no recent practice surveys to describe how critical care physicians and nurses who are familiar with ICP management provide care to their patients. A short survey was developed and disseminated electronically to the members of the Neurocritical Care Society. The summary from 241 professionals provides evidence that there is significant practice variation associated with ICP monitoring and management. The results highlight the need to develop standardized approaches to measuring, monitoring, recording, and treating ICP.


High-density lipoprotein: relations to metabolic parameters and severity of coronary artery disease.
The regulation of plasma high-density lipoprotein (HDL) cholesterol level by the joint influence of plasma lipoprotein lipids, lipoprotein lipase (LPL), hepatic lipase (HL), cholesteryl ester transfer protein (CETP), oral glucose tolerance, and postload plasma insulin and proinsulin levels was investigated in young postinfarction patients and healthy population-based control subjects. In addition, the association between HDL cholesterol and the number and severity of coronary stenoses previously reported in this cohort of young postinfarction patients was further investigated by analyzing the determinants and angiographic relations of HDL subclasses measured by gradient gel electrophoresis. The following parameters showed significant univariate relations with HDL cholesterol level in the patient group: very-low-density lipoprotein (VLDL) cholesterol and triglyceride, low-density lipoprotein (LDL) triglyceride, and postload plasma insulin concentrations, preheparin plasma LPL mass, and postheparin plasma HL activity. In the control group, significant correlations with HDL cholesterol concentration in addition to those noted among the patients were found for body mass index (BMI), LDL cholesterol level, postload plasma intact proinsulin concentration, and LPL activity in postheparin plasma. In contrast to the patients, no significant relations were noted for postload plasma insulin level and preheparin plasma LPL mass. Multiple stepwise regression analysis showed that 42% of the variability of HDL cholesterol in the patients could be accounted for by VLDL cholesterol concentration (29%), LDL triglyceride level (7%), and postheparin plasma HL activity (8%), whereas the corresponding figure in controls was 35% (VLDL cholesterol concentration [9%] and postheparin plasma HL activity [26%]. The strength of the relationships of HDL cholesterol and HDL subclasses to the coronary stenosis score was similar and statistically significant (r = .25 to .36). When the metabolic parameters that correlated with HDL cholesterol and HDL subclass concentrations in univariate analysis were used as covariates, all relations to the coronary stenosis score disappeared. This clearly indicates that the influence of triglyceride-rich lipoproteins and lipolytic enzymes needs to be considered when assessing the association between HDL cholesterol and coronary artery disease (CAD).


Animal Faux Pas: Two Legs Good Four Legs Bad for Theory of Mind, but Not in the Broad Autism Spectrum.
Research shows that the general population varies with regard to both autistic traits and theory of mind (ToM) ability. Other work has shown that autistic individuals may not underperform on ToM tests when the agent of evaluation is anthropomorphic rather than typically human. Two studies examined the relation between ToM and autistic trait profiles in over 650 adults using either the standard Faux Pas Recognition Test (FPT) or an anthropomorphized version of the FPT (FPTa). Results showed that autistic trait profiles were related to faux pas detection ability in the FPT but not the FPTa. Furthermore, while those with the broad autism phenotype scored significantly worse than those who were typically developed on the FPT, scores did not significantly differ on the FPTa. These findings add to a growing body of work suggesting that ToM ability is not at a global deficit in those on the autistic spectrum, but may relate to the mindreading of specifically human agents.


B cell depletion therapy exacerbates murine primary biliary cirrhosis.
Primary biliary cirrhosis (PBC) is considered a model autoimmune disease due to the clinical homogeneity of patients and the classic hallmark of antimitochondrial antibodies (AMAs). Indeed, the presence of AMAs represents the most highly directed and specific autoantibody in autoimmune diseases. However, the contribution of B cells to the pathogenesis of PBC is unclear. Therefore, although AMAs appear to interact with the biliary cell apotope and contribute to biliary pathology, there is no correlation of disease severity and titer of AMAs. The recent development of well-characterized monoclonal antibodies specific for the B cell populations, anti-CD20 and anti-CD79, and the development of a well-defined xenobiotic-induced model of autoimmune cholangitis prompted us to use these reagents and the model to address the contribution of B cells in the pathogenesis of murine PBC. Prior to the induction of autoimmune cholangitis, mice were treated with either anti-CD20, anti-CD79, or isotype-matched control monoclonal antibody and followed for B cell development, the appearance of AMAs, liver pathology, and cytokine production. Results of the studies reported herein show that the in vivo depletion of B cells using either anti-CD20 or anti-CD79 led to the development of a more severe form of cholangitis than observed in control mice, which is in contrast with results from several other autoimmune models that have documented an important therapeutic role of B cell-specific depletion. Anti-CD20/CD79-treated mice had increased liver T cell infiltrates and higher levels of proinflammatory cytokines. Our results reflect a novel disease-protective role of B cells in PBC and suggest that B cell depletion therapy in humans with PBC should be approached with caution.


Evidence-Based Screening Recommendations for Occult Cancers in the Setting of Newly Diagnosed Extramammary Paget Disease.
To identify the rates of associated and occult cancers in patients with extramammary Paget disease (EMPD) discovered using cancer screening methods at a tertiary medical center; to propose evidence-based cancer screening guidelines at the time of diagnosis of EMPD; and to clarify terminology associating EMPD with underlying malignancies. A retrospective review of patients with histologically confirmed EMPD presenting for care at our institution between January 1, 1992, and December 31, 2015, was performed. Both male and female patients were included. Descriptive analysis was performed. A total of 161 patients met the inclusion criteria. Most (59.6%) were female patients, and the mean age at the time of EMPD diagnosis was 70.8±10.1 years. Most (82%) of the 161 patients had at least 1 cancer screening test performed, though screening practices varied widely. Of those screened for an underlying malignancy, 17 distant, noncontiguous malignancies were identified in 15 patients (11.4%), with prostate (n=5), urinary tract (n=5), and breast (n=2) malignancies found most frequently. Most malignancies were identified by urine cytology, mammography, and prostate-specific antigen blood test. Of all patients, 37 (23.0%) had an underlying contiguous malignancy identified by pathology. All patients diagnosed with EMPD should undergo cancer screening. At minimum, evaluation should include age-appropriate screening and the addition of urine cytology, mammography, and prostate-specific antigen blood test-if not already performed-may be of particular use. An algorithm for evaluation of patients with newly diagnosed EMPD is proposed.


The clinical effects of combining postural exercises with chest physiotherapy in cystic fibrosis: A single-blind, randomized-controlled trial.
This study aims to investigate the effects of postural exercises as an adjunct to chest physiotherapy program on respiratory function, exercise tolerance, quality of life (QoL), and postural stability in patients with cystic fibrosis (CF). In this single-blind, randomized-controlled trial, 19 pediatric CF patients (11 males, 8 females; mean age: 9.36 years; range, 6 to 14 years) were randomly allocated to chest physiotherapy and postural exercise program (Group 1, n=10) or chest physiotherapy program alone (Group 2, n=9) between March 2017 and October 2017. Respiratory functions were assessed with pulmonary function tests, whereas exercise tolerance with the Modified Shuttle Test (MST), quality of life with the Cystic Fibrosis Questionnaire-Revised Child Version (CFQR), and postural stability with the Limits of Stability Test (LOS). All tests were performed before treatment and six weeks, three months, and six months after treatment. Respiratory functions were improved in both groups; however, these changes were not statistically significant. The MST increased after treatment in both groups (p<0.001 and p=0.003 respectively), without a significant difference between the groups. Emotional function and treatment difficulties subdomains in CFQR were significantly increased only in the group with postural exercises (p<0.05). The postural exercise program in addition to chest physiotherapy in pediatric CF patients whose postural changes were not taken place did not cause significant changes in respiratory function, exercise tolerance, and postural stability; however, it affected the emotional state well and improved the compliance with the treatment.


Effect of polyherbal formulation on experimental models of inflammatory bowel diseases.
A polyherbal ayurvedic formulation from an ancient authentic classical text of ayurveda was evaluated for its activity against inflammatory bowel disease (IBD). The polyherbal formulation contained four different drugs viz., Bilwa (Aegle marmeloes), Dhanyak (Coriandrum sativum), Musta (Cyperus rotundus) and Vala (Vetiveria zinzanioids). The formulation has been tried before in clinical practice and was found to be useful in certain number of cases of IBD (ulcerative colitis), so was tried in the same form i.e., decoction (aqueous extract) in experimental animals to revalidate the claims of the same. The formulation was tried on two different experimental animal models of inflammatory bowel disease, which are acetic acid-induced colitis in mice and indomethacin-induced enterocolitis in rats. Prednisolone was used as the standard drug for comparison. The formulation showed significant inhibitory activity against inflammatory bowel disease induced in these experimental animal models. The activity was comparable with the standard drug prednisolone. The results obtained established the efficacy of this polyherbal formulation against inflammatory bowel diseases.


Public Health Impact of Frailty: Role of Physical Therapists.
Frailty is an emerging and immediate public health concern given the growing aging population. The condition of frailty is characterized by a reduction in physiologic reserve, which places frail older adults at considerable risk for further functional decline, hospitalization, institutionalization, and death. Recent research suggests that frailty may be reversible, which could result in significant improvement in public health. Thus, a strong impetus exists to develop strategies for frail older adults that achieve the Triple Aim through better promotion of population health, optimization of patient experiences, and delivery of high-quality care at minimal cost. Physical therapists often treat frail older adults, yet how physical therapists can contribute to preventing or reversing frailty in healthcare settings has not been described, and may potentially influence patient outcomes and healthcare spending. Therefore, the purpose of this publication is to outline the potential role of physical therapists in achieving the Triple Aim for the frail older adult population.


Anthropogenic ocean acidification over the twenty-first century and its impact on calcifying organisms.
Today's surface ocean is saturated with respect to calcium carbonate, but increasing atmospheric carbon dioxide concentrations are reducing ocean pH and carbonate ion concentrations, and thus the level of calcium carbonate saturation. Experimental evidence suggests that if these trends continue, key marine organisms--such as corals and some plankton--will have difficulty maintaining their external calcium carbonate skeletons. Here we use 13 models of the ocean-carbon cycle to assess calcium carbonate saturation under the IS92a 'business-as-usual' scenario for future emissions of anthropogenic carbon dioxide. In our projections, Southern Ocean surface waters will begin to become undersaturated with respect to aragonite, a metastable form of calcium carbonate, by the year 2050. By 2100, this undersaturation could extend throughout the entire Southern Ocean and into the subarctic Pacific Ocean. When live pteropods were exposed to our predicted level of undersaturation during a two-day shipboard experiment, their aragonite shells showed notable dissolution. Our findings indicate that conditions detrimental to high-latitude ecosystems could develop within decades, not centuries as suggested previously.


Oestrogen affects the cardiovascular and central responses to isoproterenol of female rats.
This study examined the influence of oestrogen on cardiovascular responses to hypotension produced by administration of isoproterenol (Isop) and on neural activation in hindbrain nuclei mediating these responses. We first measured mean arterial pressure (MAP) and heart rate (HR) after administration of isoproterenol, a beta-adrenergic agonist that increases circulating levels of AngII, in ovariectomized (OVX) rats treated with oestradiol benzoate (EB). We then evaluated EB effects on Isop-induced Fos immunoreactivity (Fos-IR) in the hindbrain baroreflex circuit. To control for weight loss associated with oestrogen replacement in OVX rats, we food restricted a separate group of OVX rats and evaluated Isop-induced changes in MAP, HR and Fos-IR. The depressor response to Isop was significantly attenuated by EB, which also produced a disproportionate increase in HR. These effects were not secondary to loss of body weight after EB treatment, because cardiovascular responses to Isop in food restricted rats were similar to those in OVX rats treated with the oil vehicle. Isop significantly increased Fos-IR in the nucleus of the solitary tract (NTS), area postrema (AP), rostral ventrolateral medulla (RVLM), and lateral parabrachial nucleus (lPBN); however, EB significantly attenuated the increase in the AP and in the lPBN. Again, these effects were not secondary to body weight loss, because food restricted rats had the same pattern of Fos-IR as did rats treated with the oil vehicle. These results suggest that EB modifies cardiovascular responses to Isop, possibly by decreasing activation of the AP and lPBN.


Potent Activation of Indoleamine 2,3-Dioxygenase by Polysulfides.
Indoleamine 2,3-dioxygenase (IDO1) is a heme enzyme that catalyzes the oxygenation of the indole ring of tryptophan to afford N-formylkynurenine. This activity significantly suppresses the immune response, mediating inflammation and autoimmune reactions. These consequential effects are regulated through redox changes in the heme cofactor of IDO1, which autoxidizes to the inactive ferric state during turnover. This change in redox status increases the lability of the heme cofactor leading to further suppression of activity. The cell can thus regulate IDO1 activity through the supply of heme and reducing agents. We show here that polysulfides bind to inactive ferric IDO1 and reduce it to the oxygen-binding ferrous state, thus activating IDO1 to maximal turnover even at low, physiologically significant concentrations. The on-rate for hydrogen disulfide binding to ferric IDO1 was found to be >106 M-1 s-1 at pH 7 using stopped-flow spectrometry. Fe K-edge XANES and EPR spectroscopy indicated initial formation of a low-spin ferric sulfur-bound species followed by reduction to the ferrous state. The μM affinity of polysulfides for IDO1 implicates these polysulfides as important signaling factors in immune regulation through the kynurenine pathway. Tryptophan significantly enhanced the relatively lower-affinity binding of hydrogen sulfide to IDO1, inspiring the use of the small molecule 3-mercaptoindole (3MI), which selectively binds to and activates ferric IDO1. 3MI sustains turnover by catalytically transferring reducing equivalents from glutathione to IDO1, representing a novel strategy of upregulating innate immunosuppression for treatment of autoimmune disorders. Reactive sulfur species are thus likely unrecognized immune-mediators with potential as therapeutic agents through these interactions with IDO1.


Structure and properties of DOTA-chelated radiopharmaceuticals within the 225Ac decay pathway.
The successful delivery of toxic cargo directly to tumor cells is of primary importance in targeted (α) particle therapy. Complexes of radioactive atoms with the 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) chelating agent are considered as effective materials for such delivery processes. The DOTA chelator displays high affinity to radioactive metal isotopes and retains this capability after conjugation to tumor targeting moieties. Although the α-decay chains are well defined for many isotopes, the stability of chelations during the decay process and the impact of released energy on their structures remain unknown. The radioactive isotope 225Ac is an α-particle emitter that can be easily chelated by DOTA. However, 225Ac has a complex decay chain with four α-particle emissions during decay of each radionuclide. To advance our fundamental understanding of the consequences of α-decay on the stability of tumor-targeted 225Ac-DOTA conjugate radiopharmaceuticals, we performed first principles calculations of the structure, stability, and electronic properties of the DOTA chelator to the 225Ac radioactive isotope, and the initial daughters in the decay chain, 225Ac, 221Fr, 217At and 213Bi. Our calculations show that the atomic positions, binding energies, and electron localization functions are affected by the interplay between spin-orbit coupling, weak dispersive interactions, and environmental factors. Future empirical measurements may be guided and interpreted in light of these results.


An in silico prediction tool for the expansion culture of human skeletal muscle myoblasts.
Regenerative therapy using autologous skeletal myoblasts requires a large number of cells to be prepared for high-level secretion of cytokines and chemokines to induce good regeneration of damaged regions. However, myoblast expansion culture is hindered by a reduction in growth rate owing to cellular quiescence and differentiation, therefore optimization is required. We have developed a kinetic computational model describing skeletal myoblast proliferation and differentiation, which can be used as a prediction tool for the expansion process. In the model, myoblasts migrate, divide, quiesce and differentiate as observed during in vitro culture. We assumed cell differentiation initiates following cell-cell attachment for a defined time period. The model parameter values were estimated by fitting to several predetermined experimental datasets. Using an additional experimental dataset, we confirmed validity of the developed model. We then executed simulations using the developed model under several culture conditions and quantitatively predicted that non-uniform cell seeding had adverse effects on the expansion culture, mainly by reducing the existing ratio of proliferative cells. The proposed model is expected to be useful for predicting myoblast behaviours and in designing efficient expansion culture conditions for these cells.